WO2026013218A1

WO2026013218A1 - Anti-tdp-43 vectors, binding molecules and uses thereof

Info

Publication number: WO2026013218A1
Application number: PCT/EP2025/069818
Authority: WO
Inventors: Elodie CHEVALIER; Tariq AFROZ; Mickaël Marc Pascal AUDRAIN; Florence GAUYE; Romain Christian OLLIER
Original assignee: AC Immune SA
Current assignee: AC Immune SA
Priority date: 2024-07-10
Filing date: 2025-07-10
Publication date: 2026-01-15
Anticipated expiration: 2027-01-10

Abstract

The present invention is in the field of transactive response DNA binding protein with a molecular weight of 43 kDa (TARDB or also TDP-43). The invention relates to TDP-43 specific binding molecules, in particular to anti-TDP-43 antibodies or antigen-binding fragments or a derivative thereof, vectors delivering nucleic acid encoding antibodies of the invention, and uses thereof. The present invention provides means and methods to diagnose, prevent, alleviate and/or treat a disease, disorder and/or abnormality associated with TDP-43 aggregates including but not limited to Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), Chronic Traumatic Encephalopathy (CTE), and limbic-predominant age-related TDP-43 encephalopathy (LATE).

Description

Anti-TDP-43 vectors, binding molecules and uses thereof

Field of the invention

The present invention is in the field of transactive response DNA binding protein with a molecular weight of 43 kDa (TARDBP or also TDP-43). The invention relates to TDP-43 specific binding molecules, in particular to anti-TDP-43 antibodies or an antigen-binding fragment or a derivative thereof, vectors delivering nucleic acids encoding the antibodies, and uses thereof. The present invention provides means and methods to diagnose, prevent, alleviate and/or treat a disease, a disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy, including but not limited to Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), and limbic- predominant age-related TDP-43 encephalopathy (LATE).

BACKGROUND

Age-associated brain disorders characterised by pathological aggregation of proteins in the central nervous system (CNS) (proteinopathies) and peripheral organs represent one of the leading causes of disability and mortality in the world. The best characterised protein that forms aggregates is amyloid beta in Alzheimer's disease and related disorders. Other disease- associated, aggregation-prone proteins leading to neurodegeneration include but are not limited to tau, alpha-synuclein (aSyn, a-syn), huntingtin, fused in sarcoma (FUS), dipeptide repeat proteins (DPRs) produced by unconventional translation of the C9orf72 repeat expansion, superoxide dismutase 1 (SOD1), and TDP-43. Diseases involving TDP-43 aggregates are generally listed as TDP-43 proteinopathies including, but not limited to, ALS and FTD.

I. TDP-43 introduction

Transactive response (TAR) DNA binding protein 43 kDa (TDP-43) is a 414-amino acid protein encoded by the TARDBP gene on chromosome lp36.2 (ALS10). TARDBP is comprised of six exons (exon 1 is non-coding; exons 2-6 are protein-coding). TDP-43 belongs to the family of heterogeneous ribonucleoprotein (hnRNP) RNA binding proteins (Wang et al., Trends in Molecular Medicine Vol.14 No.11, 2008, 479-485; Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1 R46-R64). TDP-43 contains five functional domains (Figure 1 in Warraich et al., The International Journal of Biochemistry & Cell Biology 42 (2010) 1606-1609): two RNA recognition motifs (RRM1 and RRM2), which have two highly conserved hexameric ribonucleoprotein 2 (RNP2) and octameric ribonucleoprotein 1 (RNP1) regions, a nuclear export signal (NES) and a nuclear localization signal (NLS) enabling it to shuttle between the nucleus and the cytoplasm transporting bound mRNA, and a glycine-rich domain at the C-terminal, which mediates protein-protein interactions. TDP-43 is involved in multiple aspects of RNA processing, including transcription, splicing, transport, and stabilization (Buratti and Baralle, FEBS Journal 277 (2010) 2268-2281). It is a highly conserved, ubiquitously expressed protein with a tightly autoregulated expression level that shuttles continuously between the nucleus and cytoplasm, but is predominantly localized to the nucleus. In 2006, TDP-43 was identified as the protein that accumulates in the vast majority of cases of frontotemporal lobar degeneration (FTLD) with tau-negative, ubiquitin-positive inclusions (then referred to as FTLD-TDP), and in most cases of amyotrophic lateral sclerosis (ALS) (Arai et al., Biochemical and Biophysical Research Communications 351 (2006) 602-611; Neumann et al., Science 314, (2006), 130-133).

Thirty-eight negative-dominant mutations in TDP-43 have been identified in sporadic and familial ALS patients as well as in patients with inherited FTD mainly located in the glycine- rich domain (Figure 1 in Lagier-Tourenne and Cleveland, Cell 136, 2009, 1001-1004). TDP-43 is inherently aggregation-prone, as shown by sedimentation assays, and this propensity is further increased by some of the ALS-associated TARDBP mutations (Ticozzi et al., CNS Neurol. Disord. Drug Targets. 2010, 9(3), 285-296.) connecting TDP-43 aggregation with clinical disease manifestation.

II. TDP-43 in neurodegeneration

TDP-43 aggregates have been identified in a growing list of neurodegenerative conditions (Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1 R46-R64), including but not limited to: Frontotemporal dementia (FTD, such as sporadic or familial with or without motor-neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutations, with TARDBP mutation, with valosin-containing protein (VCP) mutation, linked to chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), Argyrophilic grain disease, Pick's disease, semantic variant Primary Progressive Aphasia (svPPA), behavioural variant FTD (bvFTD), nonfluent variant Primary Progressive Aphasia (nfvPPA) and the like), Amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD, including sporadic and familial forms of AD), Down syndrome, Familial British dementia, Polyglutamine diseases (Huntington’s disease and spinocerebellar ataxia type 3 (SC A3; also known as Machado Joseph Disease)), Hippocampal sclerosis dementia and Myopathies (sporadic inclusion body myositis, Inclusion body myopathy caused by a mutation in valosin- containing protein associated with Paget’s disease of the bone and frontotemporal dementia (IBMPFD), Oculo-pharyngeal muscular dystrophy with rimmed vacuoles, Myofibrillar myopathies with mutations in the myotilin (MYOT) gene or mutations in the gene coding for desmin (DES)), Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s Disease (PD). The term LATE is intended to encompass several previously used designations related to TDP-43 proteinopathy that may be associated with cognitive impairment, including hippocampal sclerosis, hippocampal sclerosis of ageing, hippocampal sclerosis dementia, cerebral age-related TDP-43 with sclerosis (CARTS), and TDP-43 pathologies in the elderly (for reviews see Kuslansky et al., 2004; Lippa and Dickson, 2004; Nelson et al., 2013, 2016b; Dutra et al., 2015).

Aggregated TDP-43 from patient brains shows a number of abnormal modifications, including hyperphosphorylation, ubiquitination, acetylation and C-terminal fragments through proteolytic cleavage (Arai et al., Biochemical and Biophysical Research Communications 351 (2006) 602- 611; Neumann et al., Science 314, (2006), 130-133; Neumann et al., Acta Neuropathol. (2009) 117: 137-149; Hasegawa et al., (2008) Annals of Neurology Vol 64 No 1, 60-70; Cohen et al., Nat Commun. 6: 5845, 2015). Another characteristic feature of TDP-43 pathology is redistribution and accumulation of TDP-43 from nucleus to cytoplasm. The hallmark lesions of FTLD-TDP are neuronal and glial cytoplasmic inclusions (NCI and GCI, respectively) and dystrophic neurites (DN) that are immunoreactive for TDP-43, as well as ubiquitin and p62, but negative for other neurodegenerative disease-related proteins. Differences in inclusion morphology and tissue distribution thereof are associated with specific mutations and/or clinical representations. Four types of TDP-43 pathology are described so far by histological classification (Mackenzie and Neumann, J. Neurochem. (2016) 138 (Suppl. 1), 54-70). FTLD- TDP type A cases are characterised by abundant short dystrophic neuritis (DN) and compact oval or crescentic NCI, predominantly in layer II of the neocortex (Fig. 2f in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Cases with this pathology usually present clinically with either behavioural -variant frontotemporal dementia (bvFTD) or nonfluent/agrammatic variants of Primary Progressive Aphasia (nfvPPA) and are associated with progranulin (GRN) mutations. Type B cases show moderate numbers of compact or granular NCI in both superficial and deep cortical layers with relatively few DN and Nil (neuronal intranuclear inclusions; Fig. 2g in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Most cases with coappearance of FTD and ALS symptoms are found to have FTLD-TDP type B pathology. Type C cases have an abundance of long, tortuous neurites, predominantly in the superficial cortical laminae, with few or no NCI (Fig. 2j in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). This pathology is particularly found in cases presenting with semantic variant of Primary Progressive Aphasia (svPPA). FTLD-TDP type D displays with abundant lentiform neuronal intranuclear inclusions (Nil) and short DN in the neocortex with only rare NCI (Fig. 2k in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Type E is characterised by granulofilamentous neuronal inclusions (GFNIs) and very fine, dot-like neuropil aggregates affecting all neocortical layers in addition to curvilinear oligodendroglial inclusions in the white matter (Edward B. Lee et al., Acta Neuropathol. 2017 July; 134(1): 65-78.). This pattern of pathology is only found in cases with VCP in association with inclusion body myositis.

III. TDP-43 in FTD

Frontotemporal dementia (FTD) is a clinical term that covers a wide spectrum of disorders based on the degeneration of frontal and temporal lobes - a pathological feature termed frontotemporal lobar degeneration (FTLD). FTD is the second most abundant cause of early degenerative dementias in the age group below 65 years (Le Ber, Revue Neurologique 169 (2013) 811-819). FTD is presented by several syndromes including bvFTD which is characterised by changes in personality and behaviour; semantic dementia (SD) and progressive nonfluent aphasia (PNFA) characterised by changes in the language function; corticobasal syndrome (CBS), progressive supranuclear palsy syndrome and motor neuron disease (FTD-MND) characterised by movement dysfunction. Clinical diagnosis of these syndromes is complicated and final conclusion can only be achieved through post-mortem histopathological analysis to detect aggregated protein and define affected brain regions. In terms of pathological, proteinaceous inclusions, about 45% of cases show pathological accumulation of misfolded tau, 45% of cases have pathological TDP- 43 and a smaller subgroup has aggregates of FUS and other proteins.

IV. TDP-43 in ALS

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterised by the premature loss of upper and lower motor neurons. The progression of ALS is marked by fatal paralysis and respiratory failure with a disease course from diagnosis to death of 1 to 5 years. In most cases of sporadic ALS, the neuropathology is characterised by abnormal cytoplasmic accumulations of TDP-43 in neurons and glia of the primary motor cortex, brainstem motor nuclei, spinal cord and the associated white matter tracts. ALS with dementia involves accumulation of TDP-43 in extramotor neocortex and hippocampus. The role of phosphorylation of TDP-43 in ALS patients has been explored with the help of antibodies that specifically bind to phosphorylated TDP-43 in nuclear and cytoplasmic inclusions with amino acids S379, S403, S404, S409, S410 as the major sites of phosphorylation of TDP-43 (Hasegawa et al., Ann Neurol 2008; 64: 60-70; Neumann et al., Acta Neuropathol (2009) 117: 137-149).

V. TDP-43 in AD and other diseases

TDP-43 pathology occurs in up to 57% of brains of patients with Alzheimer’s disease (Josephs KA et al., Acta Neuropathol. 2014; 127(6): 811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3): 441-450; McAleese et al., Brain Pathol. 2017 Jul; 27(4): 472-479). TDP-43 aggregation is associated with patient’s age and correlates with cognitive decline, memory loss and medial temporal atrophy in AD. It appears that in AD, TDP-43 represents a secondary or independent pathology that shares overlapping brain distribution with amyloid beta and tau pathologies in the medial temporal lobe. Pathologic TDP-43 follows a stereotypical pattern of progressive deposition that has been described by the so-called TDP-43 in AD (TAD) staging scheme: TDP- 43 first deposits in the amygdala (stage I) followed by hippocampus, limbic, temporal, and finally the frontostriatum (stage V) (Josephs KA et al., Acta Neuropathol. 2014;127(6): 811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3): 441-450).

VI. TDP-43 spreading

Although ALS and FTD onset and first symptoms vary significantly between patients, the common feature of disease progression is spreading of pathology from an initial focal area to anatomically connected brain regions. The continuous worsening of symptoms might be explained by the progressive spread of TDP-43 pathology. TDP-43 pathology in an ALS patient’s brain appears to be spreading in a four-stage process and it is believed that propagation occurs transynaptically via corticofugal axonal projections using anterograde axonal transport (Brettschneider et al., Ann Neurol. 2013 July; 74(1): 20-38.). Recent experimental evidence supports the hypothesis of protein propagation in neuronal tissue for amyloid-beta, tau, alpha- synuclein and TDP-43 by a prion-like mechanism (Hasegawa et al., 2017), with starting points and the topographical spreading patterns being distinct for the four proteins (Brettschneider J et al., Nature Rev. Neuroscience, 2015, 109). The common, disease unifying mechanism is believed to be based on the cell-to-cell spreading of pathological protein aggregates. This mechanism consists of the release of aggregates from a diseased cell, uptake by a naive cell and seeding of the pathological protein conformation by a templated conformational change of endogenous proteins. Pathological TDP-43 able to induce aggregation of physiological (i.e., non-pathological TDP-43) is defined as seeding-competent TDP-43. Indeed, TDP-43 has been found to misfold and aggregate into seeds that are propagating agents with the ability to trigger de novo misfolding. This “prion-like” paradigm is suspected to be one of the key elements in the disease progression.

TDP-43 cell-to cell spreading has been studied at a molecular level in few in vitro models, where insoluble TDP-43 preparations from patient brain are able to induce intracellular aggregate formation in receptor cells (Nonaka et al., Cell Reports 4 (2013), 124-134; Feiler et al., 2015; Porta et al., Nat. Comm., 2018). Further, it has been observed that intracellular TDP-43 aggregates are released in association with exosome prior to spreading to the next cell (Nonaka et al., Cell Reports 4 (2013, 124-134)). Similarly, adenovirus-transduced TDP-43 expression led to cytoplasmic aggregates which were phosphorylated, ubiquitinated and more importantly acted as seeds initiating cell-to-cell spreading (Ishii et al., PLoS ONE 12(6): e0179375, 2017). The patient-derived pathological TDP-43 can lead to widespread deposition of endogenous TDP-43 following intracerebral inoculation into transgenic and wildtype mice (Porta et al., Nat. Comm., 2018). The presence of TDP-43 seeding-competent species in CSF or ALS patients was recently confirmed using a TDP-43 seed amplification assay (Audrain et al., 2023). A TDP-43 mAb targeting the C-terminal domain was able to neutralize these seeding species (Audrain et al., 2023).

VII. Prevention and Treatment of TDP-43 proteinopathies

TDP-43 aggregation and spreading of pathology are major hallmarks of ALS and FTD - fatal diseases for which currently no cure is available. Therefore, there is a need for new methods for the treatment and prevention of TDP-43 proteinopathies. Mutations in TDP-43 are associated with familial cases of ALS and FTD providing a causative link between TDP-43 misfolding and disease progression.

VIII. Diagnosis of TDP-43 proteinopathies

The diagnosis of FTD based on clinical manifestations is insufficient since the clinical representation can overlap with other diseases, in particular in the earlier stages.

A number of approaches aims at the development of biochemical biomarkers to distinguish different types of FTD pathology. Development of antibodies against different conformations of TDP-43 may permit generating more sensitive and specific diagnostic tools. In parallel to biochemical biomarkers, the development of imaging biomarkers may enable early and specific detection of the pathology in TDP-43 proteinopathies. The ability to image TDP-43 deposition in the brain may be a substantial achievement for diagnosis and drug development for TDP-43 proteinopathies. Using cell permeable antibody fragments could enable such detection. The earliest event in neurodegenerative diseases based on misfolding of different proteins is the acquisition of an alternative conformation that renders the protein toxic. Moreover, this misfolded conformation can self-propagate by recruiting the endogenous, normal protein into the misfolded conformation as mechanistic basis for the observed spread through affected tissue. To develop antibodies against different conformational states of a given protein, supramolecular antigenic constructs were designed in which the conformation of the presented antigen was controlled to raise conformational-specific antibodies against a given target in a specific conformational state (WO2012/055933 and W02012/020124). Conformational-specific antibodies offer many advantages since they can discriminate between the disease-associated and the functional, endogenous conformation of these proteins. This approach offers many advantages in the therapeutic application since such antibodies are less likely to be adsorbed by the normal conformation of proteins while targeting the misfolded, disease associated isoform thereof. Similar to this, for diagnostic application such antibodies only recognize the disease- associated, structural state of a protein, which is paramount for the development of the sensitive and specific diagnostics.

The use of a TDP-43-based biomarker in TDP-43 proteinopathies still remains to be established. Such evaluation has been hindered in part due to the lack of high affinity antibodies that can be employed in a suitable immunoassay for quantification of pathological TDP-43 in biofluids (Feneberg et al., Molecular Neurobiology, 2018).

Therefore, there is a clear need for biomarkers able to detect misfolded TDP-43, in particular in a human sample, for diagnosing different types of TDP-43 proteinopathies and/or for monitoring efficacy of therapeutic drugs used for treatment of diseases, disorders and abnormalities associated with TDP-43, in particular associated with TDP-43 aggregates or TDP-43 proteinopathy. The TDP-43 proteinopathies are defined as a set of neurodegenerative disorders characterised by pathological TDP-43.

IX. Prior art

Patent application W02013/061163 describes TDP-43 specific binding molecules including polypeptides such as antibodies as well as fragments, derivatives and variants thereof.

Patent application WO2019/134981 describes TDP-43 specific binding molecules including polypeptides such as antibodies as well as fragments, derivatives and variants thereof.

Patent application WO2020/234473 describes specific binding molecules including polypeptides such as antibodies as well as fragments, derivatives and variants thereof. SUMMARY

At present there are no approved therapies on the market to treat and/or prevent TDP-43 associated diseases. There is therefore a pressing need to identify new therapies that can treat and/or prevent these diseases. Accordingly, the invention relates to binding molecules, in particular antibodies or antigen-binding fragments thereof, and to vectors comprising nucleic acid encoding said binding molecules, which at least preferentially binds misfolded TDP-43 over physiologically functional TDP-43 and preferably specifically bind misfolded TDP-43 and do not bind physiologically functional TDP-43. Within the invention, misfolded TDP-43 includes misfolded monomeric and/or misfolded oligomeric and/or misfolded aggregated and/or post- translationally modified and/or misfolded truncated TDP-43. Post-translationally modified TDP- 43 comprises phosphorylated, ubiquitylated, acetylated, sumoylated, and/or methylated TDP-43. Physiologically functional TDP-43 includes soluble nuclear TDP-43.

It is demonstrated herein that the binding molecules of the invention present advantageous binding properties for use in therapy. The binding molecules of the invention, in particular antibodies or antigen-binding fragments thereof, such as scFv, at least preferentially binds misfolded TDP-43 over physiologically functional TDP-43 and preferably specifically bind misfolded TDP-43 and do not bind physiologically functional TDP-43. For example, the binding molecules of the invention may be used as monoclonal antibodies in passive immunotherapy or encoded as nucleic acids in a vector used for in situ delivery allowing intracellular expression, i.e. as an intrabody. Expressed intracellularly, the binding molecules of the invention specifically or preferentially bind misfolded TDP-43 over physiologically functional TDP-43 which prevents further aggregation and misfolding of TDP-43, increasing the clearance of misfolded TDP-43 and reducing or preventing interaction of misfolded TDP-43 with physiologically functional TDP-43. The binding molecules of the invention can address both gain- and loss-of-function pathologies associated with TDP-43, where gain of function refers to toxicity driven by aggregation and spreading of misfolded TDP-43. The binding molecules of the invention can prevent the loss of physiological function of TDP-43 by preventing further spreading of pathology and restoring TDP-43 homeostasis and nuclear localization essential for RNA processing. Accordingly, the invention provides a vector comprising a nucleic acid encoding a TDP-43 binding molecule for targeted delivery to the CNS, preferably as an intrabody (e.g. an scFv intrabody). Without wishing to be bound by theory, the specific or preferential binding of the binding molecules of the invention to misfolded TDP-43 may promote misfolded TDP-43 degradation and/or prevent or inhibit misfolded TDP-43 aggregation without affecting TDP-43 physiological functions. The invention also provides the vectors or binding molecules, in particular antibodies or antigen-binding fragments thereof, for the prevention, alleviation, treatment and/or diagnosis of diseases, disorders and abnormalities associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy. The invention also provides the binding molecules, in particular antibodies or antigen-binding fragments thereof, for detecting and/or identifying the specific type of pathology causing neurodegeneration. Envisaged are uses as diagnostic biomarkers enabling more efficient and precise subject selection for longitudinal monitoring in clinical studies, supporting the development of novel therapeutics for TDP-43 proteinopathies.

The invention also provides the vectors or TDP-43 binding molecules, in particular antibodies or antigen-binding fragments thereof, for use as a medicine (therapeutic agent).

Without wishing to be bound by any particular theory, the present invention was developed based on the assumption that modified conformation-specific antigenic peptides and peptide fragments derived from TDP-43 protein or the whole TDP-43 protein and the antibodies obtainable or obtained by using said peptides or fragments or the whole TDP-43 protein as antigen block TDP- 43 cell-to-cell propagation, and/or disaggregate TDP-43 aggregates and/or block TDP-43 seeding and/or neutralize seeding-competent TDP-43 and/or inhibit the aggregation of TDP-43 protein or fragments thereof and/or inhibit the intracellular aggregation of TDP-43 and/or potentiate intracellular and/or extracellular clearance of pathological TDP-43 and/or restore the nuclear levels and physiological function of TDP-43. The binding molecules of the invention, in particular polypeptides, more particularly antibodies or antigen-binding fragments thereof, bind to misfolded TDP-43, particularly to cytoplasmic misfolded TDP-43 and optionally extracellular misfolded TDP-43. In one embodiment, the binding molecules of the invention, in particular polypeptides, more particularly antibodies or antigen-binding fragments thereof, specifically bind to cytoplasmic misfolded TDP-43. In one embodiment, the TDP-43 binding molecules of the invention, in particular antibodies or antigen-binding fragments thereof, inhibit the aggregation of TDP-43. The aggregation of TDP-43 may be intracellular and/or extracellular aggregation. In one embodiment, the TDP-43 binding molecules of the invention, in particular antibodies or antigen-binding fragments thereof, inhibit the intracellular aggregation of TDP-43. In one embodiment, the TDP-43 binding molecules of the invention, in particular antibodies or antigen-binding fragments thereof, potentiate intracellular and/or extracellular clearance of pathological TDP-43. In one embodiment, the TDP-43 binding molecules of the invention, in particular antibodies or antigen-binding fragments thereof, restore the nuclear levels and physiological function of TDP-43 in cells, particularly neurons, preferably in patients with a disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy.

Misfolded or pathology-associated TDP-43 is composed of TDP-43 proteins that lose their normal folding (i.e. are misfolded) and localization. Misfolded TDP-43 can be found in preinclusions and in neuronal and glial cytoplasmic inclusions (NCI and GCI, respectively), neuronal intranuclear inclusions (Nil) and dystrophic neurites (DN) that are immunoreactive for TDP-43.

Physiologically functional TDP-43 is predominantly located in the nucleus and shuttles between the nucleus and the cytoplasm, being in a state able to exhibit its desired function in an in vivo cellular environment. Thus, physiologically functional TDP-43 does not extend to purified soluble TDP-43 expressed in a heterologous host cell (e.g. recombinantly produced in E.colf), which may be useful for testing certain properties of the binding molecules in an in vitro environment (see Examples 1, 2 and 10 herein).

The TDP-43 binding molecules of the invention, in particular the anti-TDP-43 antibodies or antigen-binding fragments thereof, exhibit one or more, up to all of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof; c. blocks TDP-43 cell-to-cell propagation; d. disaggregates TDP-43 aggregates; e. blocks TDP-43 seeding; f. neutralizes seeding-competent TDP-43; g. blocks TDP-43 spreading; h. potentiates intracellular and/or extracellular clearance of pathological TDP-43; i. restore nuclear levels and physiological function of TDP-43; and j. reduces phosphorylated TDP-43 level.

