Anti-TDP-43 binding molecules and uses thereof

Abstract

To provide TDP-43-specific binding molecules for diagnosing, preventing, ameliorating, and / or treating diseases, disorders, and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathies.SOLUTION: Provided is a TDP-43 binding molecule that is an antibody or an antigen-binding fragment thereof, which binds misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, or a humanized variant thereof.SELECTED DRAWING: None

Description

Technical Field

[0001] Field of the Invention The present invention relates to the field of the transactivation response DNA-binding protein (TARDB or TDP-43) having a molecular weight of 43 kDa. The present invention relates to TDP-43 specific binding molecules, in particular anti-TDP-43 antibodies or antigen-binding fragments or derivatives thereof and their use. The present invention relates 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), including but not limited to, diseases, disorders and / or abnormalities associated with TDP-43, in particular TDP-43 aggregates, or methods and ways for diagnosing, preventing, reducing, and / or treating TDP-43 proteinopathies.

Background Art

[0002] Age-related brain diseases characterized by pathological aggregation of proteins (proteinopathies) in the central nervous system (CNS) and peripheral organs are one of the leading causes of disability and mortality worldwide. The most characterized protein that forms aggregates is amyloid-beta in Alzheimer's disease and related disorders. Aggregation-prone proteins associated with other neurodegenerative diseases include, but are not limited to, Tau, alpha-synuclein (aSyn, a-syn), huntingtin, fused in sarcoma (FUS), dipeptide repeat protein (DPR) (produced by a special translation of C9orf72 repeat expansion), superoxide dismutase 1 (SOD1), and TDP-43. Diseases related to TDP-43 aggregation are generally described as TDP-43 proteinopathies including, but not limited to, ALS and FTD.

[0003] I. Introduction to TDP-43 The transactivation response (TAR) DNA-binding protein 43 kDa (TDP-43) is a 414-amino acid protein encoded by the TARDBP gene on chromosome 1p36.2 (ALS10). TARDBP consists of six exons (exon 1 is non-coding; exons 2-6 are protein-coding). TDP-43 belongs to the heterogeneous ribonucleoprotein (hnRNP) RNA-binding protein family (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) (with two highly conserved hexamer ribonucleoprotein 2 (RNP2) and octamer ribonucleoprotein 1 (RNP1) regions), a nuclear export signal (NES) and a nuclear localization signal (NLS) (which enable the transport of bound mRNA back and forth between the nucleus and the cytoplasm), and a glycine-rich domain at the C-terminus (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 that shuttles intermittently between the nucleus and the cytoplasm, but is mostly localized in the nucleus and has a tightly self-regulated expression level.In 2006, TDP-43 was identified as the protein that accumulates in most cases of frontotemporal lobar degeneration (FTLD) with tau-negative ubiquitin-positive inclusions (referred to as FTLD-TDP) and in the majority of 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).

[0004] Thirty-eight negative dominant mutations in TDP-43 have been identified in sporadic and familial ALS patients, as well as in patients with hereditary FTD in which the majority localize to the glycine-rich domain (Figure 1 in Lagier-Tourenne and Cleveland, Cell 136, 2009, 1001-1004). TDP-43 has an intrinsic tendency to aggregate, as shown by sedimentation assays, and this tendency is further increased by some of the ALS-associated TARDBP mutations that link TDP-43 aggregation to clinical disease symptoms (Ticozzi et al., CNS Neurol. Disord. Drug Targets. 2010, 9(3), 285-296.).

[0005] II. TDP-43 in Neurodegeneration TDP-43 aggregation has been implicated in frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), progranulin (GRN) mutated, C9orf72 mutated, TARDBP mutated, valosin-containing protein (VCP) mutated, chromosome 9p-related, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTL) D) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), Alexander disease (AxD), limbic dominant late-life TDP-43 encephalopathy (LATE ), chronic traumatic encephalopathy (CTE), Perry syndrome, Alzheimer's disease (AD) (including sporadic and familial AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with mutations in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy with mutations in the myotilin (MYOT) gene or mutations in the gene encoding desmin (DES)), traumatic brain injury (TBI), dementia with Lewy bodies (DLB) or Parkinson's disease (PD) (Lagier-Tourenne syndrome), et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1 R46-R64).

