[go: up one dir, main page]

WO2023175171A1 - Bk polyomavirus antibodies and uses thereof - Google Patents

Bk polyomavirus antibodies and uses thereof Download PDF

Info

Publication number
WO2023175171A1
WO2023175171A1 PCT/EP2023/056946 EP2023056946W WO2023175171A1 WO 2023175171 A1 WO2023175171 A1 WO 2023175171A1 EP 2023056946 W EP2023056946 W EP 2023056946W WO 2023175171 A1 WO2023175171 A1 WO 2023175171A1
Authority
WO
WIPO (PCT)
Prior art keywords
sequence
seq
amino acid
acid sequence
antibody
Prior art date
Application number
PCT/EP2023/056946
Other languages
French (fr)
Inventor
Dorian MCILROY
Xavier SAULQUIN
Ngoc Khanh Nguyen
Céline BRESSOLLETTE-BODIN
Laetitia GAUTREAU-ROLLAND
Cynthia Fourgeux
Marie-Claire DEVILDER
Original Assignee
Inserm (Institut National De La Sante Et De La Recherche Medicale)
Centre National De La Recherche Scientifique
Nantes Université
Chu De Nantes
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Inserm (Institut National De La Sante Et De La Recherche Medicale), Centre National De La Recherche Scientifique, Nantes Université, Chu De Nantes filed Critical Inserm (Institut National De La Sante Et De La Recherche Medicale)
Publication of WO2023175171A1 publication Critical patent/WO2023175171A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
    • C07K16/084—Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • This application includes a sequence listing in computer readable form in a file named S3_SATT OUEST_PR94463_sequence listing created on March 7, 2023 (161 458 bytes), which is incorporated by reference herein.
  • the present disclosure relates to antibodies, and combinations thereof, targeting BK polyomaviruses (BKPyV), and methods using those antibodies.
  • BKPyV BK polyomaviruses
  • BK polyomavirus is a small double-stranded DNA virus with a non-enveloped icosahedral capsid, first described in 1971.
  • BKPyV has been divided into four major subtypes (genotypes) I, II, III and IV, further divided into subgroups, that are characterized by sequence variation in the major capsid protein VP1 and correspond to distinct neutralizing virus serotypes (Knowles et al. J. Med. Virol. 1989, 28, 118-123 - Pastrana et al. J. Virol. 2013, 87, 10105-10113).
  • Genotype I is the most prevalent BKPyV genotype worldwide, followed by genotype IV in East Asia and Europe, while genotypes II and III are rare throughout the world (Zhong et al. J. Gen. Virol. 2009, 90, 144-152).
  • the distribution of the molecular subgroups of the virus is associated with the region of origin, for instance, BKPyV subtype la is most common in Africa, Ibl in Southeast Asia, and Ib2 in Europe, while Ic is the most common variant in Northeast Asia (Furmaga, Jacek et al. Viruses vol. 13,8 1502. 30 Jul. 2021).
  • BKPyV genotype I-E61K or BKPyV genotype IV-E327K in vitro selected genetic variants of some genotypes of the BKPyV were characterized, such as BKPyV genotype I-E61K or BKPyV genotype IV-E327K (Lindner et al. Immunity. 2019;50:668- 676.e5).
  • the virus capsid is composed of 72 pentamers of the major capsid protein VP1, and each pentamer is associated with one copy of either VP2 or VP3, which are the minor capsid proteins.
  • the circular double-stranded DNA genome is packaged inside the capsid in a condensed form associated with histones from the host cell.
  • the exterior of the virus particle is composed entirely of the VP1 protein, and virus like particles (VLP) composed of VP1 are structurally and antigenically identical to infectious virus particles (Hurdiss et al. Structure 2018, 26, 839-847.e3).
  • BKPyV is a typical opportunistic virus, which provokes overt pathology almost exclusively in the context of immunosuppression. It is estimated that 80% of the adult population is latently infected with BKPyV. After kidney transplantation (KTx), reactivation of BKPyV from sites of persistent infection (mainly kidney epithelial cells) frequently occurs, leading to virus detection in urine (Bressollette-Bodin et al. Am. J. Transplant. 2005, 5, 1926-1933).
  • KTx recipients with persistent BKPyV viremia or PyVAN were found to have an 11 -fold greater risk of bladder carcinoma compared to KTx recipients without evidence of BKPyV replication (Liu et al.
  • BKPyV hemorrhagic cystitis
  • HCT allogeneic hematopoietic cell transplantations
  • BK polyoma virus-associated nephropathy relies on modification or dose reduction of immunosuppressive therapy, with the aim of inducing host antiviral immune responses that may then control viral replication.
  • This strategy is applied in the case of histologically confirmed BK polyomavirus-associated nephropathy (curative treatment), as well as in cases of “presumptive BK PyVAN”, defined by a high viruria and DNAemia (Zhong et al. J. Gen. Virol. 2009, 90, 144-152).
  • BKPyV-HC symptoms can persist for several weeks in subjects with high plasma DNAemia (Imlay et al. Blood Adv 2020, 4, 617-628), and their resolution is thought to depend on the reconstitution of the BK-specific immune response that follows successful engraftment (Espada et al. Blood Adv 2020, 4, 1881-1893). In both clinical situations, patients would benefit from effective antiviral therapy.
  • the incidence of BK PyVAN is higher in kidney transplantation recipients with low neutralizing antibody titers (Solis et al. J. Am. Soc. Nephrol.
  • IVIG intravenous immunoglobulin
  • BKPyV-specific neutralizing antibodies Velay et al. Antimicrob Agents Chemother 2019, 63
  • IVIG intravenous immunoglobulin
  • BKPyV-specific neutralizing antibodies Velay et al. Antimicrob Agents Chemother 2019, 63
  • BK polyomavirus-associated nephropathy Matsumura et al. Ther Clin Risk Manag 2020, 16, 947-952; Pibum et al. Pediatric Transplantation 2020, 24, el3600; Hwang et al. Transplant Proc 2018, 50, 2575-2578.
  • IVIG is a blood product that contains a complex mix of antibody specificities, and although rare, acute antibody-mediated rejection after infusion of IVIG containing donorspecific antibodies has been reported in the literature (Mainra, et al., Transpl Immunol 2013, 28, 145-147).
  • antiviral T-cell therapy has been developed for BKPyV-HC, with reported clinical response rates greater than 80% (Nelson et al. Curr. Opin. Infect. Dis., 2021, 34, 627- 634).
  • at least one fatal outcome associated with cytokine reconstitution syndrome has been reported in a BKPyV-HC patient treated with T-cell therapy.
  • the broadly neutralizing monoclonal antibody 41F17 has been identified as an anti-BKPyV antibody efficiently for binding and neutralizing all of the four genotypes of the BKPyV wildtype (Lindner et al. Immunity. 2019; 50:668-676.e5).
  • BKPyV has the ability to escape selection pressure by antibodies by developing BKPyV variants containing mutations on its VP1 capsid protein.
  • the anti-BKPyV 41F17 antibody may not be enough to efficiently neutralize all the BKPyV variants.
  • Broadly neutralizing antibodies capable of inhibiting the four wildtype BKPyV genotypes and its variants would be potential candidates for therapeutics for the treatment of BKPyV infection-associated diseases, in particular PyVAN and/or BKPyV-HC.
  • BKPyV infection-associated diseases include BK polyomavirus-associated nephropathy, confirmed and presumptive, BKPyV hemorrhagic cystitis, as well as pneumonitis, retinitis, and meningoencephalitis in immunocompromised subjects, and the likely long-term consequences of BKPyV infection-associated diseases such as reno-urinary tumors, for example bladder carcinoma.
  • the present disclosure aims at satisfying all or parts of those needs.
  • the present disclosure relates to an isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising:
  • CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
  • CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
  • CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
  • CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
  • CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution
  • CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
  • the present disclosure relates to an isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising:
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution
  • a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution
  • a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • the isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof may comprise:
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution
  • a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or an amino acid sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or an amino acid sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or an amino acid sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution
  • a CDR2-L comprising an amino acid sequence of DTS or an amino acid sequence differing from DTS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
  • the isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof may comprise:
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or an amino acid sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution
  • a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or an amino acid sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or an amino acid sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or an amino acid sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution
  • a CDR2-L comprising an amino acid sequence of GAS or an amino acid sequence differing from GAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or an amino acid sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • the inventors have surprisingly identified new antibodies able to strongly neutralize BKPyV infection induced by the different wildtype and naturally occurring variant genotypes in different host cells. Furthermore, it has been shown that combinations of at least two of those antibodies surprisingly were able to neutralize variants of BKPyV escaping neutralization by the benchmark anti-BKPyV antibody 41F17.
  • the disclosed antibodies, and combinations thereof, are able to bind and neutralize BKPyV.
  • antibodies and combinations of antibodies may be implemented in prophylactic and/or therapeutic treatment of BKPyV-induced infections and diseases associated with BKPyV infections in transplanted subjects, such as kidney transplanted subjects.
  • prophylactic and/or therapeutic uses of those antibodies and combinations of antibodies may advantageously be associated with a reduced of occurrence of antibody-resistant variants of BKPyV.
  • An antibody disclosed herein may comprise:
  • an antibody as disclosed herein may comprise a variable domain of heavy chain of the amino acid sequence SEQ ID NO: 6 or an amino acid sequence at least 85% identical thereto and/or a variable domain of light chain of the amino acid sequence SEQ ID NO: 7, or an amino acid sequence at least 85% identical thereto.
  • an antibody as disclosed herein may comprise a variable domain of heavy chain of the amino acid sequence SEQ ID NO: 84 or an amino acid sequence at least 85% identical thereto and/or a variable domain of light chain of the amino acid sequence SEQ ID NO: 85, or an amino acid sequence at least 85% identical thereto.
  • An antibody disclosed herein may comprise:
  • a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or a sequence at least 85% identical thereto; or
  • an antibody as disclosed herein may comprise a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or an amino acid sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or an amino acid sequence at least 85% identical thereto.
  • an antibody as disclosed herein may comprise a heavy chain of amino acid sequence SEQ ID NO: 86 or an amino acid sequence at least 85% identical thereto and/or a light chain of amino acid sequence SEQ ID NO: 87, or an amino acid sequence at least 85% identical thereto.
  • the disclosure relates to an isolated antibody, or an antigen-binding fragment thereof, neutralizing at least one BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101, or SEQ ID NO: 106, and for example at least one BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, or SEQ ID NO: 101, with an IC50 of less than 40 nM.
  • An antibody disclosed herein bind and neutralize a BKPy virus variant selected among BKPyV-D58Del_59A, BKPyV-E61K, BKPyV-E327K and BKPyV-K172A, and for example a BKPy virus variant selected among BKPyV-D58Del_59A, BKPyV-E61K, and BKPyV-E327K.
  • Such an antibody may bind and neutralize at least two BKPy virus variants selected among BKPyV-D58Del_59A, BKPyV-E61K and BKPyV-E327K.
  • An antibody disclosed herein may be an antibody fragment.
  • An antibody fragment may be selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, and diabodies.
  • An antibody disclosed herein may be a variable heavy chain of a single domain antibody (VHH). The variable heavy chain may be as defined herein.
  • the present disclosure relates to an immunoconjugate comprising an antibody as disclosed herein conjugated or linked to a detectable probe, label or reporter molecule.
  • Such immunoconjugate may be used as a diagnostic tool for detecting and/or measuring a BKPyV infection in a subject in need thereof.
  • a detectable probe, label or reporter molecule can be a radioisotope, such as 3 H, 14 C, 32 P, 35 S, or 125 I; a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine; or an enzyme such as alkaline phosphatase, P-galactosidase, horseradish peroxidase, or luciferase.
  • a radioisotope such as 3 H, 14 C, 32 P, 35 S, or 125 I
  • a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine
  • an enzyme such as alkaline phosphatase, P-galactosidase, horseradish peroxidase, or luciferase.
  • the present disclosure relates to a combination of at least a first antibody, or an antigen-binding fragment thereof, and a second antibody, or an antigen-binding fragment thereof, wherein said first and second antibodies may be selected among the groups of antibodies as disclosed herein.
  • the present disclosure relates to an isolated multi-specific antibody comprising at least a first antibody, or an antigen-binding fragment thereof, and a second antibody, or an antigen-binding fragment thereof, joined to each other, wherein said first and second antibodies may be selected among the groups of antibodies as disclosed herein.
  • An isolated multi-specific antibody may be a bispecific antibody.
  • a combination or an isolated multi-specific antibody disclosed herein may comprise:
  • a combination or an isolated multi-specific antibody disclosed herein may comprise:
  • the present disclosure relates to a pharmaceutical composition
  • a pharmaceutical composition comprising at least an antibody as disclosed herein, at least a combination of at least a first and a second antibodies as disclosed herein, or at least a multispecific antibody as disclosed herein, and a pharmaceutically acceptable carrier.
  • the present disclosure relates to an isolated antibody as disclosed herein, at least a combination of at least a first and a second antibodies as disclosed herein, at least a multi-specific antibody as disclosed herein, or a pharmaceutical composition as disclosed herein, for use as a medicament.
  • the present disclosure relates to an isolated antibody as disclosed herein, at least a combination of at least a first and a second antibodies as disclosed herein, at least a multi-specific antibody as disclosed herein, or a pharmaceutical composition as disclosed herein, for use in the prevention and/or treatment of a BKPyV -infection and/or a BKPyV -inf ection associated disease, in a subject in need thereof.
  • a subject in need thereof may be an organ transplant recipient.
  • An organ transplant recipient may be a kidney transplant recipient.
  • a BKPyV-infection associated disease may be a BK polyomavirus-associated nephropathy (BK PyVAN), a hemorrhagic cystitis, (BK Py-HC) a granule cell neuronopathy (GCN), an interstitial kidney disease, an ureteral stenosis, a pneumonitis, a vasculitis, a colitis, a meningitis, a retinitis, a meningoencephalitis, an immune reconstitution inflammatory syndrome (IRIS), or a reno-urinary cancer.
  • BK PyVAN BK polyomavirus-associated nephropathy
  • BK Py-HC hemorrhagic cystitis
  • GCN granule cell neuronopathy
  • IRIS immune reconstitution inflammatory syndrome
  • a disorder induced by a BKPyV infection may be a hemorrhagic cystitis.
  • a disorder induced by a BKPyV infection may be a BK polyomavirus-associated nephropathy.
  • a disorder induced by a BKPyV infection may be a reno- urinary cancer, for example a bladder cancer.
  • nucleic acid sequence disclosed herein may comprise:
  • a nucleic acid sequence disclosed herein may comprise:
  • the present disclosure relates to an isolated or recombinant expression vector comprising a nucleic acid sequence disclosed herein.
  • the present disclosure relates to a recombinant host cell which has been transformed by a nucleic acid or an expression vector disclosed herein.
  • the present disclosure relates to a method for producing an antibody binding a BK polyoma virus (BKPyV) VP1 capsid protein, or an antigenbinding fragment thereof, comprising at least the steps of culturing a host cell disclosed in a culture medium under conditions suitable for producing said antibody or fragment thereof and recovering the antibody or fragment thereof from said cultured cells.
  • BKPyV BK polyoma virus
  • the present disclosure relates to an isolated BKPy VP1 protein of amino acid sequence SEQ ID NO: 96 or an isolated VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • the present disclosure relates to an isolated BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • isolated BKPy virus may be a pseudotype virus or a virus.
  • pseudotype virus and pseudovirus are used interchangeably.
  • the present disclosure relates to a use of a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence for screening a candidate antibody able to neutralize a BKPy virus.
  • the BKPy virus may be a pseudotype virus.
  • the BKPy virus pseudotype may be BKPyV g!b2-D58DelP59A.
  • the present disclosure relates to an in vitro method for screening a candidate antibody able to neutralize a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, said method comprising at least the steps of:
  • the BKPy virus may be a pseudotype virus.
  • the BKPy virus may be BKPyV g!b2-D58DelP59A.
  • the present disclosure relates to a method for manufacturing a pharmaceutical composition for treating and/or preventing a BKPyV infection or for treating and/or preventing a disorder induced by a BKPyV infection in a subject in need thereof, the method comprising at least the steps of:
  • the present disclosure relates to an in vitro method for measuring a neutralizing activity of an anti -BKPyV antibody isolated from a subject presumed to be infected with a BKPy virus, the method comprising at least the steps of:
  • FIGURES 1A-F represent the binding properties of BK120 (Fig. 1A), BK160- 1 (Fig. IB), BK198 (Fig. 1C), BK206 (Fig. ID), BK256 (Fig. IE), and BK293 (Fig. IF), antibodies measured by ELISA against the four wildtype BKPyV genotypes: gla (•), gll ( ⁇ ), gill (A) and gIVc2 ( ⁇ ).
  • FIGURES 2A-G represent the neutralization capacity of BK120 (Fig. 2A),
  • BK160-1 (Fig. 2B), BK198 (Fig. 2C), BK206 (Fig. 2D), BK256 (Fig. 2E), BK293 (Fig. 2F) and BK304 (Fig. 2G) antibodies on HEK293TT cells infected with wildtype pseudotype viruses
  • BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
  • FIGURES 3A-G represent the neutralization capacity of BK120 (Fig. 3A), BK160-1 (Fig. 3B), BK198 (Fig. 3C), BK206 (Fig. 3D), BK256 (Fig. 3E), BK293 (Fig. 3F) and BK304 (Fig. 3G) antibodies on RS cells infected with wildtype pseudotype viruses (PSV) gla (•), gll ( ⁇ ), gill (A) and gIVc2 ( ⁇ ). Each antibody was added to BKPyV PSV in serial 5- fold dilutions starting at 10 pg/mL. Abscissa: Antibody concentration in pg/ml in loglO.
  • PSV wildtype pseudotype viruses
  • FIGURES 4A-H represent the viral neutralization capacity of BK120 (Fig. 4A), BK206 (Fig. 4B), BK293 (Fig. 4C), BK304 (Fig. 4D), BK120 + BK206 (Fig. 4E), BK120 + BK293 (Fig. 4F), BK120 + BK304 (Fig. 4G), and 41F17 (Fig. 4H) on HEK293TT cells infected with wildtype and naturally occurring variant pseudotype viruses (PSV) glb2-wt (•), glb2- DNEQ ( ⁇ ), gIb2-VVAE (+), gIVc2 ( ⁇ ) and gIVc2-SDKAN (0).
  • PSV wildtype and naturally occurring variant pseudotype viruses
  • FIGURES 5A-H represent the viral neutralization capacity of BK120 (Fig. 5A), BK206 (Fig. 5B), BK293 (Fig. 5C), BK304 (Fig. 5D), BK120 + BK206 (Fig. 5E), BK120 + BK293 (Fig. 5F), BK120 + BK304 (Fig. 5G), and 41F17 (Fig. 5A), BK206 (Fig. 5B), BK293 (Fig. 5C), BK304 (Fig. 5D), BK120 + BK206 (Fig. 5E), BK120 + BK293 (Fig. 5F), BK120 + BK304 (Fig. 5G), and 41F17 (Fig. 5A), BK206 (Fig. 5B), BK293 (Fig. 5C), BK304 (Fig. 5D), BK120 + BK206 (Fig. 5E), BK120 + BK293 (Fig. 5F), BK120 + BK304 (Fig. 5G), and 41F17 (Fig.
  • FIGURES 6A-D represent the viral neutralization capacity of BK120 (o), compared to P8D11 ( ⁇ ) and 41F17 (A) antibodies on cells infected with wildtype pseudotype viruses (PSV) gl (Fig. 6A), gll (Fig. 6B), gill (Fig. 6C) and gIV (Fig. 6D).
  • P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL
  • BK120 and 41F17 antibodies were added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL .
  • Abscissa Antibody concentration in ng/ml in log 10.
  • Ordinate BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
  • FIGURES 7A-B represent the viral neutralization capacity of BK120 (o), compared to P8D11 ( ⁇ ) and 41F17 (A) on cells infected with variants pseudotype viruses (PSV) g!b2-E61K (Fig. 7A) and g!b2-K172A (Fig. 7B).
  • P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL
  • BK120 and 41F17 antibodies were added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL .
  • Abscissa Antibody concentration in ng/ml in log 10.
  • Ordinate BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
  • FIGURES 8A-B represent the viral neutralization capacity of BK120 (o), compared to P8D11 ( ⁇ ) and 41F17 (A) antibodies on cells infected with variants pseudotype viruses (PSV) g!b2-DNEQ (Fig. 8A) and gIVc2 SDKAN (Fig. 8B).
  • P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL
  • BK120 and 41F17 antibodies were added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL.
  • Abscissa Antibody concentration in ng/ml in log 10.
  • Ordinate BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
  • SEQ ID NO: 1 represents the amino acids sequence of the HCDR1 of BK120.
  • SEQ ID NO: 2 represents the amino acids sequence of the HCDR2 of BK120.
  • SEQ ID NO: 3 represents the amino acids sequence of the HCDR3 of BK120.
  • SEQ ID NO: 4 represents the amino acids sequence of the LCDR1 of BK120.
  • SEQ ID NO: 5 represents the amino acids sequence of the LCDR3 of BK120.
  • SEQ ID NO: 6 represents the amino acids sequence of the heavy chain variable domain (VH) of BK120.
  • SEQ ID NO: 7 represents the amino acids sequence of the light chain variable domain (VL) of BK120.
  • SEQ ID NO 8 represents the amino acids sequence of the heavy chain of BK120.
  • SEQ ID NO 9 represents the amino acids sequence of the light chain of BK120.
  • SEQ ID NO 10 represents the nucleic acids sequence of the VH of BK120.
  • SEQ ID NO 11 represents the nucleic acids sequence of the VL of BK120.
  • SEQ ID NO 12 represents the nucleic acids sequence of the heavy chain of the BK120.
  • SEQ ID NO 13 represents the nucleic acids sequence of the light chain of the BK120.
  • SEQ ID NO 14 represents the amino acids sequence of the HCDR1 of BK160-1.
  • SEQ ID NO 15 represents the amino acids sequence of the HCDR2 of B KI 60-1.
  • SEQ ID NO 16 represents the amino acids sequence of the HCDR3 of B KI 60-1.
  • SEQ ID NO 17 represents the amino acids sequence of the LCDR1 of BK160-1.
  • SEQ ID NO 18 represents the amino acids sequence of the LCDR3 of B KI 60-1.
  • SEQ ID NO 19 represents the amino acids sequence of the VH of B KI 60-1.
  • SEQ ID NO 20 represents the amino acids sequence of the VL of B KI 60-1.
  • SEQ ID NO 21 represents the amino acids sequence of the heavy chain of BK160-1.
  • SEQ ID NO 22 represents the amino acids sequence of the light chain of B KI 60-1.
  • SEQ ID NO 23 represents the nucleic acids sequence of the VH of B KI 60-1.
  • SEQ ID NO 24 represents the nucleic acids sequence of the VL of B KI 60-1.
  • SEQ ID NO 25 represents the nucleic acids sequence of the heavy chain of the B KI 60-1.
  • SEQ ID NO 26 represents the nucleic acids sequence of the light chain of the B KI 60-1.
  • SEQ ID NO 27 represents the amino acids sequence of the HCDR1 of BK198.
  • SEQ ID NO 28 represents the amino acids sequence of the HCDR2 of BK198.
  • SEQ ID NO 29 represents the amino acids sequence of the HCDR3 of BK198.
  • SEQ ID NO 30 represents the amino acids sequence of the LCDR1 of BK198.
  • SEQ ID NO 31 represents the amino acids sequence of the LCDR3 of BK198.
  • SEQ ID NO 32 represents the amino acids sequence of the VH of BK198.
  • SEQ ID NO 33 represents the amino acids sequence of the VL of BK198.
  • SEQ ID NO 34 represents the amino acids sequence of the heavy chain of BK198.
  • SEQ ID NO 35 represents the amino acids sequence of the light chain of BK198.
  • SEQ ID NO 36 represents the nucleic acids sequence of the VH of BK198.
  • SEQ ID NO 37 represents the nucleic acids sequence of the VL of BK198.
  • SEQ ID NO 38 represents the nucleic acids sequence of the heavy chain of the BK198.
  • SEQ ID NO 39 represents the nucleic acids sequence of the light chain of the BK198.
  • SEQ ID NO 40 represents the amino acids sequence of the HCDR1 of BK206.
  • SEQ ID NO 41 represents the amino acids sequence of the HCDR2 of BK206.
  • SEQ ID NO 42 represents the amino acids sequence of the HCDR3 of BK206.
  • SEQ ID NO 43 represents the amino acids sequence of the LCDR1 of BK206.
  • SEQ ID NO 44 represents the amino acids sequence of the LCDR3 of BK206.
  • SEQ ID NO 45 represents the amino acids sequence of the VH of BK206.
  • SEQ ID NO 46 represents the amino acids sequence of the VL of BK206.
  • SEQ ID NO 47 represents the amino acids sequence of the heavy chain of BK206.
  • SEQ ID NO 48 represents the amino acids sequence of the light chain of BK206.
  • SEQ ID NO 49 represents the nucleic acids sequence of the VH of BK206.
  • SEQ ID NO 50 represents the nucleic acids sequence of the VL of BK206.
  • SEQ ID NO 51 represents the nucleic acids sequence of the heavy chain of the BK206.
  • SEQ ID NO 52 represents the nucleic acids sequence of the light chain of the BK206.
  • SEQ ID NO 53 represents the amino acids sequence of the HCDR1 of BK256.
  • SEQ ID NO 54 represents the amino acids sequence of the HCDR2 of BK256.
  • SEQ ID NO 55 represents the amino acids sequence of the HCDR3 of BK256.
  • SEQ ID NO 56 represents the amino acids sequence of the LCDR1 of BK256.
  • SEQ ID NO 57 represents the amino acids sequence of the LCDR3 of BK256.
  • SEQ ID NO 58 represents the amino acids sequence of the VH of BK256.
  • SEQ ID NO 59 represents the amino acids sequence of the VL of BK256.
  • SEQ ID NO 60 represents the amino acids sequence of the heavy chain of BK256.
  • SEQ ID NO 61 represents the amino acids sequence of the light chain of BK256.
  • SEQ ID NO 62 represents the nucleic acids sequence of the VH of BK256.
  • SEQ ID NO 63 represents the nucleic acids sequence of the VL of BK256.
  • SEQ ID NO 64 represents the nucleic acids sequence of the heavy chain of the BK256.
  • SEQ ID NO 65 represents the nucleic acids sequence of the light chain of the BK256.
  • SEQ ID NO 66 represents the amino acids sequence of the HCDR1 of BK293.
  • SEQ ID NO 67 represents the amino acids sequence of the HCDR2 of BK293.
  • SEQ ID NO 68 represents the amino acids sequence of the HCDR3 of BK293.
  • SEQ ID NO: 69 represents the amino acids sequence of the LCDR1 of BK293.
  • SEQ ID NO: 70 represents the amino acids sequence of the LCDR3 of BK293.
  • SEQ ID NO: 71 represents the amino acids sequence of the VH of BK293.
  • SEQ ID NO: 72 represents the amino acids sequence of the VL of BK293.
  • SEQ ID NO: 73 represents the amino acids sequence of the heavy chain of BK293.
  • SEQ ID NO: 74 represents the amino acids sequence of the light chain of BK293.
  • SEQ ID NO: 75 represents the nucleic acids sequence of the VH of BK293.
  • SEQ ID NO: 76 represents the nucleic acids sequence of the VL of BK293.
  • SEQ ID NO: 77 represents the nucleic acids sequence of the heavy chain of the BK293.
  • SEQ ID NO: 78 represents the nucleic acids sequence of the light chain of the BK293.
  • SEQ ID NO: 79 represents the amino acids sequence of the HCDR1 of BK304.
  • SEQ ID NO: 80 represents the amino acids sequence of the HCDR2 of BK304.
  • SEQ ID NO: 81 represents the amino acids sequence of the HCDR3 of BK304.
  • SEQ ID NO: 82 represents the amino acids sequence of the LCDR1 of BK304.
  • SEQ ID NO: 83 represents the amino acids sequence of the LCDR3 of BK304.
  • SEQ ID NO: 84 represents the amino acids sequence of the VH of BK304.
  • SEQ ID NO: 85 represents the amino acids sequence of the VL of BK304.
  • SEQ ID NO: 86 represents the amino acids sequence of the heavy chain of BK304.
  • SEQ ID NO: 87 represents the amino acids sequence of the light chain of BK304.
  • SEQ ID NO: 88 represents the nucleic acids sequence of the VH of BK304.
  • SEQ ID NO: 89 represents the nucleic acids sequence of the VL of BK304.
  • SEQ ID NO: 90 represents the nucleic acids sequence of the heavy chain of the BK304.
  • SEQ ID NO: 91 represents the nucleic acids sequence of the light chain of the BK304.
  • SEQ ID NO: 92 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV. genotype la.
  • SEQ ID NO: 93 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype Ib2.
  • SEQ ID NO: 94 represents the amino acids sequence of the VP1 protein of the DNEQ BKPyV genotype Ib2.
  • SEQ ID NO: 95 represents the amino acids sequence of the VP1 protein of the VVAE BKPyV genotype Ib2.
  • SEQ ID NO: 96 represents the amino acids sequence of the VP1 protein of the D58Del_59A BKPyV genotype Ib2.
  • SEQ ID NO: 97 represents the amino acids sequence of the VP1 protein of the E61K BKPyV genotype Ib2.
  • SEQ ID NO: 98 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype II.
  • SEQ ID NO: 99 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype III.
  • SEQ ID NO: 100 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype IVc2.
  • SEQ ID NO: 101 represents the amino acids sequence of the VP1 protein of the E327K BKPyV genotype IVc2.
  • SEQ ID NO: 102 represents the amino acids sequence of the VP1 protein of the SDKAN BKPyV genotype IVc2.
  • SEQ ID NO: 103 represents the amino acids sequence of the VP1 protein of the wildtype MPyV.
  • SEQ ID NO: 104 represents the nucleic acids sequence of the primer CMV-F.
  • SEQ ID NO: 105 represents the nucleic acids sequence of the primer pEGFP-Nl-R.
  • SEQ ID NO: 106 represents the amino acids sequence of the VP1 protein of the K172A BKPyV genotype Ib2.
  • BKPyV BK polyomavirus
  • a or “an” entity refers to one or more of that entity; for example, “a nucleotide sequence,” is understood to represent one or more nucleotide sequences.
  • the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably herein.
  • aspects and embodiments of the present disclosure described herein include “comprising”, “having”, “consisting of’, and “consisting essentially of’ aspects and embodiments.
  • the words “have” and “comprise,” or variations such as “has,” “having”, “comprises”, or “comprising”, will be understood to imply the inclusion of the stated element(s) (such as a composition of matter or a method step) but not the exclusion of any other elements.
  • the term “consisting of’ implies the inclusion of the stated element(s), to the exclusion of any additional elements.
  • antibody As used herein the term “antibody” have the same meaning and will be used equally in the present description.
  • the term “antibody” as used herein refers to isolated or recombinant immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds an antigen, such as VP1 capsid protein.
  • the term antibody encompasses not only whole antibody molecules, but also antibody fragments as well as variants (including derivatives) of antibodies and antibody fragments.
  • two heavy chains are linked to each other by disulphide bonds and each heavy chain is linked to a light chain by a disulphide bond.
  • the light chain includes two domains, a variable domain (VL) and a constant domain (CL).
  • the heavy chain includes four domains, a variable domain (VH) and three constant domains (CHI, CH2 and CH3, collectively referred to as CH).
  • VL variable domain
  • VH variable domain
  • CH constant domain
  • the constant region domains of the light (CL) and heavy (CH) chains confer important biological properties such as antibody chain association, secretion, trans-placental mobility, complement binding, and binding to Ec receptors (EcR).
  • the Fv fragment is the N-terminal part of the Fab fragment of an immunoglobulin and consists of the variable portions of one light chain and one heavy chain.
  • the specificity of the antibody resides in the structural complementarity between the antibody combining site and the antigenic determinant.
  • Antibody combining sites are made up of residues that are primarily from the hypervariable or complementarity determining regions (CDRs). Occasionally, residues from nonhypervariable or framework regions (FR) can participate to the antibody binding site or influence the overall domain structure and hence the combining site.
  • Complementarity Determining Regions or CDRs refer to amino acid sequences which together define the binding affinity and specificity of the natural Fv region of a native immunoglobulin binding site.
  • the light and heavy chains of an immunoglobulin each have three CDRs, designated CDR1-L, CDR2-L, L- CDR3-L and CDR1-H CDR2-H, CDR3-H, respectively.
  • An antigen-binding site therefore, typically includes six CDRs, comprising the CDR set from each of a heavy and a light chain V region.
  • Framework Regions refer to amino acid sequences interposed between CDRs.
  • an antibody includes, but is not limited to, monoclonal antibodies, human antibodies, humanized antibodies, camelid antibodies and chimeric antibodies.
  • the antibodies can be of any isotype/class (e.g., IgG, IgE, IgM, IgD, IgA and IgY), or subclass (e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2).
  • an antibody of the present disclosure refers to an anti-BKPyV antibody.
  • an antibody of the present disclosure refers to an antibody binding a BKPyV VP1 capsid protein.
  • an antibody of the present disclosure may refer to BK120 antibody having the amino acid sequences as set forth in Table 3, BK106-1 antibody having the amino acid sequences as set forth in Table 4, BK198 antibody having the amino acid sequences as set forth in Table 5, BK206 antibody having the amino acid sequences as set forth in Table 6, BK256 antibody having the amino acid sequences as set forth in Table 7, BK293 antibody having the amino acid sequences as set forth in Table 8 and/or BK304 antibody having the amino acid sequences as set forth in Table 9.
  • the term “antibody” specifically includes an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, a combination of at least two antibodies binding a BKPyV VP1 capsid protein, or a multi-specific antibody binding a BKPyV VP1 capsid protein as described herein.
  • BKPyV BK polyomavirus
  • an "isolated" compound or entity such as an antibody or antigen-binding fragment thereof, a vector or a cell, is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the compound or entity, and may include enzymes, hormones, and other proteinaceous or non- proteinaceous solutes.
  • the compound or entity is partially or substantially purified.
  • the compound or entity may be purified to greater than about 95% by weight of antibody as determined by the Lowry method, for example, more than 95%, 96%, 97%, 98% or 99% by weight of compound or entity.
  • an isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody's natural environment is not present.
  • an isolated antibody may be prepared by at least one purification step.
  • nucleic acid sequence as applied to an antibody, or an antigen-binding fragment thereof, a nucleic acid sequence, an expression vector or a host cell means that those are the products of various combinations of in vitro cloning, restriction, ligation steps, and other genetic engineering procedures.
  • monoclonal antibody refers to a preparation of antibody molecules of single molecular composition.
  • a monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope on an antigen, in particular VP1 capsid protein of BKPyV.
  • CDRs complementarity-determining regions
  • the expression “complementarity-determining regions” or “CDRs” interchangeably refers to the hypervariable regions of VL and VH.
  • the CDRs are the target protein-binding site of the antibody chains that harbors specificity for such target protein, in particular BKPyV VP1 capsid protein.
  • CDRs can be referred to by their region and order.
  • VHCDR1”, “CDRLH” or “H-CDR1” refer to the first CDR of the heavy chain variable region.
  • the CDRs are structurally complementary to the epitope of the target protein, such as BKPyV VP1 protein, and are thus directly responsible for the binding specificity.
  • the positions of the CDRs and framework regions can be determined using various well-known definitions in the art, e.g., Kabat, Chothia, IMGT, and AbM.
  • the amino acid residues of the antibody of the disclosure are numbered according to the IMGT numbering system.
  • the IMGT unique numbering has been defined to compare the variable domains whatever the antigen receptor, the chain type, or the species (Lefranc M.-P., Immunology Today, 18, 509 (1997) ; Lefranc et al. Dev. Comp. Immunol., 27, 55-77 (2003)).
  • the conserved amino acids always have the same position, for instance cysteine 23, tryptophan 41, hydrophobic amino acid 89, cysteine 104, phenylalanine or tryptophan 118.
  • the IMGT unique numbering provides a standardized delimitation of the framework regions (FR1-IMGT: positions 1 to 26, FR2- IMGT: 39 to 55, FR3-IMGT: 66 to 104 and FR4-IMGT: 118 to 128) and of the complementarity determining regions: CDR1-IMGT: 27 to 38, CDR2-IMGT: 56 to 65 and CDR3-IMGT: 105 to 117. If the CDR3-IMGT length is less than 13 amino acids, gaps are created from the top of the loop, in the following order 111, 112, 110, 113, 109, 114, etc.
  • CDR3-IMGT length is more than 13 amino acids
  • additional positions are created between positions 111 and 112 at the top of the CDR3-IMGT loop in the following order 112.1,111.1, 112.2, 111.2, 112.3, 111.3, etc. (http://www.imgt.org/IMGTScientificChart/Nomenclature/ IM GT -FRCDRdefinition.html).
  • variable domains of both the light (VL) and heavy (VH) chain portions determine antigen recognition and specificity.
  • the constant domains of the light chain (CL) and the heavy chain (CHI, CH2, or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, and the like.
  • the numbering of the constant region domains increases as they become more distal from the antigen binding site or amino-terminus of the antibody.
  • the N-terminus is a variable region and at the C-terminus is a constant region; the CH3 and CL domains actually comprise the carboxy-terminal domains of the heavy and light chain, respectively.
  • identity may mean the percentage of nucleotide bases or amino acid residues in the candidate sequence that are identical with the residue of a corresponding sequence to which it is compared, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent identity for the entire sequence, and not considering any conservative amino acid substitutions as part of the sequence identity. Neither N-terminal or C-terminal extensions nor insertions shall be construed as reducing identity or homology. Methods and computer programs for the alignment are available and well known in the art. Sequence identity may be measured using sequence analysis software. Two examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms (the National Center for Biotechnology Information). The BLASTN program is use for nucleotide sequences, whereas the BLASTP program is used for amino acid sequences.
  • antigen binding site or “antigen binding region” as used in the present disclosure refers to the part of the antibody which comprises the area which specifically binds to and is complementary to part or all of an antigen. Where an antigen is large, an antibody may only bind to a particular part of the antigen, which part is termed on epitope.
  • An antigen binding region may be provided by one or more antibody variable domains. Preferably, an antigen binding region is made of the association of an antibody light chain variable domain (VL) and an antibody heavy chain variable domain (VH).
  • VL antibody light chain variable domain
  • VH antibody heavy chain variable domain
  • an antigen can have one or more than one epitope.
  • an antigen may be a VP1 capsid protein of a human BKPyV.
  • an antigen of the disclosure may be a VP1 capsid protein selected among BKPyV VP1 gl wt, BKPyV VP1 gll wt, BKPyV VP1 gill wt, BKPyV VP1 gIV wt, BKPyV glb2 wt, BKPyV VP1 glb2 DNEQ, BKPyV glb2 VVAE, BKPyV VP1 gIVc2 wt, BKPyV VP1 gIV c2 SDKAN, BKPyV VP1 glb2 D58delP59A, BKPyV VP1 glb2 E61K, BKPyV
  • affinity means the strength of the binding of an antibody to an epitope presented on an antigen.
  • the affinity of an antibody is given by the dissociation constant KD, defined as [Ab] x [Ag] I [Ab-Ag], where [Ab-Ag] is the molar concentration of the antibody-antigen complex, [Ab] is the molar concentration of the unbound antibody and [Ag] is the molar concentration of the unbound antigen.
  • KD dissociation constant
  • Ka is defined by 1/Kd.
  • Ka and KD values determined by such standard methods measure the functional affinity, or avidity of the interaction.
  • an antibody is considered to bind an antigen when a functional binding affinity (KD), preferably measured by surface plasmon resonance, is of 10’ 8 mol/1 (M) or less, preferably 10’ 9 M to 10 12 M.
  • KD functional binding affinity
  • parenteral administration designates administration routes other than enteral and topical administration, generally by injection, by intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, epidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal routes, including, without limitation, injections, and perfusions.
  • treat refers to the administration or consumption of a compound or composition according to the disclosure with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a disorder, the symptoms of the condition, the likelihood of the disorder, or to prevent or delay the onset of the symptoms, complications, or otherwise arrest or inhibit further development of the disorder in a statistically significant manner. More particularly, “treating” or “treatment” includes any approach for obtaining beneficial or desired results in a subject’s BKPyV infection condition.
  • Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions of BKPyV infection-associated disease, diminishment or reduction of the extent of a BKPyV infection-associated disease or of a symptom-associated BKPyV infection, stabilizing, i.e., not worsening, the state of a BKPyV infection-associated disease or of a symptom-associated BKPyV infection prevention of a BKPyV infection-associated disease, delay or slowing of BKPyV infection-associated disease or symptom-associated BKPyV infection progression, amelioration or palliation of the BKPyV infection-associated disease state, diminishment of the reoccurrence of BKPyV infection- associated disease, and remission, whether partial or total and whether detectable or undetectable.
  • treatment includes any cure, amelioration, or reduction of a BKPyV infection-associated disease or related symptom.
  • a “reduction” of a symptom or a disease means decreasing of the severity or frequency of the disease or symptom, or elimination of the disease or symptom.
  • the terms “prevent” or “preventing” (and grammatical variants thereof) with respect to a disease or disorder relate to prophylactic treatment of a disease, e.g., in a subject suspected to have the disease, or at risk for developing the disease. Prevention may include, but is not limited to, preventing, or delaying onset or progression of the disease and/or maintaining one or more symptoms of the disease at a desired or sub-pathological level.
  • the term “prevent” does not require the 100% elimination of the possibility or likelihood of occurrence of the event. Rather, it denotes that the likelihood of the occurrence of the event has been reduced in the presence of a composition or method as described herein.
  • prevention refers to a decrease in the risk of occurrence of a BKPyV infection- associated disease or symptom in a subject.
  • the prevention may be complete, i.e., no detectable symptoms or disease, or partial, such that fewer symptoms or less severity of the disease are observed than would likely occur absent treatment.
  • disorders and “disease” are used interchangeably herein and comprise any undesired physiological change in a subject or a cell.
  • the term “infection” refers to the presence of BKPyV in a subject, whether the infected subject presents clinical signs of the infection, in particular by the manifestation of a disorder, or not.
  • Subjects that are infected with BKPyV but have not become clinically manifest are called asymptomatic subjects.
  • subjects that are infected with BKPyV may be asymptomatic, i.e., subjects have no clinical signs of a BKPyV infection- associated disease.
  • the terms “therapeutically effective amount” and “prophylactically effective amount” refer to an amount that provides a therapeutic benefit in the treatment, prevention, or management of pathological processes considered.
  • the specific amount that is therapeutically effective can be readily determined by an ordinary medical practitioner and may vary depending on factors such as the type and stage of pathological processes considered, the subject’s medical history and age, and the administration of other therapeutic agents.
  • the term “patient” or “subject” is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats).
  • the subject may be a human.
  • a subject may be an immunocompromised subject.
  • immunocompromised subject or “immunosuppressed subject” refers to a subject having a weakened immune system.
  • the immune system of an immunocompromised subjects has a reduced ability to fight virus or bacterial infections and other diseases.
  • An immunocompromised subject includes subjects suffering from a Primary Immune Deficiency Diseases, subjects suffering from a Secondary Immune Deficiency Diseases, subjects being or has been treated with an immunosuppressive therapy; subjects being or has been treated with a chemotherapeutic agent or subjects who has been transplanted.
  • Primary Immune Deficiency Diseases refers to an inherited immune disorders resulting from genetic mutations, usually present at birth and diagnosed in childhood.
  • Secondary Immune Deficiency Diseases refers to an acquired immunodeficiency as a result of disease or environmental factors, such as AIDS, cancer, diabetes, malnutrition, or medical treatment, such as anticancer drugs, radiation therapy, or stem cell or organ transplant.
  • nucleic acid sequence is used herein interchangeably with the term “polynucleotide” and refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or doublestranded form.
  • the term encompasses isolated or recombinant nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides.
  • Examples of such analogs include, without limitation, phosphorothioates, phosphor amidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
  • PNAs peptide-nucleic acids
  • polypeptide and protein are used interchangeably herein to refer to a sequence of amino acid residues.
  • the terms apply to amino acid sequences in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid sequences and non-naturally occurring amino acid sequence. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.
  • ICso half-maximal inhibitory concentration
  • IC50 is the concentration of antibody at which 50% of the available binding sites on the VP1 antigen are occupied.
  • target cell herein means any cell that may be infected by a BKPyV.
  • a target cell also includes a cell expressing a target antigen that can be recognized by an anti- BKPyV antibody according to the present disclosure.
  • a target cell may be a kidney cell.
  • a target cell used as cellular model in the binding and neutralization assays may be a 293TT cell line or an RS cell line.
  • virus-like particles are an assembly of VP1 pentamers into viral capsids. VLPs are composed of 72 VP1 pentamers. VLPs are structurally very similar to actual virus without the minor capsid proteins (VP2 and VP3) as well as the viral DNA genome, and therefore are non-infectious. VLPs are useful as viral epitopes are presented in a similar conformation to the actual virus.
  • BK polyomavirus BKPyV
  • the antibodies described herein specifically bind to the capsid protein VP1 of different BK polyomaviruses and BK pseudotype polyoma viruses. Further, the antibodies neutralize BKPy virus and BKPy pseudotype viruses. They can therefore be effectively used for preventing and treating an infection with BKPyV and/or a disorder induced by a BKPyV infection.
  • a pseudovirus or a pseudotype virus is a recombinant viral particle which in addition to its core/backbone, may incorporate envelope proteins derived from different viruses; moreover, the genes inside the pseudovirus are usually altered or modified so that they are unable to produce the surface protein on their own. As such, an additional plasmid or stable cell line expressing the surface proteins is needed to make the pseudovirus.
  • Pseudoviruses are capable of infecting susceptible cells, but they only replicate for 1 round in the infected host cells. Compared with wild-type viruses, pseudoviruses can be safely handled in biosafety level (BSL)- 2 laboratories and are usually easier to manipulate experimentally. Nevertheless, the conformational structure of pseudoviral surface proteins bears high similarity to that of the native viral proteins, and these surface proteins can effectively mediate viral entry into host cells.
  • BK polyoma virus (BKPyV” and “BK virus” are herein used interchangeably and intend to refer to a member of the family Polyomaviridae, genus Betapolyomavirus (Helle, Francois et al. Viruses vol. 9,11 327. 3 Nov. 2017).
  • BK polyomaviruses are icosahedral, non-enveloped, circular double-stranded DNA viruses with a genome of around 5,300 base pairs. They measure approximately 40-45 nM in diameter (Bennett et al., Microbes and Infection. 2012:14(9):672-683). Further, the BKPyVs have been associated with nephropathy, renal dysfunction such as hemorrhagic cystitis, pneumonitis, retinitis, and meningoencephalitis.
  • the BKPyV may be a whole BKPyV, BKPyV virus-like particles (VLPs) composed only of VP1 protein, or pseudotype virus that are incorporate all capsid proteins and are able to infect susceptible target cells. Up to 90% of the population as a whole is persistently infected with BKPyV. The virus reactivates and is asymptomatically shed into the urine in 5-10% of immunocompetent adults at any given time.
  • VLPs BKPyV virus-like particles
  • genotypes Based on the sequence of the VP1 gene, four genotypes have been described, corresponding to the four serologically differentiated subtypes I-IV, with different prevalence and geographic distribution. In each genotype, numerous serotype variants exist. The most common genotype worldwide is genotype I, with a frequency of about 80%, followed by genotype IV (about 15%), while genotypes II and III are isolated only sporadically. The distribution of the molecular variants of the virus is associated with the region of origin. BKPyV subtype la is most common in Africa, Ib-1 in Southeast Asia, and Ib-2 in Europe, while Ic is the most common variant in Northeast Asia.
  • the antibodies or antigen-binding fragments thereof are able to specifically bind and neutralize the BKPyV genotype I wildtype (la, Ib-1, Ib- 2 and Ic), as well as the newly identified variant BKPyV genotype Ib2-D58Del_59A and the known variants BKPyV genotype Ib2-E61K, BKPyV genotype Ib2-DNEQ and BKPyV genotype Ib2-VVAE, the BKPyV genotype II wildtype, the BKPyV genotype III wildtype, the BKPyV genotype IV wildtype (IVa-1, IVa-2, IVb-1, IVb-2, IVc-1 and IVc-2), as well as the variants BKPyV genotype IVc2-E327K and BKPyV genotype IVc2-SDKAN.
  • the BK120 antibody or antigenbinding fragments thereof is also able to specifically bind and/or neutralize the variant BKPyV genotype Ib2-K172A.
  • BKPyV variant refers to a wild-type BKPy virus wherein some mutations are present on the VP1 capsid proteins of the wild-type BKPyV.
  • BKPyV variant encompasses naturally occurring BKPyV variants and in vitro selected BKPyV variants.
  • the expression “naturally occurring BKPyV” intends to refer to variants of BKPyV obtained from subjects, such as genotype Ib2-DNEQ, the BKPyV genotype Ib2-VVAE, and the BKPyV genotype IVc2-SDKAN (McIlroy et al. Viruses 2020).
  • the term 'in vitro selected BKPyV refers to variants of BKPyV obtained by passaging viruses in vitro cell culture under suitable conditions, such as the BKPyV genotype Ia-D58Del_59A, and also the BKPyV genotype Ib2-E61K, the BKPyV genotype Ib2-K172A and the BKPyV genotype IVc2-E327K (Lindner et al. Immunity. 2019;50:668-676.e5 and McIlroy et al. Viruses. 2020 Jul 29; 12(8): 824.).
  • VP1 or “VP1 capsid protein” refers to the major polyoma virus capsid subunit protein. In the virus capsid, the monomers of VP1 oligomerize in pentamers. Expression of the VP1 protein alone is sufficient to produce VLPs that have the same antigenic properties as the infectious virus, since the minor capsid proteins VP2 and VP3 are not exposed on the surface of the virion.
  • VLPs Virus-like particles
  • viruses are molecules that closely resemble viruses but are non-infectious because they contain no viral genetic material. They can be naturally occurring or synthesized through the individual expression of viral structural proteins, which can then selfassemble into the virus-like structure.
  • the antibodies of the disclosure, or the antigen-binding fragments thereof, specifically bind to the VP1 sequences of the BKPyV of Table 1.
  • the VP1 coding region shows very high similarity (over 95%) in all genotypes of BKPyV, but the similarity between the amino acid residues from 61E to 83R is only 61-70%
  • the antibodies of the disclosure, or the antigen-binding fragments thereof specifically may bind to variants VP1 sequences comprising an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequences of the VP1 of BKPyV of Table 1.
  • the antibodies of the disclosure, or the antigen-binding fragments thereof neutralize BKPyV comprising VP1 sequences comprising an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequences of the VP1 of BKPyV of Table 1.
  • variants and/or mutants are described by reference to the amino acid sequences of the VP1 of BKPyV of Table 1 which are amino acid sequences of reference. Such a description by reference is based on the prerequisite of optimal sequence alignment in order to determine the amino acid in the variant sequence that corresponds to the amino acid defined as being in a specific position in the amino acid of reference.
  • variants and/or mutants are also described by percent identity with a sequence of reference. Percent identity between two amino acid sequences or two nucleotide sequences is determined with standard alignment algorithms as those described below.
  • a sequence identity refers to the similarity between amino acid sequences expressed in terms of the similarity between the sequences. Sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar the two sequences are. Variants of a polypeptide will possess a relatively high degree of sequence identity when aligned using standard methods.
  • sequence alignment can be achieved and percent identity can be determined Basic Local Alignment Tools (BLASTs, including BLASTP for amino acid sequence alignment and BLASTN for nucleotide sequence alignment; described in Altschul et al., (1990) J. Mol. Biol., 215: 403) available on the National Center for the Biotechnology Information (NCBI) web site at http://www.ncbi.nlm.nih.gov/BLAST and may be used using the default parameters [BLASTP: Expect value E: 10; Word size: 3; Matrix: BLOSUM62; Cost gap: Existence 11, Extension 1; and BLASTN: algorithm by default: Megablast; Expect value E: 10; Word size: 28; Match scores: 1; Mismatch score: -2;
  • variant and mutant amino acid sequences include amino acid sequences that have at least about 90% sequence identity with an amino acid sequence defined herein.
  • a variant or mutant amino acid sequence may have at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to an amino acid sequence as defined herein.
  • Amino acid sequence identity is defined as the percentage of amino acid residues in the variant sequence that are identical with the amino acid residues in the reference sequence, after aligning the sequences and if necessary, introducing gaps, to achieve the maximum percent sequence identity, and not considering any conservative substitution as part of the sequence identity. Standard alignment algorithms cited above are useful in this regard.
  • the antibodies of the disclosure, or the antigen-binding fragments thereof specifically may bind to variants VP1 sequences of Table 1.
  • the antibodies of the disclosure, or the antigen-binding fragments thereof neutralize BKPyV comprising VP1 sequences of Table 1.
  • An object of the disclosure is an isolated, or recombinant, BKPy VP1 protein of amino acid sequence SEQ ID NO: 96 or an isolated, or recombinant, VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • BKPy VP1 protein may be useful for producing VLPs and BK PyV pseudotypes usable in neutralizing assay, antibody screening, in diagnosing methods for measuring an immune response to a BK PyV infection in a patient in need thereof, such as KTx patient.
  • the present disclosure also relates to an in vitro selected BKPyV variant having a VP1 capsid protein having an amino acid sequence as set forth in SEQ ID NO: 96.
  • the disclosure relates to an isolated or recombinant BKPy virus comprising a
  • Recombinant BKPyV virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may be obtained by any known techniques in the art.
  • a recombinant BKPyV virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may be a virus, a virus-like particle (VLP), or a pseudotype virus.
  • VLP virus-like particle
  • a recombinant BKPyV may be a pseudotype virus.
  • a recombinant BKPyV virus may be prepared in any suitable cells or cell lines such as human embryonic kidney cells (HEK 293TT cells).
  • a nucleic acid sequence encoding a VP1 protein of amino acid sequence SEQ ID NO: 96 or a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may be inserted in any expression vector known in the art, for example an expression plasmid such as pCAG.
  • the minor capsid proteins VP2 and VP3 may be provided in separate plasmids, such as ph2b and ph3b plasmids, or may be inserted in the same expression plasmid used for the VP1 protein.
  • the vector expression may be transfected in suitable cells, such as HEK 293TT cells, using any nucleic acid cell delivery methods.
  • the viral particles may be harvested.
  • Harvest may be carried out by trypsinization, centrifugation and then lysis of the cells in a hypotonic lysis buffer, for example consisting of 25 mM Sodium Citrate pH 6.0, 1 mM CaC12, 1 mM MgC12 and 5mM KC1. Further to exposure to the lysis buffer, the cells may be sonicated.
  • a hypotonic lysis buffer for example consisting of 25 mM Sodium Citrate pH 6.0, 1 mM CaC12, 1 mM MgC12 and 5mM KC1. Further to exposure to the lysis buffer, the cells may be sonicated.
  • the lysate may be clarified, for example by centrifugation.
  • Purification of the viral particles may be carried on density gradient medium, by any known techniques in the art.
  • An object of the invention relates to antibodies, or fragment thereof, binding to the VP1 capsid protein of BKPy viruses.
  • antibody and “antigen-binding fragment” are used interchangeably.
  • the antibodies described herein may be isolated antibodies.
  • the antibodies described herein may be recombinant antibodies.
  • the antibodies may be monoclonal antibodies.
  • the antibodies may be human antibodies or human engineered antibodies.
  • Antigen-binding fragments may be Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, or diabodies.
  • binding fragment refers to a fragment of an intact antibody that retain the ability to specifically binds to a given antigen/ligand.
  • binding fragments include a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; a Fab’ fragment, a monovalent fragment consisting of the VL, VH, CL, CHI domains and hinge region; a E(ab’)2 fragment, a bivalent fragment comprising two Lab’ fragments linked by a disulphide bridge at the hinge region; an Ed fragment consisting of VH domains of a single arm of an antibody; a single domain antibody (sdAb) fragment (Ward et al., 1989 Nature 341:544-546), which consists of a VH domain or a VL domain; and an isolated complementary determining region (CDR).
  • Fab fragment a monovalent fragment consisting of the VL, VH, CL and CHI domains
  • a Fab’ fragment a monovalent fragment consisting of the VL,
  • the two domains of the Fv fragment, VL and VH are coded for by separate genes, they can be joined, using recombinant methods, by an artificial peptide linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (ScFv); see, e.g., Bird et al., 1989 Science 242:423-426; and Huston et al., 1988 proc. Natl. Acad. Sci. 85:5879-5883).
  • dsFv is a VH::VL heterodimer stabilized by a disulfide bond.
  • Divalent and multivalent antibody fragments can form either spontaneously by association of monovalent scFvs or can be generated by coupling monovalent scFvs by a peptide linker, such as divalent sc(Fv)2.
  • Such single chain antibodies include one or more antigen biding portions or fragments of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as are intact antibodies.
  • a unibody is another type of antibody fragment lacking the hinge region of IgG4 antibodies. The deletion of the hinge region results in a molecule that is essentially half the size of traditional IgG4 antibodies and has a univalent binding region rather than the bivalent biding region of IgG4 antibodies.
  • UniBodies may be obtained by reference to WO 2007/059782, which is incorporated by reference in its entirety.
  • Antigen binding fragments can be incorporated into single domain antibodies, SMIP, maxibodies, minibodies, intrabodies, diabodies, triabodies and tetrabodies (see, e.g., Hollinger and Hudson, 2005, Nature Biotechnology, 23, 9, 1126-1136).
  • diabodies tribodies or tetrabodies” refers to small antibody fragments with multivalent antigen-binding sites (2, 3 or four), which fragments comprise a heavy -chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL).
  • VH heavy -chain variable domain
  • VL light-chain variable domain
  • Antigen binding fragments can be incorporated into single chain molecules comprising a pair of tandem Fv segments (VH-CH1- VH-CH1) Which, together with complementary light chain polypeptides, form a pair of antigen binding regions (Zapata et al., 1995 Protein Eng. 8(10); 1057-1062 and U.S. Pat. No. 5,641,870).
  • Fab fragments can be obtained by treating an antibody with a protease, papain. Also, the Fab can be produced by inserting DNA encoding Fab of the antibody into a vector for prokaryotic expression system, or for eukaryotic expression system, and introducing the vector into a procaryote or eucaryote (as appropriate) to express the Fab.
  • F(ab’)2 can be obtained by treating an antibody with a protease, pepsin. Also, the F(ab’)2 can be produced by binding Fab’ described below via a thioether bond or a disulphide bond.
  • Fab’ can be obtained by treating F(ab’)2 with a reducing agent, dithiothreitol. Also, the Fab’ can be produced by inserting DNA encoding Fab’ fragment of the antibody into an expression vector for prokaryote, or an expression vector for eukaryote, and introducing the vector into a prokaryote or eukaryote (as appropriate) to perform its expression.
  • An antigen-binding fragment may be variable heavy chain of a single domain antibody (VHH).
  • binding or “specifically binding” are used interchangeably and refer to the ability of an antibody or antigen-binding fragment thereof to detectably bind an epitope present on a specific antigen, such as a BKPyV VP1 capsid protein, while having relatively no or little detectable reactivity with different antigen, for example non- BKPyV VP1 capsid proteins or structures, such as the VP1 of the MPyV (Murine polyoma virus).
  • a specific antigen such as a BKPyV VP1 capsid protein
  • a VP1 of the MPyV may be of SEO ID NO: 103:
  • the antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiologic conditions.
  • Specific binding can be characterized by an equilibrium dissociation constant (KD) of at least about IxlO -6 M or less (e.g., a smaller KD denotes a tighter binding).
  • KD equilibrium dissociation constant
  • Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. Moreover, multi-specific antibodies that bind to the VP1 protein of a BKPyV and one or more additional antigens or a bi-specific that binds to two different regions of the VP1 protein of a BKPyV are nonetheless considered antibodies that “specifically bind” the VP1 protein of a BKPyV, as used herein.
  • the binding affinity of the antibodies may range from about 10 -12 M to about 5xl0 -8 M, as measured by surface plasmon resonance, e.g., BIACORETM or solution-affinity ELISA, for example as described in the Examples section.
  • the binding affinity of the antibodies on the VP1 capsid protein of gl-IV BKPyV may be of about 10.0xl0 -8 M or less.
  • the binding affinity of the antibodies on the VP1 capsid protein of gl-IV BKPyV may range from about l.OxlO -7 M to about l.OxlO" 10 M, or less.
  • the binding affinity of the antibodies on the VP1 capsid protein of gl-IV BKPyV may be of about l.OxlO -7 M or less, or of about 8.0xl0 -8 M or less, or of about 6.5xl0 -8 M or less, or of about 5.0xl0 -8 M or less, or of about 3.0xl0 -8 M or less, or of about 2.0xl0 -8 M or less, or of about 1.5xl0 -8 M or less, or of about l.OxlO -8 M or less, or of about 9.OxlO -10 M or less, or of about 8.0xl0 -9 M or less, or of about 5.0xl0 -9 M or less, or of about 2.5xl0 -9 M or less, or of about 2.0xl0 -9 M or less, or of about 1.5xl0 -9 M or less, or of about l.OxlO -9 M or less, or of about 8.0xl0 -1 ° M
  • a BKPyV neutralizing antibody intends to refer to an antibody which is capable of specifically binding to an antigen of BKPyV and substantially inhibiting or eliminating infection of a target cell by the BKPyV, or reducing the infectious titer, as demonstrated for example by the absence of viral gene expression of the BKPyV.
  • a neutralizing antibody will inhibit infection activity of a BKPyV, or reduce an infection titer, by at least about 50%, or at least about 60%, or at least about 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%.
  • the neutralizing antibody of the invention is especially useful in therapeutic applications: to prevent or treat infections with BKPyV and/or disorders induced by a BKPyV infection.
  • the ability of an antibody for neutralizing a BKPyV may be determined by the measurement of the IC50, i.e., the concentration of an antibody reducing by 50% the infection of cells by the virus.
  • a neutralization assay may be carried out with a virus or a pseudotype virus, as described in the Examples section.
  • a serial dilution of a predetermined amount of infecting viral particles (virus or pseudotype virus) is used to infect cultured cells.
  • useable cells may be HEK293 TTT cells or RS cells.
  • a 2-, 3-, 4-, 5-, 6-, 7-fold dilution may be used.
  • the antibody to be assayed may be placed in contact with the viral particles before adding the viral particles to the cultured cells.
  • antibody may be placed in contact with the cultured cells before addition of the viral particles.
  • the antibody may be placed in contact with the cultured cells after addition of the viral particles to the cells.
  • the infected cells may be numbered, and the obtained value is compared with, or normalized with, the value obtained with cultured cells infected by viral particles in the absence of the antibody to be assayed .
  • the neutralization activity of the antibodies may be expressed as the IC50.
  • the neutralization activity of the antibodies on the gl-IV BKPyV expressed as the concentration inhibiting at least 50% of the infection, IC50, measured, for example on HEK293 TTT or RS cells, as described in the Examples section, may be of about 50 nM or less.
  • the neutralization activity of the antibodies on the gl-IV BKPyV may range from about 0.01 nM to about 50 nM.
  • the antibodies described herein may neutralize the gl-IV BKPyV in a target cell with a IC50 of about 50.0 nM or less, of about 45.0 nM or less, of about 40.0 nM or less, or of about 35.0 nM or less, or of about 30.0 nM or less, or of about 25.0 nM or less, or of about 20.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 5.0 or less, or of about 3.5 nM or less, or of about 2.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.15 nM or less, or of about 0.10 nM or less, or of about 0.08 nM or less, or of about
  • the antibodies may have a neutralization activity on BKPy virus comprising a VP1 amino acid sequence SEQ ID NO: 97, SEQ ID NO: 98 or SEQ ID NO: 99, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of less than about 40 nM, or less than 30 nM, or less than 15 nM, or less than 10 nM, or less than 5 nM, or less than 1 nM.
  • the antibodies may have a neutralization activity on BKPy virus comprising a VP1 amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101 or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said amino acid sequences, with an IC50 of less than about 40 nM, or less than 30 nM, or less than 15 nM, or less than 10 nM, or less than 5 nM, or less than 1 nM.
  • the antibodies may have a neutralization activity on BKPy virus comprising a VP1 amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101 or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of less than about 35 nM, or less than about 30 nM, or less than about 20 nM, or less than about 15 nM, or less than about 10 nM, or less than about 5 nM, or less than about 4 nM, or less than about 2.5 nM, or less than about 2 nM, or less than about 1.5 nM, or less than about 1 nM, or less than about 0.8 nM, or less than about 0.5 nM, or less than about 0.3
  • An antibody may bind and neutralize at least two BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101, or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
  • An antibody may bind and neutralize at least two BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101, or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of less than about 35 nM, or less than about 30 nM, or less than about 20 nM, or less than about 15 nM, or less than about 10 nM, or less than about 5 nM, or less than about 4 nM, or less than about 2.5 nM, or less than about 2 nM, or less than about 1.5 nM, or less than about 1 nM, or less than about 0.8 nM, or less than about 0.5 nM, or less than
  • the BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 101, or SEQ ID NO: 106 may be selected among BKPyV- D58Del_59A, BKPyV-E61K, BKPyV-E327K and BKPyV-K172A.
  • An antibody disclosed herein may bind and neutralize at least two BKPy virus selected among BKPyV-D58Del_59A, BKPyV-E61K, BKPyV-E327K and BKPyV-K172A.
  • An antibody disclosed herein may be an antibody from group (i), (ii), (iii), (iv), (v), (vi) or (vii) defined as comprising:
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution
  • a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution
  • a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution
  • a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution
  • a CDR2- H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution
  • a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution
  • a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution
  • a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution
  • a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • a “conservative substitution” refers to the replacement of an amino acid with another amino acid of similar chemical structure, similar chemical properties or similar sidechain volume in an amino acid sequence.
  • a conservative amino acid substitution will not substantially change the functional properties of a protein.
  • the amino acids introduced may have similar polarity, hydrophilicity or hydrophobicity to the amino acids they replace.
  • Conservative amino acid changes are well known in the art. Conservative amino acid changes may also be determined by reference to the Point Accepted Mutation (PAM) or BLOcks Substitution Matrix (BLOSUM) family of scoring matrices for conservation of amino acid sequence.
  • PAM Point Accepted Mutation
  • BLOSUM BLOSUM family of scoring matrices for conservation of amino acid sequence.
  • conservative amino acid changes may be members of an equivalence group, being a set of amino acids having mutually positive scores in the similarity representation of the scoring matrix selected for use in an alignment of the reference and mutant polypeptide chains.
  • a conservative substitution may be a substitution of an amino acid of one class by an amino acid of the same class as presented in Table 2.
  • a conservative substitution may be a substitution of an amino acid of one class by an amino acid of another class but with a similar chemical structure, a similar chemical property and/or a similar side -chain volume.
  • a substitution mutation may be a non-conservative mutation, which replaces the amino acid of one class with an amino acid of non-similar chemical structure, non-similar chemical property and/or non-similar side-chain volume.
  • a CDR of an antibody, or antigen-binding fragment thereof may comprise more than one conservative amino acid substitution, for example at least two, or at least three.
  • the full-length heavy chain, the full-length light chain, VH, VL and CDRs of an antibody disclosed herein may comprise at least one conservative substitution.
  • the full-length heavy chain, the full-length light chain, the VH chain, the VL chain, and the CDRs of antibodies disclosed herein, or an antigen-binding fragments thereof may comprise:
  • amino acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the corresponding amino acid sequences set forth in any of Tables 3 to 9 and having CDRs sequences differing from the sequences CDRs set forth in any of Tables 3 to 9 by one conservative amino acid substitution, or
  • amino acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the corresponding amino acid sequences set forth in any of Tables 3 to 9 and having CDRs sequences identical to the sequences CDRs set forth in any of Tables 3 to 9, or
  • nucleic acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence as set forth in any of Tables 3 to 9, or
  • nucleic acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence as set forth in any of Tables 3 to 9, and having nucleic acid sequences coding for CDRs differing from the CDRs set forth in any of Tables 3 to 9 by one conservative amino acid substitution, or
  • nucleic acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence as set forth in any of Tables 3 to 9, and having nucleic acid sequences coding for CDRs identical to the CDRs set forth in any of Tables 3 to 9, or
  • Percent identity between amino acid sequences may be determined with standard alignment algorithms as those described herein.
  • sequence comparison For sequence comparison of nucleic acid sequences, typically one sequence acts as a reference sequence, to which test sequences are compared.
  • test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters are used.
  • Methods of alignment of sequences for comparison are well known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482, 1981, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443, 1970, by the search for similarity method of Pearson & Lipman, Proc.