Independent of the combination of one or more, up to all of the above listed characteristics, the TDP-43 binding molecules, preferably anti-TDP-43 antibodies or antigen-binding fragments thereof, of the invention may inhibit/reduce the formation of TDP-43 pathology in patients with TDP-43 pathology. The TDP-43 binding molecules bind to an epitope within amino acids 202-211, 353-373 or 370- 414 of human TDP-43 (SEQ ID NO: 1). In some embodiments, the TDP-43 binding molecules bind to an epitope consisting of amino acids residues 202-209 or 207-211 of human TDP-43 (SEQ ID NO: 1).

According to the invention, there is provided a TDP-43 binding molecule, in particular a TDP- 43 antibody or an antigen-binding fragment thereof, comprising: a) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or b) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57, or c) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or e) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or f) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

The invention is further directed, inter alia, to (i) an immunoconjugate comprising the TDP-43 binding molecule, (ii) a labelled binding molecule comprising the TDP-43 binding molecule, (iii) a nucleic acid molecule encoding the TDP-43 binding molecule, (iv) a vector comprising a nucleic acid molecule encoding the TDP-43 binding molecule, (v) a pharmaceutical composition comprising the vector, TDP-43 binding molecule, immunoconjugate, or labelled binding molecule and a pharmaceutically acceptable carrier and/or excipients and/or diluents, (vi) a vector, TDP-43 binding molecule, immunoconjugate or pharmaceutical composition for human or veterinary therapeutic use, (vii) a vector, TDP-43 binding molecule, immunoconjugate or pharmaceutical composition for use in the prevention, alleviation, treatment of diseases, disorders and/or abnormalities associated with TDP-43 or TDP-43 proteinopathy, (viii) a TDP- 43 binding molecule, immunoconjugate, labelled antibody or pharmaceutical composition for diagnostic use (in particular for in vivo diagnosis, but also for in vitro testing), (ix) a vector, TDP- 43 binding molecule, immunoconjugate, labelled binding molecule or pharmaceutical composition for research use, in particular as an analytical tool or reference molecule, (x) a TDP- 43 binding molecule, immunoconjugate, labelled binding molecule or pharmaceutical composition for use as a diagnostic tool to monitor diseases, disorders and/or abnormalities associated with TDP-43 or TDP-43 proteinopathy, (xi) a nucleic acid encoding a TDP-43 binding molecule of the invention, (xii) an expression vector comprising a nucleic acid molecule encoding a TDP-43 binding molecule of the invention, (xiii) a host cell comprising the nucleic acid and/or the expression vector encoding a TDP-43 binding molecule of the invention, (xiv) a cell-free expression system containing the expression vector of the invention, (xiv) a method for producing a TDP-43 binding molecule of the invention, (xvi) a method of detecting and/or quantifying TDP-43 in a sample obtained from a subject using a TDP-43 binding molecule of the invention, and (xvii) a kit comprising TDP-43 binding molecules of the invention and/or nucleic acids, expression vectors, host cells and/or cell free expression systems for producing the same.

In the present invention, the binding molecules, in particular antibodies or antigen-binding fragments thereof, specifically recognize TDP-43. The binding molecules of the invention include polypeptides and/or antibodies and/or antigen-binding fragments thereof specific to/for the TDP-43 protein. “Specifically recognize TDP-43” means that the binding molecules of the invention specifically, generally, and collectively, bind to TDP-43, in particular some epitopes within TDP-43, in particular an epitope exposed/accessible in one or more pathological conformation(s) of TDP-43 protein and do not appreciably bind to proteins other than TDP-43.

The binding molecules of the invention, in particular polypeptides, more particularly antibodies or antigen-binding fragments thereof, that specifically recognize TDP-43, at least preferentially bind misfolded TDP-43 over physiologically functional TDP-43 and may specifically bind misfolded TDP-43 and do not bind physiologically functional TDP-43. Preferential binding means a greater level of binding to misfolded TDP-43 over physiologically functional TDP-43 under the same experimental conditions. For example, there may be strong binding to misfolded TDP-43 and weak binding to physiologically functional TDP-43. Specific binding means binding to misfolded TDP-43 with no measured binding to physiologically functional TDP-43 under the same experimental conditions. The experimental conditions are intended to reflect in vivo conditions and may be in situ conditions. The experimental conditions typically involve testing binding in the context of a diseased biological sample in which both misfolded TDP-43 and physiologically functional TDP-43 are present. For example, specific or preferential binding may be demonstrated in brain tissue from subjects suffering from FTD/ALS. These binding properties may be demonstrated for example using immunohistochemistry. The immunohistochemistry may involve use of a labelled (e.g. fluorescently labelled) secondary antibody for detection purposes. See Example 4 herein for experimental details of how specific and preferential binding may be tested.

Thus, TDP-43 binding molecules of the invention, in particular the antibodies or antigen-binding fragments thereof, may bind specifically or preferentially to aggregated TDP-43 as compared with non-aggregated TDP-43. More particularly, TDP-43 binding molecules of the invention, in particular the antibodies or antigen-binding fragments thereof, may bind specifically or preferentially to aggregated TDP-43 in the cytoplasm as compared with non-aggregated TDP- 43 in the nucleus.

In some embodiments, the invention encompasses TDP-43 binding molecules, particularly antibodies and antigen-binding fragments thereof of the invention as described herein, vectors comprising nucleic acid encoding the binding molecules of the invention for targeted delivery to the CNS (as described herein), and the use of these binding molecules and vectors to diagnose, prevent, alleviate and/or treat a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy including, but not limited to, frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE) and limbic- predominant age-related TDP-43 encephalopathy (LATE). The methods and compositions disclosed herein have applications in diagnosing, preventing, alleviating and/or treating a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP- 43 aggregates, or TDP-43 proteinopathy including but not limited to frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS). Preferably, the use of these binding molecules to diagnose, prevent, alleviate and/or treat a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy is directed to amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD) or Frontotemporal dementia (FTD). More preferably, the use is directed to amyotrophic lateral sclerosis (ALS). More preferably, the use is directed to Alzheimer’s disease (AD). More preferably, the use is directed to Frontotemporal dementia (FTD).

In another embodiment, a TDP-43 binding molecule, particularly a TDP-43 antibody or an antigen-binding fragment thereof of the invention as described herein is contacted with a sample to detect, diagnose and/or monitor a disease, disorder and/or abnormality associated with TDP- 43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy. In an embodiment, the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy is Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic-predominant age- related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD).

In an embodiment, Frontotemporal dementia (FTD) is selected from sporadic Frontotemporal dementia, familial Frontotemporal dementia with motor-neuron disease (MND), familial Frontotemporal dementia without motor-neuron disease (MND), Frontotemporal dementia with progranulin (GRN) mutation, Frontotemporal dementia with C9orf72 mutations, Frontotemporal dementia linked to chromosome 9p, Frontotemporal dementia with TARDBP mutation, Frontotemporal dementia with valosin-containing protein (VCP) mutation, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), Argyrophilic grain disease, semantic variant Primary Progressive Aphasia (svPPA), behavioural variant FTD (bvFTD) and nonfluent variant Primary Progressive Aphasia (nfvPPA).

In an embodiment, Amyotrophic lateral sclerosis (ALS) is selected from sporadic Amyotrophic lateral sclerosis, Amyotrophic lateral sclerosis with TARDBP mutation, Amyotrophic lateral sclerosis with C9orf72 mutation, Amyotrophic lateral sclerosis with CSL5 mutation, Amyotrophic lateral sclerosis with ALS2 mutation, Amyotrophic lateral sclerosis with ANXA11 mutation, Amyotrophic lateral sclerosis with ATXN2 mutation, Amyotrophic lateral sclerosis with ATXN3 mutation, Amyotrophic lateral sclerosis with C21orf2 mutation, Amyotrophic lateral sclerosis with CAV1 mutation, Amyotrophic lateral sclerosis with CCNF mutation, Amyotrophic lateral sclerosis with CHCHD10 mutation, Amyotrophic lateral sclerosis with CHMP2B mutation, Amyotrophic lateral sclerosis with CHRNA3 mutation, Amyotrophic lateral sclerosis with DAO mutation, Amyotrophic lateral sclerosis with DCTN1 mutation, Amyotrophic lateral sclerosis with DNAJC7 mutation, Amyotrophic lateral sclerosis with ELP3 mutation, Amyotrophic lateral sclerosis with ERBB4 mutation, Amyotrophic lateral sclerosis with EWSR1 mutation, Amyotrophic lateral sclerosis with FIG4 mutation, Amyotrophic lateral sclerosis with GLE1 mutation, Amyotrophic lateral sclerosis with GLT8D1 mutation, Amyotrophic lateral sclerosis with hnRNPAl mutation, Amyotrophic lateral sclerosis with hnRNPA2Bl mutation, Amyotrophic lateral sclerosis with KANK1 mutation, Amyotrophic lateral sclerosis with KIF5A mutation, Amyotrophic lateral sclerosis with LGALSL mutation, Amyotrophic lateral sclerosis with MATR3 mutation, Amyotrophic lateral sclerosis with MOBP mutation, Amyotrophic lateral sclerosis with NEFH mutation, Amyotrophic lateral sclerosis with NEK1 mutation, Amyotrophic lateral sclerosis with NIPA1 mutation, Amyotrophic lateral sclerosis with OPTN mutation, Amyotrophic lateral sclerosis with PARK9 mutation, Amyotrophic lateral sclerosis with PFN1 mutation, Amyotrophic lateral sclerosis with P0N1 mutation, Amyotrophic lateral sclerosis with P0N2 mutation, Amyotrophic lateral sclerosis with P0N3 mutation, Amyotrophic lateral sclerosis with PRPH mutation, Amyotrophic lateral sclerosis with SARM1 mutation, Amyotrophic lateral sclerosis with SCFD1 mutation, Amyotrophic lateral sclerosis with SETX mutation, Amyotrophic lateral sclerosis with SIGMAR1 mutation, Amyotrophic lateral sclerosis with SPG11 mutation, Amyotrophic lateral sclerosis with SPTLC1 mutation, Amyotrophic lateral sclerosis with SQSTM1 mutation, Amyotrophic lateral sclerosis with TAF15 mutation, Amyotrophic lateral sclerosis with TBK1 mutation, Amyotrophic lateral sclerosis with TIA1 mutation, Amyotrophic lateral sclerosis with TUBA4A mutation, Amyotrophic lateral sclerosis with UBQLN2 mutation, Amyotrophic lateral sclerosis with VAPB mutation, Amyotrophic lateral sclerosis with VCP mutation, Amyotrophic lateral sclerosis with WDR7 mutation and Amyotrophic lateral sclerosis with ANG mutation. In a preferred embodiment, Amyotrophic lateral sclerosis (ALS) is selected from sporadic Amyotrophic lateral sclerosis, Amyotrophic lateral sclerosis with TARDBP mutation and Amyotrophic lateral sclerosis with C9orf72 mutation.

In an embodiment, Alzheimer’s disease (AD) is selected from sporadic forms of Alzheimer’s disease and familial forms of Alzheimer’s disease.

In an embodiment, the Polyglutamine disease is selected from Huntington’s disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado Joseph Disease).

In an embodiment, the Myopathy is selected from sporadic inclusion body myositis and Inclusion body myopathy caused by mutation(s) in valosin-containing protein (VCP) and associated with Paget’s disease of the bone (PDB) and Frontotemporal dementia (FTD) abbreviated IBMPFD, Oculo-pharyngeal muscular dystrophy with rimmed vacuoles, Myofibrillar myopathy with mutation(s) in the myotilin (MYOT) gene and Myofibrillar myopathy with mutation(s) in the desmin (DES) gene. In a preferred embodiment, the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), or limbic-predominant age-related TDP-43 encephalopathy (LATE).

In a preferred embodiment, the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Alzheimer’s disease (AD). In another preferred embodiment, the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD). In another preferred embodiment, the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is amyotrophic lateral sclerosis (ALS).

In one embodiment, the invention encompasses TDP-43 binding molecules, particularly antibodies or antigen-binding fragments thereof of the invention as described herein and the use of these binding molecules, particularly of these antibodies, to detect the presence of TDP-43 in a sample. Accordingly, TDP-43 binding molecules of the invention, such as anti-TDP-43 antibodies as described herein, can be used, inter alia, to screen a clinical sample, in particular human blood or human blood derivative, cerebrospinal fluid (CSF), interstitial fluid (ISF) and/or urine for the presence of TDP-43 in samples, for example, by using an Enzyme-Linked Immunosorbent Assay (ELISA)-based or surface adapted assay. Tissue samples may be used in some circumstances, such as brain tissue samples. The methods and compositions of the invention also have applications in diagnosing presymptomatic disease and/or in monitoring disease progression and/or therapeutic efficacy. According to some embodiments, an antibody specific for TDP-43 (e.g., a full-length antibody or a TDP-43 antigen-binding fragment or derivative of an antibody) is contacted with a sample (e.g., blood, urine, cerebrospinal fluid (CSF), interstitial fluid (ISF) or brain tissue) to detect, diagnose and/or monitor Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic- predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD).

In an embodiment, the Myopathy is selected from sporadic inclusion body myositis and Inclusion body myopathy caused by mutation(s) in valosin-containing protein (VCP) and associated with Paget’s disease of the bone (PDB) and Frontotemporal dementia (FTD) abbreviated IBMPFD, Oculo-pharyngeal muscular dystrophy with rimmed vacuoles, Myofibrillar myopathy with mutation(s) in the myotilin (MYOT) gene and Myofibrillar myopathy with mutation(s) in the desmin (DES) gene.

In a preferred embodiment, the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), or limbic-predominant age-related TDP-43 encephalopathy (LATE).

The TDP-43 binding molecules of the invention may be used to quantify TDP-43 in suitable samples, in particular clinical samples such as blood or blood derivative, brain tissue, CSF, ISF or urine, with relatively high TDP-43 levels, as compared to a suitable control, indicating disease and/or more advanced disease. Many suitable immunoassay formats are known. Thus, the methods (such as ELISA, MSD (Meso Scale Discovery), HTRF (Homogeneous Time Resolved Fluorescence), SIMOA® (Single Molecule Array) and AlphaLISA®) may be performed for diagnostic purposes with high levels of TDP-43 indicating disease. Alternatively, the methods may be performed for monitoring purposes. Increased levels over time may indicate progression of the disease. Decreased levels over time may indicate regression of the disease. The methods may also be used to monitor therapy, in particular to monitor the efficacy of a particular treatment. Successful therapy may be measured with reference to stable or decreasing levels of TDP-43 following treatment. It is demonstrated in Example 12 of WO2020/234473 that TDP- 43 levels were higher in CSF samples from TDP-43 proteinopathy patients than in control samples taken from healthy subjects (healthy control). The control samples may or may not be run in parallel with the test samples. In some embodiments control levels are determined from a series of control samples taken from healthy subjects under similar or the same experimental conditions and used as a comparator for levels determined in the test sample. Methods of quantifying TDP-43 in suitable samples using binding molecules of the invention may also be used to select a therapy (for further treatment of the subject). Thus, personalized treatment methods are envisaged. A sample is taken before and after treatment. If treatment using the therapy results in stable or, preferably, decreasing levels of TDP-43 following treatment, the therapy may be selected for that subject. If the therapy does not result in stable or, preferably, decreasing levels of TDP-43 following treatment, the therapy is not selected for the subject. The therapy may be any suitable candidate therapeutic agent for treatment of TDP-43 proteinopathies. In preferred embodiments, the therapy comprises a TDP-43 binding molecule of the invention, typically in the form of a pharmaceutical composition as described herein.

The TDP-43 binding molecules of the invention may also be used for disease classification into particular types or subtypes. Thus, there is provided a method for classifying a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or for classifying a TDP-43 proteinopathy comprising: a. performing the methods of the invention in which levels of TDP-43 are quantified, as compared to suitable controls, b. optionally identifying mutations in a sample from the subject including but not limited to progranulin (GRN) mutation, C9orf72 mutations, TARDBP mutation, angiogenin (ANG) mutation, mutation in the valosin-containing protein (VCP), mutation in the myotilin (MYOT) gene or mutations in the gene coding for desmin (DES), and c. classifying the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy.

Similarly, there is provided a method for classifying a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or for classifying a TDP-43 proteinopathy comprising: performing the methods of the invention in which levels of TDP-43 are quantified in a sample obtained from a subject with a disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, wherein the levels are compared with control samples taken from subjects with different types or subtypes of disease, disorder and/or abnormality (i.e. a representative set of control levels are determined for the types or subtypes of interest) associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy; and classifying the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy based on the comparison. Thus, the classification is based on determining the closest match between the test sample and one or more of the control samples. These methods may further comprise identifying mutations in the sample including but not limited to progranulin (GRN) mutation, C9orf72 mutations, TARDBP mutation, angiogenin (ANG) mutation, mutation in the valosin- containing protein (VCP), mutation in the myotilin (MYOT) gene or mutations in the gene coding for desmin (DES), wherein the identified mutations are also used to classify the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy. For the avoidance of doubt, the identification of mutations in a sample may be performed by any suitable method, for example, based on nucleic acid sequencing of nucleic acid molecules within the sample. The sample may be separate and distinct from the sample in which TDP-43 levels are determined, but is from the same subject.

In other embodiments, the invention provides methods for preventing, alleviating and/or treating a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy. According to one embodiment, the methods of the invention comprise administering an effective concentration of a TDP-43 binding molecule of the invention (particularly an antibody of the invention such as a full-length antibody or a TDP-43 antigen-binding fragment or derivative of an antibody), an immunoconjugate, a vector or a pharmaceutical composition as described herein to a subject. In another embodiment, the invention provides a method for preventing, alleviating and/or treating a TDP-43 proteinopathy. According to some embodiments, a TDP-43 binding molecule (particularly an antibody or antigen-binding fragment thereof), an immunoconjugate, a vector or a pharmaceutical composition as described herein is administered to treat, alleviate and/or prevent frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS). In another embodiment, a TDP-43 binding molecule (particularly an antibody or antigen-binding fragment thereof), an immunoconjugate, a vector or a pharmaceutical composition as described herein specific for TDP-43 is administered to prevent, alleviate and/or treat a neurodegenerative disease selected from frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD, including sporadic and familial forms of AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), limbic-predominant age-related TDP-43 encephalopathy (LATE).

In another embodiment, a TDP-43 binding molecule (particularly an antibody or antigen-binding fragment thereof), an immunoconjugate, a vector or a pharmaceutical composition as described herein is administered to prevent, alleviate and/or treat a disease selected from Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic- predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) and Parkinson’s disease (PD).

In an embodiment, Frontotemporal dementia (FTD) is selected from sporadic Frontotemporal dementia, familial Frontotemporal dementia with motor-neuron disease (MND), familial Frontotemporal dementia without motor-neuron disease (MND), Frontotemporal dementia with progranulin (GRN) mutation, Frontotemporal dementia with C9orf72 mutations, Frontotemporal dementia linked to chromosome 9p, Frontotemporal dementia with TARDBP mutation, Frontotemporal dementia with valosin-containing protein (VCP) mutation, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), Argyrophilic grain disease, semantic variant Primary Progressive Aphasia (svPPA), behavioural variant FTD (bvFTD) and nonfluent variant Primary Progressive Aphasia (nfvPPA). In an embodiment, Amyotrophic lateral sclerosis (ALS) is selected from sporadic Amyotrophic lateral sclerosis, Amyotrophic lateral sclerosis with TARDBP mutation, Amyotrophic lateral sclerosis with C9orf72 mutation, Amyotrophic lateral sclerosis with CSL5 mutation, Amyotrophic lateral sclerosis with ALS2 mutation, Amyotrophic lateral sclerosis with ANXA11 mutation, Amyotrophic lateral sclerosis with ATXN2 mutation, Amyotrophic lateral sclerosis with ATXN3 mutation, Amyotrophic lateral sclerosis with C21orf2 mutation, Amyotrophic lateral sclerosis with CAV1 mutation, Amyotrophic lateral sclerosis with CCNF mutation, Amyotrophic lateral sclerosis with CHCHD10 mutation, Amyotrophic lateral sclerosis with CHMP2B mutation, Amyotrophic lateral sclerosis with CHRNA3 mutation, Amyotrophic lateral sclerosis with DAO mutation, Amyotrophic lateral sclerosis with DCTN1 mutation, Amyotrophic lateral sclerosis with DNAJC7 mutation, Amyotrophic lateral sclerosis with ELP3 mutation, Amyotrophic lateral sclerosis with ERBB4 mutation, Amyotrophic lateral sclerosis with EWSR1 mutation, Amyotrophic lateral sclerosis with FIG4 mutation, Amyotrophic lateral sclerosis with GLE1 mutation, Amyotrophic lateral sclerosis with GLT8D1 mutation, Amyotrophic lateral sclerosis with hnRNPAl mutation, Amyotrophic lateral sclerosis with hnRNPA2Bl mutation, Amyotrophic lateral sclerosis with KANK1 mutation, Amyotrophic lateral sclerosis with KIF5A mutation, Amyotrophic lateral sclerosis with LGALSL mutation, Amyotrophic lateral sclerosis with MATR3 mutation, Amyotrophic lateral sclerosis with MOBP mutation, Amyotrophic lateral sclerosis with NEFH mutation, Amyotrophic lateral sclerosis with NEK1 mutation, Amyotrophic lateral sclerosis with NIPA1 mutation, Amyotrophic lateral sclerosis with OPTN mutation, Amyotrophic lateral sclerosis with PARK9 mutation, Amyotrophic lateral sclerosis with PFN1 mutation, Amyotrophic lateral sclerosis with P0N1 mutation, Amyotrophic lateral sclerosis with P0N2 mutation, Amyotrophic lateral sclerosis with P0N3 mutation, Amyotrophic lateral sclerosis with PRPH mutation, Amyotrophic lateral sclerosis with SARM1 mutation, Amyotrophic lateral sclerosis with SCFD1 mutation, Amyotrophic lateral sclerosis with SETX mutation, Amyotrophic lateral sclerosis with SIGMAR1 mutation, Amyotrophic lateral sclerosis with SPG11 mutation, Amyotrophic lateral sclerosis with SPTLC1 mutation, Amyotrophic lateral sclerosis with SQSTM1 mutation, Amyotrophic lateral sclerosis with TAF15 mutation, Amyotrophic lateral sclerosis with TBK1 mutation, Amyotrophic lateral sclerosis with TIA1 mutation, Amyotrophic lateral sclerosis with TUBA4A mutation, Amyotrophic lateral sclerosis with UBQLN2 mutation, Amyotrophic lateral sclerosis with VAPB mutation, Amyotrophic lateral sclerosis with VCP mutation, Amyotrophic lateral sclerosis with WDR7 mutation and Amyotrophic lateral sclerosis with ANG mutation. In a preferred embodiment, Amyotrophic lateral sclerosis (ALS) is selected from sporadic Amyotrophic lateral sclerosis, Amyotrophic lateral sclerosis with TARDBP mutation and Amyotrophic lateral sclerosis with C9orf72 mutation.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

X. DEFINITIONS

An “antigen-binding molecule,” as used herein, is any molecule that can specifically or selectively bind to an antigen, in particular TDP-43. A binding molecule may include or be an antibody or a fragment thereof. An anti-TDP-43 binding molecule is a molecule that binds to the TDP-43 protein, such as an anti-TDP-43 antibody or fragment thereof, at a specific recognition site, epitope. That is, antigen-binding molecules of the invention bind to an epitope within the amino acid sequence of SEQ ID NO: 1. The antigen-binding molecules, in particular antibodies or antigen-binding fragments thereof, provided herein recognize full-length TDP-43. Other anti- TDP-43 binding molecules may also include multivalent molecules, multi-specific molecules (e.g., diabodies), fusion molecules, aptamers, avimers, or other naturally occurring or recombinantly created molecules. Illustrative antigen-binding molecules useful in the present invention include antibody-like molecules. An antibody-like molecule is a molecule that can exhibit functions by binding to a target molecule (See, e.g., Current Opinion in Biotechnology 2006, 17:653-658; Current Opinion in Biotechnology 2007, 18: 1-10; Current Opinion in Structural Biology 1997, 7:463-469; Protein Science 2006, 15: 14-27), and includes, for example, DARPins (WO 2002/020565), Affibody (WO 1995/001937), Avimer (WO 2004/044011; WO 2005/040229), Adnectin (WO 2002/032925) and fynomers (WO 2013/135588).