[0006] Aggregated TDP-43 from patient brains displays many abnormal modifications including hyperphosphorylation, ubiquitination, acetylation, and proteolytic cleavage to C-terminal fragments (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 property of TDP-43 pathology is the redistribution and accumulation of TDP-43 from the nucleus to the cytoplasm. The characteristic lesions of FTLD-TDP are neuronal and glial cytoplasmic inclusions (NCIs and GCIs, respectively) and degenerated neurites (DNs), which are immunoreactive for TDP-43, as well as ubiquitin and p62, but negative for other neurodegeneration-associated proteins. Differences in inclusion morphology and their tissue distribution are associated with specific mutations and / or clinical symptoms. Four types of TDP-43 lesions have been reported so far by histological classification (Mackenzie and Neumann, J. Neurochem. (2016) 138 (Suppl. 1), 54-70). FTLD-TDP type A cases are characterized by a large amount of short neurodegenerative processes (DNs) and small ovoid or semilunar NCIs, mainly in layer II of the neocortex (Fig. 2f in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). This condition usually presents clinically as either behaviorally distinct frontotemporal dementia (bvFTD) or primary progressive aphasia (nfvPPA) and is associated with mutations in progranulin (GRN).Type B cases show a moderate number of small or granular NCIs in both superficial and deep cortical layers with relatively few DNs and NIIs (neuronal intranuclear inclusions; Fig. 2g in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Many cases presenting simultaneously with FTD and ALS symptoms have been found to show FTLD-TDP Type B pathology. Type C cases have abundant long serpentine neurites, especially in superficial cortical layers, with little or no NCIs (Fig. 2j in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). This pathology is seen especially in cases presenting with primary progressive aphasia (svPPA). FTLD-TDP type D shows abundant neuronal intranuclear inclusions (NIIs) and short DNs in the neocortex with extremely rare NCIs (Fig. 2k in Mackenzie et al., 2016 J. Neurochem. 138 (Suppl. 1), 54-70). Type E is characterized by granular fibrous neuronal inclusions (GFNIs) and very fine punctate neuropil aggregates affecting all neocortical layers, including curvilinear oligodendrocyte inclusions in the white matter (Edward B. Lee et al., Acta Neuropathol. 2017 July ; 134(1): 65-78.). This lesion pattern is seen only in cases of VCP associated with inclusion body myositis.

[0007] III. TDP-43 in FTD Frontotemporal dementia (FTD) is a clinical term that includes a wide range of diseases based on the degeneration of the frontal and temporal lobes, a pathological feature called frontotemporal lobar degeneration (FTLD). FTD is the second most common case of early degenerative dementia in people under 65 years of age (Le Ber, Revue Neurologique 169 (2013) 811-819). FTD is manifested by several symptoms, including bvFTD, characterized by changes in personality and behavior; semantic dementia (SD) and progressive non-fluent aphasia (PNFA), characterized by changes in language function; and corticobasal syndrome (CBS), progressive supranuclear palsy syndrome and motor neuron disease (FTD-MND), characterized by motor dysfunction. The clinical diagnosis of these conditions is complex, and a final conclusion can only be made by postmortem histopathological analysis to detect aggregated proteins and identify the affected brain regions. From a pathological point of view, among the protein inclusions, approximately 45% of cases show pathological accumulation of misfolded tau, 45% of cases show pathological TDP-43, and a smaller population shows aggregation of FUS and other proteins.

[0008] IV. TDP-43 in ALS Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by early loss of upper and lower motor neurons. ALS progression follows disease progression from 1–5 years from diagnosis to death, marked by fatal motor paralysis and respiratory failure. In most cases of sporadic ALS, the neuropathology is characterized by abnormal cytoplasmic accumulation of TDP-43 in neurons and glia of the primary motor cortex, brainstem motor nuclei, spinal cord, and associated white matter tracts. ALS with dementia is associated with accumulation of TDP-43 in the extramotor neocortex and hippocampus. The role of TDP-43 phosphorylation in ALS patients has been studied with the help of antibodies that specifically bind to phosphorylated TDP-43 in nuclear and cytoplasmic inclusions with amino acids S379, S403, S404, S409, and S410 as the main sites of TDP-43 phosphorylation (Hasegawa et al., Ann Neurol 2008; 64: 60-70; Neumann et al., Acta Neuropathol (2009) 117: 137-149).

[0009] V. TDP-43 in AD and other diseases TDP-43 pathology occurs in up to 57% of brains of Alzheimer's disease patients (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 age and correlates with cognitive decline, memory loss, and medial temporal lobe atrophy in AD. TDP-43 in AD appears to represent a secondary or independent pathology that shares an overlapping brain distribution with amyloid beta and tau pathology in the medial temporal lobe. Lesion TDP-43 follows a uniform pattern of progressive deposition reported by the so-called TDP-43 in AD (TAD) staging scheme: initially, TDP-43 deposition in the amygdala (stage I), followed by the hippocampus, limbic system, temporal regions, and finally, the anterior striatum (stage V) (Josephs KA et al., Acta Neuropathol. 2014; 127(6): 811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3): 441-450).

[0010] VI. TDP-43 Propagation Although the onset and initial symptoms of ALS and FTD vary significantly between patients, a common feature of disease progression is the spread of lesions from the initial focal area to many neurons. The intermittent worsening of symptoms may be explained by the progressive spread of TDP-43 pathology. TDP-43 pathology in the brains of ALS patients may spread in a four-step process, spreading transsynaptically via corticosubgrade axonal projections using anterograde axonal migration (Brettschneider et al., Ann Neurol. 2013 July; 74(1): 20-38.). Recent experimental evidence supports the hypothesis of protein proliferation in neural tissues for amyloid beta, Tau, alpha-synuclein, and TDP-43 by a prion-like mechanism (Hasegawa et al., 2017), with the initiation point and tissue distribution spreading pattern differing for the four proteins (Brettschneider J et al., Nature Rev. Neuroscience, 2015, 109). This common disease integration mechanism is thought to be based on the intercellular spread of the lesion protein aggregates, which consists of release of aggregates from lesion cells, uptake by endogenous cells, and seeding of the lesion protein conformation by templated conformational changes of endogenous proteins.