  • PILEUP uses a simplification of the progressive alignment method of Feng & Doolittle, J. Mol. Evol. 35:351-360, 1987. The method used is similar to the method described by Higgins & Sharp, CABIOS 5:151-153, 1989.
  • a reference sequence is compared to other test sequences to determine the percent sequence identity relationship using the following parameters: default gap weight (3.00), default gap length weight (0.10), and weighted end gaps.
  • PILEUP can be obtained from the GCG sequence analysis software package, e.g., version 7.0 (Devereaux et al., Nuc. Acids Res. 12:387- 395, 1984).
  • BLAST Altschul et al., J. Mol. Biol. 215:403-410, 1990 and Altschul et al., Nucleic Acids Res. 25:3389-3402, 1977.
  • Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov).
  • the BLASTP program (for amino acid sequences) uses as defaults a word length (W) of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915, 1989).
  • an antibody disclosed herein may comprise:
  • a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
  • a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2,
  • a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3,
  • CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution
  • an antibody disclosed herein may comprise:
  • an antibody disclosed herein may comprise:
  • an antibody disclosed herein may comprise:
  • CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70.
  • an antibody disclosed herein may comprise:
  • a heavy chain comprising an amino acid sequence selected from SEQ ID NO: 8, 21, 34, 47, 60, 73and 86, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto; and/or
  • an antibody disclosed herein may comprise:
  • a heavy chain comprising an amino acid sequence selected from SEQ ID NO: 8, 21, 34, 47, 60, 73and 86, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and/
  • a light chain comprising an amino acid sequence from SEQ ID NO: 9, 22, 35, 48, 61, 74 and 87, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
  • an antibody disclosed herein may comprise:
  • a heavy chain comprising an amino acid sequence selected SEQ ID NO: 8, 21, 34, 47, 60, 73and 86, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80; and a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81; and/or
  • a light chain comprising an amino acid sequence from SEQ ID NO: 9, 22, 35, 48, 61, 74 and 87, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70.
  • Group (i) of antibodies comprising an amino acid sequence from SEQ ID NO: 9, 22, 35, 48, 61, 74 and 87, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,
  • the group (i) of antibodies is illustrated by the antibody B KI 20 of the Example section.
  • an antibody of group (i) may efficiently neutralize BKPyV escaping neutralization by both the antibodies 41F17 and P8D11.
  • the antibodies of group (i) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
  • the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
  • the conservative amino acid substitution may be as above described.
  • the antibodies of group (i) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 6.
  • the antibodies of group (i) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 6 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
  • the antibodies of group (i) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 6 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 6.
  • the antibodies of group (i) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 6.
  • the antibodies of group (i) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 7.
  • the antibodies of group (i) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 7 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
  • the antibodies of group (i) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 7 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 7.
  • the antibodies of group (i) may comprise a variable domain of light chain of sequence SEQ ID NO: 7.
  • the antibodies of group (i) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 8.
  • the antibodies of group (i) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 8 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
  • the antibodies of group (i) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 8 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 8.
  • the antibodies of group (i) may comprise a heavy chain of sequence SEQ ID NO: 8.
  • the antibodies of group (i) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 9.
  • the antibodies of group (i) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 9 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
  • the antibodies of group (i) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 9 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 9.
  • the antibodies of group (i) may comprise a light chain of sequence SEQ ID NO: 9.
  • an antibody of group (i) may be the antibody referred to as antibody BK120 in the Examples section.
  • variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 10.
  • a variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 10 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 10 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
  • variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 10.
  • variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 10.
  • variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 11.
  • a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 11 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 11 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
  • variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 11.
  • variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 11.
  • a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 12.
  • a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 12 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 12 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
  • a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 12.
  • a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 12.
  • a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 13.
  • a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 13 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
  • a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 13 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
  • a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 13.
  • a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 13.
  • the antibodies of group (i) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about l.OxlO -9 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 2.0 nM or less.
  • KD binding affinity
  • the antibodies of group (i) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about l.OxlO -9 mol/1 (M) or less, or of about 9.OxlO -10 M or less, or of about 5.0xl0 -1 ° M or less, or of about 2.5xlO -10 M or less, or of about 2.OxlO -10 M or less, or of about 1.5xl0 -10 M or less, or of about l.lxlO" 10 M or less, or of about l.OxlO 10 M or less, or of about 0.5 xlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0 -10 M or less, or of about l.lxlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.OxlO -10 M or less, or of about 1.6xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xlO -10 M or less, or of about 2.1xlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 9.OxlO 10 M or less, or of about 8.6xlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (i) may neutralize a BKPyV with an IC50 of about 2 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.2 or less, or of about 0.1 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less, or of about 0.05 nM or less, or about 0.01 nM or less.
  • the antibodies of group (i) may neutralize a BKPyV of genotype I with an IC50 of about 2 nM or less, or of about 1.5 nM or less, or of about 1 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less, or of about 0.05 nM or less, or of about 0.01 nM or less.
  • the antibodies of group (i) may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97 or 106, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of about 2 nM or less, or of about 1.5 nM or less, or of about 1.1 nM or less, or of about 0.13 nM or less, or of about 0.08 nM or less, or of about 0.01 nM or less.
  • the antibodies of group (i) may neutralize a BKPyV of genotype II with an IC50 of about 1 nM or less, or of about 0.5 nM or less, or of about 0.27 nM or less, or of about 0.23 nM or less.
  • the antibodies of group (i) may neutralize a BKPyV of genotype III with an IC50 of about 1 nM or less, or of about 0.6 nM or less, or of about 0.2 nM or less, or of about 0.15 nM or less.
  • the antibodies of group (i) may neutralize a BKPyV of genotype IV with an IC50 of about 1 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.15 nM or less, or of about 0.14 nM or less.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • the group (ii) of antibodies is illustrated by the antibody B KI 60-1 of the Example section.
  • the antibodies of group (ii) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
  • the antibodies of group (ii) may comprise
  • the conservative amino acid substitution may be as above described.
  • the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 19.
  • the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 19 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
  • the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 19 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 19.
  • the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 19.
  • the antibodies of group (ii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 20.
  • the antibodies of group (ii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 20 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
  • the antibodies of group (ii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 20 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 20.
  • the antibodies of group (ii) may comprise a variable domain of light chain of sequence SEQ ID NO: 20.
  • the antibodies of group (ii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 21.
  • the antibodies of group (ii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 21 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
  • the antibodies of group (ii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 21 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 21.
  • the antibodies of group (ii) may comprise a heavy chain of sequence SEQ ID NO: 21.
  • the antibodies of group (ii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 22.
  • the antibodies of group (ii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 22 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
  • the antibodies of group (ii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 22 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 22.
  • the antibodies of group (ii) may comprise a light chain of sequence SEQ ID NO: 22.
  • an antibody of group (ii) may be the antibody referred to as antibody B KI 60-1 in the Examples section.
  • variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 23.
  • a variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 23 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 23 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
  • variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 23.
  • variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 23.
  • variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 24.
  • a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 24 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 24 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
  • variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 24.
  • variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 24.
  • a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 25.
  • a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 25 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 25 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
  • a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 25.
  • a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 25.
  • a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 26.
  • a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 26 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
  • a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 26 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
  • a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 26.
  • a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 26.
  • the antibodies of group (ii) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 10 nM or less.
  • KD binding affinity
  • the antibodies of group (ii) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 M or less, or of about 4.OxlO 10 M or less, or of about 3xlO 10 M or less, or of about 2.5xlO 10 M or less, or of about 2.OxlO 10 M or less, or of about 1.8xlO 10 M or less, or of about 1.7xlO 10 M or less, or of about 1.5xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance of about 2.5xlO 10 M or less, or of about 2.3xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.8xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.OxlO 10 M or less, or of about 1.7xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.7xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (ii) may neutralize a BKPyV with an IC50 of about 10 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 or less, or of about 1.0 nM or less, or of about 0.80 nM or less, or of about 0.50 nM or less, or of about 0.30 nM or less, or of about 0.25 nM or less, or of about 0.10 nM or less, or of about 0.05 nM or less.
  • the antibodies of group (ii) may neutralize a BKPyV of genotype I with an IC50 of about 0.5 nM or less, or of about 0.44 nM or less, of about 0.2 nM or less, or of about 0.15 nM or less, or of about 0.12 nM or less.
  • the antibodies of group (ii) may neutralize a BKPyV of genotype II with an IC50 of about 5.5 nM or less, or of about 4.9 nM or less.
  • the antibodies of group (ii) may neutralize a BKPyV of genotype III with an IC50 of about 1 nM or less, or of about 0.25 nM or less.
  • the antibodies of group (ii) may neutralize a BKPyV of genotype IV with an IC50 of about 1 nM or less, or of about 0.6 nM or less, or about 0.2 nM or less.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • the antibodies of group (iii) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
  • the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
  • the conservative amino acid substitution may be as above described.
  • the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 32.
  • the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 32 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
  • the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 32 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 32.
  • the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 32.
  • the antibodies of group (iii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 33.
  • the antibodies of group (iii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 33 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
  • the antibodies of group (iii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 33 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 33.
  • the antibodies of group (iii) may comprise a variable domain of light chain of sequence SEQ ID NO: 33.
  • the antibodies of group (iii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 34.
  • the antibodies of group (iii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 34 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
  • the antibodies of group (iii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 34 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 34.
  • the antibodies of group (iii) may comprise a heavy chain of sequence SEQ ID NO: 34.
  • the antibodies of group (iii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 35.
  • the antibodies of group (iii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 35 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
  • the antibodies of group (iii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 35 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 35.
  • the antibodies of group (iii) may comprise a light chain of sequence SEQ ID NO: 35.
  • an antibody of group (iii) may be the antibody referred to as antibody BK198 in the Examples section.
  • variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 36.
  • a variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 36 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 36 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
  • variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 36.
  • variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 36.
  • variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 37.
  • a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 37 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 37 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
  • variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 37.
  • variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 37.
  • a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 38.
  • a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 38 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 38 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
  • a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 38.
  • a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
  • a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 39.
  • a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 39 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
  • a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 39 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
  • a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 39.
  • a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
  • the antibodies of group (iii) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 10 nM or less.
  • KD binding affinity
  • the antibodies of group (iii) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0 -9 M or less, or of about 2.5xl0 -9 M or less, or of about l.OxlO -9 M or less, of about 5.OxlO 10 M or less, or of about 4.OxlO 10 M or less, or of about 3xlO 10 M or less, or of about 2.OxlO 10 M or less, or of about 1.8xlO 10 M or less, or of about 1.5xlO 10 M or less, or about l.lxlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0xl0 -9 M or less, or of about 1.6xl0 -9 M or less.
  • KD binding affinity
  • the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xl0 -9 M or less, or of about 2.3xl0 -9 M or less.
  • KD binding affinity
  • the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xlO 10 M or less, or of about l.lxlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.8xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (iii) may neutralize a BKPyV with an IC50 of about 10 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 or less, or of about 1.0 nM or less, or of about 0.80 nM or less, or of about 0.50 nM or less, or of about 0.30 nM or less, or of about 0.25 nM or less.
  • the antibodies of group (iii) may neutralize a BKPyV of genotype I with an IC50 of about 2 nM or less, or of about 1.3 nM or less, or of about 1 nM or less, or of about 0.8 nM or less, or of about 0.79 nM or less.
  • the antibodies of group (iii) may neutralize a BKPyV of genotype II with an IC50 of about 10 nM or less, or of about 9 nM or less, or of about 5 nM or less, or of about 3.9 nM or less.
  • the antibodies of group (iii) may neutralize a BKPyV of genotype III with an IC50 of about 3 nM or less, or of about 2.8 nM or less.
  • the antibodies of group (iii) may neutralize a BKPyV of genotype IV with an IC50 of about 4 nM or less, or of about 3.5 nM or less, or of about 3.3 nM or less, or of about 1.7 nM or less.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • the antibodies of group (iv) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
  • the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
  • the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 45.
  • the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 45 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
  • the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 45 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 45.
  • the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 45.
  • the antibodies of group (iv) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 46.
  • the antibodies of group (iv) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 46 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
  • the antibodies of group (iv) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 46 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 46.
  • the antibodies of group (iv) may comprise a variable domain of light chain of sequence SEQ ID NO: 46.
  • the antibodies of group (iv) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 47.
  • the antibodies of group (iv) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 47 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
  • the antibodies of group (iv) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 47 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 47.
  • the antibodies of group (iv) may comprise a heavy chain of sequence SEQ ID NO: 47.
  • the antibodies of group (iv) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 48.
  • the antibodies of group (iv) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 48 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
  • the antibodies of group (iv) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 48 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 48.
  • the antibodies of group (iv) may comprise a light chain of sequence SEQ ID NO: 48.
  • an antibody of group (iv) may be the antibody referred to as antibody BK206 in the Examples section.
  • variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 49.
  • a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 49 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 49 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
  • variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 49.
  • a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 49.
  • variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 50.
  • a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 50 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 50 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
  • variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 50.
  • variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 50.
  • a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 51.
  • a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 51 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 51 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
  • a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 51.
  • a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 51.
  • a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 52.
  • a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 52 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
  • a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 52 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
  • a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 52.
  • a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 52.
  • the antibodies of group (iv) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3. Ox 10’ 9 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 10.0 nM or less.
  • KD binding affinity
  • the antibodies of group (iv) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.0xl0 -9 mol/1 (M) or less, or of about 2.5xl0 -9 M or less, or of about 2.0xl0 -9 M or less, or of about 1.8xl0 -9 M or less, or of about 1.5xl0 -9 M or less, or of about l.OxlO’ 9 M or less, or of about 8.OxlO 10 M or less, or of about 5.OxlO 10 M or less, or of about 2.5xlO 10 M or less, or of about 2.OxlO 10 M or less, or of about 1.5xl0’ 10 M or less, or of about 1.2xlO’ 10 M or less, or of about l.lxlO 10 M or less, or of about l.OxlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xlO 10 M or less, or of about 2.OxlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xl0 -9 M or less, or of about 2.2xl0 -9 M or less.
  • KD binding affinity
  • the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 4xl0 -9 M or less, or of about 3.1xl0 -9 M or less.
  • KD binding affinity
  • the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.5xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (iv) may neutralize a BKPyV with an IC50 of about 40.0 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 3.5 nM or less, or of about 3.0 nM or less, or of about 2.8 or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.2 nM or less, or about 0.16 nM or less.
  • the antibodies of group (iv) may neutralize a BKPyV of genotype I with an IC50 of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.18 nM or less, or of about 0.16 nM or less.
  • the antibodies of group (iv) may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of about 3 nM or less, or of about 2.64 nM or less, or of about 1 nM or less, or of about 0.96 nM or less, or of about 0.5 nM or less.
  • the antibodies of group (iv) may neutralize a BKPyV of genotype II with an IC50 of about 35 nM or less, or of about 34 nM or less.
  • the antibodies of group (iv) may neutralize a BKPyV of genotype III with an IC50 of about 40 nM or less, or of about 35 nM or less.
  • the antibodies of group (iv) may neutralize a BKPyV of genotype IV with an IC50 of about 0.3 nM or less, or of about 0.24 nM or less, or of about 0.19 nM or less.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • the antibodies of group (v) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
  • the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
  • the conservative amino acid substitution may be as above described.
  • the antibodies of group (v) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 58.
  • the antibodies of group (v) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 58 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
  • the antibodies of group (v) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 58 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 58.
  • the antibodies of group (v) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 58.
  • the antibodies of group (v) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 59.
  • the antibodies of group (v) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 59 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
  • the antibodies of group (v) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 59 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 59.
  • the antibodies of group (v) may comprise a variable domain of light chain of sequence SEQ ID NO: 59.
  • the antibodies of group (v) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 60.
  • the antibodies of group (v) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 60 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
  • the antibodies of group (v) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 60 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 60.
  • the antibodies of group (v) may comprise a heavy chain of sequence SEQ ID NO: 60.
  • the antibodies of group (v) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 61.
  • the antibodies of group (v) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 61 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
  • the antibodies of group (v) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 61 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 61.
  • the antibodies of group (v) may comprise a light chain of sequence SEQ ID NO: 61.
  • an antibody of group (v) may be the antibody referred to as antibody BK256 in the Examples section.
  • an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (v) of antibodies.
  • variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 62.
  • a variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 62 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 62 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
  • variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 62.
  • variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 62.
  • variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 63.
  • a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 63 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 63 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
  • variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 63.
  • variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 63.
  • a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 64.
  • a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 64 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 64 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
  • a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 64.
  • a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
  • a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 65.
  • a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 65 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
  • a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 65 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
  • a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 65.
  • a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
  • the antibodies of group (v) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5. Ox 10’ 8 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 30.0 nM or less.
  • KD binding affinity
  • the antibodies of group (v) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0 -8 mol/1 (M) or less, or of about 3.0xl0 -8 M or less, or of about 2.0xl0 -8 M or less, or of about l.OxlO -8 M or less, or of about 9.5xl0 -9 M or less, or of about 9.0xl0 -9 M or less, or of about 8.0xl0 -9 M or less, or of about 5.0xl0 -9 M or less, or of about 2.5xl0 -9 M or less, or of about 2.0xl0 -9 M or less, or of about l.OxlO -9 M or less, or of about 8.OxlO 10 M or less, or of about 5.OxlO 10 M or less, or of about 2.5xlO -10 M or less, or of about 2.Ox
  • KD binding affinity
  • the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0 -10 M or less, or of about 1.2xlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0 -10 M or less, or of about 1.2xlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0 -8 M or less, or of about 2.0xl0 -8 M or less.
  • KD binding affinity
  • the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about l.OxlO -8 M or less, or of about 9.5xl0 -9 M or less, or of about 9.3 xlO -9 M or less.
  • KD binding affinity
  • the antibodies of group (v) may neutralize a BKPyV with an IC50 of about 30.0 nM or less, or of about 25.0 nM or less, or of about 20.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 5.0 or less, or of about 2.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.25 nM or less, or of about 0.15 nM or less, or of about 0.10 nM or less.
  • the antibodies of group (v) may neutralize a BKPyV of genotype I with an IC50 of about 1 nM or less, or of about 0.23 nM or less, or of about 0.11 nM or less.
  • the antibodies of group (v) may neutralize a BKPyV of genotype II with an IC50 of about 8 nM or less, or of about 1 nM or less, or of about 0.79 nM or less.
  • the antibodies of group (v) may neutralize a BKPyV of genotype III with an IC50 of about 30 nM or less, or of about 23.5 nM or less.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • the antibodies of group (vi) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
  • the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
  • the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 71.
  • the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 71 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
  • the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 71 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 71.
  • the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 71.
  • the antibodies of group (vi) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 72.
  • the antibodies of group (vi) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 72 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
  • the antibodies of group (vi) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 72 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 72.
  • the antibodies of group (vi) may comprise a variable domain of light chain of sequence SEQ ID NO: 72.
  • the antibodies of group (vi) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 73.
  • the antibodies of group (vi) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 73 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
  • the antibodies of group (vi) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 73 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 73.
  • the antibodies of group (vi) may comprise a heavy chain of sequence SEQ ID NO: 73.
  • the antibodies of group (vi) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 74.
  • the antibodies of group (vi) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 74 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
  • the antibodies of group (vi) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 74 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 74.
  • the antibodies of group (vi) may comprise a light chain of sequence SEQ ID NO: 74.
  • an antibody of group (vi) may be the antibody referred to as antibody BK293 in the Examples section.
  • variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 75.
  • a variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 75 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 75 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
  • variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 75.
  • variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 75.
  • variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 76.
  • a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 76 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 76 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
  • variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 76.
  • variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 76.
  • a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 77.
  • a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 77 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 77 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
  • a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 77.
  • a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 77.
  • a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 78.
  • a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 78 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
  • a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 78 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
  • a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 78.
  • a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 78.
  • the antibodies of group (vi) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about lO.OxlO -8 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 40.0 nM or less.
  • KD binding affinity
  • the antibodies of group (vi) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about lO.OxlO -8 M or less, or of about 8.0xl0 -8 M or less, or of about 6.5xl0 -8 M or less, or of about 5.0xl0 -8 M or less, or of about 3.0xl0 -8 M or less, or of about 2.0xl0 -8 M or less, or of about 1.5xl0 -8 M or less, or of about l.OxlO -8 M or less, or of about 8.0xl0 -9 M or less, or of about 5.0xl0 -9 M or less, or of about 2.5xl0 -9 M or less, or of about 2.0xl0 -9 M or less, or of about 1.5xl0 -9 M or less, or of about l.OxlO -9 M or less, or of about 8.5xl
  • KD binding affinity
  • the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 9.0xl0 -8 M or less, or of about 7.0xl0 -8 M or less or of about 6.5xl0 -8 M or less.
  • KD binding affinity
  • the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xl0 -9 M or less, or of about 2.0xl0 -9 M or less, or of about 1.6xl0 -9 M or less.
  • KD binding affinity
  • the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0 -9 M or less, or of about l.OxlO -9 M or less, or of about 8.5xlO -10 M or less.
  • KD binding affinity
  • the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0xl0 -8 M or less, or about 1.5xl0 -8 M or less, or of about 1.3xl0 -8 M or less.
  • KD binding affinity
  • the antibodies of group (vi) may neutralize a BKPyV with an IC50 of about 40.0 nM or less, or of about 35.0 nM or less, or of about 30.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 5.0 or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.2 or less, or of about 0.15 nM or less, or of about 0.10 nM or less, or of about 0.08 nM or less, or of about 0.05 nM or less, or of about 0.03 nM or less, or of about 0.01 nM or less.
  • the antibodies of group (vi) may neutralize a BKPyV of genotype II with an IC50 of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.24 nM or less, or of about 0.05 nM or less, or of about 0.03 nM or less.
  • the antibodies of group (vi) may neutralize a BKPyV of genotype III with an IC50 of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.27 nM or less, or of about 0.25 nM or less.
  • the antibodies of group (vi) may neutralize a BKPyV of genotype IV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, with an IC50 of about 40 nM or less, or of about 35.0 nM or less, or of about 32.0 nM or less.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • the group (vii) of antibodies is illustrated by the antibody BK304 of the Example section.
  • the antibodies of group (vii) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • the conservative amino acid substitution may be as above described.
  • the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 84.
  • the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 84 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
  • the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 84 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
  • the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 84.
  • the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 84.
  • the antibodies of group (vii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 85.
  • the antibodies of group (vii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 85 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • the antibodies of group (vii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 85 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
  • the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 85.
  • the antibodies of group (vii) may comprise a variable domain of light chain of sequence SEQ ID NO: 85.
  • the antibodies of group (vii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 86.
  • the antibodies of group (vii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 86 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
  • the antibodies of group (vii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 86 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
  • the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 86.
  • the antibodies of group (vii) may comprise a heavy chain of sequence SEQ ID NO: 86.
  • the antibodies of group (vii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 87.
  • the antibodies of group (vii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 87 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L Ill comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • the antibodies of group (vii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 87 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
  • the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 87.
  • the antibodies of group (vii) may comprise a light chain of sequence SEQ ID NO: 87.
  • an antibody of group (vii) may be the antibody referred to as antibody BK304 in the Examples section.
  • variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 88.
  • a variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 88 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
  • variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 88 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
  • variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 88.
  • variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 88.
  • variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 89.
  • a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 89 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 89 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
  • variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 89.
  • variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 89.
  • a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 90.
  • a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 90 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
  • a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 90 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
  • a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 90.
  • a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 90.
  • a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 91.
  • a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 91 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
  • a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 91 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
  • a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 91.
  • a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 91.
  • the antibodies of group (vii) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0x1 O’ 9 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 25.0 nM or less.
  • KD binding affinity
  • the antibodies of group (vii) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0 -9 M or less, or of about 2.5xl0 -9 M or less, or of about 2.0xl0 -9 M or less, or of about 1.5xl0 -9 M or less, or of about l.OxlO -9 M or less, or of about 9.OxlO 10 M or less, or of about 8.OxlO 10 M or less, or of about 5.OxlO 10 M or less, or of about 4.OxlO 10 M or less, or of about 3.OxlO 10 M or less, or of about 2.OxlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 M or less, or of about 3.2xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0xl0 -9 M or less, or of about 1.3xl0 -9 M or less.
  • the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2xl0 -9 M or less, or of about 1.6xl0 -9 M or less.
  • the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 9.OxlO 10 M or less, or of about 7.6xlO 10 M or less.
  • KD binding affinity
  • the antibodies of group (vii) may neutralize a BKPyV with an IC50 of about 25.0 nM or less, or of about 22.5 nM or less, or of about 20.0 nM or less, or of about 18.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.80 nM or less, or of about 0.50 nM or less, or of about 0.35 nM or less, or of about 0.25 nM or less, or of about 0.12 nM or less, or of about 0.10 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less, or of about 0.04 nM
  • the antibodies of group (vii) may neutralize a BKPyV of genotype I with an IC50 of about 15.3 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 6.2 nM or less, or of about 6.0 nM or less, or of about 2.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.25 nM or less, or of about 0.10 nM or less, or of about 0.05 nM or less, or of about 0.03 nM or less, or of about 0.02 nM or less, or of about 0.01 nM or less.
  • the antibodies of group (vii) may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of about 0.5 nM or less, or of about 0.25 nM or less, or of about 0.23 nM or less, or of about 0.1 nM or less.
  • the antibodies of group (vii) may neutralize a BKPyV of genotype II with an IC50 of about 25 nM or less, or of about 21 nM or less.
  • the antibodies of group (vii) may neutralize a BKPyV of genotype III with an IC50 of about 15 nM or less, or of about 13.5 nM or less, or about 13 nM or less.
  • the antibodies of group (vii) may neutralize a BKPyV of genotype IV with an IC50 of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.8 nM or less, or of about 1.6 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.28 nM or less, or about 0.2 nM or less, or of about 0.10 nM or less.
  • the antibodies of group (vii) may neutralize a BKPyV of genotype IV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, with an IC50 of about 1 nM or less, or of about 0.5 nM or less, or of about 0.1 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less.
  • the IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
  • IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
  • the inventors have surprisingly demonstrated that a combination of at least two of the antibodies described herein induced an improve neutralizing activity against the BKPyV.
  • the combinations of antibodies may efficiently neutralize BKPyV escaping neutralization by the benchmark antibody 41F17.
  • a combination of antibodies may comprise at least a first antibody and a second antibody, or antigen-binding fragments thereof.
  • the first antibody, or antigen-binding fragment thereof, and the second antibody, or antigen-binding fragment thereof may be selected among the antibodies of groups (i) to (vii) described herein.
  • a first antibody is selected from a first group of antibodies selected from groups (i) to (vii) of antibodies and a second antibody is selected from a second group of antibodies selected from groups (i) to (vii) of antibodies, the first and second groups of antibodies being different.
  • the antibodies of a combination of antibodies may bind to distinct VP1 capsid proteins from, for example, from at least two distinct BKPyV genotypes, serotypes, or variants.
  • the antibodies of a combination of antibodies may bind to a same VP1 capsid protein, but on distinct epitopes of a same VP1 capsid protein, for example on at least two distinct epitopes of a same VP1 capsid protein.
  • a combination of at least two antibodies disclosed herein may neutralize a
  • a combination of at least two antibodies disclosed herein may neutralize a
  • a combination of at least two antibodies disclosed herein may neutralize a
  • a combination of at least two antibodies disclosed herein may neutralize a
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV of genotype I-IV with an IC50 of about 70 nM or less, or of about 68 nM or less, or of about 65 nM or less, or of about 62 nM or less, or of about 60 nM or less, or of about 58 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less, or of about 40 nM or less, or of about 30 nM or less, or of about 20 nM or less, or of about 15 nM or less, or of about 10 nM or less, or of about 8 nM or less, or of about 5 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 n
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV of genotype IV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 70 nM or less to about 0.1 nM or less.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less, or of about 68 nM or less, or of about 65 nM or less, or of about 62 nM or less, or of about 60 nM or less, or of about 58 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less, or of about 40 nM or less, or of about 30 nM or less, or of about 20 nM or less, or of about 15 nM or less, or of about 10 nM or less, or of about 8 nM or less, or of about 6 nM or less, or
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 6 nM or less, or of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less.
  • a combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, with an ICsoof about 70 nM or less, or of about 68 nM or less, or of about 65 nM or less, or of about 62 nM or less, or of about 60 nM or less, or of about 58 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less, or of about 40 nM or less, or of about 30 nM or less, or of about 20 nM or less, or of about 15 nM or less, or of about 10 nM or less, or of about 8 nM or less, or of about 6 nM or less, or of about 4 nM or less
  • a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101 may be selected among BKPyV g!b2-D58delP59A, BKPyV g!b2-E61K and BKPyV gIVc2-E327K.
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 60 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 60 nM or less to about 0.01 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 1 nM or less to about 0.01 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 60 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 60 nM or less to about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2.27 nM or less, or of about 1 nM or less, or of about 0.5 nM or less, or of about 0.36 nM or less, or of about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 3 nM or less, or of about 2.5 nM or less, or of about 2.2 nM or less, or of about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 60 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less.
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • an antibody selected from group (iv) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 45 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 46, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise:
  • an antibody selected from group (iv) of antibodies comprising a heavy chain of sequence SEQ ID NO: 47 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 48, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may comprise as a first antibody, the antibody BK120, and as a second antibody, the antibody BK206.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 70 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 70 nM or less to about 0.01 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 1 nM or less to about 0.01 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 70 nM or less to about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 3 nM or less, or of about 4 nM or less, or of about 1 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 4 nM or less, or of about 3.4 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less, or of about 65 nM or less, or of about 60 nM or less, or of about 55 nM or less.
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • an antibody selected from group (vi) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 71 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 72, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise:
  • an antibody selected from group (vi) of antibodies comprising a heavy chain of sequence SEQ ID NO: 73 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 74, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may comprise as a first antibody, the antibody BK120, and as a second antibody, the antibody BK293.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 1 nM or less to about 0.01 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 0.5 nM or less to about 0.01 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 1 nM or less to about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less, or of about 0.8 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.39 nM or less, or of about 0.3 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.16 nM or less, or of about 0.1 nM or less, or of about 0.05 nM or less.
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • an antibody selected from group (vii) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 84 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 85, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise:
  • an antibody selected from group (vii) of antibodies comprising a heavy chain of sequence SEQ ID NO: 86 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 87, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may comprise as a first antibody, the antibody BK120, and as a second antibody, the antibody BK304.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 40 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 40 nM or less to about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 5 nM or less to about 0.1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 40 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 40 nM or less to about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an ICsoof about 10 nM or less, or of about 9 nM or less, or of about 8 nM or less, or of about 7 nM or less, or of about 6 nM or less, or of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2 nM or less, or of about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 8 nM or less, or of about 7 nM or less, or of about 6 nM or less, or of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2 nM or less, or of about 1 nM or less.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 40 nM or less, or of about 38 nM or less, or of about 35 nM or less, or of about 32 nM or less, or of about 30nM or less, or of about 25 nM or less.
  • a combination of antibodies may comprise: [0819] - as a first antibody, an antibody selected from group (iv) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO
  • a combination of antibodies may comprise:
  • an antibody selected from group (vi) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 71 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 72, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise: [0825] - as a first antibody, an antibody selected from group (iv) of antibodies comprising a heavy chain of sequence SEQ ID NO: 47 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 48, or a sequence at least 85% identical thereto, and
  • an antibody selected from group (vi) of antibodies comprising a heavy chain of sequence SEQ ID NO: 73 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 74, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may comprise as a first antibody, the antibody BK206, and as a second antibody, the antibody BK293.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV of genotype I-IV.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • a combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • a combination of antibodies may comprise:
  • a combination of antibodies may comprise:
  • an antibody selected from group (iii) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 32 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 33, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise:
  • an antibody selected from group (iii) of antibodies comprising a heavy chain of sequence SEQ ID NO: 34 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 35, or a sequence at least 85% identical thereto.
  • a combination of antibodies may comprise as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may comprise as a first antibody, the antibody B KI 60-1, and as a second antibody, the antibody BK198.
  • the antibodies may be provided as separate antibodies or as multi-specific antibodies where the antibodies are joined together.
  • Multi-specific antibodies refers to monoclonal antibodies that have binding specificities for at least two different epitopes of one antigen or for at least two distinct antigens.
  • a multi-specific antibody may be a bispecific antibody.
  • multi-specific antibodies are described in US20090232811, US9382323, US9890204 or US8796424.
  • a multi-specific antibodies may be a bispecific antibodies.
  • bispecific antibodies refers to molecules which combine the antigen-binding sites of two antibodies within a single molecule. Thus, a bispecific antibody is able to bind two different antigens or to two distinct epitopes of a same antigen, simultaneously. The distinct epitope may be overlapping epitopes or non-overlapping epitopes.
  • Multi-specific or bispecific antibodies can be prepared as full-length antibodies or antibody fragments.
  • the multi-specific or bispecific antibody described herein may comprise a combination of antibodies as described herein.
  • the multi-specific or bispecific antibody may be an isolated or a recombinant bispecific antibody.
  • a multi-specific or a bispecific antibody may comprise at least a first antibody, or an antigen-binding fragment thereof, and at least a second antibody, or an antigen-binding fragment thereof, joined to each other, wherein said first and second antibodies may be selected among the antibodies of groups (i) to (vii) described herein.
  • a multi-specific or a bispecific may comprise:
  • a multi-specific or a bispecific may comprise:
  • a multi-specific antibody is a bispecific antibody.
  • An antibody of the multi-specific or a bispecific antibody can be linked to or coexpressed with another antibody by any known techniques in the art.
  • an antibody or fragment thereof can be functionally linked, e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise, to one or more other antibody or antibody fragment to produce a bispecific or a multispecific antibody.
  • bispecific formats that can be used include, without limitation, e.g., scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, Quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEED)body, leucine zipper, Duobody, IgGl/IgG2, dual acting Fab (DAF)-IgG, and Mab2 bispecific formats.
  • Immunoconj ugates e.g., scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, Quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEED)body, leu
  • An immunoconjugate comprising an antibody, or a fragment thereof, as disclosed herein may prepared according to any known methods in the art.
  • An antibody, or a fragment thereof, may be conjugated or linked to detectable label or reporter molecule.
  • Such immunoconjugate may be used as a diagnostic tool for detecting and/or measuring a BKPyV infection in a subject in need thereof.
  • a detectable label or reporter molecule can be a radioisotope, such as 3 H, 14 C, 32 P, 35 S, or 125 I; a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine; or an enzyme such as alkaline phosphatase, P-galactosidase, horseradish peroxidase, or luciferase.
  • a radioisotope such as 3 H, 14 C, 32 P, 35 S, or 125 I
  • a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine
  • an enzyme such as alkaline phosphatase, P-galactosidase, horseradish peroxidase, or luciferase.
  • the term “labeled”, with regard to the antibody according to the disclosure, is intended to encompass direct labeling of the antibody by coupling (i.e., physically linking) a detectable substance, such as a radioactive agent or a fluorophore (e.g. fluorescein isothiocyanate (FITC) or phycoerythrin (PE) or Indocyanine (Cy5)) to the polypeptide, as well as indirect labeling of the polypeptide by reactivity with a detectable substance.
  • a detectable substance such as a radioactive agent or a fluorophore (e.g. fluorescein isothiocyanate (FITC) or phycoerythrin (PE) or Indocyanine (Cy5)
  • radioactive molecules include but are not limited radioactive atom for scintigraphic studies such as 3 H, 14 C, 32 P, 35 S, I 123 , I 124 , or 125 I, In 111 , Re 186 , Re 188 , Tc".
  • Polypeptides of the invention may be also labelled with a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as iodine- 123, indium-ill, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron.
  • NMR nuclear magnetic resonance
  • MRI magnetic resonance imaging
  • immunoconjugates according to the present invention can be prepared as described in the application W02004/091668.
  • the antibodies may be covalently attached, directly or via a cleavable or non- cleavable linker, to at least one detectable probe.
  • Linker means a chemical moiety comprising a covalent bond or a chain of atoms that covalently attaches a polypeptide to a detectable probe.
  • the conjugates may be prepared by in vitro methods.
  • a linking group is used. Suitable linking groups are well known in the art and include disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups.
  • Conjugation of an antibody of the invention with a detectable probe may be made using a variety of bifunctional protein coupling agents including but not limited to N-succinimidyl pyridyldithiobutyrate (SPDB), butanoic acid 4-[(5- nitro-2-pyridinyl)dithio]-2,5-dioxo-l-pyrrolidinyl ester (nitro-SPDB), 4-(Pyridin-2- yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), N-succinimidyl (2-pyridyldithio) propionate (SPDP), succinimidyl (N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimid
  • a ricin immunotoxin can be prepared as described in Vitetta et al (1987).
  • Carbon labeled 1- isothiocyanatobenzyl methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody (WO 94/11026).
  • the linker may be a “cleavable linker” or a “non-cleavable linker” (for example SMCC linker).
  • a fusion protein comprising an antibody disclosed herein and a detectable probe may be made, by recombinant techniques or peptide synthesis.
  • the length of DNA may comprise respective regions encoding the two portions of the conjugate either adjacent one another or separated by a region encoding a linker peptide which does not destroy the desired properties of the conjugate.
  • a conjugate can be obtained by a process comprising the steps of:
  • the aqueous solution of cell-binding agent can be buffered with buffers such as, e.g. potassium phosphate, acetate, citrate or N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes buffer).
  • buffers such as, e.g. potassium phosphate, acetate, citrate or N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes buffer).
  • the reaction temperature is usually comprised between 20 and 40° C.
  • the reaction time can vary from 1 to 24 hours.
  • the reaction between the antibody and the detectable probe can be monitored by size exclusion chromatography (SEC) with a refractometric and/or UV detector. If the conjugate yield is too low, the reaction time can be extended.
  • SEC size exclusion chromatography
  • the conjugate can be purified e.g. by SEC, adsorption chromatography (such as ion exchange chromatography, IEC), hydrophobic interaction chromatograhy (HIC), affinity chromatography, mixed-support chromatography such as hydroxyapatite chromatography, or high performance liquid chromatography (HPLC). Purification by dialysis or diafiltration can also be used.
  • SEC adsorption chromatography
  • IEC hydrophobic interaction chromatograhy
  • HPLC high performance liquid chromatography
  • the term “aggregates” means the associations which can be formed between two or more antibodies, being modified or not by conjugation.
  • the aggregates can be formed under the influence of a great number of parameters, such as a high concentration of cell-binding agent in the solution, the pH of the solution, high shearing forces, the number of bonded dimers and their hydrophobic character, the temperature (see Wang & Gosh, 2008, J. Membrane Sci., 318: 311-316, and references cited therein); note that the relative influence of some of these parameters is not clearly established.
  • the person skilled in the art will refer to Cromwell et al. (2006, AAPS Journal, 8(3): E572-E579).
  • the content in aggregates can be determined with techniques well known to the skilled person, such as SEC (see Walter et al., 1993, Anal. Biochem., 212(2): 469-480).
  • step (i) or (ii) the conjugate-containing solution can be submitted to an additional step (iii) of chromatography, ultrafiltration and/or diafiltration.
  • the present disclosure relates to a use of a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence for screening a candidate antibody able to neutralize a BKPy virus.
  • the present disclosure relates to in vitro methods for screening a candidate antibody or a candidate combination of antibodies able to neutralize a BKPy.
  • An in vitro method for screening a candidate antibody may be used to screen antibodies able to neutralize a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • An in vitro method for screening a candidate antibody able to neutralize a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may comprise at least the steps of:
  • the infected cells may be counted, and the obtained value is compared to, or normalized with, the value obtained with cultured cells infected with said virus in the absence of the candidate antibody.
  • An in vitro method for screening a candidate combination of antibodies able to neutralize a BKPy virus may comprise at least the steps of:
  • a number of infected cells obtained at step (a) lower than the number of cells obtained at step (b) may indicative of a neutralizing activity of said candidate combination.
  • the virus may be a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
  • a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 may be BKPyV-D58DelP59A.
  • the methods may be carried out with a virus, or a pseudotype virus (generally referred as viral particles). Methods for preparing virus-like particles and pseudotype virus are known in the art. [0899]
  • the antibodies to be screened may be placed in contact with the viral particles before adding the viral particles and the antibody to the cultured cells.
  • the antibodies to be screened may be placed in contact with the cultured cells before addition of the viral particles.
  • the antibodies to be screened may be placed in contact with the cultured cells after addition of the viral particles to the cells.
  • a serial dilution of a predetermined amount of infecting viral particles may be used to infect cultured cells.
  • a 2-, 3-, 4-, 5-, 6- , 7-fold dilution may be used.
  • Useable cells may be HEK293 TTT cells or RS cells.
  • the neutralization activity of the antibodies may be expressed as an IC50.
  • virus infection of target cells in vitro may be measured by immunofluorescence analysis using markers for intracellular BKPyV antigen expression as disclosed in, for example, Moriyama T, Sorokin A. Curr Protoc Cell Biol. 2009 Mar;Chapter 26:Unit 26.2.
  • An antibody disclosed herein may be produced by any technique known in the art, such as, without limitation, any chemical, biological, genetic, or enzymatic technique, either alone or in combination.
  • an antibody can be synthesized by recombinant DNA techniques well-known in the art.
  • an antibody can be obtained as DNA expression products after incorporation of DNA sequences encoding the antibody into expression vectors and introduction of such vectors into suitable eukaryotic or prokaryotic host cells that will express the desired antibody, from which it can be later isolated using well-known techniques, such as described in the Examples.
  • Another aspect of the present disclosure relates to a method for producing an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising at least the steps of culturing a host cell transformed by a nucleic acid sequence encoding an antibody disclosed herein, or by an expression vector comprising such nucleic acid, in a culture medium under conditions suitable for producing said antibody or fragment thereof and recovering the antibody or fragment thereof from said cultured cells.
  • a method for producing an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof implements culture medium suitable for producing said antibody or fragment.
  • the cell culture medium under conditions suitable for producing said antibody or antigen-binding fragment thereof typically contains a large number of ingredients, which are necessary to support maintenance of the cultured host cells. Suitable combinations of ingredients can readily be formulated by the skilled person.
  • a culture medium to be used herein will generally be a nutrient solution comprising standard cell culture ingredients, such as amino acids, vitamins, inorganic salts, a carbon energy source, and a buffer, as described in the art.
  • a cell culture medium useable may be formulated in deionized, distilled water.
  • a culture medium may be sterilized prior to use to prevent contamination, e.g., by ultraviolet light, heating, gamma-irradiation, or filtration.
  • a cell culture medium may be frozen (e.g., at - 20°C or -80°C) for storage or transport.
  • the medium may contain one or more antibiotics to prevent contamination.
  • a cell culture medium may be a defined synthetic medium that is buffered at a pH of 7.4 (preferably with a pH 7.2 - 7.6 or at least 7.2 and not higher than 7.6) with a carbonate -based buffer, while the cells are cultured in an atmosphere comprising between 5 % and 10% CO2, or at least 5% and not more than 10% CO2, for example 5% CO2.
  • a potentially suitable cell culture medium may be available commercially, and includes, but is not limited to, Dulbecco's Modified Eagle Media (DMEM), Minimal Essential Medium (MEM), Knockout-DMEM (KO-DMEM), Glasgow Minimal Essential Medium (G- MEM), Basal Medium Eagle (BME), DMEM/Ham’s F12, Advanced DMEM/Ham’s F12, Iscove’s Modified Dulbecco’s Media (IMDM), Ham's F-10, Ham’s F-12, Medium 199, RPMI 1640 Media, and Hepatocyte cell culture media.
  • DMEM Dulbecco's Modified Eagle Media
  • MEM Minimal Essential Medium
  • Knockout-DMEM KO-DMEM
  • Glasgow Minimal Essential Medium G- MEM
  • Basal Medium Eagle BME
  • DMEM/Ham’s F12 DMEM/Ham’s F12
  • Advanced DMEM/Ham’s F12 Iscove’s Modified Dulbecco’s Media
  • a nucleic acid sequence may be used to produce an antibody, or an antigenbinding fragment thereof as described in the disclosure.
  • the nucleic acid sequence may be a DNA or a RNA molecule, which may be included in any suitable expression vector, such as a plasmid, cosmid, episome, artificial chromosome, phage, or a viral vector.
  • a nucleic acid sequence encoding an antibody, or an antigen-binding fragment thereof, as described may be an isolated nucleic acid sequence or a recombinant nucleic acid sequence.
  • a nucleic acid sequence usable for producing an antibody, or an antigen-binding fragment thereof, may be as disclosed herein.
  • a nucleic acid as described herein encodes the heavy chain (VH), the light chain (VL), the full-length heavy chain, and the full-length light chain of an antibody of the present disclosure for the groups (i) to (vii) of antibodies.
  • a nucleic acid sequence can be produced by de novo solid-phase DNA synthesis or by PCR mutagenesis of an existing sequence encoding an antibody as described herein or its antigen-binding fragment thereof.
  • Direct chemical synthesis of nucleic acids can be accomplished by methods known in the art, such as the phosphotriester method of Narang et al, Meth. Enzymol. 68:90, 1979; the phosphodiester method of Brown / al, Meth. Enzymol. 68: 109, 1979; the diethylphosphoramidite method of Beaucage et al , Tetra. Lett., 22: 1859, 1981; and the solid support method of U.S. Patent No. 4,458,066.
  • An expression vector may be an isolated or a recombinant expression vector.
  • a host cell may be an isolated or a recombinant host cell.
  • An expression vector may comprise a nucleic acid sequence as described herein.
  • Such a vector may comprise regulatory elements, such as a promotor, enhancer, terminator, and the like, to cause or direct expression of said antibody upon transfection into the host cell.
  • vectors will depend upon a variety of factors such as the type of host cell in which propagation is desired and the purpose of propagation. Some vectors are useful for amplifying and making large amounts of the desired DNA sequence. Other vectors are suitable for expression in cells in culture. Still other vectors are suitable for transfer and expression in cells in a whole animal. The choice of appropriate vector is well within the skill of the art. Many such vectors are available commercially.
  • Any expression vector for animal cell can be used, so long as a gene encoding the human antibody C region can be inserted and expressed.
  • suitable vectors include pAGE107, pAGE103, pHSG274, pKCR, pSGl, pCAG and the like.
  • Other examples of vectors include replicating plasmids comprising an origin of replication, or integrative plasmids, such as for instance pUC, pcDNA, pBR, and the like.
  • Other examples of viral vector include adenoviral, retroviral, herpes virus and AAV vectors. Such recombinant viruses may be produced by techniques known in the art, such as by transfecting packaging cells or by transient transfection with helper plasmids or viruses.
  • virus packaging cells include PA317 cells, PsiCRIP cells, Gpenv+ cells, 293 cells, RS cells, etc.
  • a detailed protocol for producing such replication-defective recombinant viruses may be found for instance in Pastrana et al. (PloS Pathog. 2012, 8, el002650).
  • promoters and enhancers used in the expression vector for animal cell include early promoter and enhancer of SV40, LTR promoter and enhancer of Moloney mouse leukemia virus, promoter and enhancer of immunoglobulin H chain and the like.
  • Vectors can be prepared by, for example, inserting a polynucleotide of interest into a construct backbone, typically by means of DNA ligase attachment to a cleaved restriction enzyme site in the vector.
  • the desired nucleotide sequence can be inserted by homologous recombination or site-specific recombination.
  • homologous recombination is accomplished by attaching regions of homology to the vector on the flanks of the desired nucleotide sequence, while site-specific recombination can be accomplished through use of sequences that facilitate site-specific recombination (e.g., ere -lox, att sites, etc.).
  • Nucleic acid containing such sequences can be added by, for example, ligation of oligonucleotides, or by polymerase chain reaction using primers comprising both the region of homology and a portion of the desired nucleotide sequence.
  • Expression vectors generally have convenient restriction sites located near the promoter sequence to provide for the insertion of nucleic acid sequences encoding proteins of interest.
  • a selectable marker operative in the expression host may be present to facilitate selection of cells containing the vector.
  • the expression construct may include additional elements.
  • the expression vector may have one or two replication systems, thus allowing it to be maintained in organisms, for example in mammalian or insect cells for expression and in a prokaryotic host for cloning and amplification.
  • the expression construct may contain a selectable marker gene to allow the selection of transformed host cells. Selection genes are well known in the art and will vary with the host cell used.
  • vectors encoding both the heavy and light chains may be co-expressed in the host cell for expression of the entire antibody.
  • the expression vector is transferred to a host cell by conventional techniques and the transfected cells then are cultured by conventional techniques to produce an antibody or an antigen-binding fragment thereof of the present disclosure.
  • Isolated or recombinant host cells may be transfected, infected, or transformed by a nucleic acid sequence and/or an expression vector as disclosed herein.
  • Such host cells may be used for the production of an antibody as described herein.
  • transformation means the introduction of a “foreign” (i.e., extrinsic, or extracellular) gene, DNA or RNA sequence to a host cell, so that the host cell will express the introduced gene or sequence to produce a desired substance, typically a protein or enzyme coded by the introduced gene or sequence.
  • a host cell that receives and expresses introduced DNA or RNA bas been “transformed”.
  • the present disclosure further provides a recombinant, or isolated, host cell transformed with a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof as disclosed herein, or with an expression vector comprising such nucleic acid sequence.
  • any of a number of suitable host cells can be used in the production of an antibody or antigen-binding fragment thereof.
  • the antibody or antigen-binding fragment thereof described herein may be expressed in prokaryotes or eukaryotes (eukaryotic unicellular organism), e.g., bacteria such as Escherichia coli, yeasts, such as Saccharomyces sp., or mammalian cell lines, such as Vero cells, HEK293, or CHO, in accordance with conventional techniques.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Virology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The present disclosure relates to isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising specific H- and L- CDR1, CDR2 and CDR3. The disclosure also relates to nucleic acid, expression vectors and host cells comprising sequences encoding the antibodies. The antibodies and combinations thereof can be used for the prevention and/or treatment of BKPyV induced infection and/or a disorder induced by a BKPyV infection.