The terms "anti-TDP-43 antibody" and "an antibody that binds to TDP-43" or simply “antibody” as used herein refer to an antibody that is capable of binding TDP-43 with sufficient affinity such that the antibody is considered for further assessment as a potential diagnostic and/or therapeutic agent in targeting TDP-43. In general, the term "antibody" is used herein in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multi-specific antibodies (e.g., bispecific or biparatopic antibodies), fully-human antibodies and antibody fragments so long as they exhibit the desired antigen-binding activity. Antibodies within the present invention may also be chimeric antibodies, recombinant antibodies, antigen-binding fragments of recombinant antibodies, humanized antibodies or antibodies displayed upon the surface of a phage or displayed upon the surface of a chimeric antigen receptor (CAR) T cell.

An "antigen-binding fragment" of an antibody, or “functional fragment thereof’ refers to a molecule other than an intact, or full-length, antibody that comprises a portion of an intact, or full-length, antibody and that binds (fully or partially) the antigen to which the intact, or full- length, antibody binds. Examples of antibody fragments include but are not limited to Fv, Fab, Fab', Fab' -SH, F(ab')2, diabodies, linear antibodies, single-chain antibody molecules (e.g. scFv) and multi-specific antibodies formed from antibody fragments. Antigen-binding fragments may also be referred to as “functional fragments” as they retain the binding function of the original antibody from which they are derived.

An "antibody that binds to an epitope" within a defined region of a protein is an antibody that requires the presence of one or more of the amino acids within that region for binding to the protein.

In certain embodiments, an "antibody that binds to an epitope" within a defined region of a protein is identified by mutation analysis, in which amino acids of the protein are mutated, and binding of the antibody to the resulting altered protein (e.g., an altered protein comprising the epitope) is determined to be at least 20% of the binding to unaltered protein. In some embodiments, an "antibody that binds to an epitope" within a defined region of a protein is identified by mutation analysis, in which amino acids of the protein are mutated, and binding of the antibody to the resulting altered protein (e.g., an altered protein comprising the epitope) is determined to be at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the binding to unaltered protein. In certain embodiments, binding of the antibody is determined by fluorescence-activated cell sorting (FACS), western blot (WB) or by a suitable binding assay such as ELISA.

The term “binding to” as used in the context of the present invention defines a binding (interaction) of at least two “antigen-interaction-sites” with each other. The term “antigen- interaction-site” defines, in accordance with the present invention, a motif of a polypeptide, i.e., a part of the antibody or antigen-binding fragment of the present invention, which shows the capacity of specific interaction with a specific antigen or a specific group of antigens of TDP- 43. Said binding/interaction is also understood to define a “specific recognition”. The term “specific recognition” means in accordance with this invention that the antibody is capable of specifically interacting with and/or binding to at least two amino acids of TDP-43 as defined herein, in particular interacting with/binding to at least two amino acids within amino acid residues 202-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1), even more particularly interacting with/binding to at least two amino acids within amino acid residues 202- 209, 207-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

The term “specific interaction” as used in accordance with the present invention means that the antibody or antigen-binding fragment thereof of the invention interacts with a particular (poly)peptide target and does not or essentially does not cross-react with other (poly)peptides of similar, but different, structure. Accordingly, the antibody or antigen-binding fragment thereof of the invention specifically interacts with structures of TDP-43 formed by particular amino acid sequences within amino acids residues 202-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1), more particularly interacts with structures of TDP-43 formed by particular amino acid sequences within amino acids residues 202-209, 207-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1). Whether or not such epitopes are available to bind depends on the state of TDP-43. Thus, misfolded TDP-43 may present the epitopes in a manner such that the binding molecules of the invention can bind, but physiologically functional TDP-43 may not.

Cross-reactivity of antigen-binding molecules, in particular a panel of antibodies or antigenbinding fragments thereof under investigation may be tested, for example, by assessing binding of said panel of antibodies or antigen-binding fragments thereof under conventional conditions (see, e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1988) and Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, (1999)) to the (poly)peptide of interest as well as to a number of more or less (structurally and/or functionally) closely related (poly)peptides. Only those constructs (i.e. antibodies, antigen-binding fragments thereof and the like) that bind to the certain structure of TDP-43 as defined herein, e.g., a specific epitope or (poly)peptide/protein of TDP-43 as defined herein but do not or essentially do not bind to any of the other epitope or (poly)peptides of the same TDP- 43, are considered specific for the epitope or (poly)peptide/protein of interest and selected for further studies in accordance with the method provided herein. These methods may comprise, inter alia, binding studies, blocking and competition studies with structurally and/or functionally closely related molecules. These binding studies also comprise FACS analysis, surface plasmon resonance (SPR, e.g. with BIACORE™), analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy or by radiolabelled ligand binding assays.

Accordingly, specificity can be determined experimentally by methods known in the art and methods as described herein. Such methods comprise, but are not limited to Western Blots, ELISA-, RIA-, ECL-, IRMA-tests and peptide scans.

The term “monoclonal antibody” as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Monoclonal antibodies are advantageous in that they may be synthesized by a hybridoma culture, essentially uncontaminated by other immunoglobulins. The modified "monoclonal" indicates the character of the antibody as being amongst a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. As mentioned above, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method described by Kohler, Nature 256 (1975), 495.

The term “polyclonal antibody” as used herein, refers to an antibody which was produced among or in the presence of one or more other, non-identical antibodies. In general, polyclonal antibodies are produced from a B-lymphocyte in the presence of several other B-lymphocytes which produced non-identical antibodies. Usually, polyclonal antibodies are obtained directly from an immunized animal.

The term “fully-human antibody” as used herein refers to an antibody which comprises human immunoglobulin protein sequences only. A fully human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “murine antibody” refers to an antibody which comprises mouse/murine immunoglobulin protein sequences only. Alternatively, a “fully-human antibody” may contain rat carbohydrate chains if produced in a rat, in a rat cell, in a hybridoma derived from a rat cell. Similarly, the term “rat antibody” refers to an antibody that comprises rat immunoglobulin sequences only. Fully-human antibodies may also be produced, for example, by phage display which is a widely used screening technology which enables production and screening of fully human antibodies. Phage antibodies can also be used in context of this invention. Phage display methods are described, for example, in US 5,403,484, US 5,969,108 and US 5,885,793. Another technology which enables development of fully-human antibodies involves a modification of mouse hybridoma technology. Mice are made transgenic to contain the human immunoglobulin locus in exchange for their own mouse genes (see, for example, US 5,877,397). The term “chimeric antibodies”, refers to an antibody which comprises a variable region of the present invention fused or chimerized with an antibody region (e.g., constant region) from another, human or non-human species (e.g., mouse, horse, rabbit, dog, cow, chicken).

The term antibody also relates to recombinant human antibodies, heterologous antibodies and heterohybrid antibodies. The term "recombinant (human) antibody" includes all human sequence antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes; antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant, combinatorial human antibody library, or antibodies prepared, expressed, created or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions (if present) derived from human germline immunoglobulin sequences. Such antibodies can, however, be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.

A "heterologous antibody" is defined in relation to the transgenic non-human organism producing such an antibody. This term refers to an antibody having an amino acid sequence or an encoding nucleic acid sequence corresponding to that found in an organism not consisting of the transgenic non-human animal, and generally from a species other than that of the transgenic non-human animal.

The term "heterohybrid antibody" refers to an antibody having light and heavy chains of different organismal origins. For example, an antibody having a human heavy chain associated with a murine light chain is a heterohybrid antibody. Examples of heterohybrid antibodies include chimeric and humanized antibodies.

The term antibody also relates to humanized antibodies. "Humanized" forms of non-human (e.g. murine or rabbit) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Often, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibody may comprise residues, which are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and optimize antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see: Jones et al., Nature 321 (1986), 522-525; Reichmann Nature 332 (1998), 323-327 and Presta Curr Op Struct Biol 2 (1992), 593-596.

A popular method for humanization of antibodies involves CDR grafting, where a functional antigen-binding site from a non-human ‘donor’ antibody is grafted onto a human ‘acceptor’ antibody. CDR grafting methods are known in the art and described, for example, in US 5,225,539, US 5,693,761 and US 6,407,213. Another related method is the production of humanized antibodies from transgenic animals that are genetically engineered to contain one or more humanized immunoglobulin loci which are capable of undergoing gene rearrangement and gene conversion (see, for example, US 7,129,084).

Accordingly, in the context of the present invention, the term “antibody” relates to full immunoglobulin molecules as well as to parts of such immunoglobulin molecules (i.e., “antigenbinding fragment thereof’). Furthermore, the term relates, as discussed above, to modified and/or altered antibody molecules. The term also relates to recombinantly or synthetically generated/ synthesized antibodies. The term also relates to intact antibodies as well as to antibody fragments thereof, like, separated light and heavy chains, Fab, Fv, Fab’, Fab’-SH, F(ab’)2. The term antibody also comprises but is not limited to fully-human antibodies, chimeric antibodies, humanized antibodies, CDR-grafted antibodies and antibody constructs, like single chain Fvs (scFv) or antibody-fusion proteins. “Single-chain Fv” or “scFv” antibody fragments have, in the context of the invention, the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Generally, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding. The VH and VL domains may be in any order to form a scFv (the VL may be in N-terminal and the VH in C- terminal of the amino acid sequence or vice-versa). Techniques described for the production of single chain antibodies are described, e.g., in Pliickthun in The Pharmacology of Monoclonal Antibodies, Rosenburg and Moore eds. Springer-Verlag, N.Y. (1994), 269-315.

A "Fab fragment" contains one light chain (VL+CL), a portion of one heavy chain that contains the heavy variable domain (VH), the constant CHI domain, and a short part of the hinge region between the CHI and CH2 domains, such that an interchain disulfide bond can be formed between the two chains.

An "Fc" region contains two heavy chain fragments comprising the CH2 and CH3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domains.

A "F(ab')2 fragment" contains two light chains (VL+ CL), a portion of two heavy chains (VH+CH1) and the hinge region between the CHI and CH2 domains, such that interchain disulfide bonds can be formed between 1) the light chain and heavy chain and 2) the two heavy chains. A F(ab')2 fragment is thus composed of two Fab fragments held together by disulfide bonds.

The "Fv region" comprises the variable regions from both the heavy and light chains but lacks the constant regions.

The term “intrabody” (or “intracellular antibody”) relates to an antibody, antibody fragment, or any non-antibody scaffold-based formats, not secreted by the cell in which it is expressed and able to reach subcellular compartments, including but not limited to, the nucleus, the endoplasmic reticulum, the cytoplasm, the Golgi apparatus and the mitochondria. The intrabodies of the invention may incorporate a suitable signal peptide and/or retention sequence to enable targeting to specific subcellular compartments (see Marschall et al., 2015 for examples of signal peptides suitable for use with an intrabody). Thus, the vectors of the invention may encode such signal peptides and/or retention sequences. An intrabody binds, and typically specifically binds, to one or more intracellular target antigens and may be used to inhibit, promote or modulate various intracellular processes or pathways linked to the targeted antigen(s). Non-limiting examples of intracellular processes or pathways an intrabody may inhibit, promote or modulate include transcription, translation, cell survival, cell proliferation, cell metabolism, gene replacement, oxidative stress, inflammation, aging or protein aggregation. An intrabody may be used to inhibit, promote or modulate pathological cellular processes or pathways. Non-limiting examples comprise binding to misfolded and/or aggregated proteins to prevent, block or inhibit aggregation or further aggregation and cytotoxicity, or promote degradation of the target. In particular, an intrabody may be used as a therapeutic agent for the treatment of conditions, diseases or disorders in a subject in need thereof. Intrabodies can take many forms, including but not limited to scFv, scFab, bifunctional and multispecific scFv, any non-antibody scaffold-based formats, single-domain antibody-based formats (including VH only, VL only, VHH and VNARS), any of these formats fused together and/or fused to an immunoglobulin Fc domain, and/or fused to a non-immunoglobulin fusion protein.

Antibodies, antibody constructs, antibody fragments, antibody derivatives (all being Ig-derived) to be employed in accordance with the invention or their corresponding immunoglobulin chain(s) can be further modified using conventional techniques known in the art, for example, by using amino acid deletion(s), insertion(s), substitution(s), addition(s), and/or recombination(s) and/or any other modification(s) known in the art either alone or in combination. Methods for introducing such modifications in the DNA sequence underlying the amino acid sequence of an immunoglobulin chain are well known to the person skilled in the art; see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, 2^nd edition (1989) and 3^rd edition (2001). The term “Ig-derived domain” particularly relates to (poly)peptide constructs comprising at least one CDR. Fragments or derivatives of the recited Ig-derived domains define (poly)peptides which are parts of the above antibody molecules and/or which are modified by chemical/biochemical or molecular biological methods. Corresponding methods are known in the art and described inter alia in laboratory manuals (see Sambrook et al., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, 2nd edition (1989) and 3rd edition (2001); Gerhardt et al., Methods for General and Molecular Bacteriology ASM Press (1994); Lefkovits, Immunology Methods Manual: The Comprehensive Sourcebook of Techniques; Academic Press (1997); Golemis, Protein-Protein Interactions: A Molecular Cloning Manual Cold Spring Harbor Laboratory Press (2002)). The term “CDR” as employed herein relates to “complementary determining region”, which is well known in the art. The CDRs are parts of immunoglobulins that determine the specificity of said molecules and make contact with a specific ligand. The CDRs are the most variable part of the molecule and contribute to the diversity of these molecules. There are three CDR regions CDR1, CDR2 and CDR3 in each V domain. CDR-H depicts a CDR region of a variable heavy chain and CDR-L relates to a CDR region of a variable light chain. VH means the variable heavy chain and VL means the variable light chain. The CDR regions of an Ig-derived region may be determined as described in Kabat “Sequences of Proteins of Immunological Interest”, 5th edit. NIH Publication no. 91-3242 U.S. Department of Health and Human Services (1991). CDR sequences provided herein are defined according to Kabat. However, it will be understood by the skilled person that the invention is intended to encompass binding molecules in which the CDR sequences are defined according to any useful identification/numbering scheme. For example, Chothia (Canonical structures for the hypervariable regions of immunoglobulins. Chothia C, Lesk AM. J Mol Biol. 1987 Aug 20; 196(4):901-17), IMGT (IMGT, the international ImMunoGeneTics database. Giudicelli V, Chaume D, Bodmer J, Muller W, Busin C, Marsh S, Bontrop R, Marc L, Malik A, Lefranc MP. Nucleic Acids Res. 1997 Jan 1; 25(l):206-l 1 and Unique database numbering system for immunogenetic analysis. Lefranc MP. Immunol Today. 1997 Nov; 18(11):509), MacCallum (MacCallum RM, Martin AC, Thornton JM, J Mol Biol. 1996 Oct 11; 262(5):732-45) and Martin (Abhinandan KR, Martin ACR. Analysis and improvements to Kabat and structurally correct numbering of antibody variable domains. Mol Immunol. (2008) 45:3832-9. 10.1016/j.molimm.2008.05.022) numbering schemes may be adopted in order to define the CDRs.

Accordingly, in the context of the present invention, the antibody molecule described herein above may be selected from the group consisting of a full antibody (immunoglobulin, like an IgGl, an IgG2, an IgAl, an IgGA2, an IgG3, an IgG4, an IgA, an IgM, an IgD or an IgE), F(ab)- , Fab’-SH-, Fv-, Fab’-, F(ab’)2- fragment, a chimeric antibody, a CDR-grafted antibody, a fully human antibody, a bivalent antibody-construct, an antibody-fusion protein, a synthetic antibody, bivalent single chain antibody, a trivalent single chain antibody and a multivalent single chain antibody.

“Humanization approaches” are well known in the art and in particular described for antibody molecules, e.g. Ig-derived molecules. The term “humanized” refers to humanized forms of non- human (e.g., murine) antibodies or fragments thereof (such as Fv, Fab, Fab’, F(ab’), scFvs, or other antigen-binding partial sequences of antibodies) which contain some portion of the sequence derived from non-human antibody. Humanized antibodies include human immunoglobulins in which residues from a complementary determining region (CDR) of the human immunoglobulin are replaced by residues from a CDR of a non-human species such as mouse, rat or rabbit having the desired binding specificity, affinity and capacity. In general, the humanized antibody will comprise substantially all of at least one, and generally two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin; see, inter alia, Jones et al., Nature 321 (1986), 522-525, Presta, Curr. Op. Struct. Biol. 2 (1992), 593-596. Methods for humanizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acids introduced into it from a source which is non-human still retain the original binding activity of the antibody. Methods for humanization of antibodies/antibody molecules are further detailed in Jones et al., Nature 321 (1986), 522-525; Reichmann et al., Nature 332 (1988), 323-327; and Verhoeyen et al., Science 239 (1988), 1534-1536. Specific examples of humanized antibodies, e.g. antibodies directed against EpCAM, are known in the art (see e.g. LoBuglio, Proceedings of the American Society of Clinical Oncology Abstract (1997), 1562 and Khor, Proceedings of the American Society of Clinical Oncology Abstract (1997), 847).

Accordingly, in the context of this invention, antibody molecules or antigen-binding fragments thereof are provided, which can be humanized and can successfully be employed in immunoconjugates, vectors or pharmaceutical compositions.

The invention provides humanized binding molecules, in particular humanized antibodies or antigen-binding fragments thereof. In specific embodiments, the humanized binding molecules, in particular humanized antibodies or antigen-binding fragments thereof of the invention comprise a human heavy chain variable domain subfamily 3 framework sequence. More specifically, the humanized binding molecules, in particular humanized antibodies or antigenbinding fragments thereof of the invention may comprise an IGHV3-23 (IMGT accession number M99660 and Z 12347) VH framework sequence. In specific embodiments, the humanized binding molecules, in particular humanized antibodies or antigen-binding fragments thereof of the invention comprise a human light chain variable domain kappa subfamily 2 framework sequence, a light chain variable domain kappa subfamily 1 framework, or a light chain variable domain kappa subfamily 4 framework. More specifically, the humanized binding molecules, in particular humanized antibodies or antigen-binding fragments thereof of the invention may comprise an IGKV2-28 (IMGT accession numbers X63397 and X93632), IGKV4-1 (IMGT accession numbers Z00023 and X93640) or IGKV1-39 (IMGT accession number X59315 and X93627) VL framework sequence, preferably a IGKV4- 1 VL framework sequence.

As would be understood by one skilled in the art, the heavy and light chains are both typically humanized and thus the embodiments above are preferably combined in the same humanized binding molecule. Thus, the humanized binding molecules, in particular humanized antibodies or antigen-binding fragments thereof of the invention may comprise a human heavy chain variable domain subfamily 1 framework sequence and a human light chain variable domain kappa subfamily 2 framework sequence, a light chain variable domain kappa subfamily 1 framework, or a light chain variable domain kappa subfamily 4 framework. More specifically, the humanized binding molecules, in particular humanized antibodies or antigen-binding fragments thereof of the invention may comprise an IGHV3-23 (IMGT accession number M99660) VH framework sequence and an IGKV2-28 (IMGT accession numbers X63397 and X93632), IGKV4-1 (IMGT accession numbers Z00023 and X93640) or IGKV1-39 (IMGT accession number X59315 and X93627) VL framework sequence, preferably a IGKV4-1 VL framework sequence. In one embodiment, the humanized binding molecules of the invention, in particular humanized antibodies or antigen-binding fragments thereof of the invention comprise IGHV3-23 and IGKV4-1 framework sequences.

In one embodiment, the IGHV3-23 framework comprises S49A and/or K94R mutations. In one embodiment, the IGKV2-28, IGKV4-1 or IGKV1-39 framework comprises Y36L and/or L46R mutations. In one embodiment, the IGKV4-1 human framework comprises one or more conservative substitutions, for example D55E. Mutations are according to Kabat numbering.

The chosen framework sequences may be mutated at specific positions. In some embodiments such mutations are made to positively influence CDR loop conformation and/or variable domain packing between VH and VL domains. It may be understood by a person skilled in the art that the epitopes may be comprised in the TDP-43 protein, but may also be comprised in a degradation product thereof or may be a chemically synthesized peptide. The amino acid positions are only indicated to demonstrate the position of the corresponding amino acid sequence in the sequence of the TDP-43 protein. The invention encompasses all peptides comprising the epitope. The peptide may be a part of a polypeptide of more than 100 amino acids in length or may be a small peptide of less than 100, preferably less than 50, more preferably less than 25 amino acids, even more preferably less than 16 amino acids. The amino acids of such peptide may be natural amino acids or nonnatural amino acids (e.g., beta-amino acids, gamma-amino acids, D-amino acids) or a combination thereof. Further, the present invention may encompass the respective retro-inverso peptides of the epitopes. The peptide may be unbound or bound. It may be bound, e.g., to a small molecule (e.g., a drug or a fluorophore), to a high-molecular weight polymer (e.g., polyethylene glycol (PEG), polyethylene imine (PEI), hydroxypropylmethacrylate (HPMA), etc.) or to a protein, a fatty acid, a sugar moiety or may be inserted in a membrane.

In order to test whether an antibody in question and the antibody of the present invention recognize the same epitope, the following competition study may be carried out: Vero cells infected with 3 MOI (multiplicity of infection) are incubated after 20 h with varying concentrations of the antibody in question as the competitor for 1 hour. In a second incubation step, the antibody of the present invention is applied in a constant concentration of 100 nM and its binding is flow-cytometrically detected using a fluorescence-labelled antibody directed against the constant domains of the antibody of the invention. Binding that conducts antiproportional (inversely proportional) to the concentration of the antibody in question is indicative that both antibodies recognize the same epitope. However, many other assays are known in the art which may be used.

The present invention also relates to the production of specific antibodies against native polypeptides and recombinant polypeptides of TDP-43. This production is based, for example, on the immunization of animals, like mice. However, also other animals for the production of antibody/antisera are envisaged within the present invention. For example, monoclonal and polyclonal antibodies can be produced by rabbit, mice, goats, donkeys and the like. The polynucleotide encoding a correspondingly chosen polypeptide of TDP-43 can be subcloned into an appropriate vector, wherein the recombinant polypeptide is to be expressed in an organism capable of expression, for example in bacteria. Thus, the expressed recombinant protein can be intra-peritoneally injected into mice and the resulting specific antibody can be, for example, obtained from the mice serum being provided by intra-cardiac blood puncture. The present invention also envisages the production of specific antibodies against native polypeptides and recombinant polypeptides by using a DNA/RNA vaccine strategy as exemplified in the appended examples. DNA vaccine strategies are well-known in the art and encompass liposome-mediated delivery, by gene gun or jet injection and intramuscular or intradermal injection. Thus, antibodies directed against a polypeptide or a protein or an epitope of TDP-43, in particular the epitope of the antibodies provided herein, can be obtained by directly immunizing the animal by directly injecting intramuscularly the vector expressing the desired polypeptide or a protein or an epitope of TDP-43, in particular the epitope of the antibodies of the invention, which lies within amino acid residues 202-211, 353-373 or 370-414 of SEQ ID NO: 1, more particularly which lies within amino acid residues 202-209, 207-211, 353-373 or 370-414 of SEQ ID NO: 1. The amount of obtained specific antibody can be quantified using an ELISA, which is also described herein below. Further methods for the production of antibodies are well known in the art, see, e.g. Harlow and Lane, "Antibodies, A Laboratory Manual", CSH Press, Cold Spring Harbor, 1988.