[0011] The cell-to-cell spread of TDP-43 has been studied at the molecular level in several in vitro models, where insoluble TDP-43 preparations derived from patient brains are able to induce intracellular aggregate formation in reporter cells (Nonaka et al., Cell Reports 4 (2013), 124-134; Feiler et al., 2015; Porta et al., Nat. Comm., 2018). Furthermore, it was shown that intracellular TDP-43 aggregates are bound to and released from exosomes before spreading to their neighboring cells (Nonaka et al., Cell Reports 4 (2013, 124-134)). Similarly, adenoviral-transduced TDP-43 expression leads to cytoplasmic aggregates that are phosphorylated, ubiquitous, and most importantly act as seeds to initiate cell-to-cell spread (Ishii et al., PLoS ONE 12(6): e0179375, 2017). Patient-derived pathological TDP-43 can cause widespread distribution of endogenous TDP-43 following intracerebral inoculation into transgenic and wild-type mice (Porta et al., Nat. Comm., 2018).

[0012] VII. Prevention and Treatment of TDP-43 Proteinopathy Aggregation and propagation of the lesion TDP-43 is a cardinal feature of ALS and FTD, fatal diseases for which no treatment is currently available. Mutations in TDP-43 have been associated with familial cases of ALS and FTD providing a causal link between TDP-43 misfolding and disease progression.

[0013] VIII. Diagnosis of TDP-43 proteinopathy Diagnosis of FTD based on clinical symptoms is poor, especially in the early stages, as its clinical symptoms can overlap with other diseases.

[0014] Many approaches aim at the development of biochemical biomarkers to distinguish between different types of FTD pathology. The development of antibodies against different conformations of TDP-43 may allow the creation of more sensitive and specific diagnostic tools. In parallel with biochemical biomarkers, the development of imaging biomarkers may allow early and specific detection of pathology in TDP-43 proteinopathies. The ability to image TDP-43 deposits in the brain could be a major achievement for the diagnosis and drug development of TDP-43 proteinopathies. The use of cell-permeable antibody fragments may enable such detection.

[0015] The earliest event in neurodegenerative diseases based on the misfolding of different proteins is the appearance of an alternative conformation that renders the protein toxic. Moreover, this misfolded conformation can self-propagate by recruiting endogenous normal proteins as the misfolded conformation, a fundamental mechanism of propagation observed throughout the affected tissue.

[0016] To develop antibodies against different conformational states of a given protein, supramolecular antigen constructs have been designed in which the conformation of the presented antigen is adjusted to generate conformation-specific antibodies against a given target in a particular conformational state (WO2012 / 055933 and WO2012 / 020124). Conformation-specific antibodies offer many advantages since they can distinguish between disease-associated conformations and functional, endogenous conformations of these proteins. This approach offers many advantages in therapeutic applications since such antibodies are unlikely to be adsorbed to the normal conformation of the protein, while targeting its misfolded disease-associated isoforms. Similarly in diagnostic applications, such antibodies recognize only the conformational state of the protein that is associated with the disease, which is paramount for the development of sensitive and specific diagnostics.

[0017] The use of TDP-43-based biomarkers in TDP-43 proteinopathies has yet to be established, and such assessment is hampered in part by the lack of high affinity antibodies that can be used in suitable immunoassays to quantify pathological TDP-43 in biological fluids (Feneberg et al., Molecular Neurobiology, 2018).

[0018] Thus, there is a clear need for biomarkers that can detect misfolded, aggregated TDP-43 and non-aggregated, physiological TDP-43, particularly in human samples, to diagnose different types of TDP-43 proteinopathies and / or to monitor the effectiveness of therapeutic agents used to treat diseases, disorders, and disorders associated with TDP-43, particularly TDP-43 aggregates or TDP-43 proteinopathies.

[0019] TDP-43 proteinopathies are defined as a group of neurodegenerative diseases characterized by lesions of TDP-43.

[0020] IX. Prior art Patent application WO2008 / 151055 discloses methods and materials for determining whether a mammal is suffering from a neurodegenerative disease using levels of TDP-43 polypeptide and / or TDP-43 polypeptide cleavage products (e.g., 25 kD and 35 kD TDP-43 polypeptide cleavage products) in biological fluids.

[0021] Patent application WO2013 / 061163 discloses TDP-43 specific binding molecules including polypeptides, such as human, antibodies, and fragments, derivatives and variants thereof.

[0022] In view of the above, there is a need for anti-TDP-43 binding molecules that bind to misfolded, aggregated TDP-43 and nonaggregated physiological TDP-43, especially human TDP-43. Furthermore, there is an urgent need to develop sensitive and specific biomarkers that allow differentiation between lesion types within the FTD spectrum.

[0023] The technical problem is solved by the embodiments provided herein.