Description

TITLE
BK POLYOMA VIRUS ANTIBODIES AND USES THEREOF
[0001] This application includes a sequence listing in computer readable form in a file named S3_SATT OUEST_PR94463_sequence listing created on March 7, 2023 (161 458 bytes), which is incorporated by reference herein.
TECHNICAL FIELD
[0002] The present disclosure relates to antibodies, and combinations thereof, targeting BK polyomaviruses (BKPyV), and methods using those antibodies.
BACKGROUND
[0003] The BK polyomavirus (BKPyV) is a small double-stranded DNA virus with a non-enveloped icosahedral capsid, first described in 1971. BKPyV has been divided into four major subtypes (genotypes) I, II, III and IV, further divided into subgroups, that are characterized by sequence variation in the major capsid protein VP1 and correspond to distinct neutralizing virus serotypes (Knowles et al. J. Med. Virol. 1989, 28, 118-123 - Pastrana et al. J. Virol. 2013, 87, 10105-10113). Genotype I is the most prevalent BKPyV genotype worldwide, followed by genotype IV in East Asia and Europe, while genotypes II and III are rare throughout the world (Zhong et al. J. Gen. Virol. 2009, 90, 144-152). The distribution of the molecular subgroups of the virus is associated with the region of origin, for instance, BKPyV subtype la is most common in Africa, Ibl in Southeast Asia, and Ib2 in Europe, while Ic is the most common variant in Northeast Asia (Furmaga, Jacek et al. Viruses vol. 13,8 1502. 30 Jul. 2021). Besides, in vitro selected genetic variants of some genotypes of the BKPyV were characterized, such as BKPyV genotype I-E61K or BKPyV genotype IV-E327K (Lindner et al. Immunity. 2019;50:668- 676.e5).
[0004] In structural terms, the virus capsid is composed of 72 pentamers of the major capsid protein VP1, and each pentamer is associated with one copy of either VP2 or VP3, which are the minor capsid proteins. The circular double-stranded DNA genome is packaged inside the capsid in a condensed form associated with histones from the host cell. Importantly, the exterior of the virus particle is composed entirely of the VP1 protein, and virus like particles (VLP) composed of VP1 are structurally and antigenically identical to infectious virus particles (Hurdiss et al. Structure 2018, 26, 839-847.e3).
[0005] BKPyV is a typical opportunistic virus, which provokes overt pathology almost exclusively in the context of immunosuppression. It is estimated that 80% of the adult population is latently infected with BKPyV. After kidney transplantation (KTx), reactivation of BKPyV from sites of persistent infection (mainly kidney epithelial cells) frequently occurs, leading to virus detection in urine (Bressollette-Bodin et al. Am. J. Transplant. 2005, 5, 1926-1933). Overall, 1-10% of kidney transplantation patients develop a BK polyomavirus-associated nephropathy (“PyVAN”), which emerged as an important cause of kidney allograft failure in the late 1990s (Nickeleit et al. N. Engl. J. Med. 2000, 342, 1309-1315), and remains an important cause of kidney transplant dysfunction (Hirsch et al. Am. J. Transplant. 2013, 13 Suppl 4). Furthermore, KTx recipients with persistent BKPyV viremia or PyVAN were found to have an 11 -fold greater risk of bladder carcinoma compared to KTx recipients without evidence of BKPyV replication (Liu et al. Transplantation, 2017, 101, 1488-1494), with integration of BKPyV genomes reported in some reno-urinary tumors (Papadimitriou et al. Am. J. Transplant., 2016, 16, 398-406. In addition, BKPyV is also responsible for hemorrhagic cystitis (BKPyV - HC), which arises as a complication after 5% to 25% of allogeneic hematopoietic cell transplantations (HCT) (Cesaro et al. J Antimicrob Chemother 2018, 73, 12-21). BKPyV has also been associated, less frequently, with pneumonitis, retinitis, and meningoencephalitis in immunocompromised subjects (Reploeg et al. Clin Infect Dis 2001, 33, 191-202).
[0006] There is no specific antiviral molecule licensed for the treatment of these infections, as polyomaviruses lack their own polymerase and are entirely dependent on host machinery for their replication. For this reason, the clinical management of BK polyoma virus- associated nephropathy relies on modification or dose reduction of immunosuppressive therapy, with the aim of inducing host antiviral immune responses that may then control viral replication. This strategy is applied in the case of histologically confirmed BK polyomavirus-associated nephropathy (curative treatment), as well as in cases of “presumptive BK PyVAN”, defined by a high viruria and DNAemia (Zhong et al. J. Gen. Virol. 2009, 90, 144-152). In BKPyV-HC, symptoms can persist for several weeks in subjects with high plasma DNAemia (Imlay et al. Blood Adv 2020, 4, 617-628), and their resolution is thought to depend on the reconstitution of the BK-specific immune response that follows successful engraftment (Espada et al. Blood Adv 2020, 4, 1881-1893). In both clinical situations, patients would benefit from effective antiviral therapy. [0007] The incidence of BK PyVAN is higher in kidney transplantation recipients with low neutralizing antibody titers (Solis et al. J. Am. Soc. Nephrol. 2018, 29, 326-334), and some studies and case reports have indicated that infusion of intravenous immunoglobulin (IVIG), containing BKPyV-specific neutralizing antibodies (Velay et al. Antimicrob Agents Chemother 2019, 63) can prevent active BKPyV replication in kidney transplantation recipients (Benotmane et al. Am J Transplant 2020), and successfully treat BK polyomavirus-associated nephropathy (Matsumura et al. Ther Clin Risk Manag 2020, 16, 947-952; Pibum et al. Pediatric Transplantation 2020, 24, el3600; Hwang et al. Transplant Proc 2018, 50, 2575-2578). Therefore, the existing data indicates that antibody-based therapies may be effective against BKPyV. However, IVIG is a blood product that contains a complex mix of antibody specificities, and although rare, acute antibody-mediated rejection after infusion of IVIG containing donorspecific antibodies has been reported in the literature (Mainra, et al., Transpl Immunol 2013, 28, 145-147). Similarly, antiviral T-cell therapy has been developed for BKPyV-HC, with reported clinical response rates greater than 80% (Nelson et al. Curr. Opin. Infect. Dis., 2021, 34, 627- 634). However, at least one fatal outcome associated with cytokine reconstitution syndrome has been reported in a BKPyV-HC patient treated with T-cell therapy.
[0008] The broadly neutralizing monoclonal antibody 41F17 has been identified as an anti-BKPyV antibody efficiently for binding and neutralizing all of the four genotypes of the BKPyV wildtype (Lindner et al. Immunity. 2019; 50:668-676.e5). However, BKPyV has the ability to escape selection pressure by antibodies by developing BKPyV variants containing mutations on its VP1 capsid protein. For instance, even if the anti-BKPyV 41F17 antibody is efficient against the four wildtype BKPyV genotypes, the selected BKPyV E61K variant and a newly in vitro selected BKPyV D58Del_59A variant escape, or are less neutralized, by said anti- BKPyV 41F17 antibody. Therefore, the anti-BKPyV 41F17 antibody may not be enough to efficiently neutralize all the BKPyV variants.
[0009] Broadly neutralizing antibodies, capable of inhibiting the four wildtype BKPyV genotypes and its variants would be potential candidates for therapeutics for the treatment of BKPyV infection-associated diseases, in particular PyVAN and/or BKPyV-HC.
[0010] Therefore, there is still a need to provide new antibodies and/or combinations of antibodies effective against BKPyV infection, especially for preventing and/or treating BKPyV infection-associated diseases.
[0011] BKPyV infection-associated diseases include BK polyomavirus-associated nephropathy, confirmed and presumptive, BKPyV hemorrhagic cystitis, as well as pneumonitis, retinitis, and meningoencephalitis in immunocompromised subjects, and the likely long-term consequences of BKPyV infection-associated diseases such as reno-urinary tumors, for example bladder carcinoma.
[0012] There is a need to provide antibodies and/or combination of antibodies strongly neutralizing the four wildtype BKPyV genotypes as well as naturally occurring BKPyV variants.
[0013] There is a need to provide antibodies and/or combination of antibodies able to bind and neutralize BKPyV variants escaping neutralization by the benchmark anti-BKPyV antibody 41F17.
[0014] There is a need to provide antibodies able to bind and neutralize BKPyV variants escaping neutralization by the P8D 11 anti-BKPyV antibody.
[0015] There is a need to provide antibodies able to bind and neutralize BKPyV variants escaping neutralization by both P8D11 and 41F17 anti-BKPyV antibodies.
[0016] The present disclosure aims at satisfying all or parts of those needs.
SUMMARY
[0017] According to one of its objects, the present disclosure relates to an isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising:
[0018] - a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0019] - a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0020] - a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0021] - a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0022] - a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and [0023] - a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
[0024] According to one of its objects, the present disclosure relates to an isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising:
[0025] (i) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution; or
[0026] (ii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution; or
[0027] (iii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution; or
[0028] (iv) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution; or
[0029] (v) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution; or
[0030] (vi) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution; or
[0031] (vii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0032] In some embodiments, the isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, may comprise:
[0033] a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or an amino acid sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or an amino acid sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or an amino acid sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or an amino acid sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or an amino acid sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
[0034] In some embodiments, the isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, may comprise:
[0035] a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or an amino acid sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or an amino acid sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or an amino acid sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or an amino acid sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or an amino acid sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or an amino acid sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0036] In the present description, in accordance with the context and for sake of conciseness, the above items (i) to (vii) are used to refer to (a) a group of antibodies or (b) a specific antibody comprising the indicated CDRs.
[0037] As shown in the Examples section, the inventors have surprisingly identified new antibodies able to strongly neutralize BKPyV infection induced by the different wildtype and naturally occurring variant genotypes in different host cells. Furthermore, it has been shown that combinations of at least two of those antibodies surprisingly were able to neutralize variants of BKPyV escaping neutralization by the benchmark anti-BKPyV antibody 41F17.
[0038] It has also been shown that antibody BK120 surprisingly was able to neutralize variants of BKPyV escaping neutralization by both the benchmark 41F17 and P8D11 anti- BKPyV antibodies.
[0039] The disclosed antibodies, and combinations thereof, are able to bind and neutralize BKPyV.
[0040] Advantageously those antibodies and combinations of antibodies may be implemented in prophylactic and/or therapeutic treatment of BKPyV-induced infections and diseases associated with BKPyV infections in transplanted subjects, such as kidney transplanted subjects.
[0041] Further, the prophylactic and/or therapeutic uses of those antibodies and combinations of antibodies may advantageously be associated with a reduced of occurrence of antibody-resistant variants of BKPyV.
[0042] An antibody disclosed herein may comprise:
[0043] (i) a variable domain of heavy chain of sequence SEQ ID NO: 6 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 7, or a sequence at least 85% identical thereto; or
[0044] (ii) a variable domain of heavy chain of sequence SEQ ID NO: 19 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 20, or a sequence at least 85% identical thereto; or [0045] (iii) a variable domain of heavy chain of sequence SEQ ID NO: 32 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 33, or a sequence at least 85% identical thereto; or
[0046] (iv) a variable domain of heavy chain of sequence SEQ ID NO: 45 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 46, or a sequence at least 85% identical thereto; or
[0047] (v) a variable domain of heavy chain of sequence SEQ ID NO: 58 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 59, or a sequence at least 85% identical thereto; or
[0048] (vi) a variable domain of heavy chain of sequence SEQ ID NO: 71 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 72, or a sequence at least 85% identical thereto; or
[0049] (vii) a variable domain of heavy chain of sequence SEQ ID NO: 84 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 85, or a sequence at least 85% identical thereto.
[0050] In some embodiments, an antibody as disclosed herein, may comprise a variable domain of heavy chain of the amino acid sequence SEQ ID NO: 6 or an amino acid sequence at least 85% identical thereto and/or a variable domain of light chain of the amino acid sequence SEQ ID NO: 7, or an amino acid sequence at least 85% identical thereto.
[0051] In some embodiments, an antibody as disclosed herein, may comprise a variable domain of heavy chain of the amino acid sequence SEQ ID NO: 84 or an amino acid sequence at least 85% identical thereto and/or a variable domain of light chain of the amino acid sequence SEQ ID NO: 85, or an amino acid sequence at least 85% identical thereto.
[0052] An antibody disclosed herein may comprise:
[0053] (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or a sequence at least 85% identical thereto; or
[0054] (ii) a heavy chain of sequence SEQ ID NO: 21 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 22, or a sequence at least 85% identical thereto, or
[0055] (iii) a heavy chain of sequence SEQ ID NO: 34 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 35, or a sequence at least 85% identical thereto, or [0056] (iv) a heavy chain of sequence SEQ ID NO: 47 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 48, or a sequence at least 85% identical thereto, or
[0057] (v) a heavy chain of sequence SEQ ID NO: 60 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 61, or a sequence at least 85% identical thereto, or
[0058] (vi) a heavy chain of sequence SEQ ID NO: 73 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 74, or a sequence at least 85% identical thereto, or
[0059] (vii) a heavy chain of sequence SEQ ID NO: 86 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 87, or a sequence at least 85% identical thereto.
[0060] In some embodiments, an antibody as disclosed herein, may comprise a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or an amino acid sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or an amino acid sequence at least 85% identical thereto.
[0061] In some embodiments, an antibody as disclosed herein, may comprise a heavy chain of amino acid sequence SEQ ID NO: 86 or an amino acid sequence at least 85% identical thereto and/or a light chain of amino acid sequence SEQ ID NO: 87, or an amino acid sequence at least 85% identical thereto.
[0062] In another of its objects, the disclosure relates to an isolated antibody, or an antigen-binding fragment thereof, neutralizing at least one BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101, or SEQ ID NO: 106, and for example at least one BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, or SEQ ID NO: 101, with an IC50 of less than 40 nM.
[0063] An antibody disclosed herein bind and neutralize a BKPy virus variant selected among BKPyV-D58Del_59A, BKPyV-E61K, BKPyV-E327K and BKPyV-K172A, and for example a BKPy virus variant selected among BKPyV-D58Del_59A, BKPyV-E61K, and BKPyV-E327K. Such an antibody may bind and neutralize at least two BKPy virus variants selected among BKPyV-D58Del_59A, BKPyV-E61K and BKPyV-E327K.
[0064] An antibody disclosed herein may be an antibody fragment.
[0065] An antibody fragment may be selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, and diabodies. [0066] An antibody disclosed herein may be a variable heavy chain of a single domain antibody (VHH). The variable heavy chain may be as defined herein.
[0067] According to another of its objects, the present disclosure relates to an immunoconjugate comprising an antibody as disclosed herein conjugated or linked to a detectable probe, label or reporter molecule. Such immunoconjugate may be used as a diagnostic tool for detecting and/or measuring a BKPyV infection in a subject in need thereof.
[0068] A detectable probe, label or reporter molecule can be a radioisotope, such as 3H, 14C, 32P, 35S, or 125I; a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine; or an enzyme such as alkaline phosphatase, P-galactosidase, horseradish peroxidase, or luciferase.
[0069] According to another of its objects, the present disclosure relates to a combination of at least a first antibody, or an antigen-binding fragment thereof, and a second antibody, or an antigen-binding fragment thereof, wherein said first and second antibodies may be selected among the groups of antibodies as disclosed herein.
[0070] According to another of its objects, the present disclosure relates to an isolated multi-specific antibody comprising at least a first antibody, or an antigen-binding fragment thereof, and a second antibody, or an antigen-binding fragment thereof, joined to each other, wherein said first and second antibodies may be selected among the groups of antibodies as disclosed herein.
[0071] An isolated multi-specific antibody may be a bispecific antibody.
[0072] A combination or an isolated multi-specific antibody disclosed herein may comprise:
[0073] (a) as a first antibody, an antibody selected from groups (i) or (iv) of antibodies as disclosed herein, and
[0074] (b) as a second antibody, an antibody selected from groups (i), (iv), (vi) or (vii) of antibodies as disclosed herein.
[0075] A combination or an isolated multi-specific antibody disclosed herein may comprise:
[0076] (a) as a first antibody, an antibody selected from group (i) of antibodies as disclosed herein, and as a second antibody, an antibody selected from group (iv) of antibodies as disclosed herein; or
[0077] (b) as a first antibody, an antibody selected from group (i) of antibodies as disclosed herein, and as a second antibody, an antibody selected from group (vi) of antibodies as disclosed herein; or [0078] (c) as a first antibody, an antibody selected from group (i) of antibodies as disclosed herein, and as a second antibody, an antibody selected from group (vii) of antibodies as disclosed herein; or
[0079] (d) as a first antibody, an antibody selected from group (iv) of antibodies as disclosed herein, and as a second antibody, an antibody selected from group (vi) of antibodies as disclosed herein.
[0080] According to another of its objects, the present disclosure relates to a pharmaceutical composition comprising at least an antibody as disclosed herein, at least a combination of at least a first and a second antibodies as disclosed herein, or at least a multispecific antibody as disclosed herein, and a pharmaceutically acceptable carrier.
[0081] According to another of its objects, the present disclosure relates to an isolated antibody as disclosed herein, at least a combination of at least a first and a second antibodies as disclosed herein, at least a multi-specific antibody as disclosed herein, or a pharmaceutical composition as disclosed herein, for use as a medicament.
[0082] According to another of its objects, the present disclosure relates to an isolated antibody as disclosed herein, at least a combination of at least a first and a second antibodies as disclosed herein, at least a multi-specific antibody as disclosed herein, or a pharmaceutical composition as disclosed herein, for use in the prevention and/or treatment of a BKPyV -infection and/or a BKPyV -inf ection associated disease, in a subject in need thereof.
[0083] A subject in need thereof may be an organ transplant recipient. An organ transplant recipient may be a kidney transplant recipient.
[0084] A BKPyV-infection associated disease may be a BK polyomavirus-associated nephropathy (BK PyVAN), a hemorrhagic cystitis, (BK Py-HC) a granule cell neuronopathy (GCN), an interstitial kidney disease, an ureteral stenosis, a pneumonitis, a vasculitis, a colitis, a meningitis, a retinitis, a meningoencephalitis, an immune reconstitution inflammatory syndrome (IRIS), or a reno-urinary cancer.
[0085] In one embodiment, a disorder induced by a BKPyV infection may be a hemorrhagic cystitis.
[0086] In one embodiment, a disorder induced by a BKPyV infection may be a BK polyomavirus-associated nephropathy.
[0087] In one embodiment, a disorder induced by a BKPyV infection may be a reno- urinary cancer, for example a bladder cancer.
[0088] According to another of its objects, the present disclosure relates to an isolated nucleic acid sequence comprising a sequence encoding an antibody disclosed herein. [0089] A nucleic acid sequence disclosed herein may comprise:
[0090] (i) a nucleic acid of sequence SEQ ID NO: 12 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 13 or a sequence at least 85% identical thereto; or
[0091] (ii) a nucleic acid of sequence SEQ ID NO: 25 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 26 or a sequence at least 85% identical thereto; or
[0092] (iii) a nucleic acid of sequence SEQ ID NO: 38 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 39 or a sequence at least 85% identical thereto; or
[0093] (iv) a nucleic acid of sequence SEQ ID NO: 51 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 52 or a sequence at least 85% identical thereto; or
[0094] (v) a nucleic acid of sequence SEQ ID NO: 64 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 65 or a sequence at least 85% identical thereto; or
[0095] (vi) a nucleic acid of sequence SEQ ID NO: 77 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 78 or a sequence at least 85% identical thereto; or
[0096] (vii) a nucleic acid of sequence SEQ ID NO: 90 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 91 or a sequence at least 85% identical thereto.
[0097] A nucleic acid sequence disclosed herein may comprise:
[0098] (i) a nucleic acid of sequence SEQ ID NO: 10 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 11 or a sequence at least 85% identical thereto; or
[0099] (ii) a nucleic acid of sequence SEQ ID NO: 23 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 24 or a sequence at least 85% identical thereto; or
[0100] (iii) a nucleic acid of sequence SEQ ID NO: 36 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 37 or a sequence at least 85% identical thereto; or [0101] (iv) a nucleic acid of sequence SEQ ID NO: 49 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 50 or a sequence at least 85% identical thereto; or
[0102] (v) a nucleic acid of sequence SEQ ID NO: 62 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 63 or a sequence at least 85% identical thereto; or
[0103] (vi) a nucleic acid of sequence SEQ ID NO: 75 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 76 or a sequence at least 85% identical thereto; or
[0104] (vii) a nucleic acid of sequence SEQ ID NO: 88 or a sequence at least 85% identical thereto and/or a nucleic acid of sequence SEQ ID NO: 89 or a sequence at least 85% identical thereto.
[0105] According to another of its objects, the present disclosure relates to an isolated or recombinant expression vector comprising a nucleic acid sequence disclosed herein.
[0106] According to another of its objects, the present disclosure relates to a recombinant host cell which has been transformed by a nucleic acid or an expression vector disclosed herein.
[0107] According to another of its objects, the present disclosure relates to a method for producing an antibody binding a BK polyoma virus (BKPyV) VP1 capsid protein, or an antigenbinding fragment thereof, comprising at least the steps of culturing a host cell disclosed in a culture medium under conditions suitable for producing said antibody or fragment thereof and recovering the antibody or fragment thereof from said cultured cells.
[0108] According to another of its objects, the present disclosure relates to an isolated BKPy VP1 protein of amino acid sequence SEQ ID NO: 96 or an isolated VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
[0109] According to another of its objects, the present disclosure relates to an isolated BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence. Such isolated BKPy virus may be a pseudotype virus or a virus. Herein pseudotype virus and pseudovirus are used interchangeably.
[0110] According to another of its objects, the present disclosure relates to a use of a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence for screening a candidate antibody able to neutralize a BKPy virus. [0111] The BKPy virus may be a pseudotype virus.
[0112] The BKPy virus pseudotype may be BKPyV g!b2-D58DelP59A.
[0113] According to another of its objects, the present disclosure relates to an in vitro method for screening a candidate antibody able to neutralize a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, said method comprising at least the steps of:
[0114] a) culturing cells infected with said virus (a-i) in absence or (a-ii) in presence of said candidate antibody in conditions suitable for neutralization of the pseudotype virus by the candidate antibody,
[0115] b) numbering the infected cells obtained at steps (a-i) and (a-ii),
[0116] c) comparing the number of infected cells obtained at step (a-i) with the number of infected cells obtained at step (a-ii),
[0117] wherein a number of infected cells obtained at step (a-ii) lower than the number of cells obtained at step (a-i) is indicative of a neutralizing activity of said candidate antibody. The BKPy virus may be a pseudotype virus.
[0118] In such method, the BKPy virus may be BKPyV g!b2-D58DelP59A.
[0119] According to another of its objects, the present disclosure relates to a method for manufacturing a pharmaceutical composition for treating and/or preventing a BKPyV infection or for treating and/or preventing a disorder induced by a BKPyV infection in a subject in need thereof, the method comprising at least the steps of:
[0120] a) screening an antibody according to a method disclosed herein,
[0121] b) formulating the antibody obtained at step a) with at least a pharmaceutically acceptable carrier.
[0122] According to another of its objects, the present disclosure relates to an in vitro method for measuring a neutralizing activity of an anti -BKPyV antibody isolated from a subject presumed to be infected with a BKPy virus, the method comprising at least the steps of:
[0123] a) culturing cells infected with a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, (a-i) in absence or (a-ii) in presence of said anti-BKPyV antibody in conditions suitable for neutralization of the virus by said antibody,
[0124] b) numbering the infected cells obtained at steps (a-i) and (a-ii), [0125] c) comparing the number of cells obtained at step (a-i) with the number of cells obtained at step (a-ii),
[0126] wherein a number of cells obtained at step (a-ii) lower than the number of cells obtained at step (a-i) is indicative of a neutralizing activity of the antibody.
BRIEF DESCRIPTION OF THE FIGURES
[0127] FIGURES 1A-F: represent the binding properties of BK120 (Fig. 1A), BK160- 1 (Fig. IB), BK198 (Fig. 1C), BK206 (Fig. ID), BK256 (Fig. IE), and BK293 (Fig. IF), antibodies measured by ELISA against the four wildtype BKPyV genotypes: gla (•), gll (■), gill (A) and gIVc2 (▼).
[0128] FIGURES 2A-G represent the neutralization capacity of BK120 (Fig. 2A),
BK160-1 (Fig. 2B), BK198 (Fig. 2C), BK206 (Fig. 2D), BK256 (Fig. 2E), BK293 (Fig. 2F) and BK304 (Fig. 2G) antibodies on HEK293TT cells infected with wildtype pseudotype viruses
(PSV) gla (•), gll (■), gill (A) and gIVc2 (▼). Each antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL. Abscissa: Antibody concentration in pg/ml in loglO.
BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
[0129] FIGURES 3A-G represent the neutralization capacity of BK120 (Fig. 3A), BK160-1 (Fig. 3B), BK198 (Fig. 3C), BK206 (Fig. 3D), BK256 (Fig. 3E), BK293 (Fig. 3F) and BK304 (Fig. 3G) antibodies on RS cells infected with wildtype pseudotype viruses (PSV) gla (•), gll (■), gill (A) and gIVc2 (▼). Each antibody was added to BKPyV PSV in serial 5- fold dilutions starting at 10 pg/mL. Abscissa: Antibody concentration in pg/ml in loglO.
Ordinate BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
[0130] FIGURES 4A-H represent the viral neutralization capacity of BK120 (Fig. 4A), BK206 (Fig. 4B), BK293 (Fig. 4C), BK304 (Fig. 4D), BK120 + BK206 (Fig. 4E), BK120 + BK293 (Fig. 4F), BK120 + BK304 (Fig. 4G), and 41F17 (Fig. 4H) on HEK293TT cells infected with wildtype and naturally occurring variant pseudotype viruses (PSV) glb2-wt (•), glb2- DNEQ (■), gIb2-VVAE (+), gIVc2 (▼) and gIVc2-SDKAN (0). Each antibody or combination was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL. Abscissa: Antibody or combination concentration in pg/ml in loglO. Ordinate: BKPyV infection rate in percentage relative to cells infected in the absence of antibody. [0131] FIGURES 5A-H represent the viral neutralization capacity of BK120 (Fig. 5A), BK206 (Fig. 5B), BK293 (Fig. 5C), BK304 (Fig. 5D), BK120 + BK206 (Fig. 5E), BK120 + BK293 (Fig. 5F), BK120 + BK304 (Fig. 5G), and 41F17 (Fig. 5H) on HEK293TT cells infected with naturally occurring and in vitro selected variants pseudotype viruses (PSV) g!b2-D58Del- P59A (•), gIb2-E61K (■), gIVc2-E327K (▼), gIVc2-wt (♦). Each antibody or combination was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL. Abscissa: Antibody or combination concentration in pg/ml in loglO. Ordinate: BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
[0132] FIGURES 6A-D represent the viral neutralization capacity of BK120 (o), compared to P8D11 (■) and 41F17 (A) antibodies on cells infected with wildtype pseudotype viruses (PSV) gl (Fig. 6A), gll (Fig. 6B), gill (Fig. 6C) and gIV (Fig. 6D). P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL, while BK120 and 41F17 antibodies were added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL . Abscissa: Antibody concentration in ng/ml in log 10. Ordinate: BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
[0133] FIGURES 7A-B represent the viral neutralization capacity of BK120 (o), compared to P8D11 (■) and 41F17 (A) on cells infected with variants pseudotype viruses (PSV) g!b2-E61K (Fig. 7A) and g!b2-K172A (Fig. 7B). P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL, while BK120 and 41F17 antibodies were added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL . Abscissa: Antibody concentration in ng/ml in log 10. Ordinate: BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
[0134] FIGURES 8A-B represent the viral neutralization capacity of BK120 (o), compared to P8D11 (■) and 41F17 (A) antibodies on cells infected with variants pseudotype viruses (PSV) g!b2-DNEQ (Fig. 8A) and gIVc2 SDKAN (Fig. 8B). P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL, while BK120 and 41F17 antibodies were added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL. Abscissa: Antibody concentration in ng/ml in log 10. Ordinate: BKPyV infection rate in percentage relative to cells infected in the absence of antibody.
BRIEF DESCRIPTION OF THE SEQUENCES
SEQ ID NO: 1 represents the amino acids sequence of the HCDR1 of BK120.
SEQ ID NO: 2 represents the amino acids sequence of the HCDR2 of BK120. SEQ ID NO: 3 represents the amino acids sequence of the HCDR3 of BK120.
SEQ ID NO: 4 represents the amino acids sequence of the LCDR1 of BK120.
SEQ ID NO: 5 represents the amino acids sequence of the LCDR3 of BK120.
SEQ ID NO: 6 represents the amino acids sequence of the heavy chain variable domain (VH) of BK120.
SEQ ID NO: 7 represents the amino acids sequence of the light chain variable domain (VL) of BK120.
SEQ ID NO 8 represents the amino acids sequence of the heavy chain of BK120. SEQ ID NO 9 represents the amino acids sequence of the light chain of BK120. SEQ ID NO 10 represents the nucleic acids sequence of the VH of BK120. SEQ ID NO 11 represents the nucleic acids sequence of the VL of BK120. SEQ ID NO 12 represents the nucleic acids sequence of the heavy chain of the BK120. SEQ ID NO 13 represents the nucleic acids sequence of the light chain of the BK120. SEQ ID NO 14 represents the amino acids sequence of the HCDR1 of BK160-1. SEQ ID NO 15 represents the amino acids sequence of the HCDR2 of B KI 60-1. SEQ ID NO 16 represents the amino acids sequence of the HCDR3 of B KI 60-1. SEQ ID NO 17 represents the amino acids sequence of the LCDR1 of BK160-1. SEQ ID NO 18 represents the amino acids sequence of the LCDR3 of B KI 60-1. SEQ ID NO 19 represents the amino acids sequence of the VH of B KI 60-1. SEQ ID NO 20 represents the amino acids sequence of the VL of B KI 60-1. SEQ ID NO 21 represents the amino acids sequence of the heavy chain of BK160-1. SEQ ID NO 22 represents the amino acids sequence of the light chain of B KI 60-1. SEQ ID NO 23 represents the nucleic acids sequence of the VH of B KI 60-1. SEQ ID NO 24 represents the nucleic acids sequence of the VL of B KI 60-1. SEQ ID NO 25 represents the nucleic acids sequence of the heavy chain of the B KI 60-1. SEQ ID NO 26 represents the nucleic acids sequence of the light chain of the B KI 60-1. SEQ ID NO 27 represents the amino acids sequence of the HCDR1 of BK198. SEQ ID NO 28 represents the amino acids sequence of the HCDR2 of BK198. SEQ ID NO 29 represents the amino acids sequence of the HCDR3 of BK198. SEQ ID NO 30 represents the amino acids sequence of the LCDR1 of BK198. SEQ ID NO 31 represents the amino acids sequence of the LCDR3 of BK198. SEQ ID NO 32 represents the amino acids sequence of the VH of BK198. SEQ ID NO 33 represents the amino acids sequence of the VL of BK198. SEQ ID NO 34 represents the amino acids sequence of the heavy chain of BK198. SEQ ID NO 35 represents the amino acids sequence of the light chain of BK198. SEQ ID NO 36 represents the nucleic acids sequence of the VH of BK198. SEQ ID NO 37 represents the nucleic acids sequence of the VL of BK198. SEQ ID NO 38 represents the nucleic acids sequence of the heavy chain of the BK198. SEQ ID NO 39 represents the nucleic acids sequence of the light chain of the BK198. SEQ ID NO 40 represents the amino acids sequence of the HCDR1 of BK206. SEQ ID NO 41 represents the amino acids sequence of the HCDR2 of BK206. SEQ ID NO 42 represents the amino acids sequence of the HCDR3 of BK206. SEQ ID NO 43 represents the amino acids sequence of the LCDR1 of BK206. SEQ ID NO 44 represents the amino acids sequence of the LCDR3 of BK206. SEQ ID NO 45 represents the amino acids sequence of the VH of BK206. SEQ ID NO 46 represents the amino acids sequence of the VL of BK206. SEQ ID NO 47 represents the amino acids sequence of the heavy chain of BK206. SEQ ID NO 48 represents the amino acids sequence of the light chain of BK206. SEQ ID NO 49 represents the nucleic acids sequence of the VH of BK206. SEQ ID NO 50 represents the nucleic acids sequence of the VL of BK206. SEQ ID NO 51 represents the nucleic acids sequence of the heavy chain of the BK206. SEQ ID NO 52 represents the nucleic acids sequence of the light chain of the BK206. SEQ ID NO 53 represents the amino acids sequence of the HCDR1 of BK256. SEQ ID NO 54 represents the amino acids sequence of the HCDR2 of BK256. SEQ ID NO 55 represents the amino acids sequence of the HCDR3 of BK256. SEQ ID NO 56 represents the amino acids sequence of the LCDR1 of BK256. SEQ ID NO 57 represents the amino acids sequence of the LCDR3 of BK256. SEQ ID NO 58 represents the amino acids sequence of the VH of BK256. SEQ ID NO 59 represents the amino acids sequence of the VL of BK256. SEQ ID NO 60 represents the amino acids sequence of the heavy chain of BK256. SEQ ID NO 61 represents the amino acids sequence of the light chain of BK256. SEQ ID NO 62 represents the nucleic acids sequence of the VH of BK256. SEQ ID NO 63 represents the nucleic acids sequence of the VL of BK256. SEQ ID NO 64 represents the nucleic acids sequence of the heavy chain of the BK256. SEQ ID NO 65 represents the nucleic acids sequence of the light chain of the BK256. SEQ ID NO 66 represents the amino acids sequence of the HCDR1 of BK293. SEQ ID NO 67 represents the amino acids sequence of the HCDR2 of BK293. SEQ ID NO 68 represents the amino acids sequence of the HCDR3 of BK293. SEQ ID NO: 69 represents the amino acids sequence of the LCDR1 of BK293.
SEQ ID NO: 70 represents the amino acids sequence of the LCDR3 of BK293.
SEQ ID NO: 71 represents the amino acids sequence of the VH of BK293.
SEQ ID NO: 72 represents the amino acids sequence of the VL of BK293.
SEQ ID NO: 73 represents the amino acids sequence of the heavy chain of BK293.
SEQ ID NO: 74 represents the amino acids sequence of the light chain of BK293.
SEQ ID NO: 75 represents the nucleic acids sequence of the VH of BK293.
SEQ ID NO: 76 represents the nucleic acids sequence of the VL of BK293.
SEQ ID NO: 77 represents the nucleic acids sequence of the heavy chain of the BK293.
SEQ ID NO: 78 represents the nucleic acids sequence of the light chain of the BK293.
SEQ ID NO: 79 represents the amino acids sequence of the HCDR1 of BK304.
SEQ ID NO: 80 represents the amino acids sequence of the HCDR2 of BK304.
SEQ ID NO: 81 represents the amino acids sequence of the HCDR3 of BK304.
SEQ ID NO: 82 represents the amino acids sequence of the LCDR1 of BK304.
SEQ ID NO: 83 represents the amino acids sequence of the LCDR3 of BK304.
SEQ ID NO: 84 represents the amino acids sequence of the VH of BK304.
SEQ ID NO: 85 represents the amino acids sequence of the VL of BK304.
SEQ ID NO: 86 represents the amino acids sequence of the heavy chain of BK304.
SEQ ID NO: 87 represents the amino acids sequence of the light chain of BK304.
SEQ ID NO: 88 represents the nucleic acids sequence of the VH of BK304.
SEQ ID NO: 89 represents the nucleic acids sequence of the VL of BK304.
SEQ ID NO: 90 represents the nucleic acids sequence of the heavy chain of the BK304.
SEQ ID NO: 91 represents the nucleic acids sequence of the light chain of the BK304.
SEQ ID NO: 92 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV. genotype la.
SEQ ID NO: 93 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype Ib2.
SEQ ID NO: 94 represents the amino acids sequence of the VP1 protein of the DNEQ BKPyV genotype Ib2.
SEQ ID NO: 95 represents the amino acids sequence of the VP1 protein of the VVAE BKPyV genotype Ib2.
SEQ ID NO: 96 represents the amino acids sequence of the VP1 protein of the D58Del_59A BKPyV genotype Ib2. SEQ ID NO: 97 represents the amino acids sequence of the VP1 protein of the E61K BKPyV genotype Ib2.
SEQ ID NO: 98 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype II.
SEQ ID NO: 99 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype III.
SEQ ID NO: 100 represents the amino acids sequence of the VP1 protein of the wildtype BKPyV genotype IVc2.
SEQ ID NO: 101 represents the amino acids sequence of the VP1 protein of the E327K BKPyV genotype IVc2.
SEQ ID NO: 102 represents the amino acids sequence of the VP1 protein of the SDKAN BKPyV genotype IVc2.
SEQ ID NO: 103 represents the amino acids sequence of the VP1 protein of the wildtype MPyV.
SEQ ID NO: 104 represents the nucleic acids sequence of the primer CMV-F.
SEQ ID NO: 105 represents the nucleic acids sequence of the primer pEGFP-Nl-R.
SEQ ID NO: 106 represents the amino acids sequence of the VP1 protein of the K172A BKPyV genotype Ib2.
DETAILED DESCRIPTION
[0135] The present disclosure is not limited to the particular methodology, protocols, cell lines, vectors, or reagents described herein since they may vary without departing from the spirit and scope of the disclosure. Further, the terminology used herein is for the purpose of exemplifying particular embodiments only and is not intended to limit the scope of the disclosure. Unless defined otherwise, all technical and scientific terms and any acronyms used herein have the same meanings as commonly understood by one of ordinary skill in the art in the field of the disclosure. Any method and material similar or equivalent to those described herein can be used in the practice of the present disclosure and only exemplary methods, devices, and materials are described herein.
[0136] All patents and publications mentioned herein are incorporated by reference for the purpose of describing and disclosing the antibodies, vectors, nucleic acids, host cells and methodologies reported therein that might be used with and in the present disclosure. [0137] Prior to teaching the making and using of the antibodies binding a BK polyomavirus (BKPyV) VP1 protein, or an antigen-binding fragment thereof, and combinations of such antibodies related methods and products of interest, the following non-limiting definitions of some terms and phrases are provided.
Definitions
[0138] Unless otherwise defined herein, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by those of ordinary skill in the art. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press, may provide one of skill with a general dictionary of many of the terms used in this disclosure. In case of conflict, the present specification, including definitions, will control. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention. Units, prefixes, and symbols are denoted in their International System of Units (SI) accepted form. The headings provided herein are not limitations of the various aspects of the disclosure.
[0139] Numeric ranges are inclusive of all the numbers defining the range. Unless otherwise indicated, amino acid sequences are written left to right in amino to carboxy orientation.
[0140] All publications and other references mentioned herein are incorporated by reference in their entirety.
[0141] It is to be noted that the term “a” or “an” entity refers to one or more of that entity; for example, “a nucleotide sequence,” is understood to represent one or more nucleotide sequences. As such, the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably herein.
[0142] Furthermore, “and/or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and/or” as used in a phrase such as “A and/or B” herein is intended to include “A and B”, “A or B”, “A” (alone), and “B” (alone).
[0143] It is understood that aspects and embodiments of the present disclosure described herein include “comprising”, “having”, “consisting of’, and “consisting essentially of’ aspects and embodiments. The words “have” and “comprise,” or variations such as “has,” “having”, “comprises”, or “comprising”, will be understood to imply the inclusion of the stated element(s) (such as a composition of matter or a method step) but not the exclusion of any other elements. The term “consisting of’ implies the inclusion of the stated element(s), to the exclusion of any additional elements. The term “consisting essentially of’ implies the inclusion of the stated elements, and possibly other element(s) where the other element(s) do not materially affect the characteristic(s) of the stated elements. It is understood that the different embodiments of the disclosure using the term “comprising” or equivalent cover the embodiments where this term is replaced with “consisting of’ or “consisting essentially of’.
[0144] As used herein the term "antibody" have the same meaning and will be used equally in the present description. The term "antibody" as used herein refers to isolated or recombinant immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds an antigen, such as VP1 capsid protein. As such, the term antibody encompasses not only whole antibody molecules, but also antibody fragments as well as variants (including derivatives) of antibodies and antibody fragments. In natural antibodies, two heavy chains are linked to each other by disulphide bonds and each heavy chain is linked to a light chain by a disulphide bond. There are two types of light chain, lambda (1) and kappa (k). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. Each chain contains distinct sequence domains. The light chain includes two domains, a variable domain (VL) and a constant domain (CL). The heavy chain includes four domains, a variable domain (VH) and three constant domains (CHI, CH2 and CH3, collectively referred to as CH). The variable regions of both light (VL) and heavy (VH) chains determine binding recognition and specificity to the antigen. The constant region domains of the light (CL) and heavy (CH) chains confer important biological properties such as antibody chain association, secretion, trans-placental mobility, complement binding, and binding to Ec receptors (EcR). The Fv fragment is the N-terminal part of the Fab fragment of an immunoglobulin and consists of the variable portions of one light chain and one heavy chain. The specificity of the antibody resides in the structural complementarity between the antibody combining site and the antigenic determinant. Antibody combining sites are made up of residues that are primarily from the hypervariable or complementarity determining regions (CDRs). Occasionally, residues from nonhypervariable or framework regions (FR) can participate to the antibody binding site or influence the overall domain structure and hence the combining site. Complementarity Determining Regions or CDRs refer to amino acid sequences which together define the binding affinity and specificity of the natural Fv region of a native immunoglobulin binding site. The light and heavy chains of an immunoglobulin each have three CDRs, designated CDR1-L, CDR2-L, L- CDR3-L and CDR1-H CDR2-H, CDR3-H, respectively. An antigen-binding site, therefore, typically includes six CDRs, comprising the CDR set from each of a heavy and a light chain V region. Framework Regions (FRs) refer to amino acid sequences interposed between CDRs.
[0145] The term “antibody” includes, but is not limited to, monoclonal antibodies, human antibodies, humanized antibodies, camelid antibodies and chimeric antibodies. The antibodies can be of any isotype/class (e.g., IgG, IgE, IgM, IgD, IgA and IgY), or subclass (e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2). In some embodiments, an antibody of the present disclosure refers to an anti-BKPyV antibody. In particular, an antibody of the present disclosure refers to an antibody binding a BKPyV VP1 capsid protein. Preferably, an antibody of the present disclosure may refer to BK120 antibody having the amino acid sequences as set forth in Table 3, BK106-1 antibody having the amino acid sequences as set forth in Table 4, BK198 antibody having the amino acid sequences as set forth in Table 5, BK206 antibody having the amino acid sequences as set forth in Table 6, BK256 antibody having the amino acid sequences as set forth in Table 7, BK293 antibody having the amino acid sequences as set forth in Table 8 and/or BK304 antibody having the amino acid sequences as set forth in Table 9.
[0146] In the context of the present disclosure, the term “antibody” specifically includes an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, a combination of at least two antibodies binding a BKPyV VP1 capsid protein, or a multi-specific antibody binding a BKPyV VP1 capsid protein as described herein.
[0147] It is intended that all references to the term “antibody” or “antigen-binding fragment thereof’ as used herein, referred to the antibodies or the antigen-binding fragments thereof as described in the present disclosure.
[0148] An "isolated" compound or entity, such as an antibody or antigen-binding fragment thereof, a vector or a cell, is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the compound or entity, and may include enzymes, hormones, and other proteinaceous or non- proteinaceous solutes. In some embodiments, the compound or entity is partially or substantially purified. In some embodiments, the compound or entity may be purified to greater than about 95% by weight of antibody as determined by the Lowry method, for example, more than 95%, 96%, 97%, 98% or 99% by weight of compound or entity. In particular, an isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody's natural environment is not present. Preferably, however, an isolated antibody may be prepared by at least one purification step.
[0149] The term “recombinant” as applied to an antibody, or an antigen-binding fragment thereof, a nucleic acid sequence, an expression vector or a host cell means that those are the products of various combinations of in vitro cloning, restriction, ligation steps, and other genetic engineering procedures.
[0150] The terms “monoclonal antibody”, “monoclonal Ab”, “monoclonal antibody composition”, “mAb”, or the like, as used herein refer to a preparation of antibody molecules of single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope on an antigen, in particular VP1 capsid protein of BKPyV.
[0151] The expression “complementarity-determining regions” or “CDRs” interchangeably refers to the hypervariable regions of VL and VH. The CDRs are the target protein-binding site of the antibody chains that harbors specificity for such target protein, in particular BKPyV VP1 capsid protein. There are three CDRs in each human VH (CDR1-H, CDR2-H, CDR3-H) or VL (CDR1-L, CDR2-L, CDR3-L), constituting in total about 15-20% of the variable domains. CDRs can be referred to by their region and order. For example, “VHCDR1”, “CDRLH” or “H-CDR1” refer to the first CDR of the heavy chain variable region. The CDRs are structurally complementary to the epitope of the target protein, such as BKPyV VP1 protein, and are thus directly responsible for the binding specificity. The remaining stretches of the VL or VH, i.e., framework regions, exhibit less variation in amino acid sequence. The positions of the CDRs and framework regions can be determined using various well-known definitions in the art, e.g., Kabat, Chothia, IMGT, and AbM.
[0152] In the context of the present disclosure, the amino acid residues of the antibody of the disclosure are numbered according to the IMGT numbering system. The IMGT unique numbering has been defined to compare the variable domains whatever the antigen receptor, the chain type, or the species (Lefranc M.-P., Immunology Today, 18, 509 (1997) ; Lefranc et al. Dev. Comp. Immunol., 27, 55-77 (2003)). In the IMGT unique numbering, the conserved amino acids always have the same position, for instance cysteine 23, tryptophan 41, hydrophobic amino acid 89, cysteine 104, phenylalanine or tryptophan 118. The IMGT unique numbering provides a standardized delimitation of the framework regions (FR1-IMGT: positions 1 to 26, FR2- IMGT: 39 to 55, FR3-IMGT: 66 to 104 and FR4-IMGT: 118 to 128) and of the complementarity determining regions: CDR1-IMGT: 27 to 38, CDR2-IMGT: 56 to 65 and CDR3-IMGT: 105 to 117. If the CDR3-IMGT length is less than 13 amino acids, gaps are created from the top of the loop, in the following order 111, 112, 110, 113, 109, 114, etc. If the CDR3-IMGT length is more than 13 amino acids, additional positions are created between positions 111 and 112 at the top of the CDR3-IMGT loop in the following order 112.1,111.1, 112.2, 111.2, 112.3, 111.3, etc. (http://www.imgt.org/IMGTScientificChart/Nomenclature/ IM GT -FRCDRdefinition.html).
[0153] Both the light and heavy chains are divided into regions of structural and functional homology. The terms “constant” and “variable” are used functionally. In this regard, it will be appreciated that the variable domains of both the light (VL) and heavy (VH) chain portions determine antigen recognition and specificity. Conversely, the constant domains of the light chain (CL) and the heavy chain (CHI, CH2, or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, and the like. By convention, the numbering of the constant region domains increases as they become more distal from the antigen binding site or amino-terminus of the antibody. The N-terminus is a variable region and at the C-terminus is a constant region; the CH3 and CL domains actually comprise the carboxy-terminal domains of the heavy and light chain, respectively.