Thus, under designated assay conditions, the specified antibodies and the corresponding epitope of TDP-43 bind to one another and do not bind in a significant amount to other components present in a sample. Specific binding to a target analyte under such conditions may require a binding moiety that is selected for its specificity for a particular target analyte. A variety of immunoassay formats may be used to select antibodies specifically reactive with a particular antigen. For example, solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies specifically immunoreactive with an analyte. See Shepherd and Dean (2000), Monoclonal Antibodies: A Practical Approach, Oxford University Press and/ or Howard and Bethell, for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity. Typically, a specific or selective reaction will be at least twice background signal to noise and more typically more than 10 to 100 times greater than background. The person skilled in the art is in a position to provide for and generate specific binding molecules directed against the novel polypeptides. For specific binding-assays it can be readily employed to avoid undesired cross-reactivity, for example polyclonal antibodies can easily be purified and selected by known methods (see Shepherd and Dean, loc. cit.). The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 5, a, y, and p, respectively.

In certain embodiments, amino acid sequence variants of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody. Amino acid sequence variants of an antibody may be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions from, and/or insertions into and/or substitutions of residues within the amino acid sequences of the antibody. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen-binding.

In certain embodiments, antibody variants having one or more amino acid substitutions are provided. Sites of interest for substitutional mutagenesis include the CDRs and FRs. Conservative substitutions are shown in Table 1 under the heading of "preferred substitutions." More substantial changes are provided in Table 1 under the heading of "exemplary substitutions," and as further described below in reference to amino acid side chain classes. Amino acid substitutions may be introduced into an antibody of interest and the products screened for a desired activity, e.g., retained/improved antigen binding, decreased immunogenicity, or improved Antibody-Dependent Cellular Cytotoxicity (ADCC) or Complement-Dependent Cytotoxicity (CDC).

Table 1

Amino acids may be grouped according to common side-chain properties:

(1) hydrophobic: Norleucine, Met, Ala, Vai, Leu, He;

(2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gin; (3) acidic: Asp, Glu;

(4) basic: His, Lys, Arg;

(5) residues that influence chain orientation: Gly, Pro;

(6) aromatic: Trp, Tyr, Phe.

Non-conservative substitutions will entail exchanging a member of one of these classes for another class.

One type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g. a humanized or human antibody). Generally, the resulting variant(s) selected for further study will have modifications (e.g., improvements) in certain biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parent antibody and/or will have substantially retained certain biological properties of the parent antibody. An exemplary substitutional variant is an affinity matured antibody, which may be conveniently generated, e.g., using phage di splay -based affinity maturation techniques such as those described herein. Briefly, one or more CDR residues are mutated, and the variant antibodies displayed on phage and screened for a particular biological activity (e.g. binding affinity).

Alterations (e.g., substitutions) may be made in CDRs, e.g., to improve antibody affinity. Such alterations may be made in CDR "hotspots," i.e., residues encoded by codons that undergo mutation at high frequency during the somatic maturation process (see, e.g, Chowdhury, Methods Mol. Biol. 207: 179-196 (2008)), and/or SDRs (a-CDRs), with the resulting variant VH or VL being tested for binding affinity. Affinity maturation by constructing and reselecting from secondary libraries has been described, e.g., in Hoogenboom et al., in Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001).) In some embodiments of affinity maturation, diversity is introduced into the variable genes chosen for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity. Another method to introduce diversity involves CDR- directed approaches, in which several CDR residues (e.g., 4-6 residues at a time) are randomized. CDR residues involved in antigen binding may be specifically identified, e.g., using alanine scanning mutagenesis or modelling. CDR-H3 and CDR-L3 in particular are often targeted.

In certain embodiments, substitutions, insertions, or deletions may occur within one or more CDRs so long as such alterations do not substantially reduce the ability of the antibody to bind antigen. For example, conservative alterations (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in CDRs. Such alterations may be outside of CDR "hotspots" or SDRs. In certain embodiments of the variant VH and VL sequences provided above, each CDR either is unaltered, or contains no more than one, two or three amino acid substitutions.

A useful method for identification of residues or regions of an antibody that may be targeted for mutagenesis is called “alanine scanning mutagenesis” as described by Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, a residue or group of target residues (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) are identified and replaced by a neutral or negatively charged amino acid (e.g., alanine or polyalanine) to determine whether the interaction of the antibody with antigen is affected. Further substitutions may be introduced at the amino acid locations demonstrating functional sensitivity to the initial substitutions. Alternatively, or additionally, a crystal structure of an antigen-antibody complex is used to identify contact points between the antibody and antigen. Such contact residues and neighbouring residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they contain the desired properties.

Amino acid sequence insertions include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue. Other insertional variants of the antibody molecule include the fusion to the N- or C-terminus of the antibody to an enzyme (e.g. for ADEPT) or a polypeptide which increases the serum half-life of the antibody.

In certain embodiments, an antibody provided herein is altered to increase or decrease the extent to which the antibody is glycosylated. Addition or deletion of glycosylation sites to an antibody may be conveniently accomplished by altering the amino acid sequence such that one or more glycosylation sites is created or removed.

Where the antibody comprises an Fc region, the carbohydrate attached thereto may be altered. Native antibodies produced by mammalian cells typically comprise a branched, biantennary oligosaccharide that is generally attached by an N-linkage to Asn297 of the CH2 domain of the Fc region. See, e.g., Wright et al., TIBTECH 15:26-32 (1997). The oligosaccharide may include various carbohydrates, e.g., mannose, N-acetyl glucosamine (GlcNAc), galactose, and sialic acid, as well as a fucose attached to a GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some embodiments, modifications of the oligosaccharide in an antibody of the invention may be made in order to create antibody variants with certain improved properties.

In one embodiment, antibody variants are provided having a carbohydrate structure that lacks fucose attached (directly or indirectly) to an Fc region. For example, the amount of fucose in such antibody may be from 1% to 80%, from 1% to 65%, from 5% to 65% or from 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn297, relative to the sum of all glycostructures attached to Asn297 (e. g. complex, hybrid and high mannose structures) as measured by MALDI-TOF mass spectrometry, as described in W02008/077546, for example. Asn297 refers to the asparagine residue located at about position 297 in the Fc region (Eu numbering of Fc region residues; see Edelman, G.M. et al., Proc. Natl. Acad. USA, 63, 78-85 (1969)); however, Asn297 may also be located about ± 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations in antibodies. Such fucosylation variants may have improved ADCC function. See, e.g., US Patent Publication Nos. US 2003/0157108 (Presta, L.); US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "defucosylated" or "fucose deficient" antibody variants include: US2003/0157108; W02000/61739; WO2001/29246; US2003/0115614; US2002/0164328; US2004/0093621; US2004/0132140; US2004/0110704; US 2004/0110282; US2004/0109865; W02003/085119; W02003/084570; W02005/035586; W02005/035778; W02005/053742; W02002/031140; Okazaki et al., J. Mol. Biol. 336: 1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng. 87: 614 (2004). Examples of cell lines capable of producing defucosylated antibodies include Lecl3 CHO cells deficient in protein fucosylation (Ripka et al., Arch. Biochem. Biophys. 249:533-545 (1986); US Pat Appl No US 2003/0157108 Al, Presta, L; and W02004/056312 Al, Adams et al., especially at Example 11), and knockout cell lines, such as alpha- 1,6-fucosyltransf erase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al., Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and W02003/085 107).

Antibody variants are further provided with bisected oligosaccharides, e.g., in which a biantennary oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and/or improved ADCC function. Examples of such antibody variants are described, e.g., in WO 2003/011878 (Jean-Mairet et al.); US Patent No. 6,602,684 (Umana et al.); and US 2005/0123546 (Umana et al.). Antibody variants with at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, e.g., in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO 1999/22764 (Raju, S.).

In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgGl, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.

In certain embodiments, the antibody provided herein binds to pathological TDP-43 and forms an immune complex that may be cleared by antibody-dependent cellular phagocytosis (ADCP), which as a result potentiates TDP-43 clearance. ADCP is mediated by the interaction of antibody Fc fragment with Fc receptors, such as Fc gamma receptors, expressed at the surface of innate immune cells, such as microglia or dendritic cells. Fc mediated functions may be modulated to achieve the desired effect by modifying the Fc portion of antibodies.

In certain embodiments, the invention contemplates an antibody variant that possesses some but not all effector functions, which make it a desirable candidate for applications in which the halflife of the antibody in vivo is important yet certain effector functions (such as complement activation and ADCC) are unnecessary or deleterious. In vitro and/or in vivo cytotoxicity assays can be conducted to confirm the reduction/depletion of CDC and/or ADCC activities. For example, Fc receptor (FcR) binding assays can be conducted to ensure that the antibody lacks FcyR binding (hence likely lacking ADCC activity), but retains FcRn binding ability. The primary cells for mediating ADCC, NK cells, express FcyRIII only, whereas monocytes and microglia express FcyRI, FcyRII and FcyRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest is described in U.S. Patent No. 5,500,362 (see, e.g. Hellstrom, I. et al., Proc. Nat ’I Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat’l Acad. Sci. USA 82: 1499- 1502 (1985); 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)).

Alternatively, non-radioactive assays methods may be employed (see, for example, ACTI™ non radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, CA; and CytoTox 96® non-radioactive cytotoxicity assay (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.

Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al., Proc. Nat'lAcad. sci. USA 95:652- 656 (1998).

Clq binding assays may also be carried out to confirm that the antibody is unable to bind Clq and hence lacks CDC activity. See, e.g., Clq and C3c binding ELISA in WO 2006/029879 and WO 2005/100402. To assess complement activation, a CDC assay may be performed (see, for example, Gazzano- Santoro etal., J. Immunol. Methods 202: 163 (1996); Cragg, M.S. et al., Blood 101 : 1045-1052 (2003); and Cragg, M.S. and M.J. Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance/half-life determinations can also be performed using methods known in the art (see, e.g., Petkova, S.B. et al., Int’l. Immunol. 18(12): 1759-1769 (2006)).

Antibodies with reduced effector function include those with substitution of one or more of Fc region residues 234, 235, 238, 265, 269, 270, 297, 327 and 329 (U.S. Patent No. 6,737,056). Certain antibody variants with improved or diminished binding to FcRs are described. (See, e.g., U.S. Patent No. 6,737,056; WO 2004/056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called "DANA" Fc mutant with substitution of residues 265 and 297 to alanine (US Patent No. 7,332,581) or the so-called “DANG” Fc mutant with substitution of residues 265 to alanine and 297 to glycine. Alternatively, antibodies with reduced effector function include those with substitution of one or more of Fc region residues 234, 235 and 329, so-called “PG-LALA” Fc mutant with substitution of residues 234 and 235 to alanine and 329 to glycine (Lo, M. et al., Journal of Biochemistry, 292, 3900-3908). Other known mutations at position 234, 235 and 321, the so-called TM mutant containing mutations L234F/L235E/P331S in the CH2 domain, can be used (Oganesyan et al. Acta Cryst. D64, 700-704. (2008)). Antibodies from the human IgG4 isotype include mutations S228P/L235E to stabilize the hinge and to reduce FgR binding (Schlothauer et al, PEDS, 29 (10):457-466). Numbering of the constant domain is according to the EU numbering system.

Other Fc variants include those with substitutions at one or more of Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434, e.g., substitution of Fc region residue 434 (US Patent No. 7,371,826). See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821.

In certain embodiments, the Fc region is mutated to increase its affinity to FcRn at pH 6.0 and consequently extend the antibody half-life. Antibodies with enhanced affinity to FcRn include those with substitution of one or more of Fc region residues 252, 253, 254, 256, 428, 434, including the so called YTE mutation with substitution M252Y/S254T/T256E (DalF Acqua et al, J Immunol. 169:5171-5180 (2002)) or LS mutation M428L/N434S (Zalevsky et al, Nat Biotechnol. 28(2): 157-159 (2010)).

In certain embodiments, it may be desirable to create cysteine engineered antibodies, e.g., "thioMAbs," in which one or more residues of an antibody are substituted with cysteine residues. In particular embodiments, the substituted residues occur at accessible sites of the antibody. By substituting those residues with cysteine, reactive thiol groups are thereby positioned at accessible sites of the antibody and may be used to conjugate the antibody to other moieties, such as drug moieties or linker-drug moieties, to create an immunoconjugate, as described further herein. In certain embodiments, any one or more of the following residues may be substituted with cysteine: V205 (Kabat numbering) of the light chain; Al 18 (EU numbering) of the heavy chain; and S400 (EU numbering) of the heavy chain Fc region. Cysteine engineered antibodies may be generated as described, e.g., in U.S. Patent No. 7,521,541.

In certain embodiments, an antibody provided herein may be further modified to contain additional nonproteinaceous moieties that are known in the art and readily available. The moieties suitable for derivatization of the antibody include but are not limited to water soluble polymers. Non-limiting examples of water soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1, 3 -di oxolane, poly-1, 3, 6-trioxane, ethylene/maleic anhydride copolymer, polyaminoacids (either homopolymers or random copolymers), and dextran or poly(n vinyl pyrrolidone)polyethylene glycol, propropylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer may be of any molecular weight and may be branched or unbranched. The number of polymers attached to the antibody may vary, and if more than one polymer is attached, they can be the same or different molecules. In general, the number and/or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used in a therapy under defined conditions, etc.

In another embodiment, conjugates of an antibody and nonproteinaceous moiety that may be selectively heated by exposure to radiation are provided. In one embodiment, the nonproteinaceous moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation may be of any wavelength, and includes, but is not limited to, wavelengths that do not harm ordinary cells, but which heat the nonproteinaceous moiety to a temperature at which cells proximal to the antibody-nonproteinaceous moiety are killed.

Antibodies may be produced using recombinant methods and compositions, e.g., as described in U.S. Patent No. 4,816,567.

In one embodiment, a(n isolated) nucleic acid encoding a TDP-43 antibody described herein is provided. Such nucleic acid may encode an amino acid sequence comprising the VL and/or an amino acid sequence comprising the VH of the antibody (e.g., the Light and/or Heavy Chains of the antibody). In a further embodiment, one or more vectors (e.g., recombinant expression vectors) comprising such nucleic acid are provided. In a further embodiment, a host cell comprising such nucleic acid is provided. In one such embodiment, a host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antibody, or (2) a first vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antibody and a second vector comprising a nucleic acid that encodes an amino acid sequence comprising the VH of the antibody. In one embodiment, the host cell is eukaryotic, e.g. a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., YO, NSO, Sp20). In one embodiment, a method of making an anti-misfolded TDP-43 antibody is provided, wherein the method comprises culturing a host cell comprising a nucleic acid encoding the antibody, as provided above, under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

For recombinant production of a TDP-43 antibody of the invention, nucleic acid encoding an antibody, e.g, as described above, is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell or a cell-free expression system. Such nucleic acid may be readily isolated and sequenced using conventional procedures (e.g, by using oligonucleotide probes that are capable of binding specifically to genes encoding the Heavy and Light Chains of the antibody).

In one embodiment, a method of making a TDP-43 binding molecule of the invention, in particular an antibody or antigen-binding fragment thereof, is provided, wherein the method comprises culturing a host cell or cell-free expression system comprising a nucleic acid encoding the TDP-43 binding molecule, as provided above, under conditions suitable for expression of the TDP-43 binding molecule, and isolating the TDP-43 binding molecule.

Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells described herein. For example, antibodies may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed. For expression of antibody fragments and polypeptides in bacteria, see, e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vai. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, describing expression of antibody fragments in /■/ coli.) After expression, the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.

In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors, including fungi and yeast strains whose glycosylation pathways have been "humanized," resulting in the production of an antibody with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006). Suitable host cells for the expression of glycosylated antibody are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

Plant cell cultures can also be utilized as hosts. See, e.g., US Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIES™ technology for producing antibodies in transgenic plants).

Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are macaque kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Viral. 36:59 (1977)); baby hamster kidney cells (BHK); mouse Sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod. 23:243- 251 (1980)); macaque kidney cells (CV 1); African green macaque kidney cells (VERO-76); human cervical carcinoma cells (HeLa); canine kidney cells (MDCK); buffalo rat liver cells (BRL 3 A); human lung cells (WI38); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N. Y Aead. Sei. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR CHO cells (Urlaub et al., Proc. Natl. Acad. cii. USA 77:4216 (1980)); and myeloma cell lines such as YO, NSO and Sp2/0. For a review of certain mammalian host cell lines suitable for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, VaL 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp. 255- 268 (2003).

Several art-known approaches exist for delivering molecules across the blood brain barrier (BBB) such as alteration of the administration route, disruption of the BBB and alteration of its permeability, nanoparticle delivery, Trojan horse approaches, receptor-mediated transport, and cell and gene therapy.

Alteration of the administration route can be achieved by direct injection into the brain (see, e.g., Papanastassiou et al., Gene Therapy 9: 398-406(2002)), implanting a delivery device in the brain (see, e.g., Gillet al., Nature Med. 9: 589-595 (2003); and Gliadel Wafers™, Guildford Pharmaceutical), and intranasal administration to bypass the BBB (Mittal et al, Drug Deliv.21(2):75-86. (2014))

Methods of barrier disruption include, but are not limited to, ultrasound (see, e.g., U.S. Patent Publication No.2002/0038086), osmotic pressure (e.g., by administration of hypertonic mannitol (Neuwelt, E. A., Implication of the Blood-Brain Barrier and its Manipulation, Vols 1 & 2, Plenum Press, N.Y.(1989))), permeabilization by, e.g., bradykinin or permeabilizer A- 7 (see, e.g., U.S. Patent Nos.5, 112,596, 5,268,164, 5,506,206, and 5,686,416).

Methods of altering the BBB permeability include, but are not limited to, using glucocorticoid blockers to increase permeability of the blood-brain barrier (see, e.g., U.S. Patent Application Publication Nos. 2002/0065259, 2003/0162695, and 2005/0124533); activating potassium channels (see, e.g., U.S. Patent Application Publication No. 2005/0089473), and inhibiting ABC drug transporters (see, e.g., U.S. Patent Application Publication No. 2003/0073713).

Trojan horse delivery methods of delivering the antibody or antibody fragment thereof across the blood-brain barrier include, but are not limited to, cationizing the antibodies (see, e.g., U.S. Patent No. 5,004,697), and the use of cell -penetration peptides such as Tat peptides to gain entry into the CNS. (see, e.g. Dietz et al., J. Neurochem. 104:757-765 (2008)).

Nanoparticle delivery methods of delivering the antibody or antigen-binding fragment thereof across the blood brain barrier include, but are not limited to, encapsulating the antibody or antigen-binding fragment thereof in liposomes, or extracellular vesicles such as exosomes, that are coupled to without limitation antibody or antigen-binding fragments or alternatively peptides that bind to receptors on the vascular endothelium of the blood-brain barrier (see, e.g., U.S. Patent Application Publication No. 20020025313), and coating the antibody or antigen-binding fragment thereof in low-density lipoprotein particles (see, e.g., U.S. Patent Application Publication No. 20040204354) or apolipoprotein E (see, e.g., U.S. Patent Application Publication No. 20040131692).

Antibodies of the invention can be further modified to enhance blood brain barrier penetration. The antibody or antigen-binding fragment thereof of the invention can be fused to a polypeptide binding to a blood-brain barrier receptor. BBB receptors include, but are not limited to, transferrin receptor, insulin receptor or low-density lipoprotein receptor. The polypeptide can be a peptide, a receptor ligand, a single domain antibody (VHH), a scFv or a Fab fragment.

Antibodies of the invention can be delivered as a corresponding nucleic acid molecule encoding said antibody or antigen-binding fragment. Such nucleic acid molecule is comprised in a vector, for example a viral vector such as an AAV vector. The vectors of the invention may encode signal peptides and/or retention sequences to enable targeting to specific subcellular compartments (see Marschall et al., 2015 for examples of signal peptides suitable for use with an intrabody).

The term “vector” refers to any particle used to transport (by transduction or transfection) a polynucleotide(s) into a host cell. This definition includes both non-viral and viral vectors.

The term “viral vector” includes wild-type viruses and engineered (e.g. modified) viruses. Examples of viral vectors include, but are not limited to, adeno associated virus (AAV), adenovirus, retrovirus, rhinovirus, lentivirus, hepatitis, HSV and any virus-like particle.

As used herein, an “AAV vector” refers to an adeno-associated virus suitably used to transport a polynucleotide(s) into a host cell. The AAV can be of any suitable serotypes, examples of which include, but are not limited to, AAV serotype 1 (AAV1), AAV serotype 2 (AAV2), AAV serotype 3 (AAV3), AAV serotype 4 (AAV4), AAV serotype 5 (AAV5), AAV serotype 6 (AAV6), AAV serotype 7 (AAV7), AAV serotype 8 (AAV8), AAV serotype 9 (AAV9), AAV serotype 10 (AAV10), AAV serotype 11 (AAV11), AAV serotype 12 (AAV12), AAVrhlO , or any other wild-type serotypes or engineered AAVs.

An AAV vector as described herein may specifically target cells of the central nervous system (CNS) or the blood-brain barrier (BBB). Non-limiting examples of promoters suitable with such approach include cytomegalovirus (CMV) promoter, EFl A (Human Eukaryotic translation elongation factor 1 alpha 1), CAG (CMV early enhancer fused to modified chicken P-actin promoter), CB A (CMV early enhancer fused to chicken P-actin promoter and SV40 intron), CBh (CMV early enhancer fused to modified chicken P-actin promoter), SV40 (Simian virus 40 enhancer/early promoter), GfaABCID (modified GFAP promoter), CMVe-GFAP (CMV enhancer fused to GFAP short promoter), GFAP (Human glial fibrillary acidic protein promoter), ATP1 A2 (Na, K ATPase a2), CLDN5 (Claudin 5), ADRB2 (Adrenoceptor beta 2), TNFRSF6B (TNF receptor superfamily member 6b), PDYN (prodynorphin), GH1 (Human growth hormone), OP ALES! (Opalin), SYN1 (Synapsin 1), CMVe-SYNl (CMV enhancer fused to SYN1 promoter) CAMK2A (Calcium/Calmodulin Dependent Protein Kinase II alpha), NEFH (neurofilament heavy polypeptide), NEUR0D6 (neuronal differentiation factor 6), 0LIG2 (oligodendrocyte transcription factor 2) or CMVe-0LIG2 (CMV enhancer fused to 0LIG2 promoter). However, in other instances the AAV vector does not specifically target the CNS or the BBB. For example, many wild-type viral vectors target any tissue or cell-type and thus include the CNS or the BBB. In such cases, CNS-specific or BBB-specific promoters can be used to drive the expression of the polynucleotide encoding an antibody or antibody fragment in the cells of the CNS or the BBB in a preferential or predominant manner as compared to other tissues. A suitable promoter may be the human SYN1 promoter (Glover et al., 2002).

A vector as described herein, in particular a viral vector such as an AAV vector, can be administered to the subject by any conventional route, including injection or by gradual infusion over time. The administration may be via parenteral administration. The administration may, for example, be by infusion or by intravenous, intraperitoneal, intranasal, intravitreous, subretinal, subcutaneous, intramuscular, intrathecal, intracistemal, intraparenchymal, intrastriatal or intracerebroventricular route. As further examples, suitable forms for parenteral injection (including, subcutaneous, intramuscular, intravascular or infusion) include a sterile solution, suspension or emulsion. The identification of suitable dosages is well within the routine capabilities of a person of average skill in the art. For example, the suitable dosage for a given subject will be determined by taking into consideration various factors known to modify the action of the vector for the uses according to the invention. For example, severity and type of disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, body weight, sex, diet, time and route of administration, other medications and other relevant clinical factors. The dosages and schedules may be varied according to the particular condition, disorder or symptom the overall condition of the subject. It may also be the case that there is no single accepted dose for the treatment of a given disease, but that a range of doses is considered suitable. Effective dosages may be determined by either in vitro or in vivo methods.

Cell therapy methods of delivering the antibody of the invention or antibody fragment or antibody derivatives across the blood brain barrier include, but are not limited to, the use of the homing capacity of Endothelial Progenitor Cells (EPCs) transfected ex vivo with vectors and the secretion and delivery of antibodies or antibody fragments to the brain by these cells, to overcome the powerful filtering activity of the BBB (see, e.g., Heller and al., J Cell Mol Med. 00: 1-7 (2020)), or the use of polymeric cell implant devices loaded with genetically engineered cells, to secrete antibody or antibody fragments (see, e.g. Marroquin Belaunzaran et al. PLoS ONE 6(4): el8268 (2011)).