[0024] Thus, the present invention relates to binding molecules, particularly antibodies or antigen-binding fragments thereof, that specifically recognize misfolded, aggregated TDP-43 and non-aggregated physiological TDP-43. Within the scope of the present 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 includes phosphorylated, ubiquitinated, acetylated, sumoylated, and / or methylated TDP-43. Physiological TDP-43 includes soluble nuclear TDP-43. It is shown herein that the binding molecules of the present invention, including TDP-43 aggregates and phosphorylated TDP-43, can bind to pathological TDP-43 (see Example 13). Thus, the present invention provides binding molecules, particularly antibodies or antigen-binding fragments thereof, that specifically recognize misfolded, aggregated TDP-43 and non-aggregated physiological TDP-43. Such binding molecules are referred to herein as "pan-TDP-43" binding molecules, particularly pan-TDP-43 antibodies. As described herein, the TDP-43 binding molecules of the present invention may bind equally to misfolded, aggregated TDP-43 and non-aggregated physiological TDP-43, or may specifically bind to both categories of TDP-43 but preferentially to one or the other. The present invention also provides binding molecules, particularly antibodies or antigen-binding fragments thereof, for the prevention, mitigation, treatment and / or diagnosis of diseases, disorders and disorders associated with TDP-43, particularly TDP-43 aggregates or TDP-43 proteinopathies. The present invention also provides binding molecules, particularly antibodies or antigen-binding fragments thereof, for detecting and / or recognizing (i.e., identifying) specific types of pathology that cause neurodegeneration. It is envisaged that it will be used as a diagnostic biomarker to enable more efficient and accurate subject selection for longitudinal monitoring in clinical studies, supporting the development of new therapeutics for TDP-43 proteinopathies.

[0025] The present invention also provides a TDP-43-binding molecule, particularly an antibody or an antigen-binding fragment thereof, as a pharmaceutical (therapeutic drug).

[0026] Without wishing to be bound by theory, the present invention relates to modified conformation-specific antigen peptides and peptide fragments derived from TDP-43 protein or whole TDP-43 protein, and antibodies or fragments obtainable by said peptides or fragments or whole TDP-43 protein or obtained inhibit TDP-43 cell-to-cell spread and / or disassemble TDP-43 aggregates and / or inhibit TDP-43 seeding and / or inhibit aggregation of TDP-43 protein or fragments thereof. The binding molecules of the present invention, in particular polypeptides, more particularly antibodies or antigen-binding fragments thereof, bind to misfolded aggregated TDP-43, in particular cytoplasmic and extracellular misfolded TDP-43. The binding molecules of the present invention, in particular polypeptides, more particularly antibodies or antigen-binding fragments thereof, bind to full-length TDP-43 and / or truncated TDP-43. In one embodiment, the binding molecules of the present invention, in particular polypeptides, more particularly antibodies or antigen-binding fragments thereof, specifically bind to cytoplasmic misfolded TDP-43.

[0027] Misfolded, aggregated, or pathology-associated TDP-43 consists of TDP-43 protein that has lost its normal folding (i.e., misfolded) and localization. Misfolded, aggregated TDP-43 can be found in preinclusions, neuronal and glial cytoplasmic inclusions (NCIs and GCIs, respectively), neuronal intranuclear inclusions (NIIs), and degenerating neurites (DNs), which show immunoreactivity for TDP-43.

[0028] Non-aggregated physiological TDP-43 is a physiologically functional TDP-43 protein that is primarily located in the nucleus and traffics to the cytoplasm, ready to exert its desired functions in the in vivo cellular environment.

[0029] The binding molecules of the invention, in particular antibodies or antigen-binding fragments thereof, surprisingly have at least one, preferably two, more preferably three, and even more preferably all four of the following characteristics: -Inhibits cell-to-cell spread of TDP-43; -Disintegrate TDP-43 aggregation; -Inhibits aggregation of TDP-43 protein or its fragments; -Inhibits TDP-43 seeding.

[0030] Regardless of the combination of one, two, three or four of the above-mentioned features, the binding molecules of the invention, preferably antibodies or antigen-binding fragments thereof, may ameliorate / inhibit / reduce the formation of TDP-43 pathology in in vivo models of TDP-43 proteinopathy and more importantly in patients with TDP-43 pathology.

[0031] The TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can recruit and / or activate microglia. More specifically, it is shown herein that the TDP-43 binding molecules of the present invention can affect the morphogenesis of microglia in terms of cell size and activation state (see Example 10 and Figure 5). This may contribute to the reduction of TDP-43 pathology exhibited by the TDP-43 binding molecules of the present invention.