[0154] The term “identity” or “homology” may mean the percentage of nucleotide bases or amino acid residues in the candidate sequence that are identical with the residue of a corresponding sequence to which it is compared, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent identity for the entire sequence, and not considering any conservative amino acid substitutions as part of the sequence identity. Neither N-terminal or C-terminal extensions nor insertions shall be construed as reducing identity or homology. Methods and computer programs for the alignment are available and well known in the art. Sequence identity may be measured using sequence analysis software. Two examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms (the National Center for Biotechnology Information). The BLASTN program is use for nucleotide sequences, whereas the BLASTP program is used for amino acid sequences.
[0155] The term “antigen binding site” or “antigen binding region” as used in the present disclosure refers to the part of the antibody which comprises the area which specifically binds to and is complementary to part or all of an antigen. Where an antigen is large, an antibody may only bind to a particular part of the antigen, which part is termed on epitope. An antigen binding region may be provided by one or more antibody variable domains. Preferably, an antigen binding region is made of the association of an antibody light chain variable domain (VL) and an antibody heavy chain variable domain (VH). [0156] The term “antigen” as used in the present disclosure refers to a molecule or a portion of a molecule capable of being bound by one or more antibodies. An antigen can have one or more than one epitope. For instance, in the context of the disclosure, an antigen may be a VP1 capsid protein of a human BKPyV. In some embodiments, an antigen of the disclosure may be a VP1 capsid protein selected among BKPyV VP1 gl wt, BKPyV VP1 gll wt, BKPyV VP1 gill wt, BKPyV VP1 gIV wt, BKPyV glb2 wt, BKPyV VP1 glb2 DNEQ, BKPyV glb2 VVAE, BKPyV VP1 gIVc2 wt, BKPyV VP1 gIV c2 SDKAN, BKPyV VP1 glb2 D58delP59A, BKPyV VP1 glb2 E61K, BKPyV VP1 glb2 K172A and BKPyV VP1 gIV E327K.
[0157] The term “affinity”, as used herein, means the strength of the binding of an antibody to an epitope presented on an antigen. The affinity of an antibody is given by the dissociation constant KD, defined as [Ab] x [Ag] I [Ab-Ag], where [Ab-Ag] is the molar concentration of the antibody-antigen complex, [Ab] is the molar concentration of the unbound antibody and [Ag] is the molar concentration of the unbound antigen. The affinity constant Ka is defined by 1/Kd. Methods for determining the affinity of Abs can be found in, for example, Harlow, et al., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988, Coligan et al., Current Protocols in Immunology, Greene Publishing Assoc, and Wiley Interscience, N.Y., (1992, 1993), or Muller, Methods Enzymol. 1983;92:589-601, which references are entirely incorporated herein by reference. A preferred and standard method well known in the art for determining the affinity of mAbs is the measurement of surface plasmon resonance) by using the Biacore instruments (Laure et al. Curr Protoc Protein Sci. 2006 Sep;Chapter 19:Unit 19.13). For multimeric antigens, such as virus capsids, to which both antigen binding sites of an antibody can bind simultaneously, Ka and KD values determined by such standard methods measure the functional affinity, or avidity of the interaction. Illustratively, an antibody is considered to bind an antigen when a functional binding affinity (KD), preferably measured by surface plasmon resonance, is of 10’8 mol/1 (M) or less, preferably 10’9 M to 10 12 M.
[0158] The term “parenteral administration” designates administration routes other than enteral and topical administration, generally by injection, by intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, epidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal routes, including, without limitation, injections, and perfusions.
[0159] The terms “treat”, “treatment” or “therapy” in the present disclosure refers to the administration or consumption of a compound or composition according to the disclosure with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a disorder, the symptoms of the condition, the likelihood of the disorder, or to prevent or delay the onset of the symptoms, complications, or otherwise arrest or inhibit further development of the disorder in a statistically significant manner. More particularly, “treating” or “treatment” includes any approach for obtaining beneficial or desired results in a subject’s BKPyV infection condition. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions of BKPyV infection-associated disease, diminishment or reduction of the extent of a BKPyV infection-associated disease or of a symptom-associated BKPyV infection, stabilizing, i.e., not worsening, the state of a BKPyV infection-associated disease or of a symptom-associated BKPyV infection prevention of a BKPyV infection-associated disease, delay or slowing of BKPyV infection-associated disease or symptom-associated BKPyV infection progression, amelioration or palliation of the BKPyV infection-associated disease state, diminishment of the reoccurrence of BKPyV infection- associated disease, and remission, whether partial or total and whether detectable or undetectable. In other words, “treatment” as used herein includes any cure, amelioration, or reduction of a BKPyV infection-associated disease or related symptom. A “reduction” of a symptom or a disease means decreasing of the severity or frequency of the disease or symptom, or elimination of the disease or symptom.
[0160] As used herein, the terms “prevent” or “preventing” (and grammatical variants thereof) with respect to a disease or disorder relate to prophylactic treatment of a disease, e.g., in a subject suspected to have the disease, or at risk for developing the disease. Prevention may include, but is not limited to, preventing, or delaying onset or progression of the disease and/or maintaining one or more symptoms of the disease at a desired or sub-pathological level. The term “prevent” does not require the 100% elimination of the possibility or likelihood of occurrence of the event. Rather, it denotes that the likelihood of the occurrence of the event has been reduced in the presence of a composition or method as described herein. More particular, “prevent” or “preventing” refers to a decrease in the risk of occurrence of a BKPyV infection- associated disease or symptom in a subject. As indicated above, the prevention may be complete, i.e., no detectable symptoms or disease, or partial, such that fewer symptoms or less severity of the disease are observed than would likely occur absent treatment.
[0161] As used herein, the terms “disorder” and “disease” are used interchangeably herein and comprise any undesired physiological change in a subject or a cell.
[0162] As used herein, the term “infection” refers to the presence of BKPyV in a subject, whether the infected subject presents clinical signs of the infection, in particular by the manifestation of a disorder, or not. Subjects that are infected with BKPyV but have not become clinically manifest are called asymptomatic subjects. In particular, subjects that are infected with BKPyV may be asymptomatic, i.e., subjects have no clinical signs of a BKPyV infection- associated disease.
[0163] As used herein, the terms “therapeutically effective amount” and “prophylactically effective amount” refer to an amount that provides a therapeutic benefit in the treatment, prevention, or management of pathological processes considered. The specific amount that is therapeutically effective can be readily determined by an ordinary medical practitioner and may vary depending on factors such as the type and stage of pathological processes considered, the subject’s medical history and age, and the administration of other therapeutic agents.
[0164] As used herein, the term “patient” or “subject” is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In some embodiments, the subject may be a human. In some embodiments, a subject may be an immunocompromised subject.
[0165] The expression “immunocompromised subject” or “immunosuppressed subject” refers to a subject having a weakened immune system. The immune system of an immunocompromised subjects has a reduced ability to fight virus or bacterial infections and other diseases. An immunocompromised subject includes subjects suffering from a Primary Immune Deficiency Diseases, subjects suffering from a Secondary Immune Deficiency Diseases, subjects being or has been treated with an immunosuppressive therapy; subjects being or has been treated with a chemotherapeutic agent or subjects who has been transplanted. The expression “Primary Immune Deficiency Diseases” refers to an inherited immune disorders resulting from genetic mutations, usually present at birth and diagnosed in childhood. The expression “Secondary Immune Deficiency Diseases” refers to an acquired immunodeficiency as a result of disease or environmental factors, such as AIDS, cancer, diabetes, malnutrition, or medical treatment, such as anticancer drugs, radiation therapy, or stem cell or organ transplant.
[0166] The term “nucleic acid sequence” is used herein interchangeably with the term “polynucleotide” and refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or doublestranded form. The term encompasses isolated or recombinant nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphor amidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
[0167] The terms “polypeptide” and “protein” are used interchangeably herein to refer to a sequence of amino acid residues. The terms apply to amino acid sequences in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid sequences and non-naturally occurring amino acid sequence. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.
[0168] The term “ICso” (half-maximal inhibitory concentration) refers to the concentration of a particular antibody, such as anti-BKPyV antibody, which induces a signal halfway (50%) between the baseline control and the maximum possible signal. For example, the IC50 is the concentration of antibody at which 50% of the available binding sites on the VP1 antigen are occupied.
[0169] The term “target cell” herein means any cell that may be infected by a BKPyV. A target cell also includes a cell expressing a target antigen that can be recognized by an anti- BKPyV antibody according to the present disclosure. For instance, a target cell may be a kidney cell. A target cell used as cellular model in the binding and neutralization assays may be a 293TT cell line or an RS cell line.
[0170] As used herein, “virus-like particles” or “VLP” are an assembly of VP1 pentamers into viral capsids. VLPs are composed of 72 VP1 pentamers. VLPs are structurally very similar to actual virus without the minor capsid proteins (VP2 and VP3) as well as the viral DNA genome, and therefore are non-infectious. VLPs are useful as viral epitopes are presented in a similar conformation to the actual virus.
BK polyomavirus (BKPyV)
[0171] The antibodies described herein specifically bind to the capsid protein VP1 of different BK polyomaviruses and BK pseudotype polyoma viruses. Further, the antibodies neutralize BKPy virus and BKPy pseudotype viruses. They can therefore be effectively used for preventing and treating an infection with BKPyV and/or a disorder induced by a BKPyV infection.
[0172] A pseudovirus or a pseudotype virus is a recombinant viral particle which in addition to its core/backbone, may incorporate envelope proteins derived from different viruses; moreover, the genes inside the pseudovirus are usually altered or modified so that they are unable to produce the surface protein on their own. As such, an additional plasmid or stable cell line expressing the surface proteins is needed to make the pseudovirus. Pseudoviruses are capable of infecting susceptible cells, but they only replicate for 1 round in the infected host cells. Compared with wild-type viruses, pseudoviruses can be safely handled in biosafety level (BSL)- 2 laboratories and are usually easier to manipulate experimentally. Nevertheless, the conformational structure of pseudoviral surface proteins bears high similarity to that of the native viral proteins, and these surface proteins can effectively mediate viral entry into host cells.
[0173] The expression “BK polyoma virus”, “BKPyV” and “BK virus” are herein used interchangeably and intend to refer to a member of the family Polyomaviridae, genus Betapolyomavirus (Helle, Francois et al. Viruses vol. 9,11 327. 3 Nov. 2017).
[0174] BK polyomaviruses are icosahedral, non-enveloped, circular double-stranded DNA viruses with a genome of around 5,300 base pairs. They measure approximately 40-45 nM in diameter (Bennett et al., Microbes and Infection. 2012:14(9):672-683). Further, the BKPyVs have been associated with nephropathy, renal dysfunction such as hemorrhagic cystitis, pneumonitis, retinitis, and meningoencephalitis. The BKPyV may be a whole BKPyV, BKPyV virus-like particles (VLPs) composed only of VP1 protein, or pseudotype virus that are incorporate all capsid proteins and are able to infect susceptible target cells. Up to 90% of the population as a whole is persistently infected with BKPyV. The virus reactivates and is asymptomatically shed into the urine in 5-10% of immunocompetent adults at any given time.
[0175] Based on the sequence of the VP1 gene, four genotypes have been described, corresponding to the four serologically differentiated subtypes I-IV, with different prevalence and geographic distribution. In each genotype, numerous serotype variants exist. The most common genotype worldwide is genotype I, with a frequency of about 80%, followed by genotype IV (about 15%), while genotypes II and III are isolated only sporadically. The distribution of the molecular variants of the virus is associated with the region of origin. BKPyV subtype la is most common in Africa, Ib-1 in Southeast Asia, and Ib-2 in Europe, while Ic is the most common variant in Northeast Asia.
[0176] As shown in the Examples section, the antibodies or antigen-binding fragments thereof are able to specifically bind and neutralize the BKPyV genotype I wildtype (la, Ib-1, Ib- 2 and Ic), as well as the newly identified variant BKPyV genotype Ib2-D58Del_59A and the known variants BKPyV genotype Ib2-E61K, BKPyV genotype Ib2-DNEQ and BKPyV genotype Ib2-VVAE, the BKPyV genotype II wildtype, the BKPyV genotype III wildtype, the BKPyV genotype IV wildtype (IVa-1, IVa-2, IVb-1, IVb-2, IVc-1 and IVc-2), as well as the variants BKPyV genotype IVc2-E327K and BKPyV genotype IVc2-SDKAN.
[0177] Further, as shown in the Examples section, the BK120 antibody or antigenbinding fragments thereof is also able to specifically bind and/or neutralize the variant BKPyV genotype Ib2-K172A.
[0178] The term “BKPyV variant”, or its similar forms, refers to a wild-type BKPy virus wherein some mutations are present on the VP1 capsid proteins of the wild-type BKPyV. The term “BKPyV variant” encompasses naturally occurring BKPyV variants and in vitro selected BKPyV variants.
[0179] As used herein, the expression “naturally occurring BKPyV” intends to refer to variants of BKPyV obtained from subjects, such as genotype Ib2-DNEQ, the BKPyV genotype Ib2-VVAE, and the BKPyV genotype IVc2-SDKAN (McIlroy et al. Viruses 2020).
[0180] The term 'in vitro selected BKPyV” refers to variants of BKPyV obtained by passaging viruses in vitro cell culture under suitable conditions, such as the BKPyV genotype Ia-D58Del_59A, and also the BKPyV genotype Ib2-E61K, the BKPyV genotype Ib2-K172A and the BKPyV genotype IVc2-E327K (Lindner et al. Immunity. 2019;50:668-676.e5 and McIlroy et al. Viruses. 2020 Jul 29; 12(8): 824.).
[0181] The expression “VP1” or “VP1 capsid protein” refers to the major polyoma virus capsid subunit protein. In the virus capsid, the monomers of VP1 oligomerize in pentamers. Expression of the VP1 protein alone is sufficient to produce VLPs that have the same antigenic properties as the infectious virus, since the minor capsid proteins VP2 and VP3 are not exposed on the surface of the virion.
[0182] Virus-like particles (VLPs) are molecules that closely resemble viruses but are non-infectious because they contain no viral genetic material. They can be naturally occurring or synthesized through the individual expression of viral structural proteins, which can then selfassemble into the virus-like structure.
[0183] The antibodies of the disclosure, or the antigen-binding fragments thereof, specifically bind to the VP1 sequences of the BKPyV of Table 1.
[0184] The VP1 coding region shows very high similarity (over 95%) in all genotypes of BKPyV, but the similarity between the amino acid residues from 61E to 83R is only 61-70%
[0185] In some embodiments, the antibodies of the disclosure, or the antigen-binding fragments thereof, specifically may bind to variants VP1 sequences comprising an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequences of the VP1 of BKPyV of Table 1. [0186] In some embodiments, the antibodies of the disclosure, or the antigen-binding fragments thereof, neutralize BKPyV comprising VP1 sequences comprising an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequences of the VP1 of BKPyV of Table 1.
[0187] In what follows, variants and/or mutants are described by reference to the amino acid sequences of the VP1 of BKPyV of Table 1 which are amino acid sequences of reference. Such a description by reference is based on the prerequisite of optimal sequence alignment in order to determine the amino acid in the variant sequence that corresponds to the amino acid defined as being in a specific position in the amino acid of reference.
[0188] In what follows, variants and/or mutants are also described by percent identity with a sequence of reference. Percent identity between two amino acid sequences or two nucleotide sequences is determined with standard alignment algorithms as those described below. A sequence identity refers to the similarity between amino acid sequences expressed in terms of the similarity between the sequences. Sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar the two sequences are. Variants of a polypeptide will possess a relatively high degree of sequence identity when aligned using standard methods.
[0189] Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith and Waterman, Adv. Appl. Math. 2:482, 1981; Needleman and Wunsch, J. Mol. Biol. 48:443, 1970; Pearson and Lipman, Proc. Natl. Acad. Sci. U.S.A. 85:2444, 1988; Higgins and Sharp, Gene 73:237, 1988; Higgins and Sharp, CABIOS 5:151, 1989; Corpet et al., Nucleic Acids Research 16:10881, 1988. Altschul et al., Nature Genet. 6:119, 1994, presents a detailed consideration of sequence alignment methods and homology calculations.
[0190] For example, sequence alignment can be achieved and percent identity can be determined Basic Local Alignment Tools (BLASTs, including BLASTP for amino acid sequence alignment and BLASTN for nucleotide sequence alignment; described in Altschul et al., (1990) J. Mol. Biol., 215: 403) available on the National Center for the Biotechnology Information (NCBI) web site at http://www.ncbi.nlm.nih.gov/BLAST and may be used using the default parameters [BLASTP: Expect value E: 10; Word size: 3; Matrix: BLOSUM62; Cost gap: Existence 11, Extension 1; and BLASTN: algorithm by default: Megablast; Expect value E: 10; Word size: 28; Match scores: 1; Mismatch score: -2;
[0191] Gap costs: linear (determined by the match/mismatch score) (October 2013)]. [0192] In the context of the invention, variant and mutant amino acid sequences include amino acid sequences that have at least about 90% sequence identity with an amino acid sequence defined herein. A variant or mutant amino acid sequence may have at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to an amino acid sequence as defined herein. Amino acid sequence identity is defined as the percentage of amino acid residues in the variant sequence that are identical with the amino acid residues in the reference sequence, after aligning the sequences and if necessary, introducing gaps, to achieve the maximum percent sequence identity, and not considering any conservative substitution as part of the sequence identity. Standard alignment algorithms cited above are useful in this regard. [0193] In some embodiments, the antibodies of the disclosure, or the antigen-binding fragments thereof, specifically may bind to variants VP1 sequences of Table 1.
[0194] In some embodiments, the antibodies of the disclosure, or the antigen-binding fragments thereof, neutralize BKPyV comprising VP1 sequences of Table 1.
TABLE 1: Sequences of the VP1 capsid proteins of BKPy viruses
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
[0195] An object of the disclosure is an isolated, or recombinant, BKPy VP1 protein of amino acid sequence SEQ ID NO: 96 or an isolated, or recombinant, VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence. Such BKPy VP1 protein may be useful for producing VLPs and BK PyV pseudotypes usable in neutralizing assay, antibody screening, in diagnosing methods for measuring an immune response to a BK PyV infection in a patient in need thereof, such as KTx patient.
[0196] The present disclosure also relates to an in vitro selected BKPyV variant having a VP1 capsid protein having an amino acid sequence as set forth in SEQ ID NO: 96. [0197] The disclosure relates to an isolated or recombinant BKPy virus comprising a
VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
[0198] Recombinant BKPyV virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may be obtained by any known techniques in the art.
[0199] A recombinant BKPyV virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may be a virus, a virus-like particle (VLP), or a pseudotype virus.
[0200] A recombinant BKPyV may be a pseudotype virus.
[0201] A recombinant BKPyV virus may be prepared in any suitable cells or cell lines such as human embryonic kidney cells (HEK 293TT cells).
[0202] A nucleic acid sequence encoding a VP1 protein of amino acid sequence SEQ ID NO: 96 or a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence may be inserted in any expression vector known in the art, for example an expression plasmid such as pCAG. The minor capsid proteins VP2 and VP3 may be provided in separate plasmids, such as ph2b and ph3b plasmids, or may be inserted in the same expression plasmid used for the VP1 protein.
[0203] The vector expression may be transfected in suitable cells, such as HEK 293TT cells, using any nucleic acid cell delivery methods.
[0204] After suitable culture time after transfection, the viral particles may be harvested.
[0205] Harvest may be carried out by trypsinization, centrifugation and then lysis of the cells in a hypotonic lysis buffer, for example consisting of 25 mM Sodium Citrate pH 6.0, 1 mM CaC12, 1 mM MgC12 and 5mM KC1. Further to exposure to the lysis buffer, the cells may be sonicated.
[0206] The lysate may be clarified, for example by centrifugation.
[0207] Purification of the viral particles may be carried on density gradient medium, by any known techniques in the art.
Anti-BKPyV antibodies
[0208] An object of the invention relates to antibodies, or fragment thereof, binding to the VP1 capsid protein of BKPy viruses. In the description, except otherwise indicated, “antibody” and “antigen-binding fragment” are used interchangeably.
[0209] The antibodies described herein may be isolated antibodies.
[0210] The antibodies described herein may be recombinant antibodies. [0211] The antibodies may be monoclonal antibodies.
[0212] The antibodies may be human antibodies or human engineered antibodies.
[0213] An antibody may be provided as antigen-binding fragment. Antigen-binding fragments may be Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, or diabodies.
[0214] The term “antigen-binding fragment” refers to a fragment of an intact antibody that retain the ability to specifically binds to a given antigen/ligand. Examples of binding fragments include a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; a Fab’ fragment, a monovalent fragment consisting of the VL, VH, CL, CHI domains and hinge region; a E(ab’)2 fragment, a bivalent fragment comprising two Lab’ fragments linked by a disulphide bridge at the hinge region; an Ed fragment consisting of VH domains of a single arm of an antibody; a single domain antibody (sdAb) fragment (Ward et al., 1989 Nature 341:544-546), which consists of a VH domain or a VL domain; and an isolated complementary determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by an artificial peptide linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (ScFv); see, e.g., Bird et al., 1989 Science 242:423-426; and Huston et al., 1988 proc. Natl. Acad. Sci. 85:5879-5883). “dsFv” is a VH::VL heterodimer stabilized by a disulfide bond. Divalent and multivalent antibody fragments can form either spontaneously by association of monovalent scFvs or can be generated by coupling monovalent scFvs by a peptide linker, such as divalent sc(Fv)2. Such single chain antibodies include one or more antigen biding portions or fragments of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as are intact antibodies. A unibody is another type of antibody fragment lacking the hinge region of IgG4 antibodies. The deletion of the hinge region results in a molecule that is essentially half the size of traditional IgG4 antibodies and has a univalent binding region rather than the bivalent biding region of IgG4 antibodies. Further details on UniBodies may be obtained by reference to WO 2007/059782, which is incorporated by reference in its entirety. Antigen binding fragments can be incorporated into single domain antibodies, SMIP, maxibodies, minibodies, intrabodies, diabodies, triabodies and tetrabodies (see, e.g., Hollinger and Hudson, 2005, Nature Biotechnology, 23, 9, 1126-1136). The term “diabodies” “tribodies” or “tetrabodies” refers to small antibody fragments with multivalent antigen-binding sites (2, 3 or four), which fragments comprise a heavy -chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites. Further details on domain antibodies and methods of their production are found in US 6,291,158; 6,582,915; 6,593,081; 6,172,197; and 6,696,245; US 2004/0110941; EP 1433846, 0368684 and 0616640; WO 2005/035572, 2004/101790, 2004/081026, 2004/058821, 2004/003019 and 2003/002609, each of which is herein incorporated by reference in its entirety. Antigen binding fragments can be incorporated into single chain molecules comprising a pair of tandem Fv segments (VH-CH1- VH-CH1) Which, together with complementary light chain polypeptides, form a pair of antigen binding regions (Zapata et al., 1995 Protein Eng. 8(10); 1057-1062 and U.S. Pat. No. 5,641,870).
[0215] Fab fragments can be obtained by treating an antibody with a protease, papain. Also, the Fab can be produced by inserting DNA encoding Fab of the antibody into a vector for prokaryotic expression system, or for eukaryotic expression system, and introducing the vector into a procaryote or eucaryote (as appropriate) to express the Fab.
[0216] F(ab’)2 can be obtained by treating an antibody with a protease, pepsin. Also, the F(ab’)2 can be produced by binding Fab’ described below via a thioether bond or a disulphide bond.
[0217] Fab’ can be obtained by treating F(ab’)2 with a reducing agent, dithiothreitol. Also, the Fab’ can be produced by inserting DNA encoding Fab’ fragment of the antibody into an expression vector for prokaryote, or an expression vector for eukaryote, and introducing the vector into a prokaryote or eukaryote (as appropriate) to perform its expression.
[0218] An antigen-binding fragment may be variable heavy chain of a single domain antibody (VHH).
[0219] As used herein, the term and expression “binding” or “specifically binding” are used interchangeably and refer to the ability of an antibody or antigen-binding fragment thereof to detectably bind an epitope present on a specific antigen, such as a BKPyV VP1 capsid protein, while having relatively no or little detectable reactivity with different antigen, for example non- BKPyV VP1 capsid proteins or structures, such as the VP1 of the MPyV (Murine polyoma virus).
[0220] For example, a VP1 of the MPyV may be of SEO ID NO: 103:
[0221] MAPKRKSGVSKCETKCTKACPRPAPVPKEEIKGGMEVEDEVTGPDSVT EIEAFENPRMGQPPTPESETEGGQYYGWSRGINEATSDTEDSPENNTEPTWSMAKEQE PMLNEDLTCDTLQMWEAVSVKTEVVGSGSLLDVHGFNKPTDTVNTKGISTPVEGSQY HVFAVGGEPLDLQGLVTDARTKYKEEGVVTIKTITKKDMVNKDQVLNPISKAKLDKD GMYPVEIWHPDPAKNENTRYFGNYTGGTTTPPVLQFTNTLTTVLLDENGVGPLCKGE GLYLSCVDIMGWRVTRNYDVHHWRGLPRYFKITLRKRWVKNPYPMASLISSLFNNM LPQVQGQPMEG ENTQVEEVRVYDGTEPVPGDPDMTRYVDRFGKTKTVFPGN
[0222] The antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiologic conditions. Specific binding can be characterized by an equilibrium dissociation constant (KD) of at least about IxlO-6 M or less (e.g., a smaller KD denotes a tighter binding).
[0223] Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. Moreover, multi-specific antibodies that bind to the VP1 protein of a BKPyV and one or more additional antigens or a bi-specific that binds to two different regions of the VP1 protein of a BKPyV are nonetheless considered antibodies that “specifically bind” the VP1 protein of a BKPyV, as used herein.
[0224] The binding affinity of the antibodies, expressed as KD, may range from about 10-12 M to about 5xl0-8 M, as measured by surface plasmon resonance, e.g., BIACORE™ or solution-affinity ELISA, for example as described in the Examples section.
[0225] The binding affinity of the antibodies on the VP1 capsid protein of gl-IV BKPyV may be of about 10.0xl0-8 M or less.
[0226] The binding affinity of the antibodies on the VP1 capsid protein of gl-IV BKPyV may range from about l.OxlO-7 M to about l.OxlO"10 M, or less.
[0227] The binding affinity of the antibodies on the VP1 capsid protein of gl-IV BKPyV may be of about l.OxlO-7 M or less, or of about 8.0xl0-8 M or less, or of about 6.5xl0-8 M or less, or of about 5.0xl0-8 M or less, or of about 3.0xl0-8 M or less, or of about 2.0xl0-8 M or less, or of about 1.5xl0-8 M or less, or of about l.OxlO-8 M or less, or of about 9.OxlO-10 M or less, or of about 8.0xl0-9 M or less, or of about 5.0xl0-9 M or less, or of about 2.5xl0-9 M or less, or of about 2.0xl0-9 M or less, or of about 1.5xl0-9 M or less, or of about l.OxlO-9 M or less, or of about 8.0xl0-1° M or less, or of about 5.0xl0-1° M or less, or of about 4.OxlO-10 M or less, or of about 3xl0-10 M or less, or of about 2.5xlO-10 M or less, or of about 2.OxlO-10 M or less, or of about 1.8xlO-10 M or less, or of about 1.5xlO-10 M or less, or of about 1.2xlO-10 M or less or of about l.OxlO-10 M or less.
[0228] The expression “a BKPyV neutralizing antibody” intends to refer to an antibody which is capable of specifically binding to an antigen of BKPyV and substantially inhibiting or eliminating infection of a target cell by the BKPyV, or reducing the infectious titer, as demonstrated for example by the absence of viral gene expression of the BKPyV. Typically, a neutralizing antibody will inhibit infection activity of a BKPyV, or reduce an infection titer, by at least about 50%, or at least about 60%, or at least about 70%, or at least 80%, or at least 85%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%. The neutralizing antibody of the invention is especially useful in therapeutic applications: to prevent or treat infections with BKPyV and/or disorders induced by a BKPyV infection.
[0229] The ability of an antibody for neutralizing a BKPyV may be determined by the measurement of the IC50, i.e., the concentration of an antibody reducing by 50% the infection of cells by the virus.
[0230] Methods for determining the neutralization activity of antibodies can be found in, for instance, Pastrana et al. (PloS Pathog. 2012, 8, el002650). As example of methods known in the art for determining the neutralizing activity of an antibody, one may mention fluorescencebased neutralization assay.
[0231] A neutralization assay may be carried out with a virus or a pseudotype virus, as described in the Examples section. A serial dilution of a predetermined amount of infecting viral particles (virus or pseudotype virus) is used to infect cultured cells. For BKPyV, useable cells may be HEK293 TTT cells or RS cells. For example, a 2-, 3-, 4-, 5-, 6-, 7-fold dilution may be used.
[0232] The antibody to be assayed may be placed in contact with the viral particles before adding the viral particles to the cultured cells. Alternatively, antibody may be placed in contact with the cultured cells before addition of the viral particles. Alternatively, again, the antibody may be placed in contact with the cultured cells after addition of the viral particles to the cells.
[0233] After a suitable incubation time, the infected cells may be numbered, and the obtained value is compared with, or normalized with, the value obtained with cultured cells infected by viral particles in the absence of the antibody to be assayed .
[0234] The neutralization activity of the antibodies may be expressed as the IC50.
[0235] The neutralization activity of the antibodies on the gl-IV BKPyV, expressed as the concentration inhibiting at least 50% of the infection, IC50, measured, for example on HEK293 TTT or RS cells, as described in the Examples section, may be of about 50 nM or less.
[0236] The neutralization activity of the antibodies on the gl-IV BKPyV, expressed as IC50, may range from about 0.01 nM to about 50 nM.
[0237] In some embodiments, the antibodies described herein may neutralize the gl-IV BKPyV in a target cell with a IC50 of about 50.0 nM or less, of about 45.0 nM or less, of about 40.0 nM or less, or of about 35.0 nM or less, or of about 30.0 nM or less, or of about 25.0 nM or less, or of about 20.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 5.0 or less, or of about 3.5 nM or less, or of about 2.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.15 nM or less, or of about 0.10 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less, or of about 0.05 nM or less, or of about 0.04 nM or less, or of about 0.02 nM or less, or of about 0.01 nM or less.
[0238] In some embodiments, the antibodies may have a neutralization activity on BKPy virus comprising a VP1 amino acid sequence SEQ ID NO: 97, SEQ ID NO: 98 or SEQ ID NO: 99, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of less than about 40 nM, or less than 30 nM, or less than 15 nM, or less than 10 nM, or less than 5 nM, or less than 1 nM.
[0239] In some embodiments, the antibodies may have a neutralization activity on BKPy virus comprising a VP1 amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101 or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said amino acid sequences, with an IC50 of less than about 40 nM, or less than 30 nM, or less than 15 nM, or less than 10 nM, or less than 5 nM, or less than 1 nM.
[0240] The antibodies may have a neutralization activity on BKPy virus comprising a VP1 amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101 or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of less than about 35 nM, or less than about 30 nM, or less than about 20 nM, or less than about 15 nM, or less than about 10 nM, or less than about 5 nM, or less than about 4 nM, or less than about 2.5 nM, or less than about 2 nM, or less than about 1.5 nM, or less than about 1 nM, or less than about 0.8 nM, or less than about 0.5 nM, or less than about 0.3 nM, or less than about 0.25 nM, or less than about 0.1 nM, or less than about 0.08 nM, or less than about 0.05 nM.
[0241] An antibody may bind and neutralize at least two BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101, or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
[0242] An antibody may bind and neutralize at least two BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 101, or SEQ ID NO: 106, and for example comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of less than about 35 nM, or less than about 30 nM, or less than about 20 nM, or less than about 15 nM, or less than about 10 nM, or less than about 5 nM, or less than about 4 nM, or less than about 2.5 nM, or less than about 2 nM, or less than about 1.5 nM, or less than about 1 nM, or less than about 0.8 nM, or less than about 0.5 nM, or less than about 0.3 nM, or less than about 0.25 nM, or less than about 0.1 nM, or less than about 0.08 nM, or less than about 0.05 nM.
[0243] The BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 101, or SEQ ID NO: 106 may be selected among BKPyV- D58Del_59A, BKPyV-E61K, BKPyV-E327K and BKPyV-K172A.
[0244] An antibody disclosed herein may bind and neutralize at least two BKPy virus selected among BKPyV-D58Del_59A, BKPyV-E61K, BKPyV-E327K and BKPyV-K172A.
[0245] An antibody disclosed herein may be an antibody from group (i), (ii), (iii), (iv), (v), (vi) or (vii) defined as comprising:
[0246] (i) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution; or
[0247] (ii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution; or
[0248] (iii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution; or
[0249] (iv) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution; or
[0250] (v) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution; or
[0251] (vi) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution; or
[0252] (vii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0253] A “conservative substitution” refers to the replacement of an amino acid with another amino acid of similar chemical structure, similar chemical properties or similar sidechain volume in an amino acid sequence. A conservative amino acid substitution will not substantially change the functional properties of a protein. The amino acids introduced may have similar polarity, hydrophilicity or hydrophobicity to the amino acids they replace. Conservative amino acid changes are well known in the art. Conservative amino acid changes may also be determined by reference to the Point Accepted Mutation (PAM) or BLOcks Substitution Matrix (BLOSUM) family of scoring matrices for conservation of amino acid sequence. Thus, conservative amino acid changes may be members of an equivalence group, being a set of amino acids having mutually positive scores in the similarity representation of the scoring matrix selected for use in an alignment of the reference and mutant polypeptide chains.
[0254] For example, a conservative substitution may be a substitution of an amino acid of one class by an amino acid of the same class as presented in Table 2. Alternatively, a conservative substitution may be a substitution of an amino acid of one class by an amino acid of another class but with a similar chemical structure, a similar chemical property and/or a similar side -chain volume. Alternatively, in some embodiments, a substitution mutation may be a non-conservative mutation, which replaces the amino acid of one class with an amino acid of non-similar chemical structure, non-similar chemical property and/or non-similar side-chain volume.
TABLE 2: Classes of amino acids for conservative substitution
Class Amino acids 1 -letter code
Aliphatic Glycine, Alanine, Valine, Leucine, Isoleucine G, A, V, L, I
Hydroxyl
Serine, Cysteine, Selenocysteine, Threonine, or sulfur/selenium- S, C, U, T, M
Methionine containing
Cyclic Proline P
Aromatic Phenylalanine, Tyrosine, Tryptophan F, Y, W
Basic Histidine, Lysine, Arginine H, K, R
Acidic and their
Aspartate, Glutamate, Asparagine, Glutamine D, E, N, Q amides [0255] A CDR of an antibody, or antigen-binding fragment thereof, may comprise more than one conservative amino acid substitution, for example at least two, or at least three.
[0256] In some embodiments, the full-length heavy chain, the full-length light chain, VH, VL and CDRs of an antibody disclosed herein may comprise at least one conservative substitution. [0257] In some embodiments, the full-length heavy chain, the full-length light chain, the VH chain, the VL chain, and the CDRs of antibodies disclosed herein, or an antigen-binding fragments thereof, may comprise:
[0258] - an amino acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the corresponding amino acid sequences set forth in any of Tables 3 to 9, or
[0259] - an amino acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the corresponding amino acid sequences set forth in any of Tables 3 to 9 and having CDRs sequences differing from the sequences CDRs set forth in any of Tables 3 to 9 by one conservative amino acid substitution, or
[0260] - an amino acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the corresponding amino acid sequences set forth in any of Tables 3 to 9 and having CDRs sequences identical to the sequences CDRs set forth in any of Tables 3 to 9, or
[0261] - an amino acid sequence as set forth in any of Tables 3 to 9,
[0262] or may be encoded by
[0263] - a corresponding nucleic acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence as set forth in any of Tables 3 to 9, or
[0264] - a corresponding nucleic acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence as set forth in any of Tables 3 to 9, and having nucleic acid sequences coding for CDRs differing from the CDRs set forth in any of Tables 3 to 9 by one conservative amino acid substitution, or
[0265] - a corresponding nucleic acid sequence having at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence as set forth in any of Tables 3 to 9, and having nucleic acid sequences coding for CDRs identical to the CDRs set forth in any of Tables 3 to 9, or
[0266] - a corresponding nucleic acid sequence as set forth in any of Tables 3 to 9.
[0267] Percent identity between amino acid sequences may be determined with standard alignment algorithms as those described herein.
[0268] For sequence comparison of nucleic acid sequences, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters are used. Methods of alignment of sequences for comparison are well known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482, 1981, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443, 1970, by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444, 1988, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds 1995 supplement)).
[0269] One example of a useful algorithm is PILEUP. PILEUP uses a simplification of the progressive alignment method of Feng & Doolittle, J. Mol. Evol. 35:351-360, 1987. The method used is similar to the method described by Higgins & Sharp, CABIOS 5:151-153, 1989. Using PILEUP, a reference sequence is compared to other test sequences to determine the percent sequence identity relationship using the following parameters: default gap weight (3.00), default gap length weight (0.10), and weighted end gaps. PILEUP can be obtained from the GCG sequence analysis software package, e.g., version 7.0 (Devereaux et al., Nuc. Acids Res. 12:387- 395, 1984).
[0270] Another example of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and the BLAST 2.0 algorithm, which are described in Altschul et al., J. Mol. Biol. 215:403-410, 1990 and Altschul et al., Nucleic Acids Res. 25:3389-3402, 1977. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands. The BLASTP program (for amino acid sequences) uses as defaults a word length (W) of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915, 1989).
[0271] In some embodiments, an antibody disclosed herein may comprise:
[0272] - a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; [0273] a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2,
15, 28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0274] a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3,
16, 29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0275] - a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4,
17, 30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution;
[0276] - a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and
[0277] - a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5,
18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
[0278] In some embodiments, an antibody disclosed herein may comprise:
[0279] - a variable domain of heavy chain of sequence selected from SEQ ID NO: 6, 19,
32, 45, 58, 71 and 84, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto; and/or
[0280] - a variable domain of light chain of sequence selected from SEQ ID NO: 7, 20,
33, 46, 59, 72 and 85, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto.
[0281] In some embodiments, an antibody disclosed herein may comprise:
[0282] - a variable domain of heavy chain of sequence selected from SEQ ID NO: 6, 19, 32, 45, 58, 71 and 84, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and/or [0283] - a variable domain of light chain of sequence selected from SEQ ID NO: 7, 20, 33, 46, 59, 72 and 85, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
[0284] In some embodiments, an antibody disclosed herein may comprise:
[0285] - a variable domain of heavy chain of sequence selected from SEQ ID NO: 6, 19,
32, 45, 58, 71 and 84, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80; and a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81; and/or
[0286] - a variable domain of light chain of sequence selected from SEQ ID NO: 7, 20,
33, 46, 59, 72 and 85, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70.
[0287] In some embodiments, an antibody disclosed herein may comprise:
[0288] - a heavy chain comprising an amino acid sequence selected from SEQ ID NO: 8, 21, 34, 47, 60, 73and 86, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto; and/or
[0289] - a light chain comprising an amino acid sequence from SEQ ID NO: 9, 22, 35, 48, 61, 74 and 87, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto.
[0290] In some embodiments, an antibody disclosed herein may comprise:
[0291] - a heavy chain comprising an amino acid sequence selected from SEQ ID NO: 8, 21, 34, 47, 60, 73and 86, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and/or
[0292] - a light chain comprising an amino acid sequence from SEQ ID NO: 9, 22, 35, 48, 61, 74 and 87, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
[0293] In some embodiments, an antibody disclosed herein may comprise:
[0294] - a heavy chain comprising an amino acid sequence selected SEQ ID NO: 8, 21, 34, 47, 60, 73and 86, or a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14, 27, 40, 53, 66 and 79; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15, 28, 41, 54, 67 and 80; and a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16, 29, 42, 55, 68 and 81; and/or
[0295] - a light chain comprising an amino acid sequence from SEQ ID NO: 9, 22, 35, 48, 61, 74 and 87, or a sequence at least 85%%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, and a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17, 30, 43, 56, 69 and 82; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70. Group (i) of antibodies
[0296] The group (i) of antibodies is illustrated by the antibody B KI 20 of the Example section.
[0297] The inventors have surprisingly demonstrated that antibodies of group (i) described herein induced an improved neutralizing activity against the BKPyV.
[0298] In particular, an antibody of group (i) may efficiently neutralize BKPyV escaping neutralization by both the antibodies 41F17 and P8D11.
[0299] The antibodies of group (i) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
[0300] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0301] The conservative amino acid substitution may be as above described.
[0302] In some embodiments, the antibodies of group (i) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 6.
[0303] In some embodiments, the antibodies of group (i) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 6 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
[0304] In some embodiments, the antibodies of group (i) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 6 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
[0305] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 6.
[0306] In some embodiments, the antibodies of group (i) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 6.
[0307] In some embodiments, the antibodies of group (i) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 7.
[0308] In some embodiments, the antibodies of group (i) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 7 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
[0309] In some embodiments, the antibodies of group (i) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 7 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
[0310] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 7.
[0311] In some embodiments, the antibodies of group (i) may comprise a variable domain of light chain of sequence SEQ ID NO: 7.
[0312] In some embodiments, the antibodies of group (i) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 8.
[0313] In some embodiments, the antibodies of group (i) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 8 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution. [0314] In some embodiments, the antibodies of group (i) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 8 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
[0315] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 8.
[0316] In some embodiments, the antibodies of group (i) may comprise a heavy chain of sequence SEQ ID NO: 8.
[0317] In some embodiments, the antibodies of group (i) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 9.
[0318] In some embodiments, the antibodies of group (i) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 9 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
[0319] In some embodiments, the antibodies of group (i) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 9 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
[0320] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 9.
[0321] In some embodiments, the antibodies of group (i) may comprise a light chain of sequence SEQ ID NO: 9.
[0322] In some embodiments, an antibody of group (i) may be the antibody referred to as antibody BK120 in the Examples section.
[0323] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (i) of antibodies.
[0324] In some embodiments, a variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 10. [0325] In some embodiments, a variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 10 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
[0326] In some embodiments, a variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 10 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
[0327] In some embodiments, a variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 10.
[0328] In some embodiments, a variable domain of heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 10.
[0329] In some embodiments, a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 11.
[0330] In some embodiments, a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 11 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution. [0331] In some embodiments, a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 11 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
[0332] In some embodiments, a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 11.
[0333] In some embodiments, a variable domain of light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 11.
[0334] In some embodiments, a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 12.
[0335] In some embodiments, a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 12 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution.
[0336] In some embodiments, a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 12 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3.
[0337] In some embodiments, a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 12. [0338] In some embodiments, a heavy chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 12.
[0339] In some embodiments, a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 13.
[0340] In some embodiments, a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 13 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution.
[0341] In some embodiments, a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 13 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4, a CDR2-L comprising an amino acid sequence of DTS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5.
[0342] In some embodiments, a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 13.
[0343] In some embodiments, a light chain of an antibody of group (i) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 13.
TABLE 3: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (i) of anti-BKPy V antibody
Figure imgf000059_0001
Figure imgf000060_0001
[0344] In some embodiments, the antibodies of group (i) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about l.OxlO-9 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 2.0 nM or less.
[0345] In some embodiments, the antibodies of group (i) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about l.OxlO-9 mol/1 (M) or less, or of about 9.OxlO-10 M or less, or of about 5.0xl0-1° M or less, or of about 2.5xlO-10 M or less, or of about 2.OxlO-10 M or less, or of about 1.5xl0-10 M or less, or of about l.lxlO"10 M or less, or of about l.OxlO 10 M or less, or of about 0.5 xlO-10 M or less.
[0346] In some embodiments, the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0-10 M or less, or of about l.lxlO-10 M or less.
[0347] In some embodiments, the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.OxlO-10 M or less, or of about 1.6xlO 10 M or less.