Pharmaceutically acceptable carriers, diluents, adjuvants and excipients are well known in the pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, 15th or 18th Ed. (Alfonso R. Gennaro, ed.; Mack Publishing Company, Easton, PA, 1990); Remington: the Science and Practice of Pharmacy 19th Ed. (Lippincott, Williams & Wilkins, 1995); Handbook of Pharmaceutical Excipients, 3rd Ed. (Arthur H. Kibbe, ed.; Amer. Pharmaceutical Assoc, 1999); Pharmaceutical Codex: Principles and Practice of Pharmaceutics 12th Ed. (Walter Lund ed.; Pharmaceutical Press, London, 1994); The United States Pharmacopeia: The National Formulary (United States Pharmacopeial Convention); Fiedler’s “Lexikon der Hilfstoffe” 5th Ed., Edition Cantor Verlag Aulendorf 2002; “The Handbook of Pharmaceutical Excipients”, 4th Ed., American Pharmaceuticals Association, 2003; and Goodman and Gilman's: the Pharmacological Basis of Therapeutics (Louis S. Goodman and Lee E. Limbird, eds.; McGraw Hill, 1992), the disclosures of which are hereby incorporated by reference.

The carriers, diluents, adjuvants and pharmaceutical excipients can be selected with regard to the intended route of administration and standard pharmaceutical practice. These compounds must be acceptable in the sense of being not deleterious to the recipient thereof. See Remington's Pharmaceutical Sciences, 15th or 18th Ed. (Alfonso R. Gennaro, ed.; Mack Publishing Company, Easton, PA, 1990); Remington: the Science and Practice of Pharmacy 19th Ed. (Lippincott, Williams & Wilkins, 1995); Handbook of Pharmaceutical Excipients, 3rd Ed. (Arthur H. Kibbe, ed.; Amer. Pharmaceutical Assoc, 1999); Pharmaceutical Codex: Principles and Practice of Pharmaceutics 12th Ed. (Walter Lund ed.; Pharmaceutical Press, London, 1994); The United States Pharmacopeia: The National Formulary (United States Pharmacopeial Convention); Fiedler’s “Lexikon der Hilfstoffe” 5th Ed., Edition Cantor Verlag Aulendorf 2002; “The Handbook of Pharmaceutical Excipients”, 4th Ed., American Pharmaceuticals Association, 2003; and Goodman and Gilman's: the Pharmacological Basis of Therapeutics (Louis S. Goodman and Lee E. Limbird, eds.; McGraw Hill, 1992), the disclosures of which are hereby incorporated by reference. The "effective amount" of the compound which is to be administered to a subject is the dosage which according to sound medical judgement is suitable for treating, preventing or alleviating the disease, disorder or abnormality. The specific dose level and frequency of dosage can depend, e.g., upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, mode and time of administration. The "effective amount" of the compound which is to be administered to a subject is the dosage which according to sound medical judgement is suitable for treating, preventing or alleviating the disorder, disease, disorder or abnormality. The specific dose level and frequency of dosage can depend, e.g., upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, mode and time of administration, the rate of excretion, and drug combination. Patient-specific factors such as the age, body weight, general health, sex, diet, as well as the severity of the particular condition can also influence the amount which is to be administered.

The term "clearance” (also referred as “clearance value” or “CL” or “systemic clearance”) relates to the efficiency of elimination of a substance from the body. Clearance of a substance (in this case a binding molecule of the invention) is the sum of the urinary and extrarenal clearance; for substances that are eliminated by renal and extrarenal routes, plasma clearance exceeds urinary clearance. The PK properties of mAbs are a function of their large size (150 kDa), relative polarity, Fc-receptor binding and specific binding to target antigens. The primary elimination route for mAbs is cellular uptake followed by proteolytic degradation. Low clearance of mAbs from the systemic circulation enables them to be administered less frequently than peptides or small molecules, which is often more convenient for patients (Betts et al., MABs. 2018).

XI. INVENTION EMBODIMENTS FOR TDP-43 SPECIFIC BINDING MOLECULE

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which specifically binds misfolded TDP-43 and does not bind physiologically functional TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which specifically binds misfolded aggregated TDP-43 and does not bind physiologically functional TDP-43. In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which preferentially binds misfolded TDP-43 over physiologically functional TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which preferentially binds misfolded aggregated TDP-43 over physiologically functional TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which at least preferentially binds misfolded TDP- 43 over physiologically functional TDP-43 and preferably specifically binds misfolded TDP-43 and does not bind physiologically functional TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which at least preferentially binds misfolded aggregated TDP-43 over physiologically functional TDP-43 and preferably specifically binds misfolded aggregated TDP-43 and does not bind physiologically functional TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to misfolded monomeric and/or misfolded oligomeric and/or misfolded aggregated and/or misfolded post-translationally modified and/or misfolded truncated TDP-43, preferably human TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which specifically binds misfolded human TDP- 43 and does not bind physiologically functional human TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which preferentially binds misfolded human TDP- 43 over physiologically functional human TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which specifically binds misfolded aggregated human TDP-43 and does not bind physiologically functional human TDP-43. In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which preferentially binds misfolded aggregated human TDP-43 over physiologically functional human TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope within amino acids residues 202-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope within amino acids residues 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope within amino acids residues 202-211 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope consisting of amino acids residues 202-209 or 207-211 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope consisting of amino acids residues 202-209 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope consisting of amino acids residues 207-211 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope within amino acids residues 353-373 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which binds to an epitope within amino acids residues 370-414 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; or b. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; or c. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); or d. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or b. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or c. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87. In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or b. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or c. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or e. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising: i. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; or ii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; or iii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); or iv. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and b) a Light Chain Variable Region (VL) comprising: i. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or ii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or iii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or iv. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising: i. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; or ii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; or iii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); or iv. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and b) a Light Chain Variable Region (VL) comprising: i. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or ii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or iii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or iv. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87 v. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising: i. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 41; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 42; and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43 or a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 43; or ii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 51; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 52; and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53 or a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 53; or iii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 61; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 62; and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro- Phe); or iv. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 81; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 82; and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83 or a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 83; and b) a Light Chain Variable Region (VL) comprising: i. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 45; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 46; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 47; or ii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 55; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 36; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 57; or iii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 65; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 66; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 67; or iv. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 85; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 86; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 87.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising: i. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 41; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 42; and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43 or a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 43; or ii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 51; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 52; and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53 or a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 53; or iii. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 61; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 62; and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro- Phe); or iv. a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81 or a VH- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 81; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 or a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 82; and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83 or a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 83; and b) a Light Chain Variable Region (VL) comprising: i. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 45; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 46; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 47; or ii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 55; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 36; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 57; or iii. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 65; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 66; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 67; or iv. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 85; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 86; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 87; or v. a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55 or a VL- CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 55; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 or a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 136; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57 or a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 57.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or b) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57, or c) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87.

In some embodiments, a TDP-43 binding molecule, which is a TDP-43 antibody or an antigenbinding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or b) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57, or c) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or a humanized version thereof.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or b) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57, or c) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or e) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In a preferred embodiment, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or c) a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; or b) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; or c) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; or d) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; or b) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; or c) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; or d) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; or e) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or b) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or c) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or b) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or c) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84; or e) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 94 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 94; or f) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 104 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; or g) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 114 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 114; or h) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 124 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; or i) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 134 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 134; or j) a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 144 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 144.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) selected from: i. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; or ii. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; or iii. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; or iv. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; and b) a Light Chain Variable Region (VL) selected from: i. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or ii. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or iii. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or iv. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) selected from: i. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; or ii. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; or iii. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; or iv. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; or v. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; and b) a Light Chain Variable Region (VL) selected from: i. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or ii. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or iii. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or iv. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84; or v. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 94 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 94; or vi. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 104 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; or vii. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 114 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 114; or viii. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 124 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; or ix. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 134 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 134; or x. a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 144 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 144.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or b) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or c) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or b) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or c) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84; or e) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 94 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 94; or f) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 104 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; or g) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 114 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 114; or h) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 124 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; or i) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 134 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 134; or j) a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 144 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 144.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 40 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 44; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 80 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 84.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, comprising: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 40 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 44; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 80 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 84; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94 f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or h. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or i. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or j. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which exhibits one or more, up to all of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof; c. blocks TDP-43 cell-to-cell propagation; d. disaggregates TDP-43 aggregates; e. blocks TDP-43 seeding; f. neutralizes seeding-competent TDP-43; g. blocks TDP-43 spreading; h. potentiates intracellular and/or extracellular clearance of pathological TDP-43; i. restore nuclear levels and physiological function of TDP-43; and j. reduces phosphorylated TDP-43 level.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which exhibits one or more, up to all of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof; c. blocks TDP-43 seeding; d. neutralizes seeding-competent TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which exhibits one or both of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; and b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which inhibits the aggregation of TDP-43. In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which inhibits the extracellular aggregation of TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antigen-binding fragment thereof is provided, which inhibits the intracellular aggregation of TDP-43. In an embodiment, the TDP-43 binding molecule is an intrabody. In a preferred embodiment, the TDP- 43 binding molecule is a single chain variable fragment (scFv).

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which potentiates intracellular and/or extracellular clearance of pathological TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which reduces TDP-43 pathology.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which reduces levels of misfolded aggregated TDP-43 and/or phosphorylated TDP-43.

In some embodiments, the TDP-43 binding molecule is an antibody or antigen-binding fragment thereof.

In certain embodiments, the TDP-43 binding molecule, in particular TDP-43 antibody or antigen-binding fragment thereof as provided herein has a dissociation constant (KD) of < IpM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g. 10'⁸ M or less, e.g. from 10'⁸ M to 10'¹³ M, e.g., from 10'⁹ M to 10'¹³ M), in particular with respect to binding TDP-43, in particular soluble TDP-43. For example, the TDP-43 binding molecules of the invention may have a KD for binding full-length TDP-43 of 2000 pM or less, 1500 pM or less, 1000 pM or less, or 500 pM or less. This is demonstrated for TDP-43 binding molecules of the invention in Example 2 with reference to Table 4. In one embodiment, binding affinity to full length (FL) TDP-43 may be evaluated by determining the dissociation constants (KD) using surface plasmon resonance (SPR; Biacore 8K, GE Healthcare Life Sciences). Reference may be made to Example 2 for a detailed description of suitable SPR methods that may be employed. The KD values are typically derived from in vitro testing (and may be performed using heterologously expressed TDP-43 protein, e.g. expressed in E. coli as per Examples 1 and 2). The KD values are thus derived in a different experimental setting to that used to define the functional properties of the binding molecules in terms of specific or preferential binding of misfolded TDP-43 over physiologically functional TDP-43.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which has a dissociation constant (KD) for binding TDP-43 (SEQ ID NO: 1) of 2 nM or less, 1 nM or less, 500 pM or less.

TDP-43 binding molecules of the invention, in particular the antibodies or antigen-binding fragments thereof, typically bind full length TDP-43 with high affinity. For example, they may demonstrate an EC50 value of 100 ng/ml or less, 40 ng/ml or less, 30 ng/ml or less, or 15 ng/ml or less as determined by Luminex Assay. Reference may be made to Example 1 for further details of a suitable assay.

In an embodiment, the TDP-43 binding molecule, in particular TDP-43 antibody and fragment thereof may reduce the level of pathological TDP-43 in the brain and inhibit/reduce the formation of TDP-43 pathology.

In a further embodiment, the TDP-43 binding molecule, in particular TDP-43 antibody and fragment thereof may reduce the level of phosphorylated TDP-43 in the brain and inhibit/reduce the formation of TDP-43 pathology.

In some embodiments, the TDP-43 binding molecule is an IgA, IgD, IgE, IgM, IgGl, IgG2, IgG3 or IgG4 antibody or antigen-binding fragment thereof.

In a preferred embodiment, the TDP-43 binding molecule is an IgGl or an IgG4 antibody or antigen-binding fragment thereof.

In some embodiments, a TDP-43 binding molecule, in particular a TDP-43 antibody or an antigen-binding fragment thereof is provided, which comprises an Fc mutation, preferably the S228P mutation. In some embodiments, the TDP-43 binding molecule is a single chain variable fragment (scFv).

In certain embodiments, the TDP-43 binding molecule provided herein has a dissociation constant (KD) of 150 nM or less, 100 nM or less, 55 nM or less, 50nM or less, 35nM or less, or 32 nM or less with respect to binding to TDP-43 (SEQ ID NO: 1), preferably human TDP-43. Preferably, the TDP-43 binding molecule is a scFv. The dissociation constant is demonstrated for scFvs of the invention in Example 10 with reference to Table 14. In one embodiment, binding affinity to human TDP-43 may be evaluated by determining the dissociation constants (KD) using Bio-layer interferometry (BLI). BLI may be performed for example using an Octet QKe (Sartorius). Reference may be made to Example 10 for a detailed description of suitable BLI methods that may be employed. The KD values are typically derived from in vitro testing (and may be performed using heterologously expressed TDP-43 protein, e.g. expressed in E. coif). The KD values are thus derived in a different experimental setting to that used to define the functional properties of the binding molecules in terms of specific or preferential binding of misfolded TDP-43 over physiologically functional TDP-43.

The thermostability of a protein may be determined by measuring the melting temperature (Tm) of the protein, i.e. the temperature at which 50% of the protein is unfolded or denatured.

In some embodiments, the TDP-43 binding molecule provided herein has a melting temperature of at least 56 °C, at least 58 °C, at least 60 °C, at least 62 °C or at least 63 °C. Preferably, the TDP-43 binding molecule is a scFv. The melting temperature (thermostability) is demonstrated for scFv of the invention in Example 10 with reference to Table 13. In one embodiment, the melting temperature may be evaluated by differential scanning fluorimetry (DSF). This may be performed using a QuantStudio™ 3 Real-Time PCR System (ThermoFisher scientific). Reference may be made to Example 10 for a detailed description of suitable DSF methods that may be employed.

In some embodiments, the TDP-43 binding molecule is an scFv as provided herein and has a dissociation constant (KD) of 150 nM or less and a melting temperature (Tm) of at least 62 °C.

In some embodiments, the TDP-43 binding molecule is an intrabody.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid encodes a TDP-43 binding molecule in particular a TDP-43 antibody or fragment thereof described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 48 encoding a Heavy Chain Variable Region (VH) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 49 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 58 encoding a Heavy Chain Variable Region (VH) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 59 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 68 encoding a Heavy Chain Variable Region (VH) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 69 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 88 encoding a Heavy Chain Variable Region (VH) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 89 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 98 encoding a Heavy Chain Variable Region (VH) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 99 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein. In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 109 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 119 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 129 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 139 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 149 encoding a Light Chain Variable Region (VL) of an anti-TPD-43 antibody described herein.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 58 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 59; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 48 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 49; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 68 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 69; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 88 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 89.

In some embodiments, a(n isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 58 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 59; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 48 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 49; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 68 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 69; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 88 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 89; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 99; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 109; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 119; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 129; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 139; or a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 149.

XII. INVENTION EMBODIMENTS FOR VECTOR COMPRISING A NUCLEIC ACID ENCODING A TDP-43 SPECIFIC BINDING MOLECULE

The invention also relates to vectors delivering nucleic acid encoding binding molecules of the invention (as described herein).

In an aspect of the invention, there is provided a vector for targeted delivery to the CNS comprising a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein) which at least preferentially binds misfolded TDP-43 over physiologically functional TDP-43 and preferably specifically binds misfolded TDP-43 and does not bind physiologically functional TDP-43, and which binds to an epitope within amino acid residues 202-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, the vector is for targeted delivery to neurons and/or glial cells, preferably neurons, and wherein the neurons and/or glial cells express the TDP-43 binding molecule as an intrabody.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or a humanized version thereof.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: e. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or f. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or g. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or h. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or i. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In a preferred embodiment, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84; or e. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 94 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 94; or f. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 104 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; or g. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 114 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 114; or h. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 124 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; or i. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 134 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 134; or j . a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 144 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 144.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 40 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 80 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 84.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 40 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 80 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 84; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94; or f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or h. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or i. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or j. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to an epitope consisting of amino acid residues 202-209 or 207-211 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to an epitope consisting of amino acid residues 207-211 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to an epitope consisting of amino acid residues 202-209 of human TDP-43 (SEQ ID NO: 1). In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to an epitope within amino acid residues 353-373 or within amino acid residues 370-414 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to an epitope within amino acid residues 353-373 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to an epitope within amino acid residues 370-414 of human TDP-43 (SEQ ID NO: 1).

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule binds to misfolded monomeric and/or misfolded oligomeric and/or misfolded aggregated and/or misfolded post- translationally modified and/or truncated misfolded TDP-43.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule exhibits one or more, up to all of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof; c. blocks TDP-43 cell-to-cell propagation; d. disaggregates TDP-43 aggregates; e. blocks TDP-43 seeding; f. neutralizes seeding-competent TDP-43; g. blocks TDP-43 spreading; h. potentiates intracellular and/or extracellular clearance of pathological TDP-43; i. restores nuclear levels and physiological function of TDP-43; and j. reduces phosphorylated TDP-43 level.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule is an intrabody and wherein the TDP-43 binding molecule addresses gain- and/or loss-of-function pathologies associated with TDP-43. The gain-of-function and/or loss-of-function may be from a genetic cause or non-genetic cause (e.g. post-translation modifications). Without wishing to be bound by theory, pathological gain-of-function associated with TDP-43 refers to toxicity driven by aggregation and spreading of misfolded TDP-43, while pathological loss-of-function associated with TDP-43 refers to depletion of physiological TDP-43 from the nucleus and loss of physiological function of TDP-43.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule exhibits one or both of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; and b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule inhibits the intracellular aggregation of TDP-43. In an embodiment, the TDP-43 binding molecule is an intrabody. In a preferred embodiment, the TDP-43 binding molecule is a single chain variable fragment (scFv).

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule has a dissociation constant (KD) for binding TDP-43 (SEQ ID NO: 1) of 2 nM or less, 1 nM or less, or 500 pM or less. The KD may be determined by any suitable method, such as according to the method described in Example 2.

In certain embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule as described herein, preferably wherein the TDP-43 binding molecule is a scFv as provided herein, and has a dissociation constant (KD) of 150 nM or less, 100 nM or less, 55nM or less, 50nM or less, 35nM or less or 32 nM or less with respect to binding to TDP-43 (SEQ ID NO: 1), preferably human TDP-43. This is demonstrated for scFvs of the invention in Example 10 with reference to Table 14. In one embodiment, binding affinity to human TDP-43 may be evaluated by determining the dissociation constants (KD) using Bio-layer interferometry (BLI). This may be performed using an Octet QKe (Sartorius). Reference may be made to Example 10 for a detailed description of suitable BLI methods that may be employed. The KD values are typically derived from in vitro testing (and may be performed using heterologously expressed TDP-43 protein, e.g. expressed in E. coif). The KD values are thus derived in a different experimental setting to that used to define the functional properties of the binding molecules in terms of specific or preferential binding of misfolded TDP-43 over physiologically functional TDP-43.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule provided herein (preferably a scFv as provided herein) has a melting temperature of at least 56 °C, at least 58 °C, at least 60 °C, at least 62 °C or at least 63 °C. This is demonstrated for scFvs of the invention in Example 10 with reference to Table 13. In one embodiment, the melting temperature may be evaluated by differential scanning fluorimetry (DSF). This may be performed using a QuantStudio™ 3 Real-Time PCR System (ThermoFisher scientific). Reference may be made to Example 10 for a detailed description of suitable DSF methods that may be employed.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule that is an scFv as provided herein and has a dissociation constant (KD) of 150 nM or less and a melting temperature (Tm) of at least 62 °C.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule is an antibody or antigenbinding fragment thereof. In one embodiment, the TDP-43 binding molecule is an intrabody.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule is an IgA, IgD, IgE, IgM, IgGl, IgG2, IgG3 or IgG4 antibody or antigen-binding fragment thereof.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule is an IgGl or an IgG4 antibody or antigen-binding fragment thereof.

In some embodiments, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule comprises an Fc mutation, preferably the S228P mutation.

In a preferred embodiment, the vector comprises a nucleic acid sequence encoding a TDP-43 binding molecule (as described herein), wherein the TDP-43 binding molecule is a single chain variable fragment (scFv). In an embodiment, the linker linking the VH and VL of the scFv comprises or consists of the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4. In a preferred embodiment, the linker comprises the amino acids sequence of SEQ ID NO: 3.

In some embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) for targeted delivery to the blood-brain barrier (BBB) or the central nervous system (CNS) is provided. In a preferred embodiment, the vector is for targeted delivery to the CNS.

In some embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) is provided, wherein the vector is for targeted delivery to endothelial cells of the BBB, pericytes of the BBB, neurons and/or glial cells. In a preferred embodiment, the vector is for targeted delivery to neurons and/or glial cells. In one embodiment, glial cells are astrocytes and/or oligodendrocytes. In a more preferred embodiment, the vector is for targeted delivery to neurons.

In some embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) for targeted delivery to endothelial cells of the BBB, pericytes of the BBB, neurons and/or glial cells is provided, wherein the endothelial cells of the BBB, pericytes of the BBB, neurons and/or glial cells express and secrete the binding molecule into the brain parenchyma.

In preferred embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) for targeted delivery to neurons and/or glial cells is provided, wherein the neurons and/or glial cells express the binding molecule as an intrabody. In a more preferred embodiment, the vector is for targeted delivery to neurons. In another preferred embodiment, the intrabody is a scFv (single chain variable fragment). In an embodiment, glial cells are astrocytes or oligodendrocytes. In some embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) for targeted delivery to neurons and/or glial cells is provided, wherein the TDP-43 binding molecule is expressed under the control of a CNS-specific promoter. As used herein, the term "promoter" refers to a region of DNA that generally is located upstream of a polynucleotide sequence (e.g. the polynucleotide sequence encoding the light chain and heavy chain of an anti-TDP-43 antibody) to be transcribed that is needed for transcription to occur, e.g. which initiates transcription. The promoter is operably linked to the nucleic acid encoding a TDP-43 binding molecule such that these elements are functionally connected and are able to interact with each other in the manner intended. In some embodiments, the CNS-specific promoter is CMVe-GFAP (CMV enhancer fused to GFAP short promoter), GFAP (Human glial fibrillary acidic protein promoter), GfaABCID (modified GFAP promoter), ATP1A2 (Na, K ATPase a2 promoter), CLDN5 (Claudin 5 promoter), ADRB2 (Adrenoceptor beta 2 promoter), TNFRSF6B (TNF receptor superfamily member 6b promoter), PDYN (prodynorphin promoter), GH1 (Human growth hormone promoter), OPALIN (Opalin promoter), SYN1 (Synapsin 1 promoter), CAMK2A (Calcium/Calmodulin Dependent Protein Kinase II alpha promoter), NEFH (neurofilament heavy polypeptide promoter), NEUR0D6 (neuronal differentiation factor 6 promoter), 0LIG2 (oligodendrocyte transcription factor 2 promoter), CMVe-0LIG2 (CMV enhancer fused to 0LIG2 promoter). In a preferred embodiment, the promoter is a human SYN1 promoter or CMVe-SYNl (CMV enhancer fused to human SYN1 promoter). In an embodiment, the human SYN1 promoter has the nucleic acid sequence of SEQ ID NO: 5 (CTGCAGAGGGCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCG GGGTGGGGGTGCCTACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACC CCCATTCCCCAAATTGCGCATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGC GGCGAGGCGCGTGCGCACTGCCAGCTTCAGCACCGCGGACAGTGCCTTCGCCCCC GCCTGGCGGCGCGCGCCACCGCCGCCTCAGCACTGAAGGCGCGCTGACGTCACTC GCCGGTCCCCCGCAAACTCCCCTTCCCGGCCACCTTGGTCGCGTCCGCGCCGCCGC CGGCCCAGCCGGACCGCACCACGCGAGGCGCGAGATAGGGGGGCACGGGCGCGA CCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTCAGTCTGCGGTGGGCAGC GGAGGAGTCGTGTCGTGCCTGAGAGCGCAG).