[0032] In the present invention, the binding molecules, particularly antibodies or antigen-binding fragments thereof, specifically recognize TDP-43. The binding molecules of the present invention include polypeptides, antibodies, and / or antigen-binding fragments thereof specific to the TDP-43 protein. "Specifically recognize TDP-43" means that the binding molecules of the present invention specifically, generally, and collectively bind to TDP-43, particularly to some epitopes within TDP-43, particularly epitopes exposed / accessible in the conformation of one or more lesions of the TDP-43 protein, with higher affinity than other epitopes. The binding molecules of the present invention, particularly polypeptides, more particularly antibodies or antigen-binding fragments thereof, that specifically bind to TDP-43 specifically recognize misfolded aggregated TDP-43 and non-aggregated physiological TDP-43. In a preferred embodiment, the full-length human TDP-43 preferably comprises the sequence of SEQ ID NO:1. In another preferred embodiment of the present invention, the binding molecule, in particular an antibody or antigen-binding fragment thereof, specifically binds to a defined binding region in full-length and / or truncated TDP-43, said binding region being preferably comprised within amino acids 181-195, 199-213, 307-321, 352-366, 389-411, 397-411 or 140-200 of full-length human TDP-43 having the sequence of SEQ ID NO: 1, more preferably said binding region being comprised within amino acids 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406 or 400-412. Thus, the binding molecule, particularly the antibody or antigen-binding fragment thereof, preferably specifically binds to a peptide comprising a binding region consisting of amino acids 181 to 195, 199 to 213, 307 to 321, 352 to 366, 389 to 411, 397 to 411 or 140 to 200 of full-length human TDP-43 having the sequence of SEQ ID NO:1, preferably consisting of said binding region.In another preferred embodiment of the present invention, the binding molecule, particularly an antibody or an antigen-binding fragment thereof, preferably specifically binds to a peptide comprising a binding region consisting of amino acids 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406 or 400-412 of human TDP-43 (SEQ ID NO: 1), preferably consisting of said binding region. In certain embodiments, the TDP-43 binding molecule, particularly an antibody or an antigen-binding fragment thereof, binds within the C-terminal region of TDP-43. This can be effective, for example, because the C-terminal fragment of TDP-43 is found in the insoluble fraction and may thereby be pathologically relevant. More specifically, the TDP-43 binding molecule, particularly an antibody or an antigen-binding fragment thereof, may bind to an epitope within amino acid residues 400-405, 400-406 or 400-412 of human TDP-43 (SEQ ID NO: 1). In certain embodiments of the present invention, the antibody is a monoclonal antibody. In certain embodiments, the antibody is a mouse, murine, human, humanized, or chimeric antibody. It is understood that equivalent binding regions exist in non-human TDP-43. Thus, for example, the mouse TDP-43 amino acid sequence (see Uniprot accession number Q921F2) is also 414 amino acids in length and 96% identical (398 / 414 residues) to the human sequence. The present invention encompasses binding molecules, particularly antibodies or antigen-binding fragments thereof, that bind to regions / peptides equivalent to the regions / peptides identified above with reference to SEQ ID NO: 1 in non-human TDP-43, particularly mouse TDP-43.

[0033] In particular, the present invention is summarized in the following embodiments. 1. A TDP-43 binding molecule that binds to misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, particularly human TDP-43.

[0034] 2. The TDP-43 binding molecule according to the above embodiment, which binds to an epitope within amino acid residues 181-195, 199-213, 307-321, 352-366, 389-411, 397-411 or 140-200 of the human TDP-43 binding molecule (SEQ ID NO: 1).

[0035] 3. The TDP-43 binding molecule according to the above embodiment, which binds to an epitope within amino acid residues 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406 or 400-412 of human TDP-43 (SEQ ID NO: 1).

[0036] 4. The binding molecule according to any one of the above embodiments, which is an antibody or an antigen-binding fragment thereof.

[0037] 5. a) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 11; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 12; VH-CDR3 containing the amino acid sequence ES (Glu-Ser); VL-CDR1 containing the amino acid sequence of SEQ ID NO: 15; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 17; b) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 21; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 22; VH-CDR3 containing the amino acid sequence ES (Glu-Ser); VL-CDR1 containing the amino acid sequence of SEQ ID NO: 25; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 27; c) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 31; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 32; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 33; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 35; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 36; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 37; d) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 41; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 42; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 43; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 45; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 46; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 47; e) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 61; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 62; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 63; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 65; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 66; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 67; f) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 71; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 72; VH-CDR3 containing the amino acid sequence 73; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 75; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 77; g) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 81; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 82; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 83; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 85; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 86; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 87; h) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 101; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 102; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 103; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 105; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 106; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 107; i) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 121; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 122; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 123; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 125; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 127; j) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 146; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 147; or k) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 151; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 152; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 153; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 155; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 156; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 157 The binding molecule or TDP-43 binding molecule according to any one of the above embodiments, comprising

[0038] 6. a. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 10 or having at least 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 14 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14; b. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 20 or having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 20; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 24 or having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 24; c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 30 or having at least 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 30; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 34 or having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 34; d. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 40 or having at least 90%, 91%, 92%, 93%, 94%, 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; e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 60 or having at least 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 having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; f. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 70 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 70; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 74 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 74; g. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 80 or having at least 90%, 91%, 92%, 93%, 94%, 95%, 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 having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 84; h. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 100 or having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the amino acid sequence of SEQ ID NO: 100; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 104 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; i. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 120 or having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 120; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 124 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; j. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 140 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 140; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 144; or k. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 150 or having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 150; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 154 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 154 The binding molecule or TDP-43 binding molecule according to any one of the preceding embodiments, which is an antibody or an antigen-binding fragment thereof.

[0039] 7. a. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 10 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 14; c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 20 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 24; d. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 30 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 34; e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 40 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 44; f. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 60 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 64; g. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 70 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 74; h. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 80 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 84; i. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 100 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 104; j. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 120 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 124; k. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 140 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 144; or l. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 150 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 154 The binding molecule or TDP-43 binding molecule according to any one of the above embodiments, which is an antibody or an antigen-binding fragment thereof.

[0040] In certain embodiments, the antibody is: a) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 11; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 12; VH-CDR3 containing the amino acid sequence ES (Glu-Ser); VL-CDR1 containing the amino acid sequence of SEQ ID NO: 15; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 17; b) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 21; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 22; VH-CDR3 containing the amino acid sequence ES (Glu-Ser); VL-CDR1 containing the amino acid sequence of SEQ ID NO: 25; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 27; c) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 31; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 32; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 33; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 35; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; d) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; e) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 63; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; f) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 71; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 72; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 73; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 75; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 77; g) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; h) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 101; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 102; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 103; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 105; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 106; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 107; i) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 121; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 122; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 123; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 125; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 127; j) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 146; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 147; or k) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 151; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 152; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 153; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 155; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 156; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 157 comprising.

[0041] In certain embodiments, the antibody is: a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12; and VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); b) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 21; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22; and VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); c) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 31; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 32; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 33; d) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; e) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 63; f) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 71; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 72; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 73; g) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; h) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 101; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 103; i) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 121; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 122; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 123; j) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143; or k) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 151; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 152; and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 153 comprising.

[0042] In certain embodiments, the antibody is: a) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17; b) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 25; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; c) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 35; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; e) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 75; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 77; f) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87; g) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 105; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 106; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 107; h) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 125; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 127; or i) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 155; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 156; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 157 comprising.

[0043] In certain embodiments, the antibody is: a) VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 11; VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 12; VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17; b) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 21; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 22; a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 25; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; c) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 31; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 32; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 33; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 35; 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: 37; d) 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 an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 42; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 43; 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; e) 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 an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 62; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 63; 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; f) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 71; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 72; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 73; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 75; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 77; g) 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 an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 82; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 83; 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; h) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 101; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 102; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 103; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 105; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 106; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 107; i) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 121; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 122; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 123; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 125; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 127; j) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 141; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 142; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 143; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 146; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 147; or k) A VH-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 151; a VH-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 152; a VH-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 153; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 155; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 156; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 157 comprising.

[0044] In certain embodiments, the antibody is: a) A VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12; a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); a VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 15; a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 16; and a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 17; b) A VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 21; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22; a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); a VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 25; a VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 16; and a VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 27; c) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 31; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 32; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 33; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 35; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 36; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 37; d) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 45; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 46; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 47; e) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 63; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 65; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 66; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 67; f) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 71; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 72; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 73; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 75; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 16; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 77; g) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 85; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 86; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 87; h) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 101; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 102; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 103; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 105; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 106; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 107; i) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 121; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 122; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 123; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 125; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 16; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 127; j) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 145; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 146; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 147; or k) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 151; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 152; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 153; VL-CDR1 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 155; VL-CDR2 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 156; VL-CDR3 comprising an amino acid sequence having at least 80%, 90%, 95% or 100% sequence identity to SEQ ID NO: 157 comprising

[0045] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12; (c) VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17.

[0046] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27.

[0047] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 31; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 32; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 33; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 35; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 37.

[0048] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, or 3 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43.

[0049] In certain embodiments, the TDP-43 antibody comprises at least 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47.

[0050] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 63; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67.

[0051] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 71; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 72; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 73; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 75; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 77.

[0052] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 81; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 82; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 85; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 86; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 87.

[0053] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 101; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 102; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 103; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 105; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 106; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 107.

[0054] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 121; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 122; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 123; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 125; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 127.

[0055] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, or 3 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; and (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143.

[0056] In certain embodiments, the TDP-43 antibody comprises at least 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 146; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 147.

[0057] In one embodiment, the TDP-43 antibody comprises at least 1, 2, 3, 4, 5, or 6 CDRs selected from: (a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 151; (b) VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 152; (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 153; (d) VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 155; (e) VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 156; and (f) VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 157.

[0058] In another embodiment, the TDP-43 antibody comprises a heavy chain variable domain (VH) selected from SEQ ID NOs: 10, 20, 30, 40, 60, 70, 80, 100, 120, 140, 150 (including post-translational modifications of said sequences). In certain embodiments, the heavy chain variable domain (VH) comprises at least 1, 2, or 3 CDRs selected from: (a) VH-CDR1 comprising an amino acid sequence selected from SEQ ID NOs: 11, 21, 31, 41, 61, 71, 81, 101, 121, 141, 151; (b) VH-CDR2 comprising an amino acid sequence selected from SEQ ID NOs: 12, 22, 32, 42, 62, 72, 82, 102, 122, 142, 152; (c) VH-CDR3 comprising an amino acid sequence selected from SEQ ID NOs: 33, 43, 63, 73, 89, 103, 123, 143, 153, and ES (Glu-Ser).

[0059] In another embodiment, the TDP-43 antibody comprises a light chain variable domain (VL) selected from SEQ ID NOs: 14, 24, 34, 64, 74, 84, 104, 124, 154 (including post-translational modifications of said sequences). In certain embodiments, the light chain variable domain (VL) comprises at least 1, 2, or 3 CDRs selected from: (a) VL-CDR1 comprising an amino acid sequence selected from SEQ ID NOs: 15, 25, 35, 65, 75, 85, 105, 125, 155; (b) VL-CDR2 comprising an amino acid sequence selected from SEQ ID NOs: 16, 36, 66, 86, 106, 156; (c) VL-CDR3 comprising an amino acid sequence selected from SEQ ID NOs: 17, 27, 37, 67, 77, 87, 107, 127, 157, and ES (Glu-Ser).

[0060] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, or 3 CDRs selected from (a) VH-CDR1 comprising an amino acid sequence selected from SEQ ID NO: 11, 21, 31, 41, 61, 71, 81, 101, 111, 121, 141, 151; (b) VH-CDR2 comprising an amino acid sequence selected from SEQ ID NO: 12, 22, 32, 42, 62, 72, 82, 102, 122, 142, 152; (c) VH-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 33, 43, 63, 73, 83, 103, 123, 143, 153, and ES (Glu-Ser).

[0061] In certain embodiments, the TDP-43 antibody comprises at least 1, 2, or 3 CDRs selected from (a) VL-CDR1 comprising an amino acid sequence selected from SEQ ID NO: 15, 25, 35, 65, 75, 85, 105, 125, 155; (b) VL-CDR2 comprising an amino acid sequence selected from SEQ ID NO: 16, 36, 66, 86, 106, 156; (c) VL-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 17, 27, 37, 67, 77, 87, 107, 127, 157.

[0062] In certain embodiments, the light chain variable domain (VL) comprises at least 1, 2, or 3 CDRs selected from (a) VL-CDR1 comprising an amino acid sequence selected from SEQ ID NO: 15, 25, 35, 45, 65, 75, 85, 105, 125, 145, 155; (b) VL-CDR2 comprising an amino acid sequence selected from SEQ ID NO: 16, 36, 66, 86, 106, 156; (c) VL-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 17, 27, 37, 67, 77, 87, 107, 127, 157.

[0063] In certain embodiments, the present invention relates to an antibody derived from hybridoma clones 631B2A2, 633B12C8, 634H10H7, 636E5B8, 641H1E7, 642A10B11, 642D12B4, 646B7F7, 712A6B10, 809D9C2, or 809F12D8.

[0064] In one embodiment, the present invention relates to an antibody selected from ACI-7069-631B2-Ab1, ACI-7069-633B12-Ab1, ACI-7069-634H10-Ab2, ACI-7069-636E5-Ab1, ACI-7069-641H1-Ab2, ACI-7069-642A10-Ab1, ACI-7069-642D12-Ab1, ACI-7069-646B7-Ab1, ACI-7071-712A6-Ab1, ACI-7071-809D9-Ab2, and ACI-7071-809F12-Ab1.

[0065] In certain embodiments, the binding molecules or antibodies provided herein have a dissociation constant of ≤ 1 μM, ≤ 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., 10 -8 M to 10 -13 M, e.g., 10 -9 M to 10 -13 M) with respect to binding to TDP-43, particularly to soluble TDP-43, aggregated TDP-43, and / or oligomeric TDP-43. In one embodiment, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, may have a lower KD for aggregated TDP-43 than for soluble TDP-43. For example, the TDP-43 binding molecules of the present invention may have a KD for aggregated TDP-43 of 30 nM or less, and in certain embodiments, 1 nM or less, and a KD for soluble TDP-43 of 500 nM or less. This is shown for the TDP-43 binding molecules of the present invention in Example 8A with reference to Table 8.

[0066] In one embodiment, the binding affinity to soluble or aggregated FL TDP-43 can be evaluated by examining the dissociation constant (KD) using surface plasmon resonance (SPR; Biacore T200, GE Healthcare Life Sciences). For a detailed description of suitable SPR methods that can be used, Examples 8A and 8B can be referred to.

[0067] The TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, typically bind to TDP-43 with high affinity. For example, they can exhibit an EC50 value of 200 pM or less, more preferably 20 pM or less, even more preferably 10 pM or less, as determined by a Luminex assay. For further details of suitable assays, Example 3 can be referred to. Similarly, they can exhibit an EC50 value of 1600 ng / ml or less, more preferably 120 ng / ml or less, even more preferably 60 ng / ml or less, as determined by indirect ELISA. For further details of suitable assays, Example 4 can be referred to.

[0068] The TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, bind to both non-aggregated physiological TDP-43 and aggregated TDP-43. Thus, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can bind to soluble and aggregated TDP-43 almost equally well. The TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can bind to aggregated TDP-43 almost equally well compared to non-aggregated TDP-43. More specifically, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can bind to aggregated TDP-43 in the cytoplasm almost equally well compared to non-aggregated TDP-43 in the nucleus. In other embodiments, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, bind to both but can preferentially bind to aggregated TDP-43 compared to non-aggregated TDP-43. More specifically, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, bind to both but can preferentially bind to aggregated TDP-43 in the cytoplasm compared to non-aggregated TDP-43 in the nucleus. Alternatively, in other embodiments, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, bind to both but can preferentially bind to non-aggregated TDP-43 compared to aggregated TDP-43. More specifically, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, bind to both but can preferentially bind to non-aggregated TDP-43 in the nucleus compared to aggregated TDP-43 in the cytoplasm. These binding properties may be demonstrated, for example, using immunohistochemistry. A suitable method is described herein with reference to Example 6 in which relevant controls are provided. The results are shown in Table 7.

[0069] The present invention also relates to a composition comprising a binding molecule of the invention described herein, particularly an antibody or an antigen-binding fragment thereof (including a TDP-43 binding antibody fragment and derivative). Further, the present invention relates to an immunotherapy and / or immunodiagnostic method of using such a composition in the prevention, diagnosis, and / or treatment of TDP-43 proteinopathy, wherein an effective amount of the composition is administered to a subject in need thereof.

[0070] In certain embodiments, the invention encompasses the use of a binding molecule of the invention described herein that specifically binds to TDP-43, particularly an antibody and an antigen-binding fragment thereof, for diagnosing, preventing, reducing, and / or treating diseases, disorders, and / or abnormalities associated with TDP-43 (particularly 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 described herein are applicable to the diagnosis, prevention, reduction, and / or treatment of diseases, disorders, and / or abnormalities associated with TDP-43 (particularly TDP-43 aggregates), or TDP-43 proteinopathy, including but not limited to FTD and ALS. Preferably, the use of these binding molecules for diagnosing, preventing, reducing, and / or treating diseases, disorders, and / or abnormalities associated with TDP-43 (particularly TDP-43 aggregates), or TDP-43 proteinopathy, relates to ALS, AD, or FTD. More preferably, the use relates to ALS. More preferably, the use relates to ALS. More preferably, the use relates to AD. More preferably, the use relates to FTD.

[0071] In another embodiment, the binding molecules of the invention described herein that are specific for TDP-43, particularly antibodies and antigen-binding fragments thereof, upon contact with a sample, detect, diagnose, and / or monitor a disease, disorder, and / or abnormality associated with TDP-43 (particularly TDP-43 aggregates), or TDP-43 proteinopathy, selected from frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), having a progranulin (GRN) mutation, having a C9orf72 mutation, having a TARDBP mutation, having a valosin-containing protein (VCP) mutation, associated with chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, having a TARDBP mutation, having an 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 AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia, and myopathy (sporadic inclusion body myositis, inclusion body myopathy with a mutation in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia)), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy having a mutation in the myotilin (MYOT) gene or a gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

[0072] In one embodiment, the invention encompasses the binding molecules of the invention described herein that specifically bind to TDP-43, particularly antibodies or antigen-binding fragments thereof, and the use of these molecules, particularly these antibodies, to detect the presence of TDP-43 in a sample. Thus, the TDP-43 binding molecules of the invention, such as the anti-TDP43 antibodies described herein, can be used, for example, to screen clinical samples, particularly human blood, CSF, interstitial fluid (ISF) and / or urine, to confirm the presence of TDP-43 in a sample by using, for example, an ELISA-based assay or a surface plasmon assay. Tissue samples may be used in certain situations, such as brain tissue samples. The methods and compositions of the invention are also applicable to diagnosing pre-symptomatic diseases and / or monitoring disease progression and / or treatment efficacy.According to one embodiment, an antibody specific for TDP-43 (e.g., a full-length antibody or a TDP-43 binding fragment or derivative of an antibody) can be contacted with a sample (e.g., blood, cerebrospinal fluid (CSF), interstitial fluid, or brain tissue) to detect, diagnose, and / or monitor a disease, disorder, and / or abnormality associated with TDP-43 (especially TDP-43 aggregates), or TDP-43 proteinopathy, selected from frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), having a progranulin (GRN) mutation, having a C9orf72 mutation, having a TARDBP mutation, having a valosin-containing protein (VCP) mutation, associated with chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, having a TARDBP mutation, having an 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 AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with a mutation in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia)), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy having a mutation in the myotilin (MYOT) gene or a gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB) or Parkinson's disease (PD). The TDP-43 binding molecule of the present invention can be used to quantify TDP-43 in clinical samples such as blood, CSF, ISF or urine, with a relatively high TDP-43 level compared to an appropriate control indicating a disease and / or a more advanced disease.Many suitable immunoassay methods are known. Thus, the methods (e.g., ELISA, MSD (Meso Scale Discovery), HTRF (Homogeneous Time Resolved Fluorescence), and AlphaLISA, etc.) can be performed for diagnosis using the high levels of TDP-43 indicative of the disease. Alternatively, the methods may be performed for monitoring. An increase in levels over time may indicate disease progression. A decrease in levels over time may indicate disease regression. The methods may also be used to monitor treatment, particularly to monitor the effectiveness of a particular treatment. The success of treatment can be measured with reference to the stabilization or decrease of TDP-43 levels after treatment. In this specification (Example 12), it has been shown that when TDP-43 levels were measured using the antibodies of the present invention, they were higher in CSF samples from patients with TDP-43 proteinopathy than in control samples taken from healthy subjects (healthy controls). Control samples may be tested in parallel with the test samples or may not be tested. In certain embodiments, control levels are determined from a series of test samples taken from healthy subjects under similar or identical experimental conditions and are used as a comparison for the levels examined in the test samples. Methods for quantifying TDP-43 in a suitable sample using the binding molecules of the present invention can also be used to select a treatment (for further treatment of a subject). Thus, an individualized treatment approach is envisioned. Samples are taken before and after treatment. If the treatment using the treatment method results in stabilization or preferably a decrease in TDP-43 levels after treatment, the treatment method may be selected for that subject. If the treatment does not result in stabilization or preferably a decrease in TDP-43 levels after treatment, the treatment method is not selected for that subject. The treatment may be a suitable candidate therapeutic agent for the treatment of TDP-43 proteinopathy. In a preferