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BR112021010306A2 - Método com base em células para determinar uma atividade de toxina botulínica - Google Patents

Método com base em células para determinar uma atividade de toxina botulínica

Info

Publication number
BR112021010306A2
BR112021010306A2 BR112021010306A BR112021010306A BR112021010306A2 BR 112021010306 A2 BR112021010306 A2 BR 112021010306A2 BR 112021010306 A BR112021010306 A BR 112021010306A BR 112021010306 A BR112021010306 A BR 112021010306A BR 112021010306 A2 BR112021010306 A2 BR 112021010306A2
Authority
BR
Brazil
Prior art keywords
cell
botulinum toxin
antibody
activity
present
Prior art date
Application number
BR112021010306A
Other languages
English (en)
Inventor
Gun Lee Chee
Hyun Oum Ji
Vijayakumar Ajay
Xiangai Gui
Original Assignee
Hugel Inc
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
Priority claimed from KR1020180150640A external-priority patent/KR101983216B1/ko
Priority claimed from KR1020180150997A external-priority patent/KR101940500B1/ko
Priority claimed from KR1020180159701A external-priority patent/KR102251096B1/ko
Application filed by Hugel Inc filed Critical Hugel Inc
Publication of BR112021010306A2 publication Critical patent/BR112021010306A2/pt

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1267—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-positive bacteria
    • C07K16/1282—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-positive bacteria from Clostridium (G)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1203—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria
    • C07K16/1246—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria from Rickettsiales (O)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0618—Cells of the nervous system
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5058—Neurological cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • 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
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • 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
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    • C12N2500/00—Specific components of cell culture medium
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    • C12N2503/00—Use of cells in diagnostics
    • C12N2503/02—Drug screening
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/33—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Clostridium (G)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/42—Poisoning, e.g. from bites or stings
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/70—Mechanisms involved in disease identification
    • G01N2800/709—Toxin induced

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
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  • Biomedical Technology (AREA)
  • Immunology (AREA)
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  • Hematology (AREA)
  • Urology & Nephrology (AREA)
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  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Toxicology (AREA)
  • Genetics & Genomics (AREA)
  • Food Science & Technology (AREA)
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  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Pathology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

método com base em células para determinar uma atividade de toxina botulínica. a presente invenção se refere a uma célula e um anticorpo para determinar a atividade de toxina botulínica e a um método para determinar a atividade de toxina botulínica com o uso do mesmo. atualmente, existe a necessidade de desenvolver um ensaio de potência com base em células (cbpa) para substituir o bioensaio ld50 de camundongo (mld50) no campo de medição da potência de toxina botulínica. a célula e o anticorpo para medir a atividade de toxina botulínica de acordo com a presente invenção são uma célula e um anticorpo para cpba que devem substituir o mld50. a linha celular tem um tempo de duplicação significativamente curto em comparação com as células sima convencionais que são usadas para determinar a atividade de toxina botulínica e também tem uma sensibilidade significativamente alta à toxina botulínica em comparação com sua linha celular parental. assim, a linha celular é muito adequada não apenas para a determinação da atividade de toxina botulínica com base em células, mas também para a detecção de toxina botulínica com base em células. ademais, a linha celular de acordo com a presente invenção pode ser ligada e cultivada de forma estável em uma placa de cultura revestida com poli- d-lisina (pdl) e, portanto, pode ser usado de forma muito eficaz para a determinação da atividade de toxina botulínica com base em células ou para detecção de toxina botulínica com base em células. também, o anticorpo é um anticorpo monoclonal que tem uma afinidade de ligação e especificidade significativamente altas para snap25, supera as limitações do cpba convencional e torna possível desenvolver um cbpa mais eficaz. assim, espera-se que o anticorpo seja usado ativamente nos campos farmacêutico e cosmético. além disso, a presente invenção se refere a um ensaio de potência com base em células ideal (cbpa) que usa células n2-42f e um anticorpo monoclonal que tem uma afinidade de ligação e especificidade significativamente altas para snap25, e esse cbpa pode medir a potência de 0,5 u ou menos de toxina botulínica. espera-se que o cbpa que emprega a célula e o anticorpo da presente invenção se torne um ensaio de potência com base em células altamente confiável e reproduzível para a toxina botulínica.
BR112021010306A 2018-11-29 2019-08-19 Método com base em células para determinar uma atividade de toxina botulínica BR112021010306A2 (pt)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR1020180150640A KR101983216B1 (ko) 2018-11-29 2018-11-29 보툴리눔 독소 활성을 결정하기 위한 항체, 및 이를 이용한 활성 측정방법
KR1020180150997A KR101940500B1 (ko) 2018-11-29 2018-11-29 보툴리눔 독소 활성을 결정하기 위한 세포주 및 이를 이용한 활성 결정 방법
KR1020180159701A KR102251096B1 (ko) 2018-12-12 2018-12-12 보툴리눔 독소 활성을 결정하는 세포 기반 방법
PCT/KR2019/010467 WO2020111449A1 (ko) 2018-11-29 2019-08-19 보툴리눔 독소 활성을 결정하는 세포 기반 방법

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BR112021010306A2 true BR112021010306A2 (pt) 2021-11-16

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US (8) US10823725B2 (pt)
EP (2) EP3660509B1 (pt)
CN (4) CN118754976A (pt)
AU (1) AU2019388130B2 (pt)
BR (1) BR112021010306A2 (pt)
CA (1) CA3119962A1 (pt)
LT (1) LT3660509T (pt)
MX (1) MX2021005902A (pt)
WO (1) WO2020111449A1 (pt)

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KR101940500B1 (ko) * 2018-11-29 2019-01-21 주식회사 에이비바이오 보툴리눔 독소 활성을 결정하기 위한 세포주 및 이를 이용한 활성 결정 방법

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