In some embodiments, the vector is a viral vector. In some embodiments, the vector is a wild type or engineered AAV vector. In some embodiments, the vector is a wild-type AAV vector. In some embodiments, the vector is an engineered AAV vector. In an embodiment, the AAV vector is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh.8, AAVrh.9, AAVrh.10, AAVrh.11, AAVrh.12, AAVrh.13, AAVrh.14, AAVrh.15, AAVrh.16, AAVrh.17 or AAV.Hu68. In another embodiment, the AAV vector is an engineered AAV1, engineered AAV2, an engineered AAV3, an engineered AAV4, an engineered AAV5, an engineered AAV6, an engineered AAV7, an engineered AAV8, an engineered AAV9, an engineered AAV10, an engineered AAV11, an engineered AAV12, an engineered AAVrh.8, an engineered AAVrh.9, engineered AAVrh.10, an engineered AAVrh.11, an engineered AAVrh.12, an engineered AAVrh.13, an engineered AAVrh.14, an engineered AAVrh.15, an engineered AAVrh.16, an engineered AAVrh.17 or an engineered AAV.Hu68. In an embodiment, the AAV vector is an AAV2-BR1, AAV-S, AAV-F, AAV.PHP.eB, AAV9.PHP.V1, AAV1RX, AAV1R6, AAV1R7, AAV.CAP-B10, AAV.CAP-B22, AAV.CAP- Mac, AAV VCAP-103, AAV bCap 1, V-CAP102, AAV-CGN1, AAV-STRV5, AAV-PAL2, AAV-MDV1 A, AAV-MDV1B, AAV-MaCPNSl, AAV-MaCPNS2 or AAV VCAP-100.

In some embodiments, an AAV vector comprising a nucleic acid encoding a TDP-43 binding molecule for targeted delivery to neurons and/or glial cells is provided, wherein the TDP-43 binding molecule is a single chain variable fragment (scFv) comprising: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67 c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL- CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

In some embodiments, the single chain variable fragment (scFv) comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or h. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

In some embodiments, the single chain variable fragment (scFv) comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

In a preferred embodiment, the single chain variable fragment (scFv) comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134.

In some embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) is provided, wherein the nucleic acid sequences are set forth as: a. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 58 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 59; or b. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 48 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 49; or c. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 68 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 69; or d. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 88 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 89; and wherein the vector is for targeted delivery to the CNS. In a preferred embodiment, the vector is for targeted delivery to neurons.

In some embodiments, a vector comprising a nucleic acid encoding a TDP-43 binding molecule (as described herein) is provided, wherein the nucleic acid sequences are set forth as: a. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 58 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 59; or b. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 48 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 49; or c. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 68 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 69; or d. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 88 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 89; or e. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 99; or f. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 109; or g. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 119; or h. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 129; or i. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 139; or j. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 149; and wherein the vector is for targeted delivery to the CNS. In a preferred embodiment, the vector is for targeted delivery to neurons. XIII. COMPOSITIONS AND METHODS

In some embodiments, a conjugated binding molecule, in particular antibody or antigen-binding fragment thereof, is provided, comprising a binding molecule, in particular an antibody or antigen-binding fragment thereof, described herein and a conjugated molecule. Conjugates of the invention may be referred to as immunoconjugates. Any suitable conjugated molecule may be employed according to the invention. Suitable examples include, but are not limited to enzymes (e.g. alkaline phosphatase or horseradish peroxidase), avidin, streptavidin, biotin, Protein A/G, magnetic beads, fluorophores, radioactive isotopes (i.e., radioconjugates), nucleic acid molecules, detectable labels, therapeutic agents, toxins and blood brain barrier penetration moieties. Conjugation methods are well known in the art and several technologies are commercially available for conjugating antibodies to a label or other molecule, Conjugation is typically through amino acid residues contained within the binding molecules of the invention (such as lysine, histidine or cysteine). They may rely upon methods such as the NHS (Succinimidyl) ester method, isothiocyanate method, carbodiimide method and periodate method. Conjugation may be achieved through creation of fusion proteins for example. This is appropriate where the binding molecule is conjugated with another protein molecule. Thus, suitable genetic constructs may be formed that permit the expression of a fusion of the binding molecule of the invention with the label or other molecule. Conjugation may be via a suitable linker moiety to ensure suitable spatial separation of the antibody and conjugated molecule, such as detectable label. However, a linker may not be required in all instances. In some embodiments the TDP-43 specific binding molecule of the present invention is linked to a detectable label.

The invention also relates to an immunoconjugate comprising the TDP-43 binding molecule, provided herein conjugated to one or more therapeutic agents, such as chemotherapeutic agents or drugs, growth inhibitory agents, toxins (e.g., protein toxins, enzymatically active toxins of bacterial, fungal, plant, or animal origin, or fragments thereof), radioactive isotopes (i.e., a radioconjugate), blood-brain barrier penetration moieties or detectable labels. Various techniques exist for improving drug delivery across the blood-brain barrier (BBB) as discussed herein, which discussion applies mutatis mutandis. Non-invasive techniques include the so- called “Trojan horse approach” in which conjugated molecules deliver the binding molecules of the invention by binding to BBB receptors and mediating transport. Suitable molecules may comprise endogenous ligands or antibodies, in particular monoclonal antibodies, that bind specific epitopes on the BBB receptor. In some embodiments, an immunoconjugate is provided, wherein the immunoconjugate comprises an (isolated) antibody described herein and a therapeutic agent. In some embodiments, a labelled antibody is provided, comprising an antibody described herein and a detectable label. In some embodiments the TDP-43 specific binding molecule is part of an immunoconjugate wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent. In some embodiments the TDP-43 specific binding molecule or the immunoconjugate comprising it is present as a composition comprising a TDP-43 specific binding molecule.

In some embodiments the TDP-43 specific binding molecule is part of pharmaceutical composition comprising a TDP-43 specific binding molecule, or an immunoconjugate wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising a TDP-43 specific binding molecule combined with a pharmaceutically acceptable carrier and/or excipient and/or diluent.

In some embodiments the immunoconjugate comprising a TDP-43 binding molecule of the invention crosses the blood brain barrier using a delivery vehicle or a blood brain barrier moiety. In some embodiments the delivery vehicle comprises a liposome or extracellular vesicle. In some embodiments the TDP-43 binding molecule is linked to the blood brain barrier moiety. In some embodiments the blood brain barrier moiety is a polypeptide or a small molecule, preferably, a peptide, a receptor ligand, a single domain antibody (VHH), a scFv or a Fab fragment. In some embodiments the blood brain barrier moiety binds a blood brain barrier receptor, which may comprise one or more of a transferrin receptor, insulin receptor or low-density lipoprotein receptor.

Also provided is a labelled binding molecule, in particular a labelled antibody, comprising a TDP-43 binding molecule of the invention.

The invention also relates to pharmaceutical compositions comprising a TDP-43 binding molecule (particularly an antibody or an antigen-binding fragment thereof), a labelled antibody or a vector of the invention as described herein and a pharmaceutically acceptable carrier and/or excipient and/or diluent. In some embodiments, a pharmaceutical composition is provided, comprising an (isolated) antibody described herein and a pharmaceutically acceptable carrier. In some embodiments the TDP-43 specific binding molecule is part of a detection and/or diagnostic kit comprising a TDP-43 specific binding molecule, or an immunoconjugate wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising a TDP-43 specific binding molecule.

In some embodiments, TDP-43 binding molecules described herein are used in a pairing assay comprising the steps of a) Incubating a sample with a capture antibody and a detect antibody to produce a mixture; b) Incubating the mixture obtained in step a) with a reagent suitable for detection by the detect antibody; c) Measuring the signal emitted by the detect antibody; wherein the capture antibody and/or the detect antibody are TDP-43 binding molecules of the invention.

In some embodiments, a TDP-43 binding molecule described herein is used in a pairing assay comprising the steps of a) Incubating a sample with a capture antibody and a detect antibody to produce a mixture; b) Incubating the mixture obtained in step a) with a reagent suitable for detection by the detect antibody; c) Measuring the signal emitted by the detect antibody; wherein the capture antibody is a TDP-43 binding molecule of the invention.

In some embodiments, a TDP-43 binding molecule described herein is used in a pairing assay comprising the steps of a) Incubating a sample with a capture antibody and a detect antibody to produce a mixture; b) Incubating the mixture obtained in step a) with a reagent suitable for detection by the detect antibody; c) Measuring the signal emitted by the detect antibody; wherein the detect antibody is a TDP-43 binding molecule of the invention.

In some embodiments, the invention provides a method of detecting TDP-43 in a sample comprising the steps of a) incubating a sample with a capture antibody and a detect antibody to produce a mixture; b) incubating the mixture obtained in step a) with a reagent suitable for detection of TDP- 43 by the detect antibody; and c) measuring the signal emitted by the detect antibody.

In one embodiment of the method of detecting TDP-43 in a sample, the capture antibody is a TDP-43 binding molecule of the invention. In another embodiment of the method of detecting TDP-43 in a sample, the detect antibody is a TDP-43 binding molecule of the invention. In a further embodiment of the method of detecting TDP-43 in a sample, the capture antibody and the detect antibody are TDP-43 binding molecules of the invention. The capture antibody and the detect antibody may be the same or different antibodies of the invention.

In some embodiments, provided is a pairing assay kit for detecting TDP-43 in a sample. The pairing assay kit comprises one or more TDP-43 binding molecules of the invention. The kit may be an Enzyme-Linked Immunosorbent Assay (ELISA) kit. The kit may be a Single Molecule Array (Simoa®) kit. The kit comprises a capture agent and/or a detect agent. The kit optionally further comprises a detection reagent. The TDP-43 binding molecule of the invention may be provided in the kit as the capture agent, such as the capture antibody and/or as the detect agent, such as a detect antibody.

In some embodiments, one or more TDP-43 binding molecules described herein is/are used in a pairing assay comprising the step of incubating a sample with a capture and a detect antibody, wherein the sample is blood or a blood derivative, cerebrospinal fluid (CSF), interstitial fluid (ISF) and/or urine, preferably CSF. In an embodiment, the blood derivative is plasma, plateletrich plasma or platelet cytosol fraction.

Kits containing the binding molecules of the invention are also provided. In particular, such kits may be useful for performing the diagnostic methods of the invention (which include classification, monitoring and therapy selection methods). Thus, a kit for diagnosis of a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or a TDP-43 proteinopathy, or for use in a method of the invention is provided comprising a TDP-43 specific binding molecule of the invention. Such kits may comprise all necessary components for performing the herein provided methods. Typically, each component is stored separately in a single overall packaging. Suitable additional components for inclusion in the kits are, for example, buffers, detectable dyes, laboratory equipment, reaction containers, instructions and the like. Instructions for use may be tailored to the specific method for which the kit is to be employed. Suitably labelled TDP-43 binding molecules of the invention are also provided, which may be included in such kits.

In some embodiments, the TDP-43 specific binding molecule is used in an immunodiagnostic method for use in the prevention, diagnosis or treatment of a TDP-43 proteinopathy.

In some embodiments, a TDP-43 specific binding molecule as described herein, or an immunoconjugate as described herein wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a vector comprising a nucleic acid encoding a TDP-43 specific binding molecule as described herein, or a pharmaceutical composition comprising a TDP-43 specific binding molecule as described herein, is administered to a subject in need thereof or is used to diagnose, prevent, alleviate or treat a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy including but not limited to frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), limbic-predominant age-related TDP-43 encephalopathy (LATE).

In some embodiments, a TDP-43 specific binding molecule as described herein, or an immunoconjugate as described herein wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a vector comprising a nucleic acid encoding a TDP-43 specific binding molecule as described herein, or a pharmaceutical composition comprising a TDP-43 specific binding molecule as described herein, is administered to a subject in need thereof or is used in a method for diagnosing or monitoring a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy selected from Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD). In other embodiments, the invention relates to any methods for detecting, diagnosing or monitoring a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy that is selected from frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), and limbic-predominant age-related TDP-43 encephalopathy (LATE).

Preferably, the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathies selected from amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD) and Frontotemporal dementia (FTD). More preferably, the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS). More preferably, the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy is Alzheimer’s disease (AD). More preferably, the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy is Frontotemporal dementia (FTD).

In some embodiments, the TDP-43 specific binding molecule is used in a method for diagnosing presymptomatic disease or for monitoring disease progression and therapeutic efficacy, or for predicting responsiveness, or for selecting subjects which are likely to respond to the treatment with a TDP-43 specific binding molecule. Said method is preferably performed using a sample of human blood or urine. Most preferably the method involves an ELISA-based or surface adapted assay.

In some embodiments, the TDP-43 specific binding molecule is used in a method wherein a TDP-43 specific binding molecule of the present invention is contacted with a sample (e.g., blood or blood derivative, urine, cerebrospinal fluid, interstitial fluid (ISF), or brain tissue) to detect, diagnose or monitor frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, limbic- predominant age-related TDP-43 encephalopathy (LATE) and/or Parkinson’s disease (PD).

In some embodiments, the TDP-43 specific binding molecule is used in a method wherein a TDP-43 specific binding molecule of the present invention is contacted with a sample (e.g., blood or blood derivative, urine, cerebrospinal fluid, interstitial fluid (ISF), or brain tissue) to detect, diagnose or monitor a disease selected from Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD).

In some embodiments, the TDP-43 specific binding molecule, or an immunoconjugate wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a vector comprising a nucleic acid encoding a TDP-43 specific binding molecule as described herein, or a pharmaceutical composition comprising a TDP-43 specific binding molecule, is administered to a subject in need thereof or is used for preventing, alleviating or treating a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP- 43 aggregates, or TDP-43 proteinopathies, or frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD, including sporadic and familial forms of AD), Chronic Traumatic Encephalopathy (CTE), Perry syndrome and limbic- predominant age-related TDP-43 encephalopathy (LATE) and/or Parkinson’s disease (PD).

In some embodiments, a TDP-43 specific binding molecule as described herein, or an immunoconjugate as described herein wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a vector comprising a nucleic acid encoding a TDP-43 specific binding molecule as described herein, or a pharmaceutical composition comprising a TDP-43 specific binding molecule as described herein, is administered to a subject in need thereof or is used for treating a disease selected from: Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic- predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD). Preferably said disease treatment helps to retain or increase mental recognition and or reduces the level of TDP-43 aggregates in the brain.

In some embodiments, a TDP-43 specific binding molecule as described herein, or an immunoconjugate as described herein wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a vector comprising a nucleic acid encoding a TDP-43 specific binding molecule as described herein, or a pharmaceutical composition comprising a TDP-43 specific binding molecule as described herein, is administered to a subject in need thereof or is used for manufacturing a medicament for treating a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathies or frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD, including sporadic and familial forms of AD), Chronic Traumatic Encephalopathy (CTE), Perry syndrome and limbic-predominant age-related TDP-43 encephalopathy (LATE), and/or Parkinson’s disease (PD).

Pharmaceutical formulations of an anti-TDP-43 antibody or antigen-binding fragment thereof, immunoconjugate or vector as described herein are prepared by mixing such antibody, immunoconjugate or vector having the desired degree of purity with one or more optional pharmaceutically acceptable carriers and/or excipients and/or diluents (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Typically, the antibody or fragment thereof is prepared as a lyophilized formulation or aqueous solution. Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed, and include, but are not limited to: buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g. Zn protein complexes); and/or non-ionic surfactants such as polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers herein further include interstitial drug dispersion agents such as soluble neutral -active hyaluronidase glycoproteins (sHASEGP), for example, human soluble PH-20 hyaluronidase glycoproteins, such as rHuPH20 (HYLENEX®, Baxter International, Inc.). Certain exemplary sHASEGPs and methods of use, including rHuPH20, are described in US Patent Publication Nos. 2005/0260186 and 2006/0104968. In one aspect, a sHASEGP is combined with one or more additional glycosaminoglycanases such as chondroitinases. Pharmaceutically acceptable excipients that may be used to formulate the compositions include, but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances (for example sodium carboxymethylcellulose), polyethylene glycol, polyacrylates, waxes, polyethylene- polyoxypropylene- block polymers, polyethylene glycol and lanolin. Diluents may be buffers. They may comprise a salt selected from the group consisting of phosphate, acetate, citrate, succinate and tartrate, and/or wherein the buffer comprises histidine, glycine, TRIS glycine, Tris, or mixtures thereof. It is further envisaged in the context of the present invention that the diluent is a buffer selected from the group consisting of potassium phosphate, acetic acid/sodium acetate, citric acid/sodium citrate, succinic acid/sodium succinate, tartaric acid/sodium tartrate, and histidine/histidine HCI or mixtures thereof.

Exemplary lyophilized antibody or immunoconjugate formulations are described in US Patent No. 6,267,958. Aqueous antibody or immunoconjugate formulations include those described in US Patent No. 6,171,586 and W02006/044908, the latter formulations including a histidineacetate buffer.

The formulation herein may also contain more than one active ingredient as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other.

Active ingredients may be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatinmicrocapsules and poly-(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody or immunoconjugate, which matrices are in the form of shaped articles, e.g. films, or microcapsules. The formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, e.g., by filtration through sterile filtration membranes.

Any of the antigen-binding molecules, anti-TDP-43 antibodies, immunoconjugates, vectors delivering anti-TDP-43 antibody or pharmaceutical compositions provided herein may be used in methods, e.g., therapeutic methods.

In another aspect, an anti-TDP-43 antibody or antigen-binding fragment, immunoconjugate or vector as described herein for use as a medicament is provided. In further aspects, an antimisfolded TDP-43 antibody or antigen-binding fragment, immunoconjugate or vector for use in a method of treatment is provided. In certain embodiments, an anti-TDP-43 antibody or antigenbinding fragment, immunoconjugate or vector for use in the prevention, diagnosis and/or treatment of a TDP-43 proteinopathy is provided. In a preferred embodiment of the invention, an anti-TDP-43 antibody or antigen-binding fragment, immunoconjugate or vector is provided for use in the prevention, diagnosis and/or treatment of a disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy including but not limited to frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), and/or limbic-predominant age-related TDP-43 encephalopathy (LATE).

In a further aspect, the invention provides for the use of an anti-TDP-43 antibody or antigenbinding fragment, immunoconjugate or vector in the manufacture or preparation of a medicament. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, e.g., as described below. A “subject” or an "individual" according to any of the embodiments may be an animal, a mammal, and is preferably a human.

In a further aspect, the invention provides pharmaceutical formulations comprising any of the TDP-43 antibodies or antigen-binding fragments, immunoconjugates or vectors provided herein, e.g., for use in any of the therapeutic methods. In one embodiment, a pharmaceutical formulation comprises any of the TDP-43 antibodies or antigen-binding fragments, immunoconjugates or vectors provided herein and a pharmaceutically acceptable carrier and/or excipients and/or diluents (as discussed elsewhere herein). In another embodiment, a pharmaceutical formulation comprises any of the TDP-43 antibodies or antigen-binding fragments, immunoconjugates or vectors provided herein and at least one additional therapeutic agent, e.g., as described below.

Antibodies or antigen-binding fragments, immunoconjugates or vectors of the invention can be used either alone or in combination with other agents in a therapy. For instance, an antibody or antigen-binding fragment, immunoconjugate or vector of the invention may be co-administered with at least one additional therapeutic agent targeting alpha-synuclein, BACE1, tau, betaamyloid, TDP-43 or a neuroinflammation protein.

For instance, an antibody or antigen-binding fragment, immunoconjugate or vector of the invention may be co-administered with at least one additional therapeutic agent which is selected from, but not limited to, neurological drugs, anti-amyloid beta antibodies, anti-tau antibodies, tau aggregation inhibitors (including small molecules), beta-amyloid aggregation inhibitors (including small molecules), anti-BACEl antibodies, BACE1 inhibitors, anti-alpha-synuclein inhibitors, anti-alpha-synuclein antibodies and neuroinflammation inhibitors.

Such combination therapies noted above encompass combined administration (where two or more therapeutic agents are included in the same or separate formulations), and separate administration, in which case, administration of the antibody or antigen-binding fragment, immunoconjugate or vector of the invention can occur prior to, simultaneously, and/or following, administration of the additional therapeutic agent and/or adjuvant. Antibodies or antigen-binding fragment, immunoconjugates or vectors of the invention can also be used in combination with radiation therapy.

An antibody or antigen-binding fragment, immunoconjugate or vector of the invention (and any additional therapeutic agent) can be administered by any suitable means. Suitable means for the administration of an antibody or antigen-binding fragment, or immunoconjugate includes but is not limited to parenteral, intrapulmonary, intranasal, and, if desired for local treatment, intralesional, intrauterine or intravesical administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Dosing can be by any suitable route, e.g. by injections, such as intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic. Various dosing schedules including but not limited to single or multiple administrations over various time-points, bolus administration, and pulse infusion are contemplated herein. Suitable means for the administration of a vector (for example an AAV vector) includes but is not limited to parenteral route (e.g. intravenous, intraperitoneal, intranasal, intravitreous, subretinal, subcutaneous, intramuscular, intrathecal, intracistemal (intracistema magna), intraparenchymal, intrastriatal or intracerebroventricular route). Preferably, a vector (for example an AAV vector) is administered by intravenous, intravitreous, subretinal, intrathecal, intracerebroventricular, intracistemal (intraci sterna magna), intrastriatal or intraparenchymal route. Various dosing schedules including but not limited to single or multiple administrations over various time-points are contemplated herein.

For example, an AAV vector administration may comprise delivery of from 1 • IO¹⁰ vector genomes (vg)/kg to 5 10¹⁴ vg/kg of the AAV vector to the subject per dose.

Antibodies or antigen-binding fragments, immunoconjugates or vectors of the invention are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy being treated, the particular mammal being treated, the clinical condition of the individual subject, the cause of the disease, a disorder and/or abnormality associated with TDP- 43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The antibody or immunoconjugate need not be, but is optionally, formulated with one or more agents currently used to prevent or treat the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy in question. The effective amount of such other agents depends on the amount and/or efficiency of antibody or antigen-binding fragment, immunoconjugate or vector present in the formulation, the type of disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates or TDP- 43 proteinopathy or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as described herein, or about from 1 to 99% of the dosages described herein, or in any dosage and by any route that is empirically/clinically determined to be appropriate.

For the prevention or treatment of disease, the appropriate dosage of an antibody or antigenbinding fragment, immunoconjugate or vector of the invention (when used alone or in combination with one or more other additional therapeutic agents) will depend on the type of disease to be treated, the type of antibody or antigen-binding fragment, immunoconjugate or vector, the severity and course of the disease, whether the antibody or antigen-binding fragment, immunoconjugate or vector is administered for preventive or therapeutic purposes, previous therapy, the subject's clinical history and response to the antibody or antigen-binding fragment, immunoconjugate or vector, and the discretion of the attending physician. The antibody or antigen-binding fragment, or immunoconjugate is suitably administered to the subject at one time or over a series of treatments. Depending on the type and severity of the disease, about 1 pg/kg to 15 mg/kg (e.g. 0.1 mg/kg-10 mg/kg) of antibody, antigen-binding fragment or immunoconjugate can be an initial candidate dosage for administration to the subject, whether, for example, by one or more separate administrations, or by continuous infusion. One typical daily dosage might range from about 1 pg/kg to 100 mg/kg or more, depending on the factors mentioned above. For repeated administrations over several days or longer, depending on the condition, the treatment would generally be sustained until a desired suppression of disease symptoms occurs. One exemplary dosage of the antibody or immunoconjugate would be in the range from about 0.05 mg/kg to about 10 mg/kg. Thus, one or more doses of about 0.5 mg/kg, 2.0 mg/kg, 4.0 mg/kg or 10 mg/kg (or any combination thereof) may be administered to the subject. Such doses may be administered intermittently, e.g. every week or every three weeks (e.g. such that the subject receives from about two to about twenty, or e.g. about six doses of the antibody). An initial higher loading dose, followed by one or more lower doses may be administered. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays. It is understood that any of the above formulations or therapeutic methods may be carried out using both an immunoconjugate of the invention and an anti-TDP-43 antibody.

In another aspect of the invention, an article of manufacture containing materials useful for the treatment, prevention and/or diagnosis of the diseases, disorders or abnormalities associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP-43 proteinopathy, described above is provided. The article of manufacture comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds a composition which is by itself or combined with another composition effective for treating, preventing and/or diagnosing the disease, disorder and/or abnormality associated with TDP-43, in particular associated with TDP-43 aggregates, or TDP- 43 proteinopathy, and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is an antibody or antigen-binding fragment, immunoconjugate or vector of the invention. The label or package insert indicates that the composition is used for treating the condition of choice.

Moreover, the article of manufacture may comprise (a) a first container with a composition contained therein, wherein the composition comprises an antibody or antigen-binding fragment, immunoconjugate or vector of the invention; and (b) a second container with a composition contained therein, wherein the composition comprises a further therapeutic agent. The article of manufacture in this embodiment of the invention may further comprise a package insert indicating that the compositions can be used to treat a particular condition. Alternatively, or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution or dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

In a further embodiment, the invention relates to a method of retaining or increasing cognitive memory capacity, movement and language function or preventing and/or slowing decline of cognitive memory capacity, movement and language function in a subject, comprising administering the binding molecule of the invention, the immunoconjugate of the invention, the vector of the invention, the composition of the invention or the pharmaceutical composition of the invention.

In a further embodiment, the invention relates to a method of reducing the level of TDP-43, comprising administering the binding molecule of the invention, the immunoconjugate of the invention, the vector of the invention, the composition of the invention or the pharmaceutical composition of the invention.

The methods of the invention may comprise administering at least one additional therapy, preferably wherein the additional therapy is selected from, but not limited to, antibodies or small molecules targeting alpha-synuclein, BACE1, tau, beta-amyloid, TDP-43 or a neuroinflammation protein, in particular neurological drugs, anti-beta-amyloid antibodies, anti- tau antibodies, tau aggregation inhibitors, beta-amyloid aggregation inhibitors, anti-BACEl antibodies, BACE1 inhibitors, anti-alpha-synuclein antibodies and neuroinflammation inhibitors.

The invention furthermore relates to a method of detecting TDP-43, comprising contacting a sample with the binding molecule of the invention, preferably an antibody of the invention wherein the sample is a brain sample, a cerebrospinal fluid sample, an interstitial fluid (ISF) sample, urine sample or a blood or blood derivative sample.

In further embodiment, the invention relates to a method of detecting and/or measuring the level of TDP-43, comprising contacting a sample with the binding molecule of the invention, preferably an antibody of the invention, using Single Molecule Array (SIMOA®) technology, wherein the sample is a blood or blood derivative sample, a cerebrospinal fluid sample (CSF), an interstitial fluid (ISF) sample or a urine sample, preferably a CSF sample.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1. Representative immunostainings of rat primary neurons transduced with an AAV vector expressing either ACI-8062-11 Al-Abl scFv or a negative control scFv detected with an anti -FLAG antibody. Scale bar is 100 pm.

Figure 2. Graphical representation of the number (Nb) of TDP-43 aggregates per cell in primary rat neurons co-transduced with an AAV vector expressing EGFP-TDP-43-NLSm and an AAV vector expressing either ACI-8062-11 Al-Abl scFv or a negative control scFv.

Figure 3. Representative immunostainings of rat primary neurons transduced with an AAV vector expressing either ACI-8062-11 Al-Abl scFv, ACI-8062-11 Al -Ab 1 H2L3 scFv or ACI- 8062-11 Al-Abl_H2L5 scFv detected with an anti-FLAG antibody. Scale bar is 100 pm.

Figure 4. Graphical representation of the integrated intensity of pTDP-43 immunoreactive area per cell in primary rat neurons co-transduced with an AAV vector expressing EGFP-TDP-43- NLSm and an AAV vector expressing either the murine ACI-8062-1 lAl-Abl scFv, the hACI- 8062-1 lAl-Abl_H2L3 scFv, the hACI-8062-1 lAl-Abl_H2L5 scFv, or anegative control scFv. EXAMPLES

Example 1. Generation of anti-TDP-43 antibodies

The liposome-based vaccines were prepared according to the protocols published in WO2012/055933. Vaccines containing full length TDP-43 (FL TDP-43) or the low complexity domain (LCD) of TDP-43 (LCD TDP-43) as antigen were used for antibody generation and are summarized in Table 2.

Table 2. TDP-43 protein and peptide antigen description First immunization

Female C57BL/6J01aHsd (C57BL/6) wild-type mice (Harlan, USA) were received at 9 weeks of age. Vaccinations started at 10 weeks. Mice were vaccinated with the low complexity domain (LCD, a. a. 274-414) of TDP-43 protein presented on the surface of liposomes in the presence of Monophosphoryl Hexa-acyl Lipid A as adjuvant. Mice were vaccinated by 200 pl subcutaneous injection (s.c.) on days 0, 14, 28. Mice were bled and heparinized plasma prepared 7 days before immunization (pre-immune plasma) and on days 21 and 35 after the first immunization. A second immunization was performed with different antigens as summarized below.

Second immunization with aggregated LCD TDP-43 and hybridoma selection

Mice were vaccinated with the aggregated low complexity domain (LCD, a.a. 274-414) of TDP- 43 protein in presence of CpG-Alum as adjuvant. Mice were vaccinated by 200 pl subcutaneous injection (s.c.) on days 84 and 98. Mice were bled and heparinized plasma prepared at day 105 after the first immunization. Plasma immunoreactivity to aggregated LCD was tested by ELISA (data not shown).

Mice used for myeloma fusion were additionally vaccinated with aggregated LCD 3 days and 1 day prior to fusion. Mice were euthanized and fusion with myeloma cells was performed.

Screening for antibodies from the successfully fused hybridoma supernatant was performed by ELISA using two coated antigens in parallel, FL TDP-43 and aggregated LCD TDP-43. Seven clones with binding to FL TDP-43 and aggregated LCD TDP-43 were selected for further subcloning. After subcloning, 4 subclones secreting antibody were selected for characterization, which led to the identification of ACI-8062-3D4-Abl antibody.

Second immunization with aggregated FL TDP-43

Mice were vaccinated with full length aggregated TDP-43 protein in presence of Sigma adjuvant system as adjuvant. Mice were vaccinated by 200 pl subcutaneous injection (s.c.) on days 79 and 93. Mice were bled and heparinized plasma prepared at day 100 after the first immunization. Plasma immunoreactivity to aggregated TDP-43 was tested by ELISA (data not shown).

The 2 mice selected for myeloma fusion were additionally vaccinated 3 days and 1 day prior to fusion. Mice were euthanized and fusion with myeloma cells was performed. Screening for antibodies from the successfully fused hybridoma supernatant was performed by ELISA using two coated antigens in parallel, FL TDP-43 and aggregated LCD TDP-43. Ten clones with binding to FL TDP-43 and aggregated LCD TDP-43 were selected for further subcloning. After 2 rounds of subcloning, 12 subclones secreting antibody were selected for characterization, which led to the identification of ACI-8062-9A9-Abl, ACI-8062-1 lAl-Abl and ACI-8062-12Bl-Abl antibodies.

Binding to TDP-43

The binding of ACI-8062-3D4-Abl, ACI-8062-9A9-Abl, ACI-8062-11 Al- Ab 1 and ACI-8062- 12B1-Abl recombinant antibody to full length TDP-43 was determined by ELISA (Table 3).

Table 3. ECso values determined by ELISA

Example 2. Characterization of antibodies by Surface Plasmon resonance (SPR) Measurements were performed on a Biacore 8K instrument (GE Healthcare Life Sciences) by immobilizing soluble TDP-43 on a CM5 Series S sensor chip (GE Healthcare, BR-1005-30).

KD determination for soluble TDP-43 by SPR

The instrument was primed with running buffer PBS-P+ and flow cells (Fc) 1 and 2 of channels 1-8 were activated with a fresh solution of EDC/NHS (Amine Coupling Kit, 1 : 1 ratio of both reagents, GE Healthcare Life Sciences, BR-1006-33) at 10 pL/min for 420 sec. Soluble TDP-43 (Selvita) was diluted in sodium acetate pH 4.5 to a final concentration of 5 pg/mL and injected for 80 sec with a flow rate of 10 pL/min on Fc 2. All flow cells were quenched with 1 M ethanolamine (GE Healthcare Life Sciences, BR-1006-33), at 10 pL/min for 420 sec. Immobilization levels after ethanolamine quenching were around 680 RU on all eight channels. Prior to analysis, three Startup cycles were run. Increasing mAbs concentrations were injected in single-cycle kinetics ranging from 1.2 to 100 nM, prepared from a 3-fold serial dilution in running buffer, with a contact time of 300 sec and a dissociation time of 900 sec at a flow rate of 30 pL/min. Each cycle was followed by one regeneration using 10 mM Glycine-HCl pH 1.7 with a contact time of 30 sec at 10 pL/min, followed by a stabilization period of 180 sec. Results obtained from single-cycle kinetics were double-referenced using the blank Fc 1 and buffer cycles and evaluated using Biacore 8K evaluation software using the 1 : 1 kinetic fit model with RI and global Rmax. The following kinetic parameters were obtained (Table 4): association rate constant (k_a), dissociation rate constant (ka), affinity constant (KD).

Table 4. k_a, ka, KD value of antibodies on soluble TDP-43

All four antibodies demonstrated a high binding affinity to TDP-43 ranging from 1.91 nM to 408 pM.

Example 3. Determination of binding regions

By peptide array

Epitope mapping was confirmed using a custom-made peptide array library (Pepscan, Netherlands). Briefly, arrays of overlapping linear peptides covering the entire TDP-43 were used to define epitopes.

Synthesis of peptides

To reconstruct epitopes of the target molecule, a library of peptide-based mimics was synthesized using Fmoc-based solid-phase peptide synthesis. An amino functionalized polypropylene support was obtained by grafting with a proprietary hydrophilic polymer formulation, followed by reaction with t-butyloxycarbonyl-hexamethylenediamine (BocHMDA) using dicyclohexylcarbodiimide (DCC) with N-hydroxybenzotriazole (HOBt) and subsequent cleavage of the Boc-groups using trifluoroacetic acid (TFA). Standard Fmoc-peptide synthesis was used to synthesize peptides on the amino-functionalized solid support by custom modified JANUS liquid handling stations (Perkin Elmer).

ELISA screening

The binding of antibody to each of the synthesized peptides was tested in a pepscan-based ELISA. The peptide arrays were incubated with primary antibody solution (overnight at 4°C). After washing, the peptide arrays were incubated with a 1/1000 dilution of an appropriate antibody peroxidase conjugate for one hour at 25°C. After washing, the peroxidase substrate 2,2’-azino-di-3- ethylbenzthiazoline sulfonate (ABTS) and 20 pl/ml of 3 percent H2O2 were added. After one hour, the colour development was measured. The colour development was quantified with a charge coupled device (CCD) camera and an image processing system. Determined epitopes are provided in Table 5.

Table 5. Epitopes/Binding regions for tested antibodies

By ELISA

Recombinantly produced antibody _ACI-8062-9A9-Abl was screened in an indirect ELIS A assay to determine binding region using linear peptides TP-51 and TDP-3 (Table 6). 96-well plates were coated with 100 nM of peptides overnight in carbonate buffer at 4°C. Plates were washed with 0.05% Tween-20/PBS and then blocked with 1% bovine serum albumin (BSA) in 0.05% Tween-20/PBS for 1 hour at 37°C. The antibody was then added at 1 pg/ml and incubated for 2 hours at 37°C after which the plates were washed. An AP-conjugated anti-mouse IgG secondary antibody was added at 1/1000 dilution in 0.05% Tween-20/PBS for 1 hour at 37°C. After the final wash, plates were incubated with pNPP (Sigma-Aldrich, Switzerland) phosphatase substrate solution and read at 405 nm using an ELISA plate reader (Tecan, Switzerland).

Table 6. Peptides used for determination of binding regions by ELISA

ACI-8062-9A9-Abl antibody was found to bind TP-51 but not TDP-3 suggesting binding to an epitope within amino acids 370-414 of TDP-43.

Example 4. Detection of TDP-43 in brain tissues from FTLD-TDP subjects by immunohistochemistry

Target engagement was evaluated in immunohistochemistry experiments on tissues from FTLD- TDP subject brains. Human FTLD-TDP brain tissues were obtained from UCSF Neurodegenerative Disease Brain Bank. All material has been collected from donors from whom a written informed consent for brain autopsy and the use of the material and clinical information for research purposes has been obtained by the brain bank. Immunohistochemistry was performed on 10 pm thick frozen sections using fluorescently labelled secondary antibody for detection. The rat monoclonal anti -phospho TDP-43 p409/410 antibody (Biolegend, 829901) was used as a positive control to detect phosphorylated TDP-43 and a secondary antibody without primary antibody (No 1° Ab) was used as negative control to detect non-specific background.

ACI-8062-l lAl-Abl, ACI-8062-12Bl-Abl and ACI-8062-3D4-Abl specifically bound misfolded and aggregated TDP-43 and did not recognize non-aggregated physiological nuclear TDP-43. ACI-8062-9A9-Abl recognized both non-aggregated physiological nuclear TDP-43 and misfolded and aggregated TDP-43, but showed preferential binding to misfolded and aggregated TDP-43. The detailed evaluation of binding characteristics is summarized in Table 7.

Table 7. Detection of TDP-43 in brain tissues from FTLD-TDP subjects

NA data not available; - absent; + weak; ++ medium; +++ abundant Example 5. In vitro functionality in recombinant TDP-43 aggregation assay

To evaluate the functionality of antibodies in vitro, the ability of ACI-8062-11 Al-Abl and ACI- 8O62-12B1-Abl to inhibit TDP-43 aggregation was tested. FL TDP-43 was fused at C-terminus to maltose binding protein (MBP) which was separated by a Tobacco Etch Virus (TEV) protease cleavage site and produced recombinantly. Aggregation of 2.5 pM TDP-43 -TEV-MBP fusion protein in 30 mM Tris, 150 mM NaCl, pH 7.4 in the presence of 600 nM of each anti-TDP-43 antibody or negative control mAb that does not bind to TDP-43 was induced by addition of TEV protease (AcTEV, Invitrogen) and absorbance was monitored in a pClear® 96 well plate (Greiner) at 600 nm over 20 h. For evaluation, end points were normalized to the negative control mAb and the percentage of TDP-43 aggregation inhibition compared to the negative control mAb was calculated for each antibody.

ACI-8062-12Bl-Abl inhibited TDP-43 aggregation by 72% while ACI-8062-11 Al-Abl inhibited TDP-43 aggregation by 95% compared to the negative control mAb. This data confirmed the therapeutic potential of the antibodies of the invention.

Example 6. Inhibition of TDP-43 aggregation in neurons in vitro by AAV-delivered intrabody

To evaluate the functionality of antibodies in an intrabody format, ACI-8062-11 Al-Abl antibody was vectorized as an scFv in an AAV6 under the control of a SYN1 promoter. The scFv consisted of the VL of ACI-8062-11 Al-Abl antibody (SEQ ID NO: 54) linked to the VH (SEQ ID NO: 50) by a peptide linker having the amino acid sequence (G4EsS)4 (SEQ ID NO: 3). The vectorized intrabody was transduced in an in vitro model of TDP-43 aggregation consisting of rat primary neurons expressing human TDP-43 with a mutated nuclear localisation signal (NLSm) and fused to an EGFP. The decrease in TDP-43 aggregation was assessed by immunofluorescent staining and fluorescent puncta quantification.

Briefly, rat primary neuron cultures were prepared from rat pup brains. The pups were dissected in HBSS under a binocular microscope at room temperature. The brains were kept in MEM (Gibco, 12360-038) before cutting and transfer into a digestion medium (30 mM K2SO4, 90 mM Na2SO4, 5.8 mM MgC12, 0.25 mM CaC12, 1.6 mM HEPES, 0.2 mM NaOH, 0.5% phenol red, 20 U/mL Papain, 0.1 % (w/v) L-Cysteine, 0.36 % (w/v) Glucose) for 30 minutes at 37°C. Supernatant was removed and 10 mL of protease inhibitor medium was added to the tissue (30 mM K2SO4, 90 mM Na2SO4, 5.8 mM MgC12, 0.25 mM CaC12, 1.6 mM HEPES, 0.2 mM NaOH, 0.5% phenol red, 0.36 % (w/v) Glucose, 0.1 % (w/v) trypsin inhibitor (Sigma, T9253)), incubated for 10 minutes, centrifuged at 300 g for 2 min and the supernatant discarded. Tissue was resuspended in 5 mL of trituration medium (10 % horse serum (Sigma, Hl 138), 1% L- Glutamine (Gibco, 25030-025), 0.36% (w/v) Glucose in MEM) and triturated using serological pipette. Once all the tissue was dissolved, the tube was centrifuged at 300 g for 2 min, the supernatant discarded, the cells resuspended in 10 mL of adhesion medium (10% horse serum, 1% L-Glutamine, 1% Pen/Strep, 0.36% (w/v) glucose in MEM) and passed through a 70 pm cell strainer. Cells were counted and plated at 25,000 cells/well in 96-well Poly-D-Lysin-coated plates in 100 pL and incubated at 37°C and 5% CO2. After 4 h, the medium was changed for a growth medium (1% L-Glutamine, 1% Pen/Strep, 2% B27 (Gibco, 17504-044) in Neurobasal medium (Gibco, 12348-017)). After 4 days in vitro, cells were treated with 2.5 pM AraC (Sigma, Cl 768) diluted at 10X in growth medium to promote astrocyte cell death. After 5 days of culture, cells were co-transduced with an AAV6 vector expressing EGFP-TDP-43-NLSm under the SYN1 promoter at 8,000 genome copies/cell and an AAV6 vector expressing either ACI-8062- 11 Al-Abl scFv or a negative control scFv (that does not bind TDP-43) as intrabodies under the SYN1 promoter at 32,000 genome copies/cell and incubated for 13 days. Both scFvs comprised a FLAG tag in C-terminal to assess their proper expression and localization in cell (e.g., nuclear or cytosolic). Cells were fixed for 20 min using 4% PFA in PBS and washed three times with PBS before immunostaining with anti-FLAG (Sigma F3165) and anti-MAP2 (abeam ab5392) primary antibodies followed by anti-mouse Alexa-Fluor 647 (Invitrogen A21237) and antichicken Alexa Fluor 555 (Invitrogen A21437) secondary antibodies and DAPI counterstaining. Image acquisition was performed using an IN Cell Analyzer 2200 (GE Healthcare) with a Nikon 20X/0.75, Plan Apo, CFE60 objective. An analysis pipeline was generated using Cell Profiler v4.2.1 consisting of image segmentation based on recognition of nuclei (DAPI) to count cells, and recognition of neuronal network using MAP2 (Cy5). Cell body overexpressing EGFP were segmented out of the MAP2 mask and EGFP positive aggregates were counted in the neurite mask (MAP2 total - cell body) to obtain the number of aggregates per cell. Statistical analysis was performed on 4 independent experiments with 6 technical replicates using Two-way ANOVA followed by multiple comparisons with Dunnett’s correction (**** = p-value<0.0001).

Immunostaining with the anti-FLAG antibody showed that both the negative control scFv and ACI-8062-11 Al-Abl were properly expressed in the rat primary neurons (Figure 1). Importantly, ACI-8062-11 Al-Abl expression is restricted to the cell cytosol, with no expression detected in the nucleus where physiological TDP-43 maintains essential functions (Figure 1). ACI-8062-1 lAl-Abl was able to significantly inhibit TDP-43 aggregation in rat primary neurons compared to the EGFP-TDP-43NLSm control while the negative control scFv did not (Figure 2). This result confirmed the therapeutic potential of the antibodies of the invention as an scFv intrabody. Example 7. Antibody sequencing

Clonal hybridoma cell lysates were used for gene sequencing of the variable region. Mouse hybridomas were harvested and lysed using a lysis buffer containing guanidinium salts to deactivate RNases. cDNA was obtained by reverse transcription of total mRNA. DNA fragments coding for antibody variable region were amplified by RACE-PCR (Takara Bio, cat# 634839) using specific primer annealing in the antibody constant region. PCR products were gel purified and cloned into shuttle vector for Sanger sequencing. Sequencing was carried out in both directions to provide overlap at both ends. The sequences were analysed using multiple sequence alignment (Clustal tool) and annotated using the algorithm of Kabat as described in Kabat et al., Sequences of Proteins of Immunological Interest, 91-3242 (1991). Nucleotide sequences of the Heavy Chain (VH) and Light Chain (VL) Variable Regions are shown in Table 8. Translated protein sequences for selected Heavy (VH) and Light (VL) Chain Variable Regions, and their complementarity-determining regions (CDRs) are shown in Table 9.

Table 8. Nucleotide sequence of the Heavy Chain (VH) and Light Chain (VL) Variable Regions

Table 9. Amino acid sequence of the Heavy Chain (VH) and Light Chain (VL) Variable Regions and their CDRs

Example 8. Humanization of ACI-8062-llAl-Abl antibody

The use of scFv can be associated with challenges related to stability, expression and loss of affinity upon reformatting from standard IgG (Worn and Pliickthun, 2001). Hence, these considerations were taken into account during the humanization process of the ACI-8062- HAl-Abl antibody.

The selection of human germlines for humanization of ACI-8062-1 lAl-Abl was based on sequence identity with the mouse sequence, putative stability and isoelectric point (pl). Databases of human and mouse germline variable genes such as the IMGT database (Ehrenmann et al., 2010) or IgBlast (Ye et al., 2013) were used to identify the closest human variable domain subfamilies to the murine heavy and light chain variable regions of ACI-8062- 11 Al-Abl antibody. IGHV3-23 (77.6% sequence identity with parental mouse VH and a pl of 8.1) was selected as VH framework for humanization, while IGKV2-28 (71% sequence identity with parental mouse VL, pl of 4.7), IGKV4-1 (59.4% sequence identity with parental mouse VL, pl of 4.7) and IGKV1-39 (51% sequence identity with parental mouse VL, pl of 7.3) were selected as VL frameworks.

As a starting point for the humanization process, murine CDRs were grafted on IGHV3-23, IGKV2-28, IGKV4-1 and IGKV1-39 human acceptor frameworks. To retain the conformation of CDRs, key positions in the human acceptor frameworks were modified in some constructs by substituting human to mouse residues, and these backmutations are listed in Table 10.

Table 10: Backmutations introduced in the VH and VL human frameworks

In the VL-CDR2, a conservative substitution, D55E, was introduced in some constructs to replace the murine residue (D) with the human counterpart (E) to increase the percentage sequence identity to the IGKV4-1 human framework.

Resulting humanized ACI-8062-11 Al-Abl variants are listed in Table 11 (amino acid sequences) and 12 (nucleotide sequences). Table 11. Amino acid sequence of the ACI-8062-llAl-Abl humanized variants Heavy Chain (VH) and Light Chain (VL) Variable Regions and their CDRs

Table 12. Nucleotide sequence of the ACI-8062-llAl-Abl humanized variants Heavy Chain (VH) and Light Chain (VL) Variable Regions

Example 9. Generation of humanized ACI-8062-llAl-Abl scFv antibodies

The variable domains of the murine parent ACI-8062-1 lAl-Abl antibody, as well as the variable domains of the humanized variants of Example 8 were fused with a standard glycineserine linker GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 4) and tagged with a cMyc, Flag and 6xHis tags in C-terminal resulting in the following scFv construct: VH-(G4S)4-VL-cMyc- Flag-6xHis. The amino acid and nucleotide sequences of the scFv constructs are shown in Table 13.

The scFvs were recombinantly produced in CHO cells and affinity purified from cell culture supernatant before characterization. Briefly, DNA fragments coding for each of the scFv constructs were synthesized and individually cloned in pcDNA 3.4 mammalian expression vectors using a Gibson assembly method (NEB). Plasmids were amplified and produced from single positive colonies using midiprep kits (Zymoresearch). The scFvs were transiently expressed in ExpiCHO-S cells (ThermoFisher scientific) by transfecting the scFv coding plasmid using the ExpiCHO™ Expression System Kit (ThermoFisher scientific). Cells were cultured for 7 days at 37 °C under 120 rpm agitation and 8% CO2 level. Supernatants were harvested and recombinant scFvs were affinity purified by adding Ni-Sepharose excel resin (Cytivia) (1 mL of resin per 10 mg of tagged protein). After 30 minutes under agitation at room temperature, the mixture was added into a gravity flow chromatography column (BioRad), the bound antibodies were washed with 20 column volume (CV) of lx PBS, 300 mM NaCl, 25 mM imidazole, pH 8 and eluted with 5 CV of lx PBS, 300 mM NaCl, 250 mM imidazole, pH 8. Samples were then buffer-exchanged in PBS buffer by dialysis (Sigma-Aldrich) and stored at 4 °C.

Example 10. Characterization of humanized ACI-8062-1 lAl-Abl scFv antibodies

All humanized antibodies were characterized for thermostability and affinity binding to TDP- 43.

Expression and thermostability

Thermostability was evaluated by differential scanning fluorimetry (DSF) using a QuantStudio™ 3 Real-Time PCR System (ThermoFisher scientific). Briefly, 5 pg of protein, diluted in thermal shift assay buffer were mixed with 2 pL of 5*Protein Thermal Shift Dye (ThermoFisher Scientific) in a final volume of 20 pl in each well of a 96 well plate (ThermoFisher Scientific). Samples were scanned from 25 °C to 100 °C at a rate of 0.05 °C/s. The melting temperature (Tm) was calculated using the derivative method and are reported in Table 13.

Table 13. Expression yield and thermostability of humanized ACI-8062-llAl-Abl scFvs

All humanized scFv were expressed in CHO cells. Although all scFv constructs comprise the same VH, various expression level and thermostability were measured highlighting the critical role of the VL on the humanized ACI-8062-11 Al-Abl scFvs stability.

Binding affinity

Binding affinities were measured by Bio-layer interferometry (BLI) using an Octet QKe (Sartorius). Human TDP-43 was diluted at 50 nM in 10 mM acetate pH 4 and immobilized by amine coupling on preactivated Octet® AR2G Biosensors (Sartorius). After 300 seconds of equilibration in assay buffer (lx PBS, 0.1% BSA, 0.01% Tween-20), immobilized TDP-43 was tested against purified scFv ranging from 4.69-300 nM prepared from serial 2-fold dilutions in assay buffer with an association of 300 seconds and a dissociation in assay buffer of 600 seconds. Each serial dilutions included a reference sample containing assay buffer only. Between measurements, biosensors were regenerated for 30 seconds in 10 mM Glycine pH 2 and equilibrated in assay buffer for 120 seconds. Resulting sensorgrams were referenced subtracted using the dedicated reference sample and analyzed using the Octet Data Analysis HT software with 1 : 1 kinetic fitting model. The following kinetic parameters were obtained: association rate constant (k_a), dissociation rate constant (ka), affinity constant (KD) and are reported in Table 14. Table 14. Binding affinity to human TDP-43 of humanized ACI-8062-llAl-Abl scFvs

The H2L3, H2L4, H2L5 and H2L6 scFvs (humanized using the VH/VL frameworks IGHV3- 23/IGKV4-1) demonstrated similar binding affinities to human TDP-43 ranging from 27 to 55 nM, while the H2L1 scFv (humanized using the VH/VL frameworks IGHV3-23/IGKV4-1) showed a lower binding affinity to human TDP-43.

According to the tests performed in the examples described above, hACI-8062-11A1- Abl_H2L3 and hACI-8062-1 lAl-Abl_H2L5 scFv displayed excellent biophysical properties with the overall highest expression yield, thermostability and binding affinity to TDP-43.

Example 11. Inhibition of TDP-43 aggregation in neurons in vitro by AAV-delivered humanized ACI-8062-llAl-Abl scFv intrabody

To evaluate the intracellular expression and functionality of humanized ACI-8062-1 lAl-Abl scFv variants as an intrabody format, the parent murine ACI-8062-1 lAl-Abl and humanized variants H2L3 and H2L5 scFv were vectorized in an AAV6 under the control of a human SYN 1 promoter. The scFv constructs were designed with the VH-VL orientation linked by a (G4EsS)4 linker (SEQ ID NO: 3) and fused to a cMyc and FLAG tag in C-terminal. The vectorized intrabodies were transduced in an in vitro model of TDP-43 pathology consisting of rat primary neurons expressing human TDP-43 with a mutated nuclear localization signal (NLSm) and fused to an eGFP. The decrease in TDP-43 aggregation was assessed by quantifying pTDP-43 by immunofluorescent staining and immunoreactive area quantification.

Briefly, rat primary neuron cultures were prepared as in Example 6, except that cells were plated at 30,000 cells/well. After 5 days of culture, cells were co-transduced with an AAV6 vector expressing eGFP-TDP-43-NLSm under the human SYN1 promoter at 10,000 genome copies/cell and an AAV6 vector expressing either the murine ACI-8062-11 Al- Ab 1 scFv, the hACI-8062-1 lAl-Abl_H2L3 scFv, the hACI-8062-1 lAl-Abl_H2L5 scFv, or a negative control scFv (that does not bind TDP-43) as intrabodies under the human SYN1 promoter at 60,000 genome copies/cell (for expression analysis) or 30,000 genome copies/cell (for efficacy assessment) and incubated for 6 days. Cells were treated with 5 pM sodium arsenite for 18 hours to induce stress prior to fixation and immunofluorescence staining as described in Example 6 or with the addition of an anti-pTDP-43 antibody (Biolegend 829901) followed by anti-rat Alexa-Fluor 633 (Invitrogen A21094). Image acquisition was performed using an INCell Analyzer 2200 (GE Healthcare) with a Nikon 20X/0.75, Plan Apo, CFV60 objective. An analysis pipeline was generated using Cell Profiler v4.2.1 consisting of image segmentation based on recognition of nuclei (Hoechst 33342) to count cells, and recognition of neuronal network using MAP2 (Cy5). Cell bodies were segmented using MAP2 staining and pTDP-43 positive immunoreactive area was quantified in the MAP2 positive neuronal mask. To quantify intrabodies expression, the integrated intensity of the FLAG immunoreactive area was reported as a percentage increase over the murine ACI-8062-11 Al-Abl scFv. For efficacy, the mean of 3 replicates were reported and a Kruskal-Wallis test with Dunn’s correction for multiple comparisons was performed, *p>0.05.

Immunostaining with the anti -FLAG antibody and quantification of the FLAG immunoreactive area confirmed the improved intracellular expression of humanized hACI-8062-11A1- Abl_H2L3 (19.2% increase in expression) and hACI-8062-11 Al-Abl_H2L5 (30.2% increase in expression) scFvs compared to the parental murine ACI-8062-11 Al -Ab 1 scFv in rat primary neurons (Figure 3).

Murine ACI-8062-11 Al-Abl scFv and humanized hACI-8062-1 lAl-Abl_H2L3 and hACI- 8062-1 lAl-Abl_H2L5 scFvs were able to inhibit pTDP-43 aggregates formation in rat primary neurons compared to the negative control scFv (Figure 4). The hACI-8062-11A1- Abl_H2L3 and hACI-8062-1 lAl-Abl_H2L5 scFvs were more potent at inhibiting pTDP-43 aggregates than the parental murine ACI-8062-11 Al-Abl scFv. These results confirmed the therapeutic potential of the antibodies of the invention. REFERENCES

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Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications and patents specifically mentioned herein are incorporated by reference in their entirety for all purposes in connection with the invention.

The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims. Moreover, all aspects and embodiments of the invention described herein are considered to be broadly applicable and combinable with any and all other consistent embodiments, including those taken from other aspects of the invention (including in isolation) as appropriate.

Claims

1. A vector for targeted delivery to the CNS comprising a nucleic acid sequence encoding a TDP-43 binding molecule which at least preferentially binds misfolded TDP-43 over physiologically functional TDP-43 and preferably specifically binds misfolded TDP-43 and does not bind physiologically functional TDP-43, and which binds to an epitope within amino acid residues 202-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

2. The vector of claim 1, wherein the vector is a viral vector.

3. The vector of claim 1 or 2, wherein the vector is for targeted delivery to neurons and/or glial cells, preferably neurons, and wherein the neurons and/or glial cells express the TDP- 43 binding molecule as an intrabody.

4. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or e. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL- CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

5. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84; or d. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 94 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 94; or e. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 104 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; or f. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 114 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 114; or g. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 124 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; or h. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 134 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 134; or i. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 144 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 144.

6. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 40 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 80 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 84; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or h. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or i. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

7. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule binds to an epitope consisting of amino acid residues 202-209 or 207-211 of human TDP-43 (SEQ ID NO: 1).

8. The vector of any one of claims 1 to 6, wherein the TDP-43 binding molecule binds to an epitope within amino acid residues 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

9. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule binds to misfolded monomeric and/or misfolded oligomeric and/or misfolded aggregated and/or misfolded post-translationally modified and/or truncated misfolded TDP-43.

10. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule exhibits one or more, up to all of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof; c. blocks TDP-43 cell-to-cell propagation; d. disaggregates TDP-43 aggregates; e. blocks TDP-43 seeding; f. neutralizes seeding-competent TDP-43; g. blocks TDP-43 spreading; h. potentiates intracellular and/or extracellular clearance of pathological TDP-43; i. restores nuclear levels and physiological function of TDP-43; and j. reduces phosphorylated TDP-43 level.

11. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule exhibits one or both of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; and b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof.

12. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule has a dissociation constant (KD) for binding TDP-43 (SEQ ID NO: 1) of 2 nM or less, 1 nM or less, or 500 pM or less.

13. The vector of any one of claims 1 to 11, wherein the TDP-43 binding molecule has a dissociation constant (KD) for binding TDP-43 (SEQ ID NO: 1) of 150 nM or less, 100 nM or less, 50nM or less, or 35nM or less.

14. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule has a melting temperature of at least 56 °C, at least 58 °C, at least 60 °C, at least 62 °C or at least 63 °C.

15. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule is an antibody or antigen-binding fragment thereof, preferably an intrabody.

16. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule is an IgA, IgD, IgE, IgM, IgGl, IgG2, IgG3 or IgG4 antibody or antigen-binding fragment thereof.

17. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule is an IgGl or an IgG4 antibody or antigen-binding fragment thereof.

18. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule comprises an Fc mutation, preferably the S228P mutation.

19. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule is a single chain variable fragment (scFv), optionally wherein the linker linking the VH and VL of the scFv comprises the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4.

20. The vector of any one of the preceding claims, wherein the vector is a wild-type or an engineered AAV vector.

21. The vector of any one of the preceding claims, wherein the vector is a wild-type or an engineered AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh.8, AAVrh.9, AAVrh.10, AAVrh. l l, AAVrh.12, AAVrh.13, AAVrh.14, AAVrh.15, AAVrh.16, AAVrh.17 or AAV.Hu68.

22. The vector of any one of the preceding claims, wherein the vector is an AAV vector and wherein the TDP-43 binding molecule is a single chain variable fragment (scFv) comprising: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL- CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

23. The AAV vector of claim 22, wherein the single chain variable fragment (scFv) comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or h. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

24. The AAV vector of claim 22 or 23, wherein the single chain variable fragment (scFv) comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134.

25. The vector of any one of the preceding claims, wherein the TDP-43 binding molecule is expressed under the control of a CNS-specific promoter, preferably a human SYN1 promoter or a CMVe-SYNl promoter.

26. A TDP-43 binding molecule which at least preferentially binds misfolded TDP-43 over physiologically functional TDP-43 and preferably specifically binds misfolded TDP-43 and does not bind physiologically functional TDP-43, which binds to an epitope within amino acid residues 202-211, 353-373 or 370-414 of human TDP-43 (SEQ ID NO: 1).

27. The TDP-43 binding molecule of claim 26, wherein the TDP-43 binding molecule binds to an epitope consisting of amino acid residues 202-209 or 207-211 of human TDP-43 (SEQ ID NO: 1).

28. The TDP-43 binding molecule of claim 26, wherein the TDP-43 binding molecule binds to an epitope within amino acid residues 353-373 or within amino acid residues 370-414 of human TDP-43 (SEQ ID NO: 1).

29. The TDP-43 binding molecule of any one of claims 26 to 28, wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 57; or b. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or c. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62 and a VH-CDR3 comprising the amino acid sequence GPF (Gly-Pro-Phe); and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; or d. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86 and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; or e. a Heavy Chain Variable Region (VH) which comprises a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52 and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and a Light Chain Variable Region (VL) which comprises a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and a VL- CDR3 comprising the amino acid sequence of SEQ ID NO: 57.

30. The TDP-43 binding molecule of any one of claims 26 or 29, wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 50 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 50; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 54 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 40 or a Heavy Chain Variable Region (VH) having at least 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 40; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 44 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 60 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 60; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 64 or a Light Chain Variable Region (VL) having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 80 or a Heavy Chain Variable Region (VH) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; and a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 84; or e. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 94 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 94; or f. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 104 or a Light Chain Variable Region (VL) having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; or g. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 114 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 114; or h. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 124 or a Light Chain Variable Region (VL) having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; or i. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 134 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 134; or j. a Heavy Chain Variable Region (VH) comprising the sequence of SEQ ID NO: 90 or a Heavy Chain Variable Region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 90; a Light Chain Variable Region (VL) comprising the sequence of SEQ ID NO: 144 or a Light Chain Variable Region (VL) having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 144.

31. The TDP-43 binding molecule of any one of claims 26 to 30, wherein the TDP-43 binding molecule comprises: a. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 50 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 54; or b. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 40 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 44; or c. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 60 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 64; or d. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 80 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 84; or e. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 94; or f. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 104; or g. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 114; or h. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 124; or i. a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 134; or j . a Heavy Chain Variable Region (VH) comprising the amino acid sequence of SEQ ID NO: 90 and a Light Chain Variable Region (VL) comprising the amino acid sequence of SEQ ID NO: 144.

32. The TDP-43 binding molecule of any one of claims 26 to 31, wherein the TDP-43 binding molecule binds to misfolded monomeric and/or misfolded oligomeric and/or misfolded aggregated and/or misfolded post-translationally modified and/or truncated misfolded TDP-43.

33. The TDP-43 binding molecule of any one of claims 26 to 32, wherein the TDP-43 binding molecule exhibits one or more, up to all of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof; c. blocks TDP-43 cell-to-cell propagation; d. disaggregates TDP-43 aggregates; e. blocks TDP-43 seeding; f. neutralizes seeding-competent TDP-43; g. blocks TDP-43 spreading; h. potentiates intracellular and/or extracellular clearance of pathological TDP-43; i. restores nuclear levels and physiological function of TDP-43; and j. reduces phosphorylated TDP-43 level.

34. The TDP-43 binding molecule of any one of claims 26 to 33, wherein the TDP-43 binding molecule exhibits one or both of the following characteristics: a. inhibits the extracellular aggregation of TDP-43 protein or fragments thereof; and b. inhibits the intracellular aggregation of TDP-43 protein or fragments thereof.

35. The TDP-43 binding molecule of any one of claims 26 to 34, wherein the TDP-43 binding molecule has a dissociation constant (KD) for binding TDP-43 (SEQ ID NO: 1) of 2 nM or less, 1 nM or less, or 500 pM or less.

36. The TDP-43 binding molecule of any one of claims 26 to 34, wherein the TDP-43 binding molecule has a dissociation constant (KD) for binding TDP-43 (SEQ ID NO: 1) of 150 nM or less, 100 nM or less, 50nM or less, or 35nM or less.

37. The TDP-43 binding molecule of any one of claims 26 to 36, wherein the TDP-43 binding molecule has a melting temperature of at least 56 °C, at least 58 °C, at least 60 °C, at least 62 °C or at least 63 °C.

38. The TDP-43 binding molecule of any one of claims 26 to 37, wherein the TDP-43 binding molecule is an antibody or antigen-binding fragment thereof.

39. The TDP-43 binding molecule of any one of claims 26 to 38, wherein the TDP-43 binding molecule is an IgA, IgD, IgE, IgM, IgGl, IgG2, IgG3 or IgG4 antibody or antigen-binding fragment thereof.

40. The TDP-43 binding molecule of any one of claims 26 to 39, wherein the TDP-43 binding molecule is an IgGl or an IgG4 antibody or antigen-binding fragment thereof.

41. The TDP-43 binding molecule of any one of claims 26 to 40, wherein the TDP-43 binding molecule comprises an Fc mutation, preferably the S228P mutation.

42. The TDP-43 binding molecule of any one of claims 26 to 41, wherein the TDP-43 binding molecule is a single chain variable fragment (scFv).

43. An immunoconjugate comprising the TDP-43 binding molecule according to any one of claims 26 to 42.

44. The immunoconjugate of claim 43, wherein the immunoconjugate crosses the blood brain barrier using a delivery vehicle or a blood brain barrier moiety.

45. The immunoconjugate of claim 44, wherein the delivery vehicle comprises a liposome or extracellular vesicle.

46. The immunoconjugate of claim 44, wherein the TDP-43 binding molecule is linked to the blood brain barrier moiety.

47. The immunoconjugate of claim 44 or 46, wherein the blood brain barrier moiety is a polypeptide or a small molecule, preferably, a peptide, a receptor ligand, a single domain antibody (VHH), a scFv or a Fab fragment.

48. The immunoconjugate of claim 44, 46 or 47, wherein the blood brain barrier moiety binds a blood brain barrier receptor.

49. The immunoconjugate of claim 48, wherein the blood brain barrier receptor comprises one or more of a transferrin receptor, insulin receptor or low-density lipoprotein receptor.

50. A labelled binding molecule, in particular a labelled antibody, comprising the TDP-43 binding molecule according to any one of claims 26 to 42.

51. A pharmaceutical composition comprising the vector of any one of claims 1 to 25, or the TDP-43 binding molecule of any one of claims 26 to 42, or the immunoconjugate of any one of claims 43 to 49 and a pharmaceutically acceptable carrier and/or excipient and/or diluent.

52. The vector of any one of claims 1 to 25, the TDP-43 binding molecule of any one of claims 26 to 42, the immunoconjugate of any one of claims 43 to 49, or the pharmaceutical composition of claim 51 for human or veterinary therapeutic use.

53. The vector of any one of claims 1 to 25, the TDP-43 binding molecule of any one of claims 26 to 42, the immunoconjugate of any one of claims 43 to 49 or the pharmaceutical composition of claim 51 for use in the prevention, alleviation, treatment of diseases, disorders and/or abnormalities associated with TDP-43 or TDP-43 proteinopathy.

54. The vector or pharmaceutical composition for use of claim 53, wherein the vector or pharmaceutical composition is administered by intravenous, intravitreous, subretinal, intrathecal, intracerebroventricular, intraci sternal (intracistema magna), intrastriatal or intraparenchymal route.

55. The TDP-43 binding molecule, immunoconjugate or pharmaceutical composition for use of claim 53, wherein the TDP-43 binding molecule, immunoconjugate or pharmaceutical composition is administered by intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

56. The TDP-43 binding molecule of any one of claims 26 to 42, the immunoconjugate of any one of claims 43 to 49, the labelled binding molecule of claim 50, or the pharmaceutical composition of claim 51 for diagnostic use.

57. The TDP-43 binding molecule, immunoconjugate, labelled binding molecule or pharmaceutical composition for diagnostic use according to claim 56 for diagnosis of diseases, disorders and/or abnormalities associated with TDP-43 or TDP-43 proteinopathy.

58. The vector of any one of claims 1 to 25, the TDP-43 binding molecule of any one of claims 26 to 42, the immunoconjugate of any one of claims 43 to 49, the labelled binding molecule of claim 50 or the pharmaceutical composition of claim 51 for research use, in particular as an analytical tool or reference molecule.

59. The TDP-43 binding molecule of any one of claims 26 to 42, the immunoconjugate of any one of claims 43 to 49, the labelled binding molecule of claim 50, or the pharmaceutical composition of claim 51 for use as a diagnostic tool to monitor diseases, disorders and/or abnormalities associated with TDP-43 or TDP-43 proteinopathy.

60. The TDP-43 binding molecule, immunoconjugate or pharmaceutical composition for use according to claim 53 or 55, or the vector or pharmaceutical composition for use according to claim 53 or 54, or the TDP-43 binding molecule, immunoconjugate, labelled binding molecule or pharmaceutical composition for use according to claim 57 or 59, wherein the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD), Amyotrophic lateral sclerosis (ALS), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), Chronic Traumatic Encephalopathy (CTE), Perry syndrome, Alzheimer’s disease (AD), Down syndrome, Familial British dementia, a Polyglutamine disease, Hippocampal sclerosis dementia, a Myopathy, Traumatic Brain Injury (TBI), Dementia with Lewy Bodies (DLB) or Parkinson’s disease (PD).

61. The TDP-43 binding molecule, immunoconjugate, labelled binding molecule, vector or pharmaceutical composition for use according to claim 60, wherein the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), Chronic Traumatic Encephalopathy (CTE), or limbic- predominant age-related TDP-43 encephalopathy (LATE).

62. The TDP-43 binding molecule, immunoconjugate, labelled binding molecule, vector or pharmaceutical composition for use according to claim 61, wherein the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is amyotrophic lateral sclerosis (ALS).

63. The TDP-43 binding molecule, immunoconjugate, labelled binding molecule, vector or pharmaceutical composition for use according to claim 61, wherein the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Alzheimer’s disease (AD).

64. The TDP-43 binding molecule, immunoconjugate, labelled binding molecule, vector or pharmaceutical composition for use according to claim 61, wherein the disease, disorder and/or abnormality associated with TDP-43, or TDP-43 proteinopathy, is Frontotemporal dementia (FTD).

65. A nucleic acid molecule encoding the TDP-43 binding molecule of any one of claims 26 to 42.

66. The nucleic acid molecule of claim 65, comprising a nucleotide sequence of SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 109, SEQ ID NO: 119, SEQ ID NO: 129, SEQ ID NO: 139 or SEQ ID NO: 149.

67. A nucleic acid molecule encoding a TDP-43 binding molecule comprising the nucleotide sequences set forth as: a. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 58 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 59; or b. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 48 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 49; or c. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 68 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 69; or d. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 88 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 89; or e. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 99; or f. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 119; or g. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 129; or h. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 139; or i. a Heavy Chain Variable Region (VH) encoded by SEQ ID NO: 98 and a Light Chain Variable Region (VL) encoded by SEQ ID NO: 149.

68. An expression vector comprising the nucleic acid of any one of claims 65 to 67.

69. A host cell comprising the nucleic acid of any one of claims 65 to 67 and/or the expression vector of claim 68.

70. A cell-free expression system containing the expression vector of claim 68.

71. A method for producing a TDP-43 binding molecule, in particular an antibody or antigenbinding fragment thereof, comprising the steps of: a. culturing the host cell of claim 69 or cell-free expression system of claim 70 under conditions suitable for producing the TDP-43 binding molecule, in particular the antibody or antigen-binding fragment thereof; and b. isolating the TDP-43 binding molecule, in particular the antibody or antigen-binding fragment thereof.

72. A method of detecting and/or quantifying TDP-43 in a sample obtained from a subject, the method comprising contacting the sample with a TDP-43 binding molecule according to any one of claims 26 to 42 and comparing the TDP-43 levels in the sample to those in a control sample or samples.

73. Use of a TDP-43 binding molecule in a pairing assay comprising the steps of: a) Incubating a sample with a capture antibody and a detect antibody to produce a mixture; b) Incubating the mixture obtained in step a) with a reagent suitable for detection by the detect antibody; c) Measuring the signal emitted by the detect antibody; wherein the detect antibody and/or the capture antibody is/are selected from an antibody as defined in any one of claims 26 to 42.

74. The method of claim 72 or the use according to claim 73, wherein the sample is blood or a blood derivative, cerebrospinal fluid (CSF), interstitial fluid (ISF) and/or urine, preferably CSF.

75. A kit for diagnosis of a disease, disorder and/or abnormality associated with TDP-43 or a TDP-43 proteinopathy, or for use according to any one of claims 56, 57 or 59, comprising a TDP-43 binding molecule according to any one of claims 26 to 42.