[0348] In some embodiments, the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xlO-10 M or less, or of about 2.1xlO-10 M or less.
[0349] In some embodiments, the antibodies of group (i) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 9.OxlO 10 M or less, or of about 8.6xlO-10 M or less.
[0350] In some embodiments, the antibodies of group (i) may neutralize a BKPyV with an IC50 of about 2 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.2 or less, or of about 0.1 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less, or of about 0.05 nM or less, or about 0.01 nM or less.
[0351] In some embodiments, the antibodies of group (i) may neutralize a BKPyV of genotype I with an IC50 of about 2 nM or less, or of about 1.5 nM or less, or of about 1 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less, or of about 0.05 nM or less, or of about 0.01 nM or less.
[0352] In some embodiments, the antibodies of group (i) may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97 or 106, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of about 2 nM or less, or of about 1.5 nM or less, or of about 1.1 nM or less, or of about 0.13 nM or less, or of about 0.08 nM or less, or of about 0.01 nM or less.
[0353] In some embodiments, the antibodies of group (i) may neutralize a BKPyV of genotype II with an IC50 of about 1 nM or less, or of about 0.5 nM or less, or of about 0.27 nM or less, or of about 0.23 nM or less.
[0354] In some embodiments, the antibodies of group (i) may neutralize a BKPyV of genotype III with an IC50 of about 1 nM or less, or of about 0.6 nM or less, or of about 0.2 nM or less, or of about 0.15 nM or less.
[0355] In some embodiments, the antibodies of group (i) may neutralize a BKPyV of genotype IV with an IC50 of about 1 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.15 nM or less, or of about 0.14 nM or less.
[0356] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
[0357] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
Group (ii) of antibodies
[0358] The group (ii) of antibodies is illustrated by the antibody B KI 60-1 of the Example section.
[0359] The antibodies of group (ii) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution. [0360] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0361] The conservative amino acid substitution may be as above described.
[0362] In some embodiments, the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 19.
[0363] In some embodiments, the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 19 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
[0364] In some embodiments, the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 19 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
[0365] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 19.
[0366] In some embodiments, the antibodies of group (ii) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 19.
[0367] In some embodiments, the antibodies of group (ii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 20.
[0368] In some embodiments, the antibodies of group (ii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 20 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
[0369] In some embodiments, the antibodies of group (ii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 20 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
[0370] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 20.
[0371] In some embodiments, the antibodies of group (ii) may comprise a variable domain of light chain of sequence SEQ ID NO: 20.
[0372] In some embodiments, the antibodies of group (ii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 21.
[0373] In some embodiments, the antibodies of group (ii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 21 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
[0374] In some embodiments, the antibodies of group (ii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 21 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
[0375] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 21.
[0376] In some embodiments, the antibodies of group (ii) may comprise a heavy chain of sequence SEQ ID NO: 21.
[0377] In some embodiments, the antibodies of group (ii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 22.
[0378] In some embodiments, the antibodies of group (ii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 22 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
[0379] In some embodiments, the antibodies of group (ii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 22 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
[0380] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 22.
[0381] In some embodiments, the antibodies of group (ii) may comprise a light chain of sequence SEQ ID NO: 22.
[0382] In some embodiments, an antibody of group (ii) may be the antibody referred to as antibody B KI 60-1 in the Examples section.
[0383] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (ii) of antibodies.
[0384] In some embodiments, a variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 23.
[0385] In some embodiments, a variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 23 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
[0386] In some embodiments, a variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 23 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16. [0387] In some embodiments, a variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 23.
[0388] In some embodiments, a variable domain of heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 23.
[0389] In some embodiments, a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 24.
[0390] In some embodiments, a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 24 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
[0391] In some embodiments, a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 24 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
[0392] In some embodiments, a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 24.
[0393] In some embodiments, a variable domain of light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 24. [0394] In some embodiments, a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 25.
[0395] In some embodiments, a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 25 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution.
[0396] In some embodiments, a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 25 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16.
[0397] In some embodiments, a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 25.
[0398] In some embodiments, a heavy chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 25.
[0399] In some embodiments, a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 26.
[0400] In some embodiments, a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 26 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution.
[0401] In some embodiments, a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 26 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18.
[0402] In some embodiments, a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 26.
[0403] In some embodiments, a light chain of an antibody of group (ii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 26.
TABLE 4: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (ii) of anti-BKPy V antibody
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
[0404] In some embodiments, the antibodies of group (ii) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 10 nM or less. [0405] In some embodiments, the antibodies of group (ii) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 M or less, or of about 4.OxlO 10 M or less, or of about 3xlO 10 M or less, or of about 2.5xlO 10 M or less, or of about 2.OxlO 10 M or less, or of about 1.8xlO 10 M or less, or of about 1.7xlO 10 M or less, or of about 1.5xlO 10 M or less.
[0406] In some embodiments, the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance of about 2.5xlO 10 M or less, or of about 2.3xlO 10 M or less.
[0407] In some embodiments, the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.8xlO 10 M or less.
[0408] In some embodiments, the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.OxlO 10 M or less, or of about 1.7xlO 10 M or less.
[0409] In some embodiments, the antibodies of group (ii) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.7xlO 10 M or less.
[0410] In some embodiments, the antibodies of group (ii) may neutralize a BKPyV with an IC50 of about 10 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 or less, or of about 1.0 nM or less, or of about 0.80 nM or less, or of about 0.50 nM or less, or of about 0.30 nM or less, or of about 0.25 nM or less, or of about 0.10 nM or less, or of about 0.05 nM or less.
[0411] In some embodiments, the antibodies of group (ii) may neutralize a BKPyV of genotype I with an IC50 of about 0.5 nM or less, or of about 0.44 nM or less, of about 0.2 nM or less, or of about 0.15 nM or less, or of about 0.12 nM or less.
[0412] In some embodiments, the antibodies of group (ii) may neutralize a BKPyV of genotype II with an IC50 of about 5.5 nM or less, or of about 4.9 nM or less.
[0413] In some embodiments, the antibodies of group (ii) may neutralize a BKPyV of genotype III with an IC50 of about 1 nM or less, or of about 0.25 nM or less.
[0414] In some embodiments, the antibodies of group (ii) may neutralize a BKPyV of genotype IV with an IC50 of about 1 nM or less, or of about 0.6 nM or less, or about 0.2 nM or less.
[0415] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section. [0416] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
Group (iii) of antibodies
[0417] The group (iii) of antibodies is illustrated by the antibody BK198 of the Example section.
[0418] The antibodies of group (iii) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
[0419] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0420] The conservative amino acid substitution may be as above described.
[0421] In some embodiments, the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 32.
[0422] In some embodiments, the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 32 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
[0423] In some embodiments, the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 32 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
[0424] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 32.
[0425] In some embodiments, the antibodies of group (iii) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 32.
[0426] In some embodiments, the antibodies of group (iii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 33.
[0427] In some embodiments, the antibodies of group (iii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 33 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
[0428] In some embodiments, the antibodies of group (iii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 33 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
[0429] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 33.
[0430] In some embodiments, the antibodies of group (iii) may comprise a variable domain of light chain of sequence SEQ ID NO: 33.
[0431] In some embodiments, the antibodies of group (iii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 34.
[0432] In some embodiments, the antibodies of group (iii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 34 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
[0433] In some embodiments, the antibodies of group (iii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 34 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
[0434] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 34.
[0435] In some embodiments, the antibodies of group (iii) may comprise a heavy chain of sequence SEQ ID NO: 34.
[0436] In some embodiments, the antibodies of group (iii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 35.
[0437] In some embodiments, the antibodies of group (iii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 35 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
[0438] In some embodiments, the antibodies of group (iii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 35 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
[0439] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 35.
[0440] In some embodiments, the antibodies of group (iii) may comprise a light chain of sequence SEQ ID NO: 35.
[0441] In some embodiments, an antibody of group (iii) may be the antibody referred to as antibody BK198 in the Examples section.
[0442] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (iii) of antibodies. [0443] In some embodiments, a variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 36.
[0444] In some embodiments, a variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 36 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
[0445] In some embodiments, a variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 36 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29.
[0446] In some embodiments, a variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 36.
[0447] In some embodiments, a variable domain of heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 36.
[0448] In some embodiments, a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 37.
[0449] In some embodiments, a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 37 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
[0450] In some embodiments, a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 37 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
[0451] In some embodiments, a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 37.
[0452] In some embodiments, a variable domain of light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 37.
[0453] In some embodiments, a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 38.
[0454] In some embodiments, a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 38 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution.
[0455] In some embodiments, a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 38 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29. [0456] In some embodiments, a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 38.
[0457] In some embodiments, a heavy chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
38.
[0458] In some embodiments, a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 39.
[0459] In some embodiments, a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 39 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
[0460] In some embodiments, a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 39 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30, a CDR2-L comprising an amino acid sequence of DAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31.
[0461] In some embodiments, a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 39.
[0462] In some embodiments, a light chain of an antibody of group (iii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
39. TABLE 5: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (iii) of anti-BKPyV antibody
Figure imgf000078_0001
Figure imgf000079_0001
Figure imgf000080_0001
[0001]
[0463] In some embodiments, the antibodies of group (iii) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 10 nM or less.
[0464] In some embodiments, the antibodies of group (iii) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0-9 M or less, or of about 2.5xl0-9 M or less, or of about l.OxlO-9 M or less, of about 5.OxlO 10 M or less, or of about 4.OxlO 10 M or less, or of about 3xlO 10 M or less, or of about 2.OxlO 10 M or less, or of about 1.8xlO 10 M or less, or of about 1.5xlO 10 M or less, or about l.lxlO 10 M or less.
[0465] In some embodiments, the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0xl0-9 M or less, or of about 1.6xl0-9 M or less.
[0466] In some embodiments, the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xl0-9 M or less, or of about 2.3xl0-9 M or less.
[0467] In some embodiments, the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xlO 10 M or less, or of about l.lxlO 10 M or less.
[0468] In some embodiments, the antibodies of group (iii) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.8xlO 10 M or less.
[0469] In some embodiments, the antibodies of group (iii) may neutralize a BKPyV with an IC50 of about 10 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 or less, or of about 1.0 nM or less, or of about 0.80 nM or less, or of about 0.50 nM or less, or of about 0.30 nM or less, or of about 0.25 nM or less. [0470] In some embodiments, the antibodies of group (iii) may neutralize a BKPyV of genotype I with an IC50 of about 2 nM or less, or of about 1.3 nM or less, or of about 1 nM or less, or of about 0.8 nM or less, or of about 0.79 nM or less.
[0471] In some embodiments, the antibodies of group (iii) may neutralize a BKPyV of genotype II with an IC50 of about 10 nM or less, or of about 9 nM or less, or of about 5 nM or less, or of about 3.9 nM or less.
[0472] In some embodiments, the antibodies of group (iii) may neutralize a BKPyV of genotype III with an IC50 of about 3 nM or less, or of about 2.8 nM or less.
[0473] In some embodiments, the antibodies of group (iii) may neutralize a BKPyV of genotype IV with an IC50 of about 4 nM or less, or of about 3.5 nM or less, or of about 3.3 nM or less, or of about 1.7 nM or less.
[0474] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
[0475] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
Group (iv) of antibodies
[0476] The group (iv) of antibodies is illustrated by the antibody BK206 of the Example section.
[0477] The antibodies of group (iv) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
[0478] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0479] The conservative amino acid substitution may be as above described. [0480] In some embodiments, the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 45.
[0481] In some embodiments, the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 45 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
[0482] In some embodiments, the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 45 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
[0483] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 45.
[0484] In some embodiments, the antibodies of group (iv) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 45.
[0485] In some embodiments, the antibodies of group (iv) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 46.
[0486] In some embodiments, the antibodies of group (iv) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 46 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
[0487] In some embodiments, the antibodies of group (iv) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 46 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44. [0488] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 46.
[0489] In some embodiments, the antibodies of group (iv) may comprise a variable domain of light chain of sequence SEQ ID NO: 46.
[0490] In some embodiments, the antibodies of group (iv) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 47.
[0491] In some embodiments, the antibodies of group (iv) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 47 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
[0492] In some embodiments, the antibodies of group (iv) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 47 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
[0493] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 47.
[0494] In some embodiments, the antibodies of group (iv) may comprise a heavy chain of sequence SEQ ID NO: 47.
[0495] In some embodiments, the antibodies of group (iv) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 48.
[0496] In some embodiments, the antibodies of group (iv) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 48 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
[0497] In some embodiments, the antibodies of group (iv) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 48 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
[0498] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 48.
[0499] In some embodiments, the antibodies of group (iv) may comprise a light chain of sequence SEQ ID NO: 48.
[0500] In some embodiments, an antibody of group (iv) may be the antibody referred to as antibody BK206 in the Examples section.
[0501] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (iv) of antibodies.
[0502] In some embodiments, a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 49.
[0503] In some embodiments, a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 49 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
[0504] In some embodiments, a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 49 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
[0505] In some embodiments, a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 49. [0506] In some embodiments, a variable domain of heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 49.
[0507] In some embodiments, a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 50.
[0508] In some embodiments, a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 50 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
[0509] In some embodiments, a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 50 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
[0510] In some embodiments, a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 50.
[0511] In some embodiments, a variable domain of light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 50.
[0512] In some embodiments, a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 51.
[0513] In some embodiments, a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 51 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution.
[0514] In some embodiments, a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 51 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42.
[0515] In some embodiments, a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 51.
[0516] In some embodiments, a heavy chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 51.
[0517] In some embodiments, a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 52.
[0518] In some embodiments, a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 52 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
[0519] In some embodiments, a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 52 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43, a CDR2-L comprising an amino acid sequence of LGS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44.
[0520] In some embodiments, a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 52.
[0521] In some embodiments, a light chain of an antibody of group (iv) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 52.
TABLE 6: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (iv) of anti-BKPyV antibody
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
[0522] In some embodiments, the antibodies of group (iv) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3. Ox 10’9 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 10.0 nM or less.
[0523] In some embodiments, the antibodies of group (iv) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.0xl0-9 mol/1 (M) or less, or of about 2.5xl0-9 M or less, or of about 2.0xl0-9 M or less, or of about 1.8xl0-9 M or less, or of about 1.5xl0-9 M or less, or of about l.OxlO’9 M or less, or of about 8.OxlO 10 M or less, or of about 5.OxlO 10 M or less, or of about 2.5xlO 10 M or less, or of about 2.OxlO 10 M or less, or of about 1.5xl0’10 M or less, or of about 1.2xlO’10 M or less, or of about l.lxlO 10 M or less, or of about l.OxlO 10 M or less. [0524] In some embodiments, the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xlO 10 M or less, or of about 2.OxlO 10 M or less.
[0525] In some embodiments, the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xl0-9 M or less, or of about 2.2xl0-9 M or less.
[0526] In some embodiments, the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 4xl0-9 M or less, or of about 3.1xl0-9 M or less.
[0527] In some embodiments, the antibodies of group (iv) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 3.OxlO 10 M or less, or of about 2.5xlO 10 M or less.
[0528] In some embodiments, the antibodies of group (iv) may neutralize a BKPyV with an IC50 of about 40.0 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 3.5 nM or less, or of about 3.0 nM or less, or of about 2.8 or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.2 nM or less, or about 0.16 nM or less.
[0529] In some embodiments, the antibodies of group (iv) may neutralize a BKPyV of genotype I with an IC50 of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.18 nM or less, or of about 0.16 nM or less.
[0530] In some embodiments, the antibodies of group (iv) may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of about 3 nM or less, or of about 2.64 nM or less, or of about 1 nM or less, or of about 0.96 nM or less, or of about 0.5 nM or less.
[0531] In some embodiments, the antibodies of group (iv) may neutralize a BKPyV of genotype II with an IC50 of about 35 nM or less, or of about 34 nM or less.
[0532] In some embodiments, the antibodies of group (iv) may neutralize a BKPyV of genotype III with an IC50 of about 40 nM or less, or of about 35 nM or less.
[0533] In some embodiments, the antibodies of group (iv) may neutralize a BKPyV of genotype IV with an IC50 of about 0.3 nM or less, or of about 0.24 nM or less, or of about 0.19 nM or less. [0534] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
[0535] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
Group (v) of antibodies
[0536] The group (v) of antibodies is illustrated by the antibody BK256 of the Example section.
[0537] The antibodies of group (v) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
[0538] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0539] The conservative amino acid substitution may be as above described.
[0540] In some embodiments, the antibodies of group (v) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 58.
[0541] In some embodiments, the antibodies of group (v) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 58 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution. [0542] In some embodiments, the antibodies of group (v) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 58 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
[0543] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 58.
[0544] In some embodiments, the antibodies of group (v) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 58.
[0545] In some embodiments, the antibodies of group (v) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 59.
[0546] In some embodiments, the antibodies of group (v) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 59 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
[0547] In some embodiments, the antibodies of group (v) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 59 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
[0548] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 59.
[0549] In some embodiments, the antibodies of group (v) may comprise a variable domain of light chain of sequence SEQ ID NO: 59.
[0550] In some embodiments, the antibodies of group (v) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 60.
[0551] In some embodiments, the antibodies of group (v) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 60 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
[0552] In some embodiments, the antibodies of group (v) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 60 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
[0553] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 60.
[0554] In some embodiments, the antibodies of group (v) may comprise a heavy chain of sequence SEQ ID NO: 60.
[0555] In some embodiments, the antibodies of group (v) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 61.
[0556] In some embodiments, the antibodies of group (v) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 61 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
[0557] In some embodiments, the antibodies of group (v) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 61 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
[0558] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 61.
[0559] In some embodiments, the antibodies of group (v) may comprise a light chain of sequence SEQ ID NO: 61.
[0560] In some embodiments, an antibody of group (v) may be the antibody referred to as antibody BK256 in the Examples section. [0561] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (v) of antibodies.
[0562] In some embodiments, a variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 62.
[0563] In some embodiments, a variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 62 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
[0564] In some embodiments, a variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 62 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
[0565] In some embodiments, a variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 62.
[0566] In some embodiments, a variable domain of heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 62.
[0567] In some embodiments, a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 63.
[0568] In some embodiments, a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 63 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
[0569] In some embodiments, a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 63 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
[0570] In some embodiments, a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 63.
[0571] In some embodiments, a variable domain of light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 63.
[0572] In some embodiments, a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 64.
[0573] In some embodiments, a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 64 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution.
[0574] In some embodiments, a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 64 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55.
[0575] In some embodiments, a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 64.
[0576] In some embodiments, a heavy chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
64.
[0577] In some embodiments, a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 65.
[0578] In some embodiments, a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 65 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution.
[0579] In some embodiments, a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 65 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57.
[0580] In some embodiments, a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 65.
[0581] In some embodiments, a light chain of an antibody of group (v) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO:
65. TABLE 7: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (v) of anti-BKPyV antibody
Figure imgf000097_0001
Figure imgf000098_0001
[0582] In some embodiments, the antibodies of group (v) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5. Ox 10’8 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 30.0 nM or less.
[0583] In some embodiments, the antibodies of group (v) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0-8 mol/1 (M) or less, or of about 3.0xl0-8 M or less, or of about 2.0xl0-8 M or less, or of about l.OxlO-8 M or less, or of about 9.5xl0-9 M or less, or of about 9.0xl0-9 M or less, or of about 8.0xl0-9 M or less, or of about 5.0xl0-9 M or less, or of about 2.5xl0-9 M or less, or of about 2.0xl0-9 M or less, or of about l.OxlO-9 M or less, or of about 8.OxlO 10 M or less, or of about 5.OxlO 10 M or less, or of about 2.5xlO-10 M or less, or of about 2.OxlO 10 M or less, or of about 1.5xl0-10 M or less, or of about 1.2xlO-10 M or less or of about l.OxlO 10 M or less.
[0584] In some embodiments, the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0-10 M or less, or of about 1.2xlO-10 M or less.
[0585] In some embodiments, the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0-10 M or less, or of about 1.2xlO-10 M or less.
[0586] In some embodiments, the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0-8 M or less, or of about 2.0xl0-8 M or less.
[0587] In some embodiments, the antibodies of group (v) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about l.OxlO-8 M or less, or of about 9.5xl0-9 M or less, or of about 9.3 xlO-9 M or less.
[0588] In some embodiments, the antibodies of group (v) may neutralize a BKPyV with an IC50 of about 30.0 nM or less, or of about 25.0 nM or less, or of about 20.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 5.0 or less, or of about 2.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.25 nM or less, or of about 0.15 nM or less, or of about 0.10 nM or less.
[0589] In some embodiments, the antibodies of group (v) may neutralize a BKPyV of genotype I with an IC50 of about 1 nM or less, or of about 0.23 nM or less, or of about 0.11 nM or less. [0590] In some embodiments, the antibodies of group (v) may neutralize a BKPyV of genotype II with an IC50 of about 8 nM or less, or of about 1 nM or less, or of about 0.79 nM or less.
[0591] In some embodiments, the antibodies of group (v) may neutralize a BKPyV of genotype III with an IC50 of about 30 nM or less, or of about 23.5 nM or less.
[0592] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
[0593] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
Group (vi) of antibodies
[0594] The group (vi) of antibodies is illustrated by the antibody BK293 of the Example section.
[0595] The antibodies of group (vi) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3- H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0596] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0597] The conservative amino acid substitution may be as above described.
[0598] In some embodiments, the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 71.
[0599] In some embodiments, the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 71 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
[0600] In some embodiments, the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 71 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
[0601] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 71.
[0602] In some embodiments, the antibodies of group (vi) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 71.
[0603] In some embodiments, the antibodies of group (vi) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 72.
[0604] In some embodiments, the antibodies of group (vi) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 72 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0605] In some embodiments, the antibodies of group (vi) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 72 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
[0606] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 72.
[0607] In some embodiments, the antibodies of group (vi) may comprise a variable domain of light chain of sequence SEQ ID NO: 72. [0608] In some embodiments, the antibodies of group (vi) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 73.
[0609] In some embodiments, the antibodies of group (vi) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 73 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
[0610] In some embodiments, the antibodies of group (vi) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 73 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
[0611] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 73.
[0612] In some embodiments, the antibodies of group (vi) may comprise a heavy chain of sequence SEQ ID NO: 73.
[0613] In some embodiments, the antibodies of group (vi) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 74.
[0614] In some embodiments, the antibodies of group (vi) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 74 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0615] In some embodiments, the antibodies of group (vi) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 74 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
[0616] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 74. [0617] In some embodiments, the antibodies of group (vi) may comprise a light chain of sequence SEQ ID NO: 74.
[0618] In some embodiments, an antibody of group (vi) may be the antibody referred to as antibody BK293 in the Examples section.
[0619] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (vi) of antibodies.
[0620] In some embodiments, a variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 75.
[0621] In some embodiments, a variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 75 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution.
[0622] In some embodiments, a variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 75 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
[0623] In some embodiments, a variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 75.
[0624] In some embodiments, a variable domain of heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 75.
[0625] In some embodiments, a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 76. [0626] In some embodiments, a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 76 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0627] In some embodiments, a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 76 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
[0628] In some embodiments, a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 76.
[0629] In some embodiments, a variable domain of light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 76.
[0630] In some embodiments, a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 77.
[0631] In some embodiments, a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 77 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution. [0632] In some embodiments, a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 77 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68.
[0633] In some embodiments, a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 77.
[0634] In some embodiments, a heavy chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 77.
[0635] In some embodiments, a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 78.
[0636] In some embodiments, a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 78 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0637] In some embodiments, a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 78 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69, a CDR2-L comprising an amino acid sequence of DQS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70.
[0638] In some embodiments, a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 78. [0639] In some embodiments, a light chain of an antibody of group (vi) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 78. TABLE 8: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (vi) of anti-BKPyV antibody
Figure imgf000106_0001
Figure imgf000107_0001
[0640] In some embodiments, the antibodies of group (vi) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about lO.OxlO-8 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 40.0 nM or less.
[0641] In some embodiments, the antibodies of group (vi) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about lO.OxlO-8 M or less, or of about 8.0xl0-8 M or less, or of about 6.5xl0-8 M or less, or of about 5.0xl0-8 M or less, or of about 3.0xl0-8 M or less, or of about 2.0xl0-8 M or less, or of about 1.5xl0-8 M or less, or of about l.OxlO-8 M or less, or of about 8.0xl0-9 M or less, or of about 5.0xl0-9 M or less, or of about 2.5xl0-9 M or less, or of about 2.0xl0-9 M or less, or of about 1.5xl0-9 M or less, or of about l.OxlO-9 M or less, or of about 8.5xlO-10 M or less, or of about 8.OxlO 10 M or less, or of about 7.OxlO 10 M or less, or of about 5.0xl0-1° M or less.
[0642] In some embodiments, the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 9.0xl0-8 M or less, or of about 7.0xl0-8 M or less or of about 6.5xl0-8 M or less.
[0643] In some embodiments, the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.5xl0-9 M or less, or of about 2.0xl0-9 M or less, or of about 1.6xl0-9 M or less.
[0644] In some embodiments, the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 1.5xl0-9 M or less, or of about l.OxlO-9 M or less, or of about 8.5xlO-10 M or less.
[0645] In some embodiments, the antibodies of group (vi) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0xl0-8 M or less, or about 1.5xl0-8 M or less, or of about 1.3xl0-8 M or less.
[0646] In some embodiments, the antibodies of group (vi) may neutralize a BKPyV with an IC50 of about 40.0 nM or less, or of about 35.0 nM or less, or of about 30.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 5.0 or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.2 or less, or of about 0.15 nM or less, or of about 0.10 nM or less, or of about 0.08 nM or less, or of about 0.05 nM or less, or of about 0.03 nM or less, or of about 0.01 nM or less.
[0647] In some embodiments, the antibodies of group (vi) may neutralize a BKPyV of genotype II with an IC50 of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.24 nM or less, or of about 0.05 nM or less, or of about 0.03 nM or less.
[0648] In some embodiments, the antibodies of group (vi) may neutralize a BKPyV of genotype III with an IC50 of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.27 nM or less, or of about 0.25 nM or less.
[0649] In some embodiments, the antibodies of group (vi) may neutralize a BKPyV of genotype IV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, with an IC50 of about 40 nM or less, or of about 35.0 nM or less, or of about 32.0 nM or less.
[0650] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
[0651] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
Group (vii) of antibodies
[0652] The group (vii) of antibodies is illustrated by the antibody BK304 of the Example section.
[0653] The antibodies of group (vii) may comprise a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution. [0654] In some embodiments, the CDRs may comprise at least two, or at least 3 conservative amino acid substitution.
[0655] The conservative amino acid substitution may be as above described.
[0656] In some embodiments, the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 84.
[0657] In some embodiments, the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 84 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
[0658] In some embodiments, the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 84 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
[0659] In some embodiments, the amino acid sequence of the variable domain of heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 84.
[0660] In some embodiments, the antibodies of group (vii) may comprise a variable domain of heavy chain of sequence SEQ ID NO: 84.
[0661] In some embodiments, the antibodies of group (vii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 85.
[0662] In some embodiments, the antibodies of group (vii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 85 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0663] In some embodiments, the antibodies of group (vii) may comprise a variable domain of light chain of sequence at least about 85% identical to sequence SEQ ID NO: 85 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
[0664] In some embodiments, the amino acid sequence of the variable domain of light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 85.
[0665] In some embodiments, the antibodies of group (vii) may comprise a variable domain of light chain of sequence SEQ ID NO: 85.
[0666] In some embodiments, the antibodies of group (vii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 86.
[0667] In some embodiments, the antibodies of group (vii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 86 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
[0668] In some embodiments, the antibodies of group (vii) may comprise a heavy chain of sequence at least about 85% identical to sequence SEQ ID NO: 86 and a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
[0669] In some embodiments, the amino acid sequence of the heavy chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 86.
[0670] In some embodiments, the antibodies of group (vii) may comprise a heavy chain of sequence SEQ ID NO: 86.
[0671] In some embodiments, the antibodies of group (vii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 87.
[0672] In some embodiments, the antibodies of group (vii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 87 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L Ill comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0673] In some embodiments, the antibodies of group (vii) may comprise a light chain of sequence at least about 85% identical to sequence SEQ ID NO: 87 and a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
[0674] In some embodiments, the amino acid sequence of the light chain may have at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to sequence SEQ ID NO: 87.
[0675] In some embodiments, the antibodies of group (vii) may comprise a light chain of sequence SEQ ID NO: 87.
[0676] In some embodiments, an antibody of group (vii) may be the antibody referred to as antibody BK304 in the Examples section.
[0677] It is further described an isolated nucleic acid sequence comprising a sequence encoding an antibody of group (vii) of antibodies.
[0678] In some embodiments, a variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 88.
[0679] In some embodiments, a variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 88 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2- H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
[0680] In some embodiments, a variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 88 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81. [0681] In some embodiments, a variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 88.
[0682] In some embodiments, a variable domain of heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 88.
[0683] In some embodiments, a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 89.
[0684] In some embodiments, a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 89 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0685] In some embodiments, a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 89 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
[0686] In some embodiments, a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 89.
[0687] In some embodiments, a variable domain of light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 89. [0688] In some embodiments, a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 90.
[0689] In some embodiments, a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 90 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution.
[0690] In some embodiments, a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 90 and nucleic acid sequences coding: a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81.
[0691] In some embodiments, a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 90.
[0692] In some embodiments, a heavy chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 90.
[0693] In some embodiments, a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 91.
[0694] In some embodiments, a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 91 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0695] In some embodiments, a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 85% identical to a nucleic acid of sequence SEQ ID NO: 91 and nucleic acid sequences coding: a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82, a CDR2-L comprising an amino acid sequence of GAS, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83.
[0696] In some embodiments, a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of a sequence at least about 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid of sequence SEQ ID NO: 91.
[0697] In some embodiments, a light chain of an antibody of group (vii) of antibodies may be encoded by a nucleic acid sequence comprising a nucleic acid of sequence SEQ ID NO: 91.
TABLE 9: CDRs, VL, VH, full-length heavy chain and full-length light chain sequences of group (vii) of anti-BKPyV antibody
Figure imgf000115_0001
Figure imgf000116_0001
Figure imgf000117_0001
[0698] In some embodiments, the antibodies of group (vii) may (a) bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0x1 O’9 mol/1 (M) or less, and (b) neutralize a BKPyV with an IC50 of about 25.0 nM or less.
[0699] In some embodiments, the antibodies of group (vii) may bind to BKPyV VP1 capsid protein with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.0xl0-9 M or less, or of about 2.5xl0-9 M or less, or of about 2.0xl0-9 M or less, or of about 1.5xl0-9 M or less, or of about l.OxlO-9 M or less, or of about 9.OxlO 10 M or less, or of about 8.OxlO 10 M or less, or of about 5.OxlO 10 M or less, or of about 4.OxlO 10 M or less, or of about 3.OxlO 10 M or less, or of about 2.OxlO 10 M or less.
[0700] In some embodiments, the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype I BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 5.OxlO 10 M or less, or of about 3.2xlO 10 M or less.
[0701] In some embodiments, the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype II BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2.0xl0-9 M or less, or of about 1.3xl0-9 M or less. [0702] In some embodiments, the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype III BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 2xl0-9 M or less, or of about 1.6xl0-9 M or less.
[0703] In some embodiments, the antibodies of group (vii) may bind to BKPyV VP1 capsid protein of genotype IV BKPyV with a binding affinity (KD), for example measured by surface plasmon resonance, of about 9.OxlO 10 M or less, or of about 7.6xlO 10 M or less.
[0704] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV with an IC50 of about 25.0 nM or less, or of about 22.5 nM or less, or of about 20.0 nM or less, or of about 18.0 nM or less, or of about 15.0 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 5.0 nM or less, or of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.80 nM or less, or of about 0.50 nM or less, or of about 0.35 nM or less, or of about 0.25 nM or less, or of about 0.12 nM or less, or of about 0.10 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less, or of about 0.04 nM or less, or of about 0.02 nM or less, or of about 0.01 nM or less.
[0705] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV of genotype I with an IC50 of about 15.3 nM or less, or of about 10.0 nM or less, or of about 8.0 nM or less, or of about 6.2 nM or less, or of about 6.0 nM or less, or of about 2.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.25 nM or less, or of about 0.10 nM or less, or of about 0.05 nM or less, or of about 0.03 nM or less, or of about 0.02 nM or less, or of about 0.01 nM or less.
[0706] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences, with an IC50 of about 0.5 nM or less, or of about 0.25 nM or less, or of about 0.23 nM or less, or of about 0.1 nM or less.
[0707] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV of genotype II with an IC50 of about 25 nM or less, or of about 21 nM or less.
[0708] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV of genotype III with an IC50 of about 15 nM or less, or of about 13.5 nM or less, or about 13 nM or less.
[0709] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV of genotype IV with an IC50 of about 2.5 nM or less, or of about 2.0 nM or less, or of about 1.8 nM or less, or of about 1.6 nM or less, or of about 1.5 nM or less, or of about 1.0 nM or less, or of about 0.8 nM or less, or of about 0.5 nM or less, or of about 0.3 nM or less, or of about 0.28 nM or less, or about 0.2 nM or less, or of about 0.10 nM or less.
[0710] In some embodiments, the antibodies of group (vii) may neutralize a BKPyV of genotype IV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, with an IC50 of about 1 nM or less, or of about 0.5 nM or less, or of about 0.1 nM or less, or of about 0.08 nM or less, or of about 0.06 nM or less.
[0711] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured on HEK293 TTT or RS cells as described in the Examples section.
[0712] The IC50 of the inhibition or neutralization effect of the antibodies disclosed herein may be measured with a neutralization assay as disclosed in the Examples section.
Combination of antibodies
[0713] The inventors have surprisingly demonstrated that a combination of at least two of the antibodies described herein induced an improve neutralizing activity against the BKPyV.
[0714] In particular, the combinations of antibodies may efficiently neutralize BKPyV escaping neutralization by the benchmark antibody 41F17.
[0715] A combination of antibodies may comprise at least a first antibody and a second antibody, or antigen-binding fragments thereof.
[0716] The first antibody, or antigen-binding fragment thereof, and the second antibody, or antigen-binding fragment thereof, may be selected among the antibodies of groups (i) to (vii) described herein.
[0717] A first antibody is selected from a first group of antibodies selected from groups (i) to (vii) of antibodies and a second antibody is selected from a second group of antibodies selected from groups (i) to (vii) of antibodies, the first and second groups of antibodies being different.
[0718] The antibodies of a combination of antibodies may bind to distinct VP1 capsid proteins from, for example, from at least two distinct BKPyV genotypes, serotypes, or variants.
[0719] The antibodies of a combination of antibodies may bind to a same VP1 capsid protein, but on distinct epitopes of a same VP1 capsid protein, for example on at least two distinct epitopes of a same VP1 capsid protein. [0720] A combination of at least two antibodies disclosed herein may neutralize a
BKPyV of genotype I-IV.
[0721] A combination of at least two antibodies disclosed herein may neutralize a
BKPyV of genotype I-IV with an IC50 of about 70 nM or less.
[0722] A combination of at least two antibodies disclosed herein may neutralize a
BKPyV of genotype I-IV with an IC50 ranging from about 70 nM or less to 0.01 nM or less.
[0723] A combination of at least two antibodies disclosed herein may neutralize a
BKPyV of wildtype genotype I-IV with an IC50 ranging from about 10 nM or less to 0.01 nM or less.
[0724] A combination of at least two antibodies disclosed herein may neutralize a BKPyV of genotype I-IV with an IC50 of about 70 nM or less, or of about 68 nM or less, or of about 65 nM or less, or of about 62 nM or less, or of about 60 nM or less, or of about 58 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less, or of about 40 nM or less, or of about 30 nM or less, or of about 20 nM or less, or of about 15 nM or less, or of about 10 nM or less, or of about 8 nM or less, or of about 5 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less, or of about 0.09 nM or less, or of about 0.08 nM or less, or of about 0.07 nM or less, or of about 0.06 nM or less, or of about 0.05 nM or less, or of about 0.04 nM or less, or of about 0.03 nM or less, or of about 0.02 nM or less, or of about 0.01 nM or less.
[0725] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
[0726] A combination of at least two antibodies disclosed herein may neutralize a BKPyV of genotype I comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
[0727] A combination of at least two antibodies disclosed herein may neutralize a BKPyV of genotype IV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence. [0728] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less.
[0729] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 70 nM or less to about 0.1 nM or less.
[0730] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less, or of about 68 nM or less, or of about 65 nM or less, or of about 62 nM or less, or of about 60 nM or less, or of about 58 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less, or of about 40 nM or less, or of about 30 nM or less, or of about 20 nM or less, or of about 15 nM or less, or of about 10 nM or less, or of about 8 nM or less, or of about 6 nM or less, or of about 4 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less.
[0731] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less.
[0732] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 6 nM or less, or of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less.
[0733] A combination of at least two antibodies disclosed herein may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, with an ICsoof about 70 nM or less, or of about 68 nM or less, or of about 65 nM or less, or of about 62 nM or less, or of about 60 nM or less, or of about 58 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less, or of about 40 nM or less, or of about 30 nM or less, or of about 20 nM or less, or of about 15 nM or less, or of about 10 nM or less, or of about 8 nM or less, or of about 6 nM or less, or of about 4 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less.
[0734] A BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101 may be selected among BKPyV g!b2-D58delP59A, BKPyV g!b2-E61K and BKPyV gIVc2-E327K.
[0735] In some embodiments, a combination of antibodies may comprise:
[0736] (a) as a first antibody, an antibody selected from groups (i), (iii), (v), or (iv) of antibodies, and
[0737] (b) as a second antibody, an antibody selected from groups (i), (ii), (iv), (vi) or (vii) of antibodies.
[0738] In some embodiments, a combination of antibodies may comprise:
[0739] (a) as a first antibody, an antibody selected from groups (i) or (iv) of antibodies, and
[0740] (b) as a second antibody, an antibody selected from groups (i), (iv), (vi) or (vii) of antibodies.
[0741] A combination of antibodies may comprise:
[0742] (a) as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies; or
[0743] (b) as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies; or [0744] (c) as a first antibody, an antibody selected from group (i) antibodies, and as a second antibody, an antibody selected from group (vii) antibodies; or
[0745] (d) as a first antibody, an antibody selected from group (iv) of anyone of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies.
[0746] In some embodiments, a combination of antibodies may comprise:
[0747] (a) as a first antibody, an antibody selected from group (ii) of antibodies, and
[0748] (b) as a second antibody, an antibody selected from group (iii) of antibodies.
[0749] In some embodiments, a combination of antibodies may comprise:
[0750] (a) as a first antibody, an antibody selected from group (v) of antibodies, and
[0751] (b) as a second antibody, an antibody selected from group (vi) of antibodies.
[0752] In some embodiments, a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies.
[0753] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 60 nM or less.
[0754] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 60 nM or less to about 0.01 nM or less.
[0755] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 1 nM or less to about 0.01 nM or less.
[0756] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 60 nM or less.
[0757] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 60 nM or less to about 0.1 nM or less.
[0758] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2.27 nM or less, or of about 1 nM or less, or of about 0.5 nM or less, or of about 0.36 nM or less, or of about 0.1 nM or less.
[0759] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 3 nM or less, or of about 2.5 nM or less, or of about 2.2 nM or less, or of about 1 nM or less.
[0760] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 60 nM or less, or of about 55 nM or less, or of about 52 nM or less, or of about 50 nM or less, or of about 45 nM or less.
[0761] A combination of antibodies may comprise:
[0762] - as a first antibody, an antibody selected from group (i) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution, and [0763] - as a second antibody, an antibody selected from group (iv) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution.
[0764] A combination of antibodies may comprise:
[0765] - as a first antibody, an antibody selected from group (i) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 6 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 7, or a sequence at least 85% identical thereto, and
[0766] - as a second antibody, an antibody selected from group (iv) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 45 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 46, or a sequence at least 85% identical thereto.
[0767] A combination of antibodies may comprise:
[0768] - as a first antibody, an antibody selected from group (i) of antibodies comprising a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or a sequence at least 85% identical thereto, and
[0769] - as a second antibody, an antibody selected from group (iv) of antibodies comprising a heavy chain of sequence SEQ ID NO: 47 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 48, or a sequence at least 85% identical thereto.
[0770] A combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies, may comprise as a first antibody, the antibody BK120, and as a second antibody, the antibody BK206. [0771] In some embodiments, a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies.
[0772] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 70 nM or less.
[0773] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 70 nM or less to about 0.01 nM or less.
[0774] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 1 nM or less to about 0.01 nM or less.
[0775] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less.
[0776] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 70 nM or less to about 0.1 nM or less.
[0777] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 3 nM or less, or of about 4 nM or less, or of about 1 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less. [0778] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 5 nM or less, or of about 4 nM or less, or of about 3.4 nM or less, or of about 2.5 nM or less, or of about 2 nM or less, or of about 1 nM or less.
[0779] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 70 nM or less, or of about 65 nM or less, or of about 60 nM or less, or of about 55 nM or less.
[0780] A combination of antibodies may comprise:
[0781] - as a first antibody, an antibody selected from group (i) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution, and
[0782] - as a second antibody, an antibody selected from group (vi) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0783] A combination of antibodies may comprise:
[0784] - as a first antibody, an antibody selected from group (i) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 6 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 7, or a sequence at least 85% identical thereto, and
[0785] - as a second antibody, an antibody selected from group (vi) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 71 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 72, or a sequence at least 85% identical thereto.
[0786] A combination of antibodies may comprise:
[0787] - as a first antibody, an antibody selected from group (i) of antibodies comprising a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or a sequence at least 85% identical thereto, and
[0788] - as a second antibody, an antibody selected from group (vi) of antibodies comprising a heavy chain of sequence SEQ ID NO: 73 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 74, or a sequence at least 85% identical thereto.
[0789] A combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may comprise as a first antibody, the antibody BK120, and as a second antibody, the antibody BK293.
[0790] In some embodiments, a combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies.
[0791] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 1 nM or less.
[0792] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 1 nM or less to about 0.01 nM or less.
[0793] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 0.5 nM or less to about 0.01 nM or less.
[0794] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less.
[0795] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 1 nM or less to about 0.1 nM or less.
[0796] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.1 nM or less.
[0797] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less, or of about 0.8 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.39 nM or less, or of about 0.3 nM or less.
[0798] A combination of antibodies comprising as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 1 nM or less, or of about 0.9 nM or less, or of about 0.8 nM or less, or of about 0.7 nM or less, or of about 0.6 nM or less, or of about 0.5 nM or less, or of about 0.4 nM or less, or of about 0.3 nM or less, or of about 0.2 nM or less, or of about 0.16 nM or less, or of about 0.1 nM or less, or of about 0.05 nM or less.
[0799] A combination of antibodies may comprise:
[0800] - as a first antibody, an antibody selected from group (i) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution, and
[0801] - as a second antibody, an antibody selected from group (vii) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
[0802] A combination of antibodies may comprise:
[0803] - as a first antibody, an antibody selected from group (i) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 6 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 7, or a sequence at least 85% identical thereto, and
[0804] - as a second antibody, an antibody selected from group (vii) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 84 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 85, or a sequence at least 85% identical thereto.
[0805] A combination of antibodies may comprise:
[0806] - as a first antibody, an antibody selected from group (i) of antibodies comprising a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or a sequence at least 85% identical thereto, and
[0807] - as a second antibody, an antibody selected from group (vii) of antibodies comprising a heavy chain of sequence SEQ ID NO: 86 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 87, or a sequence at least 85% identical thereto.
[0808] A combination of antibodies may comprise as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vii) of antibodies, may comprise as a first antibody, the antibody BK120, and as a second antibody, the antibody BK304.
[0809] In some embodiments, a combination of antibodies may comprise as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies.
[0810] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 of about 40 nM or less.
[0811] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of genotype I-IV with an IC50 ranging from about 40 nM or less to about 0.1 nM or less.
[0812] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV of wildtype genotype I-IV with an IC50 ranging from about 5 nM or less to about 0.1 nM or less. [0813] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 40 nM or less.
[0814] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 ranging from about 40 nM or less to about 1 nM or less.
[0815] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an ICsoof about 10 nM or less, or of about 9 nM or less, or of about 8 nM or less, or of about 7 nM or less, or of about 6 nM or less, or of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2 nM or less, or of about 1 nM or less.
[0816] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 8 nM or less, or of about 7 nM or less, or of about 6 nM or less, or of about 5 nM or less, or of about 4 nM or less, or of about 3 nM or less, or of about 2 nM or less, or of about 1 nM or less.
[0817] A combination of antibodies comprising as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences with an IC50 of about 40 nM or less, or of about 38 nM or less, or of about 35 nM or less, or of about 32 nM or less, or of about 30nM or less, or of about 25 nM or less.
[0818] A combination of antibodies may comprise: [0819] - as a first antibody, an antibody selected from group (iv) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution, and
[0820] - as a second antibody, an antibody selected from group (vi) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution.
[0821] A combination of antibodies may comprise:
[0822] - as a first antibody, an antibody selected from group (iv) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 45 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 46, or a sequence at least 85% identical thereto, and
[0823] - as a second antibody, an antibody selected from group (vi) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 71 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 72, or a sequence at least 85% identical thereto.
[0824] A combination of antibodies may comprise: [0825] - as a first antibody, an antibody selected from group (iv) of antibodies comprising a heavy chain of sequence SEQ ID NO: 47 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 48, or a sequence at least 85% identical thereto, and
[0826] - as a second antibody, an antibody selected from group (vi) of antibodies comprising a heavy chain of sequence SEQ ID NO: 73 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 74, or a sequence at least 85% identical thereto.
[0827] A combination of antibodies may comprise as a first antibody, an antibody selected from group (iv) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies, may comprise as a first antibody, the antibody BK206, and as a second antibody, the antibody BK293.
[0828] In some embodiments, a combination of antibodies may comprise as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies.
[0829] A combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV of genotype I-IV.
[0830] A combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, 97 or 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
[0831] A combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96 or 97, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequences.
[0832] A combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence. [0833] A combination of antibodies comprising as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may neutralize a BKPyV comprising a VP1 capsid protein of amino acid sequence SEQ ID NO: 101, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
[0834] A combination of antibodies may comprise:
[0835] - as a first antibody, an antibody selected from group (ii) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution, and
[0836] - as a second antibody, an antibody selected from group (iii) of antibodies comprising a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution.
[0837] A combination of antibodies may comprise:
[0838] - as a first antibody, an antibody selected from group (ii) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 19 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 20, or a sequence at least 85% identical thereto, and
[0839] - as a second antibody, an antibody selected from group (iii) of antibodies comprising a variable domain of heavy chain of sequence SEQ ID NO: 32 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 33, or a sequence at least 85% identical thereto.
[0840] A combination of antibodies may comprise:
[0841] - as a first antibody, an antibody selected from group (ii) of antibodies comprising a heavy chain of sequence SEQ ID NO: 21 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 22, or a sequence at least 85% identical thereto, and
[0842] - as a second antibody, an antibody selected from group (iii) of antibodies comprising a heavy chain of sequence SEQ ID NO: 34 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 35, or a sequence at least 85% identical thereto.
[0843] A combination of antibodies may comprise as a first antibody, an antibody selected from group (ii) of antibodies, and as a second antibody, an antibody selected from group (iii) of antibodies, may comprise as a first antibody, the antibody B KI 60-1, and as a second antibody, the antibody BK198.
[0844] In a combination of antibodies described herein, the antibodies may be provided as separate antibodies or as multi-specific antibodies where the antibodies are joined together.
Multi-specific antibodies
[0845] “Multi-specific antibodies” refers to monoclonal antibodies that have binding specificities for at least two different epitopes of one antigen or for at least two distinct antigens. In some embodiments, a multi-specific antibody may be a bispecific antibody. Illustratively, multi-specific antibodies are described in US20090232811, US9382323, US9890204 or US8796424.
[0846] A multi-specific antibodies may be a bispecific antibodies.
[0847] The expression “bispecific antibodies (BsAbs)” refers to molecules which combine the antigen-binding sites of two antibodies within a single molecule. Thus, a bispecific antibody is able to bind two different antigens or to two distinct epitopes of a same antigen, simultaneously. The distinct epitope may be overlapping epitopes or non-overlapping epitopes. [0848] Multi-specific or bispecific antibodies can be prepared as full-length antibodies or antibody fragments.
[0849] The multi-specific or bispecific antibody described herein may comprise a combination of antibodies as described herein.
[0850] The multi-specific or bispecific antibody may be an isolated or a recombinant bispecific antibody.
[0851] A multi-specific or a bispecific antibody may comprise at least a first antibody, or an antigen-binding fragment thereof, and at least a second antibody, or an antigen-binding fragment thereof, joined to each other, wherein said first and second antibodies may be selected among the antibodies of groups (i) to (vii) described herein.
[0852] In some embodiments, a multi-specific or a bispecific may comprise:
[0853] (a) as a first antibody, an antibody selected from groups (i) or (iv) of antibodies, and
[0854] (b) as a second antibody, an antibody selected from groups (i), (iv), (vi) or (vii) of antibodies.
[0855] A multi-specific or a bispecific may comprise:
[0856] (a) as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (iv) of antibodies; or
[0857] (b) as a first antibody, an antibody selected from group (i) of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies; or
[0858] (c) as a first antibody, an antibody selected from group (i) antibodies, and as a second antibody, an antibody selected from group (vii) antibodies; or
[0859] (d) as a first antibody, an antibody selected from group (iv) of anyone of antibodies, and as a second antibody, an antibody selected from group (vi) of antibodies.
[0860] In some embodiments, a multi-specific antibody is a bispecific antibody.
[0861] An antibody of the multi-specific or a bispecific antibody can be linked to or coexpressed with another antibody by any known techniques in the art. For example, an antibody or fragment thereof can be functionally linked, e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise, to one or more other antibody or antibody fragment to produce a bispecific or a multispecific antibody. Specific exemplary bispecific formats that can be used include, without limitation, e.g., scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, Quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEED)body, leucine zipper, Duobody, IgGl/IgG2, dual acting Fab (DAF)-IgG, and Mab2 bispecific formats. Immunoconj ugates
[0862] An immunoconjugate comprising an antibody, or a fragment thereof, as disclosed herein may prepared according to any known methods in the art.
[0863] An antibody, or a fragment thereof, may be conjugated or linked to detectable label or reporter molecule. Such immunoconjugate may be used as a diagnostic tool for detecting and/or measuring a BKPyV infection in a subject in need thereof.
[0864] A detectable label or reporter molecule can be a radioisotope, such as 3H, 14C, 32P, 35S, or 125I; a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine; or an enzyme such as alkaline phosphatase, P-galactosidase, horseradish peroxidase, or luciferase.
[0865] As used herein, the term “labeled”, with regard to the antibody according to the disclosure, is intended to encompass direct labeling of the antibody by coupling (i.e., physically linking) a detectable substance, such as a radioactive agent or a fluorophore (e.g. fluorescein isothiocyanate (FITC) or phycoerythrin (PE) or Indocyanine (Cy5)) to the polypeptide, as well as indirect labeling of the polypeptide by reactivity with a detectable substance.
[0866] An antibody may be labelled with a radioactive molecule by any method known to the art. For example, radioactive molecules include but are not limited radioactive atom for scintigraphic studies such as 3H, 14C, 32P, 35S, I123, I124, or 125I, In111, Re186, Re188, Tc". Polypeptides of the invention may be also labelled with a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as iodine- 123, indium-ill, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron.
[0867] The immunoconjugates according to the present invention can be prepared as described in the application W02004/091668.
[0868] The antibodies may be covalently attached, directly or via a cleavable or non- cleavable linker, to at least one detectable probe.
[0869] ‘ ‘Linker”, as used herein, means a chemical moiety comprising a covalent bond or a chain of atoms that covalently attaches a polypeptide to a detectable probe.
[0870] The conjugates may be prepared by in vitro methods. In order to link a a detectable probe to the antibody, a linking group is used. Suitable linking groups are well known in the art and include disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups. Conjugation of an antibody of the invention with a detectable probe may be made using a variety of bifunctional protein coupling agents including but not limited to N-succinimidyl pyridyldithiobutyrate (SPDB), butanoic acid 4-[(5- nitro-2-pyridinyl)dithio]-2,5-dioxo-l-pyrrolidinyl ester (nitro-SPDB), 4-(Pyridin-2- yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), N-succinimidyl (2-pyridyldithio) propionate (SPDP), succinimidyl (N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis (p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as l,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al (1987). Carbon labeled 1- isothiocyanatobenzyl methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody (WO 94/11026).
[0871] The linker may be a “cleavable linker” or a “non-cleavable linker” (for example SMCC linker).
[0872] Alternatively, a fusion protein comprising an antibody disclosed herein and a detectable probe may be made, by recombinant techniques or peptide synthesis. The length of DNA may comprise respective regions encoding the two portions of the conjugate either adjacent one another or separated by a region encoding a linker peptide which does not destroy the desired properties of the conjugate.
[0873] In general, a conjugate can be obtained by a process comprising the steps of:
[0874] (i) bringing into contact an optionally-buffered aqueous solution of an antibody as disclosed herein with solutions of a linker and a detectable probe;
[0875] (ii) then optionally separating the conjugate which was formed in (i) from the unreacted antibody.
[0876] The aqueous solution of cell-binding agent can be buffered with buffers such as, e.g. potassium phosphate, acetate, citrate or N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes buffer).
[0877] The reaction temperature is usually comprised between 20 and 40° C. The reaction time can vary from 1 to 24 hours. The reaction between the antibody and the detectable probe can be monitored by size exclusion chromatography (SEC) with a refractometric and/or UV detector. If the conjugate yield is too low, the reaction time can be extended.
[0878] A number of different chromatography methods can be used by the person skilled in the art in order to perform the separation of step (ii): the conjugate can be purified e.g. by SEC, adsorption chromatography (such as ion exchange chromatography, IEC), hydrophobic interaction chromatograhy (HIC), affinity chromatography, mixed-support chromatography such as hydroxyapatite chromatography, or high performance liquid chromatography (HPLC). Purification by dialysis or diafiltration can also be used.
[0879] As used herein, the term “aggregates” means the associations which can be formed between two or more antibodies, being modified or not by conjugation. The aggregates can be formed under the influence of a great number of parameters, such as a high concentration of cell-binding agent in the solution, the pH of the solution, high shearing forces, the number of bonded dimers and their hydrophobic character, the temperature (see Wang & Gosh, 2008, J. Membrane Sci., 318: 311-316, and references cited therein); note that the relative influence of some of these parameters is not clearly established. In the case of proteins and antibodies, the person skilled in the art will refer to Cromwell et al. (2006, AAPS Journal, 8(3): E572-E579). The content in aggregates can be determined with techniques well known to the skilled person, such as SEC (see Walter et al., 1993, Anal. Biochem., 212(2): 469-480).
[0880] After step (i) or (ii), the conjugate-containing solution can be submitted to an additional step (iii) of chromatography, ultrafiltration and/or diafiltration.
[0881] The conjugate is recovered at the end of these steps in an aqueous solution.
Methods for screening BKPy V neutralizing antibody
[0882] The present disclosure relates to a use of a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence for screening a candidate antibody able to neutralize a BKPy virus.
[0883] The present disclosure relates to in vitro methods for screening a candidate antibody or a candidate combination of antibodies able to neutralize a BKPy.
[0884] An in vitro method for screening a candidate antibody may be used to screen antibodies able to neutralize a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
[0885] An in vitro method for screening a candidate antibody able to neutralize a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, may comprise at least the steps of:
[0886] a) culturing cells infected with said virus (a-i) in absence or (a-ii) in presence of said candidate antibody in conditions suitable for neutralization of the virus by the candidate antibody,
[0887] b) numbering the infected cells obtained at steps (a-i) and (a-ii),
[0888] c) comparing the number of cells obtained at step (a-i) with the number of cells obtained at step (a-ii),
[0889] wherein a number of infected cells obtained at step (a-ii) lower than the number of cells obtained at step (a-i) is indicative of a neutralizing activity of said antibody candidate.
[0890] After a suitable incubation time, the infected cells may be counted, and the obtained value is compared to, or normalized with, the value obtained with cultured cells infected with said virus in the absence of the candidate antibody.
[0891] An in vitro method for screening a candidate combination of antibodies able to neutralize a BKPy virus, may comprise at least the steps of:
[0892] a) culturing cells infected with said virus in presence of said candidate combination in conditions suitable for the binding of the antibodies of the combination to the virus,
[0893] b) culturing cells infected with said virus in presence of a combination comprising a first antibody, or antigen-binding fragment thereof, and a second antibody, or antigen-binding fragment thereof, selected among the antibodies of groups (i) to (vii) described herein in conditions suitable for the binding of the antibodies to the virus,
[0894] c) comparing the number of infected cells obtained at step (a) with the number of cells obtained at step (b),
[0895] wherein a number of infected cells obtained at step (a) lower than the number of cells obtained at step (b) may indicative of a neutralizing activity of said candidate combination.
[0896] In some embodiment, the virus may be a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence.
[0897] A BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 may be BKPyV-D58DelP59A.
[0898] The methods may be carried out with a virus, or a pseudotype virus (generally referred as viral particles). Methods for preparing virus-like particles and pseudotype virus are known in the art. [0899] The antibodies to be screened may be placed in contact with the viral particles before adding the viral particles and the antibody to the cultured cells.
[0900] Alternatively, the antibodies to be screened may be placed in contact with the cultured cells before addition of the viral particles.
[0901] Alternatively, again, the antibodies to be screened may be placed in contact with the cultured cells after addition of the viral particles to the cells.
[0902] A serial dilution of a predetermined amount of infecting viral particles (virus, VLPs or pseudotype virus) may be used to infect cultured cells. For example, a 2-, 3-, 4-, 5-, 6- , 7-fold dilution may be used.
[0903] Useable cells may be HEK293 TTT cells or RS cells.
[0904] The neutralization activity of the antibodies may be expressed as an IC50.
[0905] Methods for identifying and selecting infected cells are well know in the art. Illustratively, virus infection of target cells in vitro may be measured by immunofluorescence analysis using markers for intracellular BKPyV antigen expression as disclosed in, for example, Moriyama T, Sorokin A. Curr Protoc Cell Biol. 2009 Mar;Chapter 26:Unit 26.2.
Production of antibodies
[0906] An antibody disclosed herein may be produced by any technique known in the art, such as, without limitation, any chemical, biological, genetic, or enzymatic technique, either alone or in combination.
[0907] An antibody can be synthesized by recombinant DNA techniques well-known in the art. For example, an antibody can be obtained as DNA expression products after incorporation of DNA sequences encoding the antibody into expression vectors and introduction of such vectors into suitable eukaryotic or prokaryotic host cells that will express the desired antibody, from which it can be later isolated using well-known techniques, such as described in the Examples.
[0908] Another aspect of the present disclosure relates to a method for producing an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising at least the steps of culturing a host cell transformed by a nucleic acid sequence encoding an antibody disclosed herein, or by an expression vector comprising such nucleic acid, in a culture medium under conditions suitable for producing said antibody or fragment thereof and recovering the antibody or fragment thereof from said cultured cells. [0909] A method for producing an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof implements culture medium suitable for producing said antibody or fragment.
[0910] The cell culture medium under conditions suitable for producing said antibody or antigen-binding fragment thereof typically contains a large number of ingredients, which are necessary to support maintenance of the cultured host cells. Suitable combinations of ingredients can readily be formulated by the skilled person. A culture medium to be used herein will generally be a nutrient solution comprising standard cell culture ingredients, such as amino acids, vitamins, inorganic salts, a carbon energy source, and a buffer, as described in the art.
[0911] A cell culture medium useable may be formulated in deionized, distilled water. A culture medium may be sterilized prior to use to prevent contamination, e.g., by ultraviolet light, heating, gamma-irradiation, or filtration. A cell culture medium may be frozen (e.g., at - 20°C or -80°C) for storage or transport. The medium may contain one or more antibiotics to prevent contamination.
[0912] Illustratively, a cell culture medium may be a defined synthetic medium that is buffered at a pH of 7.4 (preferably with a pH 7.2 - 7.6 or at least 7.2 and not higher than 7.6) with a carbonate -based buffer, while the cells are cultured in an atmosphere comprising between 5 % and 10% CO2, or at least 5% and not more than 10% CO2, for example 5% CO2.
[0913] A potentially suitable cell culture medium may be available commercially, and includes, but is not limited to, Dulbecco's Modified Eagle Media (DMEM), Minimal Essential Medium (MEM), Knockout-DMEM (KO-DMEM), Glasgow Minimal Essential Medium (G- MEM), Basal Medium Eagle (BME), DMEM/Ham’s F12, Advanced DMEM/Ham’s F12, Iscove’s Modified Dulbecco’s Media (IMDM), Ham's F-10, Ham’s F-12, Medium 199, RPMI 1640 Media, and Hepatocyte cell culture media.
[0914] A nucleic acid sequence may be used to produce an antibody, or an antigenbinding fragment thereof as described in the disclosure. The nucleic acid sequence may be a DNA or a RNA molecule, which may be included in any suitable expression vector, such as a plasmid, cosmid, episome, artificial chromosome, phage, or a viral vector.
[0915] A nucleic acid sequence encoding an antibody, or an antigen-binding fragment thereof, as described may be an isolated nucleic acid sequence or a recombinant nucleic acid sequence. [0916] A nucleic acid sequence usable for producing an antibody, or an antigen-binding fragment thereof, may be as disclosed herein.
[0917] More particularly, a nucleic acid as described herein encodes the heavy chain (VH), the light chain (VL), the full-length heavy chain, and the full-length light chain of an antibody of the present disclosure for the groups (i) to (vii) of antibodies.
[0918] A nucleic acid sequence can be produced by de novo solid-phase DNA synthesis or by PCR mutagenesis of an existing sequence encoding an antibody as described herein or its antigen-binding fragment thereof. Direct chemical synthesis of nucleic acids can be accomplished by methods known in the art, such as the phosphotriester method of Narang et al, Meth. Enzymol. 68:90, 1979; the phosphodiester method of Brown / al, Meth. Enzymol. 68: 109, 1979; the diethylphosphoramidite method of Beaucage et al , Tetra. Lett., 22: 1859, 1981; and the solid support method of U.S. Patent No. 4,458,066.
[0919] Introducing mutations to a polynucleotide sequence by PCR can be performed as described in, e.g., PCR Technology: Principles and Applications for DNA Amplification, H.A. Erlich (Ed.), Freeman Press, NY, NY, 1992; PCR Protocols: A Guide to Methods and Applications, Innis et al. (Ed.), Academic Press, San Diego, CA, 1990; Mattila et al , Nucleic Acids Res. 19:967, 1991; and Eckert et al, PCR Methods and Applications 1:17, 1991.
[0920] Further aspects of the present disclosure are expression vectors and host cells for producing the anti-BKPyV antibodies described herein.
[0921] An expression vector may be an isolated or a recombinant expression vector.
[0922] A host cell may be an isolated or a recombinant host cell.
[0923] An expression vector may comprise a nucleic acid sequence as described herein. Such a vector may comprise regulatory elements, such as a promotor, enhancer, terminator, and the like, to cause or direct expression of said antibody upon transfection into the host cell.
[0924] The choice of vector will depend upon a variety of factors such as the type of host cell in which propagation is desired and the purpose of propagation. Some vectors are useful for amplifying and making large amounts of the desired DNA sequence. Other vectors are suitable for expression in cells in culture. Still other vectors are suitable for transfer and expression in cells in a whole animal. The choice of appropriate vector is well within the skill of the art. Many such vectors are available commercially.
[0925] Any expression vector for animal cell can be used, so long as a gene encoding the human antibody C region can be inserted and expressed. Examples of suitable vectors include pAGE107, pAGE103, pHSG274, pKCR, pSGl, pCAG and the like. Other examples of vectors include replicating plasmids comprising an origin of replication, or integrative plasmids, such as for instance pUC, pcDNA, pBR, and the like. Other examples of viral vector include adenoviral, retroviral, herpes virus and AAV vectors. Such recombinant viruses may be produced by techniques known in the art, such as by transfecting packaging cells or by transient transfection with helper plasmids or viruses. Typical examples of virus packaging cells include PA317 cells, PsiCRIP cells, Gpenv+ cells, 293 cells, RS cells, etc. A detailed protocol for producing such replication-defective recombinant viruses may be found for instance in Pastrana et al. (PloS Pathog. 2012, 8, el002650).
[0926] Examples of promoters and enhancers used in the expression vector for animal cell include early promoter and enhancer of SV40, LTR promoter and enhancer of Moloney mouse leukemia virus, promoter and enhancer of immunoglobulin H chain and the like.
[0927] Vectors can be prepared by, for example, inserting a polynucleotide of interest into a construct backbone, typically by means of DNA ligase attachment to a cleaved restriction enzyme site in the vector. Alternatively, the desired nucleotide sequence can be inserted by homologous recombination or site-specific recombination. Typically, homologous recombination is accomplished by attaching regions of homology to the vector on the flanks of the desired nucleotide sequence, while site-specific recombination can be accomplished through use of sequences that facilitate site-specific recombination (e.g., ere -lox, att sites, etc.). Nucleic acid containing such sequences can be added by, for example, ligation of oligonucleotides, or by polymerase chain reaction using primers comprising both the region of homology and a portion of the desired nucleotide sequence.
[0928] Expression vectors generally have convenient restriction sites located near the promoter sequence to provide for the insertion of nucleic acid sequences encoding proteins of interest. A selectable marker operative in the expression host may be present to facilitate selection of cells containing the vector. In addition, the expression construct may include additional elements. For example, the expression vector may have one or two replication systems, thus allowing it to be maintained in organisms, for example in mammalian or insect cells for expression and in a prokaryotic host for cloning and amplification. In addition, the expression construct may contain a selectable marker gene to allow the selection of transformed host cells. Selection genes are well known in the art and will vary with the host cell used.
[0929] In some embodiments, vectors encoding both the heavy and light chains may be co-expressed in the host cell for expression of the entire antibody. [0930] The expression vector is transferred to a host cell by conventional techniques and the transfected cells then are cultured by conventional techniques to produce an antibody or an antigen-binding fragment thereof of the present disclosure.
[0931] Isolated or recombinant host cells may be transfected, infected, or transformed by a nucleic acid sequence and/or an expression vector as disclosed herein.
[0932] Such host cells may be used for the production of an antibody as described herein.
[0933] The term “transformation” means the introduction of a “foreign” (i.e., extrinsic, or extracellular) gene, DNA or RNA sequence to a host cell, so that the host cell will express the introduced gene or sequence to produce a desired substance, typically a protein or enzyme coded by the introduced gene or sequence. A host cell that receives and expresses introduced DNA or RNA bas been “transformed”.
[0934] Thus, the present disclosure further provides a recombinant, or isolated, host cell transformed with a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof as disclosed herein, or with an expression vector comprising such nucleic acid sequence.
[0935] Any of a number of suitable host cells can be used in the production of an antibody or antigen-binding fragment thereof. In general, the antibody or antigen-binding fragment thereof described herein may be expressed in prokaryotes or eukaryotes (eukaryotic unicellular organism), e.g., bacteria such as Escherichia coli, yeasts, such as Saccharomyces sp., or mammalian cell lines, such as Vero cells, HEK293, or CHO, in accordance with conventional techniques.
[0936] Suitable bacteria include but are not limited to BL21 Competent E. coli, BL21(DE3) Competent E. coli, NEB Express Competent E. col, NEB Express Iq Competent E. coli, T7 Express Competent E. coli, T7 Express Iq Competent E. coli, T7 Express lysY Competent E. coli, T7 Express lysY/Iq Competent E. coli, T7 Express Crystal Competent E. coli, SHuffle Express Competent E. coli, SHuffle T7 Express Competent E. coli, SHuffle T7 Express lysY Competent E. coli, SHuffle T7 Competent E. coli, NiCo21(DE3) Competent E. coli, Lemo21(DE3) Competent E. coli.
[0937] Suitable mammalian cell lines include, but are not limited to, HeLa cells (e.g., American Type Culture Collection (ATCC) No. CCL-2), CHO cells (e.g., ATCC Nos. CRL9618, CCL61, CRL9096), 293 cells (e.g., ATCC No. CRL-1573), Vero cells, NIH 3T3 cells (e.g., ATCC No. CRL-1658), Huh-7 cells, BHK cells (e.g., ATCC No. CCL10), PC12 cells (ATCC No. CRL1721), COS cells, COS-7 cells (ATCC No. CRL1651), RATI cells, mouse L cells (ATCC No. CCLI.3), human embryonic kidney (HEK) cells (ATCC No. CRL1573), HLHepG2 cells, and the like.). [0938] The antibody or antigen-binding fragment thereof can be prepared in substantially pure or substantially isolated form. Purified antibody or antigen-binding fragment thereof can be provided such that the antibody or antigen-binding fragment thereof is present in a composition that is substantially free of other contaminant components such as enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.
[0939] The present disclosure also relates to a method of producing a recombinant host cell comprising an antibody according to the disclosure, said method comprising the steps of: (i) introducing in vitro or ex vivo a recombinant nucleic acid sequence or an expression vector as described above into a competent host cell, (ii) culturing in vitro or ex vivo the recombinant host cell obtained and (iii), optionally, selecting the cells which express and/or secrete said antibody.
[0940] Antibodies of the present disclosure are suitably separated from the culture medium by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
Pharmaceutical compositions
[0941] An antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multi-specific antibody, or a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof as described herein may be formulated in pharmaceutical composition with a pharmaceutically acceptable carrier.
[0942] A pharmaceutical composition disclosed herein may be use for treating and/or preventing a BKPyV infection or for treating and/or preventing a disorder induced by a BKPyV infection in a subject in need thereof.
[0943] A pharmaceutical composition may comprise an antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multispecific antibody, or a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof disclosed herein and a pharmaceutically acceptable carrier or excipient.
[0944] A pharmaceutical composition may comprise a BKPyV-neutralizing antibody screened according to a method disclosed herein and a pharmaceutically acceptable carrier or excipient.
[0945] A pharmaceutical composition may be prepared according to a method comprising at least a step of combining an antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multi-specific antibody, or a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof disclosed herein with a pharmaceutically acceptable carrier or excipient.
[0946] A method for manufacturing a pharmaceutical composition for treating and/or preventing a BKPyV infection or for treating and/or preventing a disorder induced by a BKPyV infection in a subject in need thereof, may comprise at least the steps of:
[0947] a) screening an antibody according to a method disclosed herein,
[0948] b) formulating the antibody obtained at step a) with at least a pharmaceutically acceptable carrier.
[0949] Combination of antibodies may be formulated to be simultaneously, separately, or sequentially administered in a subject in need thereof.
[0950] Simultaneous administration intends to refer to an administration of the antibodies at the same time and at the same location of the subject’s body.
[0951] Separate administration intends to refer to an administration of the antibodies at the same time and at separate locations of the body, e.g., in the arm and in the limb, or by separate routes, e.g., intramuscular and intravenous. In a separate administration, the administration of different antibodies may be separated by a few seconds or a few minutes.
[0952] Sequential administration intends to refer to an administration of the antibodies at different times, either at the same location or at different locations, or via the same or different routes. In a sequential administration, the different antibodies may be administered with a period of time between each administration ranging, for example, from about 30 min, or 1, 2 or 3 hours to about 24 hours, or 1, 2 or 3 days.
[0953] As used herein, the expression “pharmaceutically acceptable carrier or excipient” designates any substance or composition compatible with the organism of the subject to whom an antibody of the disclosure is to be administered. A pharmaceutically acceptable carrier is a substance or composition the administration of which to a subject does not come with significant deleterious effects such as adverse, allergic or other untoward reaction when administered to a subject. It can be mentioned, for example, non-toxic solvents, dispersion media, coatings, lubricating agents, saccharides, sugar alcohols, polymers, antibacterial and antifungal agents, and the like that are physiologically compatible. Examples of suitable carriers include one or more of a sterile water, a saline aqueous solution, sucrose, saccharose, starches, sorbitol, PVP or PEG, magnesium stearate, preservatives, dyeing agents or flavors. Particularly, such a carrier or vehicle is compatible with oral, parenteral, or rectal administration, and is preferably adapted to parenteral administration. [0954] The form of the pharmaceutical compositions, the route of administration, the dosage and the regimen naturally depend upon the condition to be treated, the severity of the diseases, the age, weight, gender of the subject, etc.
[0955] The pharmaceutical compositions of the invention can be formulated for a topical, oral, parenteral, intranasal, intravenous, intramuscular, subcutaneous or intraocular administration and the like.
[0956] The pharmaceutical composition can be formulated in the form of a solution, microemulsion, solution, microemulsion, liposome or other ordered structure suited for high medicament concentration.
[0957] The pharmaceutical compositions described herein can also include a pharmaceutically acceptable antioxidant. Examples of pharmaceutically acceptable antioxidants comprise (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc., (2) oil-soluble antioxidants, oxidizing agents such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and others; and (3) metal chelators such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and others are included.
[0958] Examples of suitable aqueous or non-aqueous carriers that can be used in the pharmaceutical compositions described herein comprise water, ethanol, polyols (for example, glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil and injectable organic esters such as ethyl oleate. Suitable flowability can be maintained, for example, conserving the required particle size in the case of a dispersion and using surfactants. These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. The prevention of the presence of microorganisms can be ensured by both sterilization methods and the inclusion of several antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid and others. It can also be desirable to include isotonic agents, such as sugars, sodium chloride, in the composition. Furthermore, the inclusion of absorption-retardant agents, such as aluminum monostearate and gelatin, can delay the absorption of injectable pharmaceutical forms.
[0959] Pharmaceutically acceptable carriers comprise sterile aqueous solutions or dispersions and sterile powders for the extemporary preparation of sterile injectable solutions or dispersions. The use of such carriers and agents for pharmaceutically active substances is well known in the field. Except where a conventional medium or agent is not compatible with the antibody, the use thereof in the pharmaceutical compositions described herein is contemplated. Complementary active substances can also be incorporated in the compositions.
[0960] The sterile injectable solutions can be prepared including the antibody, in a required amount in a suitable solvent, potentially with one or a combination of ingredients listed hereinafter, then sterilizing by means of microfiltration. Generally, the dispersions are prepared incorporating the active compound in a sterile vehicle containing a basic dispersion medium and any other necessary ingredients from those listed hereinafter. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred preparation method is a method of drying under vacuum, wherein an antibody powder plus any other desired ingredient is prepared from the previously sterilized and filtered solution, and lyophilizing.
[0961] The amount of antibody which can be combined with carrier materials to prepare a unit dosage form can vary according to the subject to be treated and the particular mode of administration. The amount of antibody which can be combined with a carrier material to prepare a unit dosage form, will generally be the amount of the composition which causes a therapeutic effect.
[0962] According to some embodiments, the amount of antibody may be about 0.01% to about 99% with respect to the final pharmaceutical composition amount, preferably about 0.1% to about 70% with respect to the final pharmaceutical composition amount, in combination with a pharmaceutically acceptable carrier.
[0963] Dosage schedules are adjusted in order to obtain the desired optimal response (for example, a therapeutic response). According to some embodiments, a single bolus administration is possible, and several divided doses can be administered over a long period of time, or the dose can be proportionally reduced or increased as indicated in a situation of imminent treatment. Preferably, the formulation of parenteral compositions is in the form of unit doses, specifically in order to facilitate administration and dosage uniformity.
[0964] According to some embodiments, a unit dosage form designates a physical unit which is convenient as a unit dose for the subject to be treated; each unit, associated with the required pharmaceutical carrier, causes the desired therapeutic effect.
[0965] According to some embodiments, the antibody may be formulated within a pharmaceutical composition to be administered at the dosage ranges between about 0.0001 mg and 100 mg, more generally between 0.01 mg and 5 mg per kg of body weight of the subject. For instance, the dosage is comprised between 0.3 mg/kg of body weight, 1 mg/kg of body weight, 3 mg/kg of body weight, 5 mg/kg of body weight or 10 mg/kg of body weight, or 1-10 mg/kg. [0966] The selected dosage level depends on the particular composition being used, the route of administration, the time of administration, the elimination rate of the particular compound being used, the duration of treatment, other medicaments, compounds and/or substances used in combination with a particular composition, the age, sex, weight, condition, general health and medical history of the subject to be treated, as well as similar factors well known in the medical field. The dosage level can also vary depending on several pharmacokinetic factors.
[0967] The pharmaceutical composition may be administered by means of medical devices known in the field. According to some particular embodiments, the therapeutic compositions according to the disclosure include the devices: needleless hypodermic injection as described in US 5,399,163, pre-filled ampule and non-invasive hypodermic injection as described in US 4,941,880; a non-invasive hypodermic injection as described in US 4,790,824; or a hypodermic injection device without a needle such as the device described in US 4,596,556.
[0968] The pharmaceutical compositions described herein can also be administered in combination therapy, that is, in combination with other agents. For example, the combination therapy can include an antibody, or antigen-binding fragments thereof, or a combination of antibody described herein in combination with at least another antiviral agent.
Therapeutic methods and uses
[0969] An antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multi-specific antibody, a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof, or a pharmaceutical composition as described herein may be for use as a medicament.
[0970] An antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multi-specific antibody, a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof, or a pharmaceutical composition as described herein may be for use in the prevention and/or treatment of a BKPyV- infection and/or a BKPyV-inf ection associated disease, in a subject in need thereof.
[0971] It is also disclosed herein a use of an antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multi-specific antibody, or a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof, for the manufacture of a medicament. The medicament may be for the prevention and/or treatment of a BKPyV-infection and/or a BKPyV-infection associated disease, in a subject in need thereof.
[0972] It is also disclosed herein a method for treating and/or preventing a BKPyV- infection and/or a BKPyV-infection associated disease, in a subject in need thereof comprising administering to said subject an effective amount of an antibody, or an antigen-binding fragment thereof, a combination of antibodies, or antigen-binding fragments thereof, a multi-specific antibody, a nucleic acid sequence encoding such antibody or antigen-binding fragment thereof, or a pharmaceutical composition as described herein.
[0973] In some embodiments, a BKPyV-infection associated disease may be selected from the group comprising: BK polyomavirus-associated nephropathy (PyVAN), hemorrhagic cystitis (HC), granule cell neuronopathy (GCN), interstitial kidney disease, ureteral stenosis, pneumonitis, vasculitis, colitis, retinitis, meningitis, meningoencephalitis, reno-urinary cancer, and immune reconstitution inflammatory syndrome (IRIS).
[0974] In some embodiments of the present disclosure, a BKPyV-infection associated disease may be a BK polyomavirus-associated nephropathy (PyVAN) or a hemorrhagic cystitis (HC).
Diagnostic methods
[0975] The disclosure relates to the use of a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence as a diagnostic tool for evaluating the capacity of an immune response of a subject to neutralize a BKPyV infection.
[0976] The disclosure relates to an in vitro method for measuring a neutralizing activity of an anti -BKPyV antibody isolated from a subject presumed to be infected with a BKPy virus. Such method may be used to evaluate whether the immune response induced by the presumed infection in the subject is able to neutralize the infection. Such method may be a method for diagnosing the capacity of a subject in need thereof to develop a neutralizing immune response against BKPyV.
[0977] The method may comprise at least the steps of:
[0978] a) culturing cells infected with a BKPy pseudotype virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence, (a-i) in absence or (a-ii) in presence of said anti-BKPyV antibody in conditions suitable for neutralization of the virus by said antibody,
[0979] b) numbering the infected cells obtained at steps (a-i) and (a-ii),
[0980] c) comparing the number of infected cells obtained at step (a-i) with the number of infected cells obtained at step (a-ii),
[0981] wherein a number of infected cells obtained at step (a-ii) lower than the number of cells obtained at step (a-i) may be indicative of a neutralizing activity of the antibody.
[0982] The measure of the neutralizing activity of the antibody may be indicative of the capacity of the immune response of the subject to neutralize the BKPyV infection.
[0983] If the number of cells obtained at step (a-ii) is not significantly lower than the number of cells obtained at step (a-i), then it may be indicative that the immune response of the subject is not sufficient to neutralize the BKPyV infection.
[0984] The term “significantly” used with respect to an observed change intends to mean that the observed change is noticeable and/or it has a statistic meaning.
[0985] In case of immune response insufficient to neutralize the BKPyV infection, then the subject may be administered an antibody, or an antigen-binding fragment thereof, or a combination of antibodies, or a multi-specific antibody, or pharmaceutical composition as disclosed herein.
[0986] In some embodiment, the method may be a method for diagnosing the capacity of a subject in need thereof to develop an immune response neutralizing a BKPyV infection and for treating said subject, the method comprises:
[0987] a) culturing cells infected with a BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 or comprising a VP1 sequence comprising an amino acid sequence having at least about 90% identity with said sequence (a-i) in absence or (a-ii) in presence of an anti-BKPyV antibody isolated from said subject in conditions suitable for neutralization of the virus by said antibody,
[0988] b) numbering the infected cells obtained at steps (a-i) and (a-ii),
[0989] c) comparing the number of infected cells obtained at step (a-i) with the number of infected cells obtained at step (a-ii), and
[0990] d) administering an antibody, or an antigen-binding fragment thereof, or a combination of antibodies, or a multi-specific antibody, or pharmaceutical composition as disclosed herein to said subject where a number of infected cells obtained at step (a-ii) is not significantly lower than the number of infected cells obtained at step (a-i). [0991] A BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96 may be BKPyV-D58DelP59A.
[0992] An anti-BKPyV antibody of the subject may be isolated from a biological sample obtained from said subject. A “biological sample” encompasses a variety of sample types obtained from a subject and can be used in a diagnostic or monitoring assay. Biological samples include but are not limited to blood and other liquid samples of biological origin, solid tissue samples such as a biopsy specimen or tissue cultures or cells derived therefrom, and the progeny thereof. Therefore, biological samples encompass clinical samples, cells in culture, cell supernatants, cell lysates, serum, plasma, biological fluid, and tissue samples.
[0993] The present disclosure is further illustrated, without in any way being limited to, by the Examples hereafter.
EXAMPLES
Example 1: Materials and Methods
Labeled and non-labeled BKPyV VLP production
[0994] BKPyV virus-like particles (VLPs) were prepared in human embryonic kidney cells (HEK 293TT cells) purchased from National Cancer Institute (NCI), USA, following the procedure of Pastrana et al. (PloS Pathog. 2012, 8, el002650) using expression plasmids coding for the major capsid VP1 protein of BKPyV genotypes la, Ib2, II, III and IVc2 wildtypes (BKPyV gla, BKPyV glb2, BKPyV gll, BKPyV gill and BKPyVc2 gIV) (SEQ ID NO: 92, 93, 98, 99 and 100 - Table 1) provided by Dr C. Buck (NCI, Bethesda, USA), and expression plasmids coding for the VP1 protein of the mutated variants BKPyV genotypes Ib2- D58Del_P59A, genotype Ib2-E61K, genotype IVc2-E327K and genotype Ib2-K172A (SEQ ID NO: 96, 97, 101, and 106 respectively).
[0995] Briefly, BKPyV virus-like particles (VLPs) were produced by transfecting HEK 293TT cells, using Lipofectamine2000 (Invitrogen), with plasmids pGwf (Addgene catalog #22517), which encodes the capsid proteins VP1 of BKPyV genotypes I, II, III and IV. VP1 mutants corresponding to SEQ ID NO: 96, 97, 101 and 106 were introduced into the glb2 VP1 plasmid (SEQ ID 96, 97 and 106), or the gIVc2 VP1 plasmid (SEQ ID 101) by site-directed mutagenesis using the NEBase Changer kit (New England Biolabs). Two days after transfection, the cells were lysed with 0.5% Triton XI 00 in Dulbecco's Phosphate Buffered Saline (DPBS) supplemented with 25 mM ammonium sulfate, Benzonase (Sigma), and 1.2 U/ml neuraminidase V (Sigma #N2876). Then, the lysates were incubated at 37°C overnight, then adjusted to 0.85 M NaCl and clarified by centrifugation for 10 min at 5,000 x g. BKPyV virus-like particles were then purified out of the clarified supernatant by ultracentrifugation through a 27-33-39% iodixanol (Optiprep, Sigma) step gradient. VP1 protein content of purified BKPyV VLPs was validated by SDS-PAGE.
[0996] The control virus used in this study, mouse polyomavirus (MPyV) VLPs, were prepared following the same above-indicated protocol using an expression plasmid coding for the capsid protein VLP1 of MPyV (SEQ ID NO: 103) in the expression vector pGwf (Addgene catalog #22517). Double-stranded DNA coding for the MPyV VP1 sequence was synthesized by Eurofins Genomics, then cloned into the vector by standard methods in this field.
[0997] For measurements of antibody binding by ELISA (see hereafter) and surface plasmon resonance (see hereafter), unlabeled BKPyV VLPs were used. For the specific labeling of B -cells carrying antibodies specific for BKPyV, genotype I BKPyV VLPs were covalently labeled with Alexa Fluor 555 (AF555) or Alexa Fluor 647 (AF647) using succinimidyl ester AF555 and AF647 conjugates (ThermoFisher Scientific) following the manufacturer’ s protocol.
Patient Samples
[0998] BKPyV-specific B -cells were isolated from cryopreserved peripheral blood mononuclear cells (PBMC) from six adult kidney transplant patients. These patients were included in a prospective observational study at the CHU Hotel Dieu Hospital, Nantes, approved by the local ethics committee and declared to the French Commission Nationale de ITnformatique et des Libertes (CNIL, n°1600141). All of the six patients gave informed consent authorizing the use of archived urine and blood samples for research purposes.
[0999] All of the six patients in this study were diagnosed with BKPyV reactivation based on the detection of viruria >107 copies/mL and viremia >103 copies/mL and had a documented increase in serum neutralizing titer of more than 1 log 10, indicating that the humoral response had been mobilized in these patients.
Isolation of BKPyV-specific B cells and antibody sequencing
[1000] Peripheral blood mononuclear cell (PBMC) isolated from the patients were thawed for the study. B -cells were enriched using the human B cell isolation kit II (Miltenyi Biotech) according to the manufacturer’s instructions, then labeled with the following anti- human antibodies: anti-CD3-BV510 (Becton-Dickinson), anti-CD19-BV421 (Becton- Dickinson), BKPyV-gI-VLP-AF555, BKPyV-gI-VLP-AF647, and Hashtag-oligonucleotide antibodies (BioLegend) in PBS supplemented with 0.5% bovine serum albumin. CD19+ AF555+AF647+ B-cells were screened using a BD Aria cell sorter, then either processed as single sorted cells, or loaded directly onto a 10X Genomics Chromium A Chip.
[1001] cDNA banks were prepared using the 10X Genomics Chromium Single Cell 5’ Library & Gel Bead kit; 10X Chromium Single Cell 5' Library Construction kit; 10X Chromium Single Cell 5' Feature Barcode Library kit; 10X Chromium Single Cell V(D)J Enrichment kit, Human B Cell; 10X Chromium Single Cell A Chip kit and 10X Chromium i7 Multiplex kit according to the manufacturer’s protocols. Three libraries were prepared separately, including V(D)J enriched library, 5’ gene expression library and 5’ cell surface protein library. Libraries comprising the B -cells having the anti-BKPyV antibodies were quantified and verified using the Qubit 3.0 instrument (Thermo-Fisher) then pooled and sequenced using NextSeq550 system. Data were analyzed using the 10X Genomics VDJ browser, then the Immunarch package in R (https://immunarch.com/).
Cell culture
[1002] Vero cells and HEK 293TT cells, purchased from the National Cancer Institute’s Developmental Therapeutics Program and HEK293A cells (ThermoFisher Catalog# R70507), were maintained in DMEM High Glucose (Thermo-Fisher) containing 10% FBS (Dutscher), 100 U/ml penicillin, 100 pg/ml streptomycin (Thermo-Fisher), lx Glutamax (Thermo-Fisher) and 250 pg/mE Hygromycin (Sigma). RS cells (Evercyte, Vienna, Austria), which are immortalized human renal tubular epithelial cells, were cultured in Proxup-3 medium (Evercyte, Vienna, Austria) supplemented with 100 U/ml penicillin, 100 pg/ml streptomycin (ThermoFisher) in tissue -culture plasticware coated with 50pg/mE collagen I (Thermo-Fisher). 293TT and RS cells were grown at 37°C with 5% CO2 in a humidified incubator, and passaged at confluence by trypsinization for 10 minutes with lx Trypsin-EDTA in PBS (Thermo-Fisher). Cultures were routinely tested for mycoplasma contamination by PCR and were consistently negative.
Cloning, expression, and purification of monoclonal anti-BKPyV antibodies
[1003] Once the anti-BKPyV antibodies of the B -cells of the patients were sequenced, candidate antibody clones were selected based on clonotype expansion, number of somatic hypermutations, and proportion of IgG within the clonotype, then paired antibodies variable heavy (VH) and light chain (VE) sequences were sent to Eurofins Genomics for gene synthesis, then cloned into expression vectors containing constant regions of heavy chain (Cyl of IgGl) and light kappa chain (CK) or light lambda chain (CX). Cloned expression vectors were confirmed by Sanger sequencing, then plasmid DNA, which encodes for the antibodies from patient B -cells, was transfected using Eipofectamine2000 (Invitrogen) into human embryonic kidney (HEK 293A) cells (Thermo Fisher) in 96-well plates. After initial screening for BKPyV specificity, confirmed antibodies were scaled up for production and purification.
[1004] Briefly, the day before transfection, 6xl06 HEK 293 A cells (Thermo Fisher) were seeded into 175 cm2 flasks containing 25 mF DMEM supplemented with 10% fetal bovine serum. The following day (day of transfection), 10 |jg of vH and 10 |jg of vL expression vectors were transfected into HEK 293 A cells following the jetPEI DNA transfection protocol. 16h post transfection, medium was replaced with serum-free Pro293a medium (Lonza Bioscience) and cells were cultured for 5 days at 37°C in a humidified 5% CO2 incubator.
[1005] The P8D11 antibody was described in Table 2 of WO2017046676.
[1006] The monoclonal antibody 41F17 was described in Lindner et al. Immunity. 2019; 50:668-676.e5. The benchmark 41F17 antibody was produced after gene synthesis (Eurofins Genomics) of the variable heavy (vH) and variable light (vL) chain sequences of the 41F17 antibody (Genbank references MH975428.1 and MH975588.1, respectively), into expression vectors containing constant regions of heavy chain (Cyl of IgGl) and light kappa chain (CK). Cloned expression vectors were confirmed by Sanger sequencing, then plasmid DNA, encoding the 41F17 antibody was transfected into HEK 293A cells (Thermo Fisher) following the jetPEI DNA transfection protocol. 16h post transfection, medium was replaced with serum-free Pro293a medium (Lonza Bioscience) and cells were cultured for 5 days at 37°C in a humidified 5% CO2 incubator.
[1007] Antibodies were then purified from filtered supernatants using 1 mL HiTrap rProtein A Fast Flow columns (Sigma- Aldrich) on a fast protein liquid chromatography (FPLC) system (Bio-Rad), and antibody purity was controlled by size-exclusion chromatography using Superdex200 Increase 10/300 GL column (GE Healthcare) following the manufacturer’s instructions. Purified, filter-sterilized antibodies were used in the following analyses.
ELISA screening
[1008] To verify antibody binding specificity, antibody-containing supernatants from 96-well-plate transfection was used. Briefly, Nunc MaxiSorp 96-well plates (Sigma) were coated with 50 ng/well of BKPyV VLPs, MPyV VLP as negative control and goat anti-human IgG Fc fragment (Bethyl) as positive control in 50 pL of PBS at 37°C overnight. PBS with 5% powdered milk (blocking buffer) was used to block the coated plates for Ih at room temperature. The plates were then washed with PBS 0.05% Tween-20 and bound antibodies to VLPs were detected using goat anti-human IgG horseradish peroxidase -conjugated secondary antibody (Bethyl) diluted 1:5000 in blocking buffer. The plates were incubated with 50 p L TMB substrate, and the reaction was stopped by adding 50 pL 0.5M H2SO4. Absorbance was read at 450 nm in a TECAN plate reader. [1009] To measure antibody binding properties, Nunc MaxiSorp 96-well plates (Sigma) were coated with 100 ng/well of BKPyV VLPs in 50 pL PBS at 37°C overnight, then blocked with PBS with 5% powdered milk (blocking buffer) for Ih. Purified antibodies were assayed at 5-fold serial dilutions, starting at 4 pg/mL in 50 pL blocking buffer. The plates were then washed with PBS 0.05% Tween-20 and antibodies bound to BKPyV VLPs were detected using goat anti -human IgG horseradish peroxidase-conjugated secondary antibody (Bethyl) diluted 1:5000 in blocking buffer. The plates were incubated with 50 pL TMB substrate, and the reaction was stopped by adding 50 pL 0.5M H2SO4. Absorbance was read at 450 nm in a TECAN plate reader.
[1010] The effective concentration 50% (EC50) was calculated using GraphPad Prism software.
Neutralization assays
[1011] Pseudotype virus BKPyV (PSV) particles for the different BKPyV genotypes were prepared following the previously established procedure of Pastrana et al. (PloS Pathog. 2012, 8, el002650) using expression plasmids coding for the major capsid protein of BKPyV genotypes la, Ib2 II, III and IVc2 wildtype (SEQ ID NO: 92, 93, 98, 99 and 100), provided by Dr C. Buck (NCI, Bethesda, USA), and expression plasmids coding for the VP1 protein of BKPyV genotype Ib2-DNEQ, genotype Ib2-VVAE, genotype Ib2-D58Del_P59A, genotype Ib2-E61K, genotype Ib2-K172A, genotype IVc2-E327K, and genotype IVc2-SDKAN, (SEQ ID NO: 94, 95, 96, 97, 106, 101, and 102, respectively) prepared as above described, the ph2b and ph3b plasmids (Addgene, #32109 and #32110) coding for the minor capsid proteins VP2 and VP3, and the pEGFP-N 1 plasmid (Clontech) as a reporter.
[1012] Briefly, 293TT cells were seeded at IxlO7 cells in a 75 cm2 flask in DMEM 10% FBS without antibiotics, then co-transfected using Eipofectamine 2000 reagent (Thermo-Fisher) according to manufacturer’s instructions. A total of 36 pg plasmid DNA comprising 16pg VP1 plasmid, 4pg ph2b, 8pg ph3b and 8 pg pEGFP-Nl was mixed with 1.5 mF of Opti-MEM I (Thermo-Fisher). 85 pl of Eipofectamine 2000 was diluted in 1.5 mF of Opti-MEM I and incubated for 5 min at room temperature, then mixed with the diluted plasmid DNA. After 20 min at room temperature, DNA-Eipofectamine complexes were added to each flask containing pre -plated 293TT cells.
[1013] Producer cells were harvested by trypsinization 48h after transfection. The pellet was washed once in cold PBS then resuspended in 800 pF hypotonic lysis buffer consisting of 25 mM Sodium Citrate pH 6.0, 1 mM CaCF, 1 mM MgCI and 5mM KC1. Cells were sonicated in a Bioruptor Plus device (Diagenode) for 10 minutes at 4°C with 5 cycles of 1 min. ON 1 1 min OFF. Type V neuraminidase (Sigma) was added to a final concentration of 1 U/mL and the extract was incubated for 30 min. at 37°C. The pH was neutralized by adding 100 pL of IM HEPES buff er pH 7.4, then IpL (250 U) Pierce Nuclease (Pierce) was added followed by 2h incubation at 37°C. The lysate was clarified by centrifuging twice at 5000 g for 5 min at 4°C, then layered onto an Optiprep 27%/33%/39% step gradient prepared in DPBS/0.8M NaCl. Gradients were centrifuged overnight at 175.000 g at 4°C in an SwTi55 rotor. Tubes were punctured with a 25G syringe needle, and ten fractions of each gradient were collected into 1.5 mL microcentrifuge tubes. 8 pL of each fraction was removed for qPCR, and PBS 5% BSA was then added to each fraction to give a final concentration of 0.1% BSA as a stabilizing agent before tubes were transferred to -80°C. The two or three peak fractions from each pseudotype virus preparation were pooled and aliquoted for use in neutralization assays.
[1014] For quantification of reporter plasmid, 8 pL of each fraction was mixed with 2 pL of proteinase K buffer containing 100 mM Tris-HCl pH7.5 (Thermo-Fisher), 100 mM DTT (Sigma), 25 mM EDTA (Sigma), 1% SDS (Sigma) and 200 pg/ml proteinase K (Qiagen). This solution was incubated at 50°C for 60 min, then heated to 95°C for 10 min and diluted 80-fold in water. 1 pL of this diluted solution was used for qPCR using Applied Biosystems 2x SybrMix (Applied Biosystems). Primers were CMV-F 5’-CGC AAA TGG GCG GTA GGC GTG-3’ and pEGFP-Nl-R 5’-GTC CAG CTC GAC CAG GAT G-3’. Thermal cycling was initiated with a first denaturation step at 95°C for 10 minutes to activate the polymerase, followed by 35 cycles of 95°C for 15 sec and 55°C for 40 sec. Standard curves were constructed using serial dilutions from 102 to 107 copies of the pEGFP-Nl plasmid.
[1015] For neutralization assays, HEK 293TT (NCI, USA) or RS (Evercyte, Vienna) cells were seeded at 10 OOOcells/well in flat bottom 96-well cell culture plates (BD Falcon) and allowed to adhere at 37°C for at least one hour. In parallel, BKPyV PSV were prepared at a dose of 5xl06 EGFP-N1 copies/well for RS cells and 2xl06 EGFP-N1 copies/well for HEK 293TT cells. To test neutralization capacity, each antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL. For the test neutralization capacity of example 2 parts C.6, C.7, and C.8, P8D11 antibody was added to BKPyV PSV in serial 5-fold dilutions starting at 10 pg/mL; and BK120 or 41F17 was added to BKPyV PSV in serial 5-fold dilutions starting at 5 pg/mL. Antibody-BKPyV PSV mixes were incubated at 4°C for Ih prior to an incubation with plates HEK 293TT or RS cell cultures. Plates were kept in a humidified 5% CO2 incubator at 37°C for 72 hours (HEK 293TT cells), or 96 hours (RS cells), then fixed in PBS 1% paraformaldehyde and counterstained with 10 pg/mL of a Hoechst 33342 solution (Thermo Scientific) before quantifying the number and percentage of GFP+ cells using a Cellomics ArrayScan VTI HCS Reader (Thermo Scientific).
[1016] Neutralization curves were constructed by normalizing to the percentage of GFP+ cells observed in the absence of antibody, and IC50 values for neutralization were calculated using GraphPad Prism software.
Surface plasmon resonance (SPR)
[1017] The binding strength and kinetics of produced BKPyV-specific antibodies to each BKPyV VLP wildtype genotypes I, II, III and IV were assessed using surface plasmon resonance (SPR). Briefly, anti-IgG antibody diluted to 20 pg/mL in 10 mM sodium acetate buffer pH 5.0 was immobilized to a CM5 sensor chip (GE Healthcare) until the SPR signal reached 9.000 response units (Rus) using a Biacore T200 (GE Healthcare). BKPyV-specific monoclonal antibodies were then captured at 50 nM concentration. Different BKPyV VLPs including genotype la, II, III and IVc2 were injected with a series of 2-fold dilutions from 600 nM to 0 nM. Kinetic parameters and avidity were determined by non-linear regression analysis using Biacore Evaluation software.
Production of neutralization-resistant BKPyV capsid variants
BKPyV DNEQ, BKPyV WAE, BKPyV SDKAN variants
[1018] The previously described neutralization-resistant BKPyV VP1 capsid variants, i.e., BKPyV genotype Ib2-DNEQ (naturally occurring variant with DNEQ sequence in the VP1 capsid protein - gIb2-DNEQ), BKPyV genotype Ib2-VVAE (gIb2-VVAE) and BKPyV genotype IVc2-SDKAN (gIVc2-SDKAN), observed in patients with persistent BKPyV replication (McIlroy et al. Viruses 2020, 12 (8)), were introduced into plasmids coding for genotype Ib2 and genotype IVc2 BKPyV VP1, to give sequences VP1 mutants gIb2-DNEQ, gIb2-VVAE, and gIVc2-SDKAN (SEQ ID NO: 94, 95, and 102). The production protocol of said BKPyV pseudotype particles has been described according to the above-described method. BKPyV E61K BKPyV E327K variants
[1019] The previously described E61K and E327K mutations in BKPyV (Lindner et al. Immunity. 2019;50:668-676.e5) were introduced into genotype Ib2 and genotype IVc2 BKPyV VP1 expression plasmids to produce further 41F17-resistant capsid variants (SEQ ID NO: 97 and 101). The production protocol of BKPyV pseudotype particles has been described according to the above-described method.
BKPyV KI 72A variant
[1020] The K172A mutation was introduced into genotype Ib2 BKPyV VP1 expression plasmid to produce P8D11 -resistant capsid variant (SEQ ID NO: 106). The BKPyV pseudotype particles were obtained as previously described.
BKPyV D58Del P59A variant
[1021] New BKPyV mutants that escape neutralization by the 41F17 anti-BKPyV antibody (Lindner et al. Immunity. 2019;50:668-676.e5) were identify by infecting 2xl05 Vero cells (ATCC CCL-81) with 2xl07 BKPyV genome copies of the genotype I BK-MM reference strain (pBKV 35-1, ATCC 45026) in the presence of 50 ng/mL purified 41F17 anti-BKPyV antibodies produced as described above. After 14 days culture at 37°C in 5% CO2, DNA was extracted from infected cells by SDS-proteinase K lysis, the BKPyV VP1 capsid gene was amplified by PCR, and sequenced by the Sanger method.
[1022] BKPy virus selected in the presence of 41F17 anti-BKPyV antibody carried a three-nucleotide deletion, resulting in deletion of the D58 amino acid, and the P59A substitution. The corresponding mutations were introduced into a plasmid coding for genotype Ib2 BKPyV VP1, and BKPyV pseudotype particles were produced according to the above -described method to give the BKPyV gl D58Del_P59A variant (SEQ ID NO: 96).
Example 2: Results
A. Identification of new BKPyV-specific antibodies [1023] Seven antibodies, BK160-1, BK206, BK120, BK198, BK256, BK293 and BK304, were selected from the cDNA obtained from the patients’ PBMC.
[1024] The sequences and CDRs of the antibodies were analyzed using the IMGT® databases and tools. [1025] Full-length L and H chains, vL and vH, and complementarity-determining regions (CDRs) sequences for each of the antibodies are presented in Tables 3 to 9.
B. Identification of a new BKPyV variant resistant to 41F17 neutralization
[1026] A new BKPyV VP1 variant resistant to 41F17 neutralization was obtained. The amino acid sequence (SEQ ID NO: 96) of the VP1 capsid protein is presented in Table 1.
C. Specificity and neutralizing properties of antibodies and combinations thereof
1. Binding properties of the antibodies
[1027] The binding properties of the BK120, BK160-1, BK198, BK206, BK256, BK293 and BK304 antibodies on BKPyV gl-glV VLPs (gla, gll, gill, and gIVc2 wt) measured by ELISA assays and SPR are summarized in Table 10, and presented on Figures 1A-F.
Table 10: Binding properties of the antibodies
Figure imgf000163_0001
Figure imgf000164_0001
Table 10 continued
Figure imgf000164_0002
[1028] OD450 values > 0.1 indicate antibody binding to the coated antigen.
[1029] NT - Not tested
[1030] An irrelevant anti-HLA-A2 human monoclonal antibody (Control Mab) did not bind to BKPyV VLPs, or MPyV VLPs.
[1031] As shown with the ELISA assays, antibodies BK160-1, BK206, BK120, BK198, BK256 and BK293 bound to all four BKPyV genotypes. BK256 and BK293 antibodies strongly bound to BKPyV genotypes I/II/III and BKPyV genotypes II/III, respectively. BK304 antibody bound to BKPyV genotypes I/IV. These bindings were specific since the antibodies did not bind to MPyV VLPs.
[1032] Binding avidity measured by SPR of the BK160-1, BK206, BK120, BK198 and BK304 antibodies to BKPyV VLPs was in the range IxlO 10 to 3xl0-9 M. The BK256 antibody bound to genotype I and II BKPyV VLPs with a binding avidity of 1.2xlO 10 M, and to genotype III and IV VLPs with a weaker avidity. The BK293 antibody showed a greater binding avidity to genotype II and III BKPyV VLPs (1.6xl0-9 and 8.5xlO 10, respectively) compared to genotype I and IV VLPs.
2. Neutralization of wildtype BKPyV genotypes in HEK293TT cells
[1033] The results of the neutralizing activity of BK160-1, BK206, BK120, BK198, BK256, BK293 and BK304 antibodies on HEK293TT cells infected with pseudotype viruses (gla, gll, gill, and gIVc2 wt) are presented in Table 11 and on Figures 2A-G.
Table 11: Neutralizing titer of the antibodies
Figure imgf000165_0001
Figure imgf000166_0001
[1034] Except for the BK256 and BK293 antibodies, all the BK160-1, BK120, BK206, BK198 and BK304 antibodies neutralized infectious entry of all four BKPyV wildtype (wt) genotypes into HEK293TT cells.
[1035] Antibody BK206 efficiently neutralized wt genotype I and genotype IV BKPyV PSV and to a lesser extent, wt genotype II and III BKPyV PSV.
[1036] BK256 antibody was able to neutralize genotype I, II and to a lesser extent, genotype III BKPyV PSV. BK293 antibody specifically and efficiently neutralized wt genotype II and genotype III BKPyV PSV.
[1037] BK304 antibody efficiently neutralized wt genotype IV BKPyV, with a weaker neutralizing activity for wt genotypes I, II and III.
[1038] BK160-1, BK120, BK206 and BK198 antibodies were therefore broadly neutralizing in 293TT cells, with an IC50 < InM for all wt genotypes for antibody BK120.
3. Neutralization of wildtype BKPyV genotypes in RS cells
[1039] The results of the neutralizing activity of BK160-1, BK206, BK120, BK198, BK256, BK293 and BK304 antibodies on RS cells infected with pseudotype viruses (gla, gll, gill, and gIVc2 wt) are presented in Table 12 and on Figures 3A-G.
Table 12: Neutralizing titer of antibodies BK160-1, BK206, BK120, BK198, BK256, BK293, BK304 in RS cells
Figure imgf000166_0002
Figure imgf000167_0001
[1040] B KI 60-1 and BK206 antibodies neutralized infectious entry of wt genotype I and genotype IV BKPyV PSV into RS cells, whereas BK120 antibody neutralized infectious entry of all the four wt BKPyV genotypes into RS cells.
[1041] BK198 antibody efficiently neutralized three out of four genotypes, except for wt genotype III.
[1042] BK256 antibody neutralized wt genotype I and genotype II BKPyV PSV, but with less efficiency on genotype II. Also, antibody BK256 neutralized genotype IV and genotype I BKPyV PSV with much less efficiency on genotype I
[1043] BK293 antibody efficiently and specifically neutralized wt genotype II and genotype III BKPyV PSV.
[1044] BK304 antibody neutralized wt genotype IV and genotype I BKPyV PSV, but with less efficiency on genotype I. BK304 antibody neutralized genotype IV and genotype I BKPyV PSV, but with less efficiency on genotype I.
[1045] BK160-1, BK206, BK198 and BK304 antibodies therefore neutralized the two most frequent BKPyV genotypes (gl & gIV), while BK120 antibody was broadly neutralizing in RS cells, which are immortalized human renal epithelial cells, with neutralizing IC50 <lnM for all genotypes.
4. Neutralization of wildtype and naturally occurring variant BKPyV genotypes in HEK293TT cells
[1046] The results of the neutralizing activity of the BK120, BK206, BK293, BK304 and 41F17 antibodies used alone and of the BK120 + BK206, BK120 + BK293 and BK120 + BK304 combinations on HEK293TT cells infected with wildtype and variants pseudotype viruses (glb2-wt, g!b2-DNEQ, g!b2-VVAE, gIVc2-wt and gIVc2-SDKAN) are presented in Table 13 and on Figures 4A-H.
Table 13: Neutralizing titer of antibodies and combinations against wildtype and naturally occurring VP1 variants in 293TT cells
Figure imgf000168_0001
Table 13 continued
Figure imgf000168_0002
[1047] BK120, BK206 and BK304 antibodies neutralized infectious entry of naturally occurring genotype I and genotype IV BKPyV capsid variants, BKPyV gl DNEQ, BKPyV gl VVAE and BKPyV gIV SDKAN, associated with neutralization escape and persistent high-level virus replication. This property is shared by the combinations of antibodies BK120 + BK206, BK120 + BK 293, BK120 + BK304 and BK206 + BK293.
[1048] Surprisingly, BK293 antibody neutralized infectious entry of naturally occurring genotype IV BKPyV SDKAN variant, while it did not neutralize BKPyV gIV wildtype.
[1049] The previously published 41F17 antibody was also able to neutralize naturally occurring variants, but was less effective against the genotype I VVAE variant compared to wildtype genotype I.
[1050] Variant neutralization was only tested in HEK293TT cells, since the infectivity of some variants (notable BKPyV genotype Ib2-DNEQ) was very low in RS cells, even though they replicate efficiently in patients.
[1051] Therefore, BK120, BK304 and BK206 antibodies and BK120 + BK206, BK120 + BK293, BK120 + BK304 and BK206 + BK293 combinations were at least as effective as the reference antibody 41F17 against various genotype I and genotype IV BKPyV variants.
5. Neutralization of in vitro selected BKPyV genotypes variants in HEK293TT cells
[1052] The results of the neutralizing activity of the BK120, BK206, BK293, BK304 antibodies used alone and of the BK120 + BK206, BK120 + BK293 and BK206 + BK293 combinations on HEK293TT cells infected with wildtype and in vitro selected variants pseudotype viruses (glb2-wt, gI-D58delP59A, g!b2-E61K, gIV-wt, and gIV-E327K) are presented in Table 14 and on Figures 5A-H.
[1053] Three BKPyV variants, gl D58delP59A, gl E61K, and gIV E327K, were selected in vitro to escape neutralization by reference antibody 41F17. The process of selecting resistant variants in presence of suboptimal concentration of a neutralizing antibody allows mimicking the naturally occurring process which may occur in a patient for whom the antibody dosage is not efficient.
Table 14: Neutralizing titers of antibodies against wildtype and in vitro selected BKPyV VP1 variants in 293TT cells
Figure imgf000170_0001
Table 14 continued
Figure imgf000170_0002
[1054] As shown in Table 14, all the in vitro selected BKPyV gl and BKPyV gIV variants escaped neutralization with the antibody of reference 41F17.
[1055] BK120 and BK206 antibodies neutralized infectious entry of the in vitro selected genotype I variants E61K and D58delP59A, whereas both these variants escaped neutralization by the previously described 41F17 antibody. The in vitro selected genotype IV E327K variant escaped neutralization by the BK120 and BK206 antibodies but did not escape neutralization by BK304 antibody. BK293 had a neutralizing, albeit weak, activity against BKPyV gIV E327K (Table 14).
[1056] Surprisingly, all the tested combinations of antibodies, i.e., the BK120 + BK206, BK120 + BK304, BK206 + BK293 and BK120 + BK293 combinations, were able to neutralize the infectious entry of the in vitro selected genotypes I variants E61K and D58delP59A, and the genotype IV E327K variant.
[1057] The BK120 + BK206, BK206 + BK293 and BK120 + BK293 combinations had a weak but measurable neutralizing activity against the genotype IV E327K variant. Whereas BK120 + BK304 combination highly neutralized infectious entry of all 41F17 escaping virus variants: gl D58delP59A, gl E61K, and gIV E327K (Table 14).
6. Comparison of neutralization of BKPyV wildtypes genotypes by BK120, P8D11 and
41F17 antibodies
[1058] The three monoclonal antibodies were tested in parallel in the same experiment. The results of the neutralizing activity of BK120 on HEK293TT cells infected with pseudotype viruses (gla, gll, gill, and gIVc2 wt) compared to the P8D11 and 41F17 antibodies are presented in Table 15 and on Figures 6A-6D.
Table 15: Neutralizing titers of the antibodies BK120, P8D11 and 41F17
BK120 P8D11 41F17
PSV Neutralizing ICso BKPyV gl 0.08 nM 0.12 nM 0.05 nM eplk BKPyV gll 0.11 nM 0.09 nM 0.04 nM
Figure imgf000171_0001
ecus
BKPyV gill 0.20 nM 0.38 nM 0.21 nM
BKPyV gIV 0.16 nM 0.46 nM 0.15 nM
[1059] The neutralizing titers were broadly equivalent, but with the BK120 antibody neutralizing infectious entry of the BKPyV gIV pseudotype virus with a greater potency than P8D11 antibody.
7. Comparison of neutralization of BKPyV VP1 mutants E61K and K172A by BK120, P8D11 and 41F17 antibodies [1060] The three monoclonal antibodies were tested in parallel in the same experiment. The results of the neutralizing activity of BK120 on HEK293TT cells infected with mutants pseudotype viruses g!b2-E61K and g!b2-K172A compared to the P8D11 and 41F17 antibodies are presented in Table 16 and on Figures 7A-7B.
Table 16: Neutralizing titers of the antibody BK120, P8D11 and 41F17
BK120 P8D11 41F17
PSV Neutralizing IC5o BKPyV g!b2-E61K 0.58 nM 0.15 nM in HEK293TT cells BKPyV !b2-K172A 0.01 nM 0.51 nM 0.02 nM
[1061] Both BK120 and 41F17 antibodies neutralized the BKPyV K172A variant, whereas the BKPyV K172A variant escaped neutralization by the P8D11 antibody. In contrast, both P8D11 and BK120 antibodies neutralized the variant BKPyV E61K, which resists neutralization by the 41F17 antibody.
[1062] BK120 antibody therefore has a unique specificity compared to the two previously described broadly neutralizing 41F17 and P8D11 antibodies for neutralizing some BKPy viruses.
[1063] Thus, BK120 antibody proved to be an efficient antibody capable of neutralizing all the four wild-type genotypes BKPyV and BKPyV variants.
[1064] Mutations at positions 61 and 170 of the VP1 protein are observed in kidney transplant recipients with active BKPyV replication (Figure 2, McIlroy et al. Viruses 2020 PMID: 32751274). Therefore, mutants resistant to both P8D11 and 41F17 antibodies can arise naturally, and BK120 would be expected to have more potent antiviral activity against such variants.
8. Comparison of neutralization of BKPyV VP1 mutants DNEQ and SDKAN by BK120, P8D11 and 41F17 antibodies
[1065] The three monoclonal antibodies were tested in parallel in the same experiment. The results of the neutralizing activity of BK120 antibody on HEK293TT cells infected with mutants pseudotype viruses g!b2-DNEQ and gIVc2-SDKAN compared to the P8D11 and 41F17 antibodies are presented in Table 17 and on Figures 8A-8B.
Table 17: Neutralizing titers of the BK120, P8D11 and 41F17 antibodies
BK120 P8D11 41F17
PSV Neutralizing ICso BKPyV g!b2-DNEQ 0.40nM 0.43 nM 0.43 nM ta HEK293TT cells SlVt2’ 0 20 ”M 0 40 ”M 0 18 ”M
Figure imgf000173_0001
[1066] The neutralizing titers were broadly equivalent, but with the BK120 antibody being slightly more potent against gIVc2-SDKAN than P8D11 antibody.
Example 3: Conclusions
[1067] As shown in the Examples, the newly identified BK120, BK160-1, BK198, BK206, and BK304 anti-BKPyV antibodies were able to neutralize the 4 wildtype genotypes of the BK polyoma virus in HEK293TT cells infected with pseudotype viruses (Figures 2A, B, C, D and G). The BK256 and BK293 antibodies showed a partial effect, being efficient only on gl/II/III or gll/III wildtype genotypes (Figures 2E and F). On the RS cells model, only the BK120 antibody showed a capacity to neutralize all the four wildtype genotypes (Figures 3A- G).
[1068] BK120, BK206 and BK304 antibodies neutralized all the naturally occurring BKPyV variants tested (Figures 4A, B and D) while BK293 antibody showed a neutralization effect only on the BKPyV gIV SDKAN (Figure 4C). Advantageously, all the combinations of antibodies tested, BK120 + BK206, BK120 + BK293, BK120 + BK304 and BK206 + BK293 efficiently neutralized all the naturally occurring BKPyV evaluated (Figures 4E-G).
[1069] Surprisingly, the BK304 antibody was able to neutralize infectious entry of all the in vitro selected BKPyV variants escaping neutralization by the reference 41F17 antibody, i.e., BKPyV gl E61K, BKPyV gl D58delP59A and BKPyV gIV E327K (Figures 5D and H). Advantageously, the BK120 and BK206 antibodies were still able to neutralize infectious entry of some of the in vitro selected BKPyV variants: BKPyV gl E61K and BKPyV gl D58delP59A (Figures 5A-B).
[1070] In a surprising manner, all the tested combinations of antibodies, i.e., BK120 + BK206, BK120 + BK293, BK120 + BK304 and BK206 + BK293, effectively neutralized BKPyV variants selected in vitro for escaping neutralization by the reference 41 Fl 7 antibody (Figures 5E-G).
[1071] Finally, surprisingly, the BK120 antibody is shown to effectively neutralize the wild-type genotypes BKPyV as well as the BKPyV variants escaping neutralization by 41F17 and P8D11 antibodies (Figures 7A-7B).
REFERENCES
Altschul SF, Boguski MS, Gish W, Wootton JC. Issues in searching molecular sequence databases. Nat Genet. 1994;6(2): 119-129. doi:10.1038/ng0294-119
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990;215(3):403-410. doi:10.1016/S0022-2836(05)80360-2
Altschul SF, Madden TL, Schaffer AA, et al. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997;25(17):3389-3402. doi: 10.1093/nar/25.17.3389
Ausubel et al., 1995 Current Protocols in Molecular Biology
Benotmane, I.; Solis, M.; Velay, A.; Cognard, N.; Olagne, J.; Gautier Vargas, G.; Perrin, P.; Marx, D.; Soulier, E.; Gallais, E.; et al. Intravenous immunoglobulin as a preventive strategy against BK virus viremia and BKV-associated nephropathy in kidney transplant recipients- Results from a proof-of-concept study. Am J Transplant 2020, doi:10.1111/ajt.l6233.
Bressollette-Bodin, C.; Coste-Burel, M.; Hourmant, M.; Sebille, V.; Andre-Garnier, E.; Imbert-Marcille, B.M. A prospective longitudinal study of BK virus infection in 104 renal transplant recipients. Am. J. Transplant. 2005, 5, 1926-1933, doi:10.111 l/j.1600- 6143.2005.00934.x.
Brooks Charles L. (Editor), Martin Karplus (Editor), B. Montgomery Pettitt (Editor), Ilya Prigogine (Series Editor), Stuart A. Rice (Series Editor). Proteins: A Theoretical Perspective of Dynamics, Structure, and Thermodynamics, Volume 71. ISBN: 978-0-471-52977-4 January 1991 280 Pages
Brooks, B.R.; Bruccoleri, R. E.; Olafson, B.D.; States, D.J.; Swaminathan, S.; Karplus, M. (1983). "CHARMM: A program for macromolecular energy, minimization, and dynamics calculations". J. Comp. Chem. 4: 187-217.
Case David A., Thomas E. Cheatham III, Tom Darden, Holger Gohlke, Ray Luo, Kenneth M. Merz Jr., Alexey Onufriev, Carlos Simmerling, Bing Wang, Robert J. Woods. The Amber biomolecular simulation programs. Journal of Computational Chemistry. Volume 26, Issue 16 p. 1668-1688
Cesaro, S.; Dalianis, T.; Hanssen Rinaldo, C.; Koskenvuo, M.; Pegoraro, A.; Einsele, H.; Cordonnier, C.; Hirsch, H.H.; ECIL-6 Group ECIL guidelines for the prevention, diagnosis, and treatment of BK polyoma virus-associated haemorrhagic cystitis in haematopoietic stem cell transplant recipients. J Antimicrob Chemo ther 2018, 73, 12-21, doi:10.1093/jac/dkx324. Coligan et al., Current Protocols in Immunology, Greene Publishing Assoc, and Wiley Interscience, N.Y., (1992, 1993)
Corpet F. Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res. 1988;16(22):10881-10890. doi:10.1093/nar/16.22.10881
Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res . 1984;12(1 Pt l):387-395. doi:10.1093/nar/12.1partl.387
Espada, E.; Cheng, M.P.; Kim, H.T.; Woolley, A.E.; Avigan, J.I.; Forcade, E.; Soares, M.V.D.; Lacerda, J.F.; Nikiforow, S.; Gooptu, M.; et al. BK virus-specific T-cell immune reconstitution after allogeneic hematopoietic cell transplantation. Blood Adv 2020, 4. 1881— 1893, doi: 10.1182/bloodadvances.2019001120.
Feng DF, Doolittle RF. Progressive sequence alignment as a prerequisite to correct phylogenetic trees. J Mol Evol. 1987;25(4):351-360. doi:10.1007/BF02603120
Furmaga J, Kowalczyk M, Zapolski T, Furmaga O, Krakowski L, Rudzki G, Jaroszyhski A, Jakubczak A. BK Polyomavirus-Biology, Genomic Variation and Diagnosis. Viruses. 2021 Jul 30;13(8):1502. Doi: 10.3390/vl3081502. PMID: 34452367; PMCID: PMC8402805.
Haralambieva IH, Ovsyannikova IG, Vierkant RA, Poland GA. Development of a novel efficient fluorescence-based plaque reduction microneutralization assay for measles virus immunity. Clin Vaccine Immunol. 2008;15(7): 1054-1059. doi: 10.1128/CVI.00008-08 Harlow, et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988.
Helle F, Brochot E, Handala L, et al. Biology of the BKPyV: An Update. Viruses. 2017;9(ll):327. Published 2017 Nov 3. Doi: 10.3390/v9110327
Henikoff S, Henikoff JG. Amino acid substitution matrices from protein blocks. Proc Natl Acad Sci U SA. 1992;89(22):10915-10919. doi:10.1073/pnas.89.22.10915
Higgins DG, Sharp PM. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988;73(l):237-244. doi: 10.1016/0378-1119(88)90330- 7
Higgins DG, Sharp PM. Fast and sensitive multiple sequence alignments on a microcomputer. Comput Appl Biosci. 1989;5(2): 151-153. doi:10.1093/bioinformatics/5.2.151
Hirsch, H.H.; Randhawa, P.; AST Infectious Diseases Community of Practice BK polyomavirus in solid organ transplantation. Am. J. Transplant. 2013, 13 Suppl 4, 179-188, doi: 10.1111/ajt.12110. Hurdiss, D.L.; Frank, M.; Snowden, J.S.; Macdonald, A.; Ranson, N.A. The Structure of an Infectious Human Polyoma virus and Its Interactions with Cellular Receptors. Structure 2018, 26, 839-847.e3, doi:10.1016/j.str.2018.03.019.
Hwang, S.D.; Lee, J.H.; Lee, S.W.; Kim, J.K.; Kim, M.-J.; Song, J.H. High-Dose Intravenous Immunoglobulin Treatment of Polyomavirus Nephropathy Developing After T Cell-Mediated Rejection Treatment: A Case Report. Transplant Proc 2018, 50, 2575-2578, doi:10.1016/j.transproceed.2018.01.021.
Imlay, H.; Xie, H.; Leisenring, W.M.; Duke, E.R.; Kimball, L.E.; Huang, M.-L.; Pergam, S.A.; Hill, J.A.; Jerome, K.R.; Milano, F.; et al. Presentation of BK polyomavirus-associated hemorrhagic cystitis after allogeneic hematopoietic cell transplantation. Blood Adv 2020, 4, 617- 628, doi: 10.1182/bloodadvances.2019000802.
Knowles, W.A.; Gibson, P.E.; Gardner, S.D. Serological typing scheme for BK-like isolates of human polyomavirus. J. Med. Virol. 1989, 28, 118-123.
Kundu S, Melton JS, Sorensen DC, Phillips GN Jr. Dynamics of proteins in crystals: comparison of experiment with simple models. Biophys J. 2002;83(2):723-732. Doi:10.1016/S0006-3495(02)75203-X
Laure Jason-Moller, Murphy M, Bruno J. Overview of Biacore systems and their applications. Curr Protoc Protein Sci. 2006 Sep;Chapter 19:Unit 19.13. doi: 10.1002/0471140864.psl913s45. PMID: 18429302
Lefranc MP, Pommie C, Ruiz M, Giudicelli V, Foulquier E, Truong L, Thouvenin-Contet V, Lefranc G. IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains. Dev Comp Immunol. 2003 Jan;27(l): 55-77. Doi: 10.1016/s0145-305x(02)00039-3. PMID: 12477501.
Lefranc MP. Unique database numbering system for immunogenetic analysis. Immunol Today. 1997 Nov;18(ll):509. Doi: 10.1016/s0167-5699(97)01163-8. PMID: 9386342.
Liu S, Chaudhry MR, Berrebi AA, Papadimitriou JC, Drachenberg CB, Haririan A, Alexiev BA. Polyomavirus Replication and Smoking Are Independent Risk Factors for Bladder Cancer After Renal Transplantation. Transplantation. 2017 Jun;101(6):1488-1494. doi: 10.1097/TP.0000000000001260. PMID: 27232933.
Lindner J.M., Cornacchione V., Sathe A., Be C., Srinivas H., Riquet E., Leber X.-C., Hein A., Wrobel M.B., Scharenberg M., et al. Human Memory B Cells Harbor Diverse Cross- Neutralizing Antibodies against BK and JC Polyomaviruses. Immunity. 2019;50:668-676.e5. doi: 10.1016/j.immuni.2019.02.003 vn
MacKerell, A.D., Jr.; Brooks, B.; Brooks, C. L., Ill; Nilsson, L.; Roux, B.; Won, Y.; Karplus, M. (1998). "CHARMM: The Energy Function and Its Parameterization with an Overview of the Program". The Encyclopedia of Computational Chemistry 1: 271-277. Ed. Schley er, P.v.R.; et al. Chichester: John Wiley & Sons.
McIlroy D, Honemann M, Nguyen NK, Barbier P, Peltier C, Rodallec A, Halary F, Przyrowski E, Liebert U, Hourmant M, Bressollette-Bodin C. Persistent BK Polyomavirus Viruria is Associated with Accumulation of VP1 Mutations and Neutralization Escape. Viruses. 2020 Jul 29; 12(8): 824. doi: 10.3390/vl2080824. PMID: 32751274; PMCID: PMC7472262.
Mainra, R.; Xu, Q.; Chibbar, R.; Hassan, A.; Shoker, A. Severe antibody-mediated rejection following IVIG infusion in a kidney transplant recipient with BK-virus nephropathy. Transpl Immunol 2013, 28, 145-147, doi:10.1016/j.trim.2013.05.004.
Matsumura, S.; Kato, T.; Taniguchi, A.; Kawamura, M.; Nakazawa, S.; Namba-Hamano, T.; Abe, T.; Nonomura, N.; Imamura, R. Clinical Efficacy of Intravenous Immunoglobulin for BK Polyomavirus- Associated Nephropathy After Living Kidney Transplantation. Ther Clin Risk Manag 2020, 16, 947-952, doi:10.2147/TCRM.S273388.
Moriyama T, Sorokin A. BK virus (BKV): infection, propagation, quantitation, purification, labeling, and analysis of cell entry. Curr Protoc Cell Biol. 2009 Mar;Chapter 26:Unit 26.2. doi: 10.1002/0471143030.cb2602s42. PMID: 19283732; PMCID: PMC2818100.
Muller R. Determination of affinity and specificity of anti-hapten antibodies by competitive radioimmunoassay. Methods Enzymol. 1983;92:589-601. Doi: 10.1016/0076- 6879(83)92046-3. PMID: 6190069.
Needleman SB, Wunsch CD. A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol. 1970;48(3):443-453. doi: 10.1016/0022- 2836(70)90057-4
Nelson AS, Yalamarthi N, Yong MK, Blyth E. Beyond antivirals: virus-specific T-cell immunotherapy for BK virus haemorrhagic cystitis and JC virus progressive multifocal leukoencephalopathy. Curr Opin Infect Dis. 2021 Dec l;34(6):627-634. doi: 10.1097/QC0.0000000000000794. PMID: 34751182.
Nickeleit, V.; Klimkait, T.; Binet, I.F.; Dalquen, P.; Del Zenero, V.; Thiel, G.; Mihatsch, M.J.; Hirsch, H.H. Testing for polyomavirus type BK DNA in plasma to identify renal-allograft recipients with viral nephropathy. N. Engl. J. Med. 2000, 342, 1309-1315, doi: 10.1056/NEJM200005043421802. Pastrana, D.V.; Brennan, D.C.; Cuburu, N.; Storch, G.A.; Viscidi, R.P.; Randhawa, P.S.; Buck, C.B. Neutralization serotyping of BK polyomavirus infection in kidney transplant recipients. PloS Pathog. 2012, 8, el002650, doi:10.1371/journal.ppat.!002650.
Pastrana, D.V.; Ray, U.; Magaldi, T.G.; Schowalter, R.M.; Cuburu, N.; Buck, C.B. BK polyomavirus genotypes represent distinct serotypes with distinct entry tropism. J. Virol. 2013, 87, 10105-10113, doi:10.1128/JVI.01189-13.
Pearson WR, Lipman DJ. Improved tools for biological sequence comparison. Proc Natl Acad Sci USA. 1988;85(8):2444-2448. doi:10.1073/pnas.85.8.2444
Piburn, K.H.; ALAkash, S. Use of intravenous immunoglobulin in a highly sensitized pediatric renal transplant recipient with severe BK DNAemia and rising DS A. Pediatr Transplant 2020, 24, e!3600, doi:10.1111/petr.l3600.
Reploeg, M.D.; Storch, G.A.; Clifford, D.B. Bk virus: a clinical review. Clin Infect Dis 2001, 33, 191-202, doi:10.1086/321813.
Smith and Waterman, Adv. Appl. Math. 2:482, 1981
Smith TF, Waterman MS. Identification of common molecular subsequences. J Mol Biol. 1981;147(1):195-197. doi:10.1016/0022-2836(81)90087-5
Solis, M.; Velay, A.; Porcher, R.; Domingo-Calap, P.; Soulier, E.; Joly, M.; Meddeb, M.; Kack-Kack, W.; Moulin, B.; Bahram, S.; et al. Neutralizing Antibody-Mediated Response and Risk of BK Virus- Associated Nephropathy. J. Am. Soc. Nephrol. 2018, 29, 326-334, doi:10.1681/ASN.2017050532.
US 4,522,811
US 4,596,556
US 4,790,824
US 4,941,880
US 5,374,548
US 5,399,163
US 5,399,331
US 2009/0232811
US6548640 Bl - Altered antibodies
US8796424
US9382323
US9890204
Velay, A.; Solis, M.; Benotmane, I.; Gantner, P.; Soulier, E.; Moulin, B.; Caillard, S.; Fafi- Kremer, S. Intravenous Immunoglobulin Administration Significantly Increases BKPyV Genotype-Specific Neutralizing Antibody Titers in Kidney Transplant Recipients. Antimicrob Agents Chemother 2019, 63, doi:10.1128/AAC.00393-19.
Winter Greg and Milstein Cesar, Man-made antibodies. 1991, Nature 349:293-299
Zhong, S.; Randhawa, P.S.; Ikegaya, H.; Chen, Q.; Zheng, H.-Y.; Suzuki, M.; Takeuchi, T.; Shibuya, A.; Kitamura, T.; Yogo, Y. Distribution patterns of BK polyomavirus (BKV) subtypes and subgroups in American, European, and Asian populations suggest co-migration of BKV and the human race. J. Gen. Virol. 2009, 90, 144-152, doi:10.1099/vir.0.83611-0.

Claims

1. An isolated antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising: a CDR1-H comprising an amino acid sequence selected from SEQ ID NOs: 1, 14,
27, 40, 53, 66 and 79, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-H comprising an amino acid sequence selected from SEQ ID NOs: 2, 15,
28, 41, 54, 67 and 80, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR3-H comprising an amino acid sequence selected from SEQ ID NOs: 3, 16,
29, 42, 55, 68 and 81, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR1-L comprising an amino acid sequence selected from SEQ ID NOs: 4, 17,
30, 43, 56, 69 and 82, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; a CDR2-L comprising an amino acid sequence selected from DTS, DAS, LGS, GAS, DQS, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution; and a CDR3-L comprising an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 44, 57, 70 and 83, or an amino acid sequence differing from said sequences by at least one conservative amino acid substitution.
2. The isolated antibody according to claim 1, comprising:
(i) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 1 or a sequence differing from SEQ ID NO: 1 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 2 or a sequence differing from SEQ ID NO: 2 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 3 or a sequence differing from SEQ ID NO: 3 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 4 or a sequence differing from SEQ ID NO: 4 by one conservative amino acid substitution, a CDR2-L comprising an amino acid sequence of DTS or a sequence differing from DTS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 5 or a sequence differing from SEQ ID NO: 5 by one conservative amino acid substitution; or
(ii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 14 or a sequence differing from SEQ ID NO: 14 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 15 or a sequence differing from SEQ ID NO: 15 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 16 or a sequence differing from SEQ ID NO: 16 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 17 or a sequence differing from SEQ ID NO: 17 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 18 or a sequence differing from SEQ ID NO: 18 by one conservative amino acid substitution; or
(iii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 27 or a sequence differing from SEQ ID NO: 27 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 28 or a sequence differing from SEQ ID NO: 28 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 29 or a sequence differing from SEQ ID NO: 29 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 30 or a sequence differing from SEQ ID NO: 30 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DAS or a sequence differing from DAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 31 or a sequence differing from SEQ ID NO: 31 by one conservative amino acid substitution; or
(iv) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 40 or a sequence differing from SEQ ID NO: 40 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 41 or a sequence differing from SEQ ID NO: 41 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 42 or a sequence differing from SEQ ID NO: 42 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 43 or a sequence differing from SEQ ID NO: 43 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of LGS or a sequence differing from LGS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 44 or a sequence differing from SEQ ID NO: 44 by one conservative amino acid substitution; or
(v) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 53 or a sequence differing from SEQ ID NO: 53 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 54 or a sequence differing from SEQ ID NO: 54 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 55 or a sequence differing from SEQ ID NO: 55 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 56 or a sequence differing from SEQ ID NO: 56 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 57 or a sequence differing from SEQ ID NO: 57 by one conservative amino acid substitution; or
(vi) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 66 or a sequence differing from SEQ ID NO: 66 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 67 or a sequence differing from SEQ ID NO: 67 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 68 or a sequence differing from SEQ ID NO: 68 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 69 or a sequence differing from SEQ ID NO: 69 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of DQS or a sequence differing from DQS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 70 or a sequence differing from SEQ ID NO: 70 by one conservative amino acid substitution; or
(vii) a CDR1-H comprising an amino acid sequence of SEQ ID NO: 79 or a sequence differing from SEQ ID NO: 79 by one conservative amino acid substitution, a CDR2-H comprising an amino acid sequence of SEQ ID NO: 80 or a sequence differing from SEQ ID NO: 80 by one conservative amino acid substitution, a CDR3-H comprising an amino acid sequence of SEQ ID NO: 81 or a sequence differing from SEQ ID NO: 81 by one conservative amino acid substitution, a CDR1-L comprising an amino acid sequence of SEQ ID NO: 82 or a sequence differing from SEQ ID NO: 82 by one conservative amino acid substitution, a CDR2- L comprising an amino acid sequence of GAS or a sequence differing from GAS by one conservative amino acid substitution, and a CDR3-L comprising an amino acid sequence of SEQ ID NO: 83 or a sequence differing from SEQ ID NO: 83 by one conservative amino acid substitution.
3. The antibody according to claim 1 or 2, which comprises:
(i) a variable domain of heavy chain of sequence SEQ ID NO: 6 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 7, or a sequence at least 85% identical thereto; or
(ii) a variable domain of heavy chain of sequence SEQ ID NO: 19 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 20, or a sequence at least 85% identical thereto; or
(iii) a variable domain of heavy chain of sequence SEQ ID NO: 32 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 33, or a sequence at least 85% identical thereto; or
(iv) a variable domain of heavy chain of sequence SEQ ID NO: 45 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 46, or a sequence at least 85% identical thereto; or
(v) a variable domain of heavy chain of sequence SEQ ID NO: 58 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 59, or a sequence at least 85% identical thereto; or
(vi) a variable domain of heavy chain of sequence SEQ ID NO: 71 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 72, or a sequence at least 85% identical thereto; or
(vii) a variable domain of heavy chain of sequence SEQ ID NO: 84 or a sequence at least 85% identical thereto and/or a variable domain of light chain of sequence SEQ ID NO: 85, or a sequence at least 85% identical thereto.
4. The antibody according to anyone of claims 1 to 3, which comprises: (i) a heavy chain comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least 85% identical thereto and/or a light chain comprising an amino acid sequence of SEQ ID NO: 9, or a sequence at least 85% identical thereto; or
(ii) a heavy chain of sequence SEQ ID NO: 21 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 22, or a sequence at least 85% identical thereto, or
(iii) a heavy chain of sequence SEQ ID NO: 34 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 35, or a sequence at least 85% identical thereto, or
(iv) a heavy chain of sequence SEQ ID NO: 47 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 48, or a sequence at least 85% identical thereto, or
(v) a heavy chain of sequence SEQ ID NO: 60 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 61, or a sequence at least 85% identical thereto, or
(vi) a heavy chain of sequence SEQ ID NO: 73 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 74, or a sequence at least 85% identical thereto, or
(vii) a heavy chain of sequence SEQ ID NO: 86 or a sequence at least 85% identical thereto and/or a light chain of sequence SEQ ID NO: 87, or a sequence at least 85% identical thereto.
5. An isolated antibody, or an antigen-binding fragment thereof, neutralizing at least one BKPy virus comprising a VP1 protein of amino acid sequence SEQ ID NO: 96, SEQ ID NO: 97 or SEQ ID NO: 101 with an IC50 of less than 40 nM.
6. An immunoconjugate comprising an antibody according to any one of claims 1 to 5 conjugated or linked to a detectable probe.
7. A combination of at least a first antibody, or an antigen-binding fragment thereof, and a second antibody, or an antigen-binding fragment thereof, wherein said first and second antibodies being selected among the antibodies defined in anyone of claims 1 to 5.
8. An isolated multi-specific antibody comprising at least a first antibody, or an antigenbinding fragment thereof, and at least a second antibody, or an antigen-binding fragment thereof, joined to each other, wherein said first and second antibodies being selected among the antibodies defined in anyone of claims 1 to 5.
9. The combination according to claim 7 or the isolated multi-specific antibody according to claim 8 comprising:
(a) as the first antibody, antibody (i) or (iv) of anyone of claims 1 to 4, and
(b) as the second antibody, an antibody selected among antibodies (i), (iv), (vi) or (vii) of anyone of claims 1 to 4.
10. The combination or the isolated multi-specific antibody according to claim 9 comprising:
(a) as the first antibody, the antibody (i) of anyone of claims 1 to 4, and as the second antibody, the antibody (iv) of anyone of claims 1 to 4; or
(b) as the first antibody, the antibody (i) of anyone of claims 1 to 4, and as the second antibody, the antibody (vi) of anyone of claims 1 to 4; or
(c) as the first antibody, the antibody (i) of anyone of claims 1 to 4, and as the second antibody, the antibody (vii) of anyone of claims 1 to 4; or
(d) as the first antibody, the antibody (iv) of anyone of claims 1 to 4, and as the second antibody, the antibody (vi) of anyone of claims 1 to 4.
11. A pharmaceutical composition comprising at least an antibody according to anyone of claims 1 to 5, at least a combination of at least a first and a second antibodies according to claims 7, 9 or 10, or at least an isolated multi-specific antibody according to anyone of claims 8 to 10, and a pharmaceutically acceptable carrier.
12. An isolated antibody according to anyone of claims 1 to 5, at least a combination of at least a first and a second antibodies according to claims 7, 9 or 10, at least an isolated multispecific antibody according to anyone of claims 8 to 10, or a pharmaceutical composition according to claim 11 , for use as a medicament.
13. An isolated antibody according to anyone of claims 1 to 5, at least a combination of at least a first and a second antibodies according to claims 7, 9 or 10, at least a multi-specific antibody according to anyone of claims 8 to 10, or a pharmaceutical composition according to claim 11, for use in the prevention and/or treatment of a BKPyV -inf ection and/or a BKPyV - inf ection associated disease, in a subject in need thereof.
14. An isolated nucleic acid sequence comprising a sequence encoding an antibody according to any one of claims 1 to 5.
15. An isolated expression vector comprising the nucleic acid a sequence according to claim 14.
16. A recombinant host cell which has been transformed by a nucleic acid sequence according to claim 14 or an expression vector according to claim 15.
17. A method for producing an antibody binding a BK polyomavirus (BKPyV) VP1 capsid protein, or an antigen-binding fragment thereof, comprising at least the steps of culturing the host cell of claim 16 in a culture medium under conditions suitable for producing said antibody or fragment thereof and recovering the antibody or fragment thereof from said cultured cells.
PCT/EP2023/056946 2022-03-18 2023-03-17 Bk polyomavirus antibodies and uses thereof WO2023175171A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP22305326.5 2022-03-18
EP22305326 2022-03-18

Publications (1)

Publication Number Publication Date
WO2023175171A1 true WO2023175171A1 (en) 2023-09-21

Family

ID=81308377

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2023/056946 WO2023175171A1 (en) 2022-03-18 2023-03-17 Bk polyomavirus antibodies and uses thereof

Country Status (1)

Country Link
WO (1) WO2023175171A1 (en)

Citations (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4458066A (en) 1980-02-29 1984-07-03 University Patents, Inc. Process for preparing polynucleotides
US4596556A (en) 1985-03-25 1986-06-24 Bioject, Inc. Hypodermic injection apparatus
US4790824A (en) 1987-06-19 1988-12-13 Bioject, Inc. Non-invasive hypodermic injection device
EP0368684A1 (en) 1988-11-11 1990-05-16 Medical Research Council Cloning immunoglobulin variable domain sequences.
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
WO1994011026A2 (en) 1992-11-13 1994-05-26 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human b lymphocyte restricted differentiation antigen for treatment of b cell lymphoma
EP0616640A1 (en) 1991-12-02 1994-09-28 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
US5399163A (en) 1992-07-24 1995-03-21 Bioject Inc. Needleless hypodermic injection methods and device
US5641870A (en) 1995-04-20 1997-06-24 Genentech, Inc. Low pH hydrophobic interaction chromatography for antibody purification
US6172197B1 (en) 1991-07-10 2001-01-09 Medical Research Council Methods for producing members of specific binding pairs
US6291158B1 (en) 1989-05-16 2001-09-18 Scripps Research Institute Method for tapping the immunological repertoire
WO2003002609A2 (en) 2001-06-28 2003-01-09 Domantis Limited Dual-specific ligand and its use
US6548640B1 (en) 1986-03-27 2003-04-15 Btg International Limited Altered antibodies
US6582915B1 (en) 1991-12-02 2003-06-24 Medical Research Council Production of anti-self bodies from antibody segment repertories and displayed on phage
WO2004003019A2 (en) 2002-06-28 2004-01-08 Domantis Limited Immunoglobin single variant antigen-binding domains and dual-specific constructs
US6696245B2 (en) 1997-10-20 2004-02-24 Domantis Limited Methods for selecting functional polypeptides
EP1433846A2 (en) 1990-07-10 2004-06-30 Cambridge Antibody Technology LTD Phagemid-based method of producing filamentous bacteriophage particles displaying antibody molecules and the corresponding bacteriophage particles.
WO2004058821A2 (en) 2002-12-27 2004-07-15 Domantis Limited Dual specific single domain antibodies specific for a ligand and for the receptor of the ligand
WO2004081026A2 (en) 2003-06-30 2004-09-23 Domantis Limited Polypeptides
WO2004091668A1 (en) 2003-04-15 2004-10-28 Algeta As Thorium-227 for use in radiotherapy of soft tissue disease
WO2004101790A1 (en) 2003-05-14 2004-11-25 Domantis Limited A process for recovering polypeptides that unfold reversibly from a polypeptide repertoire
WO2007059782A1 (en) 2005-11-28 2007-05-31 Genmab A/S Recombinant monovalent antibodies and methods for production thereof
US20090232811A1 (en) 2007-12-21 2009-09-17 Christian Klein Bivalent, bispecific antibodies
US8796424B2 (en) 2009-05-27 2014-08-05 Hoffmann-La Roche Inc. Tri- or tetraspecific antibodies
US9382323B2 (en) 2009-04-02 2016-07-05 Roche Glycart Ag Multispecific antibodies comprising full length antibodies and single chain fab fragments
WO2017046676A1 (en) 2015-09-16 2017-03-23 Novartis Ag Polyomavirus neutralizing antibodies
US9890204B2 (en) 2009-04-07 2018-02-13 Hoffmann-La Roche Inc. Trivalent, bispecific antibodies
WO2019106578A2 (en) * 2017-12-01 2019-06-06 Novartis Ag Polyomavirus neutralizing antibodies
FR3096682A1 (en) * 2019-06-03 2020-12-04 Universite De Tours NEW ANTIBODIES TARGETING VP-1 PROTEIN, ITS FRAGMENTS AND THEIR USES TO DETECT POLYOMAVIRUS BK INFECTION
WO2021250097A1 (en) * 2020-06-09 2021-12-16 Memo Therapeutics Ag Anti-bk virus antibody molecules

Patent Citations (33)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4458066A (en) 1980-02-29 1984-07-03 University Patents, Inc. Process for preparing polynucleotides
US4596556A (en) 1985-03-25 1986-06-24 Bioject, Inc. Hypodermic injection apparatus
US6548640B1 (en) 1986-03-27 2003-04-15 Btg International Limited Altered antibodies
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
US4790824A (en) 1987-06-19 1988-12-13 Bioject, Inc. Non-invasive hypodermic injection device
EP0368684A1 (en) 1988-11-11 1990-05-16 Medical Research Council Cloning immunoglobulin variable domain sequences.
US20040110941A2 (en) 1988-11-11 2004-06-10 Medical Research Council Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors
US6291158B1 (en) 1989-05-16 2001-09-18 Scripps Research Institute Method for tapping the immunological repertoire
EP1433846A2 (en) 1990-07-10 2004-06-30 Cambridge Antibody Technology LTD Phagemid-based method of producing filamentous bacteriophage particles displaying antibody molecules and the corresponding bacteriophage particles.
US6172197B1 (en) 1991-07-10 2001-01-09 Medical Research Council Methods for producing members of specific binding pairs
EP0616640A1 (en) 1991-12-02 1994-09-28 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
US6582915B1 (en) 1991-12-02 2003-06-24 Medical Research Council Production of anti-self bodies from antibody segment repertories and displayed on phage
US6593081B1 (en) 1991-12-02 2003-07-15 Medical Research Council Production of anti-self antibodies from antibody segment repertoires and displayed on phage
US5399163A (en) 1992-07-24 1995-03-21 Bioject Inc. Needleless hypodermic injection methods and device
WO1994011026A2 (en) 1992-11-13 1994-05-26 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human b lymphocyte restricted differentiation antigen for treatment of b cell lymphoma
US5641870A (en) 1995-04-20 1997-06-24 Genentech, Inc. Low pH hydrophobic interaction chromatography for antibody purification
US6696245B2 (en) 1997-10-20 2004-02-24 Domantis Limited Methods for selecting functional polypeptides
WO2003002609A2 (en) 2001-06-28 2003-01-09 Domantis Limited Dual-specific ligand and its use
WO2004003019A2 (en) 2002-06-28 2004-01-08 Domantis Limited Immunoglobin single variant antigen-binding domains and dual-specific constructs
WO2004058821A2 (en) 2002-12-27 2004-07-15 Domantis Limited Dual specific single domain antibodies specific for a ligand and for the receptor of the ligand
WO2004091668A1 (en) 2003-04-15 2004-10-28 Algeta As Thorium-227 for use in radiotherapy of soft tissue disease
WO2004101790A1 (en) 2003-05-14 2004-11-25 Domantis Limited A process for recovering polypeptides that unfold reversibly from a polypeptide repertoire
WO2004081026A2 (en) 2003-06-30 2004-09-23 Domantis Limited Polypeptides
WO2005035572A2 (en) 2003-10-08 2005-04-21 Domantis Limited Antibody compositions and methods
WO2007059782A1 (en) 2005-11-28 2007-05-31 Genmab A/S Recombinant monovalent antibodies and methods for production thereof
US20090232811A1 (en) 2007-12-21 2009-09-17 Christian Klein Bivalent, bispecific antibodies
US9382323B2 (en) 2009-04-02 2016-07-05 Roche Glycart Ag Multispecific antibodies comprising full length antibodies and single chain fab fragments
US9890204B2 (en) 2009-04-07 2018-02-13 Hoffmann-La Roche Inc. Trivalent, bispecific antibodies
US8796424B2 (en) 2009-05-27 2014-08-05 Hoffmann-La Roche Inc. Tri- or tetraspecific antibodies
WO2017046676A1 (en) 2015-09-16 2017-03-23 Novartis Ag Polyomavirus neutralizing antibodies
WO2019106578A2 (en) * 2017-12-01 2019-06-06 Novartis Ag Polyomavirus neutralizing antibodies
FR3096682A1 (en) * 2019-06-03 2020-12-04 Universite De Tours NEW ANTIBODIES TARGETING VP-1 PROTEIN, ITS FRAGMENTS AND THEIR USES TO DETECT POLYOMAVIRUS BK INFECTION
WO2021250097A1 (en) * 2020-06-09 2021-12-16 Memo Therapeutics Ag Anti-bk virus antibody molecules

Non-Patent Citations (87)

* Cited by examiner, † Cited by third party
Title
"Oxford Dictionary Of Biochemistry And Molecular Biology", 2000, OXFORD UNIVERSITY PRESS
"Proteins: A Theoretical Perspective of Dynamics, Structure, and Thermodynamics", vol. 71, January 1991, pages: 280
"The Dictionary of Cell and Molecular Biology", 1999, ACADEMIC PRESS
ALTSCHUL ET AL., J. MOL. BIOL., vol. 215, 1990, pages 403 - 410
ALTSCHUL ET AL., NATURE GENET, vol. 6, 1994, pages 119
ALTSCHUL ET AL., NUCLEIC ACIDS RES., vol. 25, 1977, pages 3389 - 3402
ALTSCHUL SFBOGUSKI MSGISH WWOOTTON JC: "Issues in searching molecular sequence databases", NAT GENET, vol. 6, no. 2, 1994, pages 119 - 129, XP001006126, DOI: 10.1038/ng0294-119
ALTSCHUL SFGISH WMILLER WMYERS EWLIPMAN DJ: "Basic local alignment search tool", J MOL BIOL., vol. 215, no. 3, 1990, pages 403 - 410, XP002949123, DOI: 10.1006/jmbi.1990.9999
ALTSCHUL SFMADDEN TLSCHAFFER AA ET AL.: "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", NUCLEIC ACIDS RES., vol. 25, no. 17, 1997, pages 3389 - 3402, XP002905950, DOI: 10.1093/nar/25.17.3389
AUSUBEL ET AL., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, 1995
BEAUCAGE ET AL., TETRA. LETT., vol. 22, 1981, pages 1859
BENNETT ET AL., MICROBES AND INFECTION, vol. 14, no. 9, 2012, pages 672 - 683
BENOTMANE, I.SOLIS, M.VELAY, A.COGNARD, N.OLAGNE, J.GAUTIER VARGAS, G.PERRIN, P.MARX, D.SOULIER, E.GALLAIS, F. ET AL.: "Intravenous immunoglobulin as a preventive strategy against BK virus viremia and BKV-associated nephropathy in kidney transplant recipients-Results from a proof-of-concept study", AM J TRANSPLANT, 2020
BIRD ET AL., SCIENCE, vol. 242, 1989, pages 423 - 426
BRESSOLLETTE-BODIN, C.COSTE-BUREL, M.HOURMANT, M.SEBILLE, V.ANDRE-GARNIER, E.IMBERT-MARCILLE, B.M.: "A prospective longitudinal study of BK virus infection in 104 renal transplant recipients", AM. J. TRANSPLANT., vol. 5, 2005, pages 1926 - 1933
BROOKS, B.R.; BRUCCOLERI, R. E.; OLAFSON, B.D.; STATES, D.J.; SWAMINATHAN, S.; KARPLUS, M.: "CHARMM: A program for macromolecular energy, minimization, and dynamics calculations", J. COMP. CHEM., vol. 4, 1983, pages 187 - 217, XP008008192, DOI: 10.1002/jcc.540040211
CASE DAVID A.THOMAS E. CHEATHAM IIITOM DARDENHOLGER GOHLKERAY LUOKENNETH M. MERZ JR.ALEXEY ONUFRIEVCARLOS SIMMERLINGBING WANGROBER: "The Amber biomolecular simulation programs", JOURNAL OF COMPUTATIONAL CHEMISTRY, vol. 26, pages 1668 - 1688
CESARO, S.DALIANIS, T.HANSSEN RINALDO, C.KOSKENVUO, M.PEGORARO, A.EINSELE, H.CORDONNIER, C.HIRSCH, H.H.: "ECIL-6 Group ECIL guidelines for the prevention, diagnosis, and treatment of BK polyomavirus-associated haemorrhagic cystitis in haematopoietic stem cell transplant recipients", J ANTIMICROB CHEMOTHER, vol. 73, 2018, pages 12 - 21
CORPET ET AL., NUCLEIC ACIDS RESEARCH, vol. 16, 1988, pages 10881
CORPET F: "Multiple sequence alignment with hierarchical clustering", NUCLEIC ACIDS RES, vol. 16, no. 22, 1988, pages 10881 - 10890
CROMWELL ET AL., AAPS JOURNAL, vol. 8, no. 3, 2006, pages E572 - E579
DEVEREAUX ET AL., NUC. ACIDS RES., vol. 12, 1984, pages 387 - 395
DEVEREUX JHAEBERLI PSMITHIES O: "A comprehensive set of sequence analysis programs for the VAX", NUCLEIC ACIDS RES., vol. 12, 1984, pages 387 - 395
ECKERT ET AL., PCR METHODS AND APPLICATIONS, vol. 1, 1991, pages 17
ESPADA, E.CHENG, M.P.KIM, H.T.WOOLLEY, A.E.AVIGAN, J.I.FORCADE, E.SOARES, M.V.D.LACERDA, J.F.NIKIFOROW, S.GOOPTU, M. ET AL.: "BK virus-specific T-cell immune reconstitution after allogeneic hematopoietic cell transplantation", BLOOD ADV, vol. 4, 2020, pages 1881 - 1893
FENG DFDOOLITTLE RF: "Progressive sequence alignment as a prerequisite to correct phylogenetic trees", J MOL EVOL, vol. 25, no. 4, 1987, pages 351 - 360
FENGDOOLITTLE, J. MOL. EVOL., vol. 35, 1987, pages 351 - 360
FURMAGA JKOWALCZYK MZAPOLSKI TFURMAGA OKRAKOWSKI LRUDZKI GAROSZYNSKI AJAKUBCZAK A: "BK Polyomavirus-Biology, Genomic Variation and Diagnosis", VIRUSES, vol. 13, no. 8, 30 July 2021 (2021-07-30), pages 1502
HARALAMBIEVA IHOVSYANNIKOVA IGVIERKANT RAPOLAND GA: "Development of a novel efficient fluorescence-based plaque reduction microneutralization assay for measles virus immunity", CLIN VACCINE IMMUNOL, vol. 15, no. 7, 2008, pages 1054 - 1059, XP008149270, DOI: 10.1128/​CVI.00008-08
HELLE FBROCHOT EHANDALA L ET AL.: "Biology of the BKPyV: An Update", VIRUSES, vol. 9, no. 11, 3 November 2017 (2017-11-03), pages 327
HENIKOFF SHENIKOFF JG: "Amino acid substitution matrices from protein blocks", PROC NATL ACAD SCI U S A., vol. 89, no. 22, 1992, pages 10915 - 10919, XP002599751, DOI: 10.1073/pnas.89.22.10915
HENIKOFFHENIKOFF, PROC. NATL. ACAD. SCI. USA, vol. 89, 1989, pages 10915
HIGGINS DGSHARP PM: "CLUSTAL: a package for performing multiple sequence alignment on a microcomputer", GENE, vol. 73, no. 1, 1988, pages 237 - 244, XP025705287, DOI: 10.1016/0378-1119(88)90330-7
HIGGINS DGSHARP PM: "Fast and sensitive multiple sequence alignments on a microcomputer", COMPUT APPL BIOSCI, vol. 5, no. 2, 1989, pages 151 - 153
HIGGINSSHARP, CABIOS, vol. 5, 1989, pages 151 - 153
HIRSCH, H.H.RANDHAWA, P.: "AST Infectious Diseases Community of Practice BK polyomavirus in solid organ transplantation", AM. J. TRANSPLANT., vol. 13, 2013, pages 179 - 188
HOLLINGERHUDSON, NATURE BIOTECHNOLOGY, vol. 23, no. 9, 2005, pages 1126 - 1136
HURDISS, D.L.FRANK, M.SNOWDEN, J.S.MACDONALD, A.RANSON, N.A.: "The Structure of an Infectious Human Polyomavirus and Its Interactions with Cellular Receptors", STRUCTURE, vol. 26, 2018, pages 839 - 847
HUSTON ET AL., PROC. NATL. ACAD. SCI., vol. 85, 1988, pages 5879 - 5883
HWANG, S.D.LEE, J.H.LEE, S.W.KIM, J.K.KIM, M.-J.SONG, J.H.: "High-Dose Intravenous Immunoglobulin Treatment of Polyomavirus Nephropathy Developing After T Cell-Mediated Rejection Treatment: A Case Report", TRANSPLANT PROC, vol. 50, 2018, pages 2575 - 2578, XP085504917, DOI: 10.1016/j.transproceed.2018.01.021
IMLAY, H.XIE, H.LEISENRING, W.M.DUKE, E.R.KIMBALL, L.E.HUANG, M.-L.PERGAM, S.A.HILL, J.A.JEROME, K.R.MILANO, F. ET AL.: "Presentation of BK polyomavirus-associated hemorrhagic cystitis after allogeneic hematopoietic cell transplantation", BLOOD ADV, vol. 4, 2020, pages 617 - 628
KNOWLES, W.A.GIBSON, P.E.GARDNER, S.D.: "Serological typing scheme for BK-like isolates of human polyomavirus", J. MED. VIROL., vol. 28, 1989, pages 118 - 123, XP055491577, DOI: 10.1002/jmv.1890280212
KUNDU SMELTON JSSORENSEN DCPHILLIPS GN JR: "Dynamics of proteins in crystals: comparison of experiment with simple models", BIOPHYS J, vol. 83, no. 2, 2002, pages 723 - 732
LAURE JASON-MOLLERMURPHY MBRUNO J: "Overview of Biacore systems and their applications", CURR PROTOC PROTEIN SCI, September 2006 (2006-09-01)
LEFRANC ET AL., DEV. COMP. IMMUNOL., vol. 27, 2003, pages 55 - 77
LEFRANC M.-P., IMMUNOLOGY TODAY, vol. 18, 1997, pages 509
LEFRANC MP: "Unique database numbering system for immunogenetic analysis", IMMUNOL TODAY, vol. 18, no. 11, November 1997 (1997-11-01), pages 509, XP004093509, DOI: 10.1016/S0167-5699(97)01163-8
LEFRANC MPPOMMIE CRUIZ MGIUDICELLI VFOULQUIER ETRUONG LTHOUVENIN-CONTET VLEFRANC G: "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains", DEV COMP IMMUNOL, vol. 27, no. 1, January 2003 (2003-01-01), pages 55 - 77, XP055585227, DOI: 10.1016/S0145-305X(02)00039-3
LINDNER J.M.CORNACCHIONE V.SATHE A.BE C.SRINIVAS H.RIQUET E.LEBER X.-C.HEIN A.WROBEL M.B.SCHARENBERG M. ET AL.: "Human Memory B Cells Harbor Diverse Cross-Neutralizing Antibodies against BK and JC Polyomaviruses", IMMUNITY, vol. 50, 2019, pages 668 - 676
LIU ET AL., TRANSPLANTATION, vol. 101, 2017, pages 1488 - 1494
LIU SCHAUDHRY MRBERREBI AAPAPADIMITRIOU JCDRACHENBERG CBHARIRIAN AALEXIEV BA: "Polyomavirus Replication and Smoking Are Independent Risk Factors for Bladder Cancer After Renal Transplantation", TRANSPLANTATION, vol. 101, no. 6, June 2017 (2017-06-01), pages 1488 - 1494
MACKERELL, A.D., JR.BROOKS, B.BROOKS, C. L., IIINILSSON, L.ROUX, B.WON, Y.KARPLUS, M. ET AL.: "The Encyclopedia of Computational Chemistry", vol. 1, 1998, JOHN WILEY & SONS, article "CHARMM: The Energy Function and Its Parameterization with an Overview of the Program", pages: 271 - 277
MAINRA, R.XU, Q.CHIBBAR, R.HASSAN, A.SHOKER, A.: "Severe antibody-mediated rejection following IVIG infusion in a kidney transplant recipient with BK-virus nephropathy", TRANSPL IMMUNOL, vol. 28, 2013, pages 145 - 147
MATSUMURA, S.KATO, T.TANIGUCHI, A.KAWAMURA, M.NAKAZAWA, S.NAMBA-HAMANO, T.ABE, T.NONOMURA, N.IMAMURA, R.: "Clinical Efficacy of Intravenous Immunoglobulin for BK Polyomavirus-Associated Nephropathy After Living Kidney Transplantation", THER CLIN RISK MANAG, vol. 16, 2020, pages 947 - 952
MATTILA ET AL., NUCLEIC ACIDS RES., vol. 19, 1991, pages 967
MCILROY DHONEMANN MNGUYEN NKBARBIER PPELTIER CRODALLEC AHALARY FPRZYROWSKI ELIEBERT UHOURMANT M: "Persistent BK Polyomavirus Viruria is Associated with Accumulation of VP1 Mutations and Neutralization Escape", VIRUSES, vol. 12, no. 8, 29 July 2020 (2020-07-29), pages 824
MCILROY ET AL., VIRUSES, vol. 12, no. 8, 2020
MORIYAMA TSOROKIN A.: "BK virus (BKV): infection, propagation, quantitation, purification, labeling, and analysis of cell entry", CURR PROTOC CELL BIOL, March 2009 (2009-03-01)
MULLER R: "Determination of affinity and specificity of anti-hapten antibodies by competitive radioimmunoassay", METHODS ENZYMOL., vol. 92, 1983, pages 589 - 601
NARANG ET AL., METH. ENZYMOL., vol. 68, 1979, pages 109
NEEDLEMAN SBWUNSCH CD: "A general method applicable to the search for similarities in the amino acid sequence of two proteins", J MOL BIOL., vol. 48, no. 3, 1970, pages 443 - 453, XP024011703, DOI: 10.1016/0022-2836(70)90057-4
NEEDLEMANWUNSCH, J. MOL. BIOL., vol. 48, 1970, pages 443
NELSON ASYALAMARTHI NYONG MKBLYTH E: "Beyond antivirals: virus-specific T-cell immunotherapy for BK virus haemorrhagic cystitis and JC virus progressive multifocal leukoencephalopathy", CURR OPIN INFECT DIS, vol. 34, no. 6, 1 December 2021 (2021-12-01), pages 627 - 634
NELSON ET AL., CURR. OPIN. INFECT. DIS., vol. 34, 2021, pages 627 - 634
NICKELEIT, V.KLIMKAIT, T.BINET, I.F.DALQUEN, P.DEL ZENERO, V.THIEL, G.MIHATSCH, M.J.HIRSCH, H.H.: "Testing for polyomavirus type BK DNA in plasma to identify renal-allograft recipients with viral nephropathy", N. ENGL. J. MED., vol. 342, 2000, pages 1309 - 1315
PAPADIMITRIOU ET AL., AM. J. TRANSPLANT., vol. 16, 2016, pages 398 - 406
PASTRANA ET AL., PIOS PATHOG, vol. 8, 2012, pages el002650
PASTRANA, D.V.BRENNAN, D.C.CUBURU, N.STORCH, G.A.VISCIDI, R.P.RANDHAWA, P.S.BUCK, C.B.: "Neutralization serotyping of BK polyomavirus infection in kidney transplant recipients", PLOS PATHOG, vol. 8, no. 8, 2012, pages e1002650, XP055038943, DOI: 10.1371/journal.ppat.1002650
PASTRANA, D.V.RAY, U.MAGALDI, T.G.SCHOWALTER, R.M.QUBURU, N.BUCK, C.B.: "BK polyomavirus genotypes represent distinct serotypes with distinct entry tropism", J. VIROL., vol. 87, 2013, pages 10105 - 10113, XP055175214, DOI: 10.1128/JVI.01189-13
PEARSON WRLIPMAN DJ: "Improved tools for biological sequence comparison", PROC NATL ACAD SCI U S A., vol. 85, no. 8, 1988, pages 2444 - 2448, XP002060460, DOI: 10.1073/pnas.85.8.2444
PEARSONLIPMAN, PROC. NATL. ACAD. SCI. U.S.A., vol. 85, 1988, pages 2444
PEARSONLIPMAN, PROC. NAT'L. ACAD. SCI. USA, vol. 85, 1988, pages 2444
PIBUM ET AL., PEDIATRIC TRANSPLANTATION, vol. 24, 2020, pages e13600
PIBURN, K.H.AL-AKASH, S.: "Use of intravenous immunoglobulin in a highly sensitized pediatric renal transplant recipient with severe BK DNAemia and rising DSA", PEDIATR TRANSPLANT, vol. 24, 2020, pages e13600
REPLOEG, M.D.STORCH, G.A.CLIFFORD, D.B.: "Bk virus: a clinical review", CLIN INFECT DIS, vol. 33, 2001, pages 191 - 202, XP055173693
SATHE ATUL ET AL: "1475 Pre-Clinical Characterization of MAU868, a Novel Neutralizing Antibody Targeting BK Virus", 5 December 2020 (2020-12-05), pages 1 - 1, XP055949946, Retrieved from the Internet <URL:https://ash.confex.com/ash/2020/webprogram/Paper141134.html> *
SMITH TFWATERMAN MS: "Identification of common molecular subsequences", J MOL BIOL, vol. 147, no. 1, 1981, pages 195 - 197, XP024015032, DOI: 10.1016/0022-2836(81)90087-5
SMITHWATERMAN, ADV. APPL. MATH., vol. 2, 1981, pages 482
SOLIS, M.VELAY, A.PORCHER, R.DOMINGO-CALAP, P.SOULIER, E.JOLY, M.MEDDEB, M.KACK-KACK, W.MOULIN, B.BAHRAM, S. ET AL.: "Neutralizing Antibody-Mediated Response and Risk of BK Virus-Associated Nephropathy", J. AM. SOC. NEPHROL., vol. 29, 2018, pages 326 - 334
VELAY ET AL., ANTIMICROB AGENTS CHEMOTHER, 2019, pages 63
VELAY, A.SOLIS, M.BENOTMANE, I.GANTNER, P.SOULIER, E.MOULIN, B.CAILLARD, S.FAFI-KREMER, S.: "Intravenous Immunoglobulin Administration Significantly Increases BKPyV Genotype-Specific Neutralizing Antibody Titers in Kidney Transplant Recipients", ANTIMICROB AGENTS CHEMOTHER, vol. 2019, pages 63
WALTER ET AL., ANAL. BIOCHEM., vol. 212, no. 2, 1993, pages 469 - 480
WANGGOSH, J. MEMBRANE SCI., vol. 318, 2008, pages 311 - 316
WARD ET AL., NATURE, vol. 341, 1989, pages 544 - 546
WINTER GREGMILSTEIN CESAR: "Man-made antibodies", NATURE, vol. 349, 1991, pages 293 - 299, XP037115400, DOI: 10.1038/349293a0
ZAPATA ET AL., PROTEIN ENG, vol. 8, no. 10, 1995, pages 1057 - 1062
ZHONG, S.RANDHAWA, P.S.IKEGAYA, H.CHEN, Q.ZHENG, H.-Y.SUZUKI, M.TAKEUCHI, T.SHIBUYA, A.KITAMURA, T.YOGO, Y.: "Distribution patterns of BK polyomavirus (BKV) subtypes and subgroups in American, European, and Asian populations suggest co-migration of BKV and the human race", J. GEN. VIROL., vol. 90, 2009, pages 144 - 152, XP055491281, DOI: 10.1099/vir.0.83611-0

Similar Documents

Publication Publication Date Title
Wang et al. Importance of neutralizing monoclonal antibodies targeting multiple antigenic sites on the Middle East respiratory syndrome coronavirus spike glycoprotein to avoid neutralization escape
Maeda et al. A panel of nanobodies recognizing conserved hidden clefts of all SARS-CoV-2 spike variants including Omicron
JP7644087B2 (en) Human monoclonal antibodies against enterovirus D68
TWI700295B (en) Antibodies specific for enteroviruses that infect humans
US11834494B2 (en) Antibodies to zika virus and methods of use thereof
JP2023518849A (en) Human monoclonal antibody against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Huang et al. Antibody recognition of the Pneumovirus fusion protein trimer interface
Addetia et al. Therapeutic and vaccine-induced cross-reactive antibodies with effector function against emerging Omicron variants
US20230073067A1 (en) Humanized monoclonal antibody for 2019 novel coronavirus and use thereof
JP2023528072A (en) Therapeutic antibodies with neutralizing activity against SARS-CoV-2 glycoprotein S
US9512201B2 (en) Human binding molecules capable of binding to and neutralizing hepatitis B viruses and uses thereof
Bar-Peled et al. A potent neutralizing site III-specific human antibody neutralizes human metapneumovirus in vivo
WO2022216223A1 (en) Vaccine and/or antibody for viral infection
JP2022031634A (en) Epitope of hepatitis b virus surface antigen and binding molecule specifically binding to same for neutralizing hepatitis b virus
WO2022241200A1 (en) Cross-reactive coronavirus antibodies
CN116514960A (en) A fully human monoclonal antibody against respiratory syncytial virus and its application
WO2022069232A1 (en) Single domain antibodies against the nucleoprotein of sars-cov-2
TW202144405A (en) Novel monoclonal antibodies against sars-cov-2 and uses thereof
Sun et al. Development of AAV-delivered broadly neutralizing anti-human ACE2 antibodies against SARS-CoV-2 variants
Capella-Pujol et al. Signatures of VH 1-69-derived hepatitis C virus neutralizing antibody precursors defined by binding to envelope glycoproteins
Behzadi et al. A cross-reactive mouse monoclonal antibody against rhinovirus mediates phagocytosis in vitro
WO2023175171A1 (en) Bk polyomavirus antibodies and uses thereof
CN115087667B (en) Antigen binding proteins that specifically bind SARS-CoV-2
Iankov et al. Neutralization capacity of measles virus H protein specific IgG determines the balance between antibody-enhanced infectivity and protection in microglial cells
Guo et al. A SARS-CoV-2 neutralizing antibody with exceptional spike binding coverage and optimized therapeutic potentials

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23712018

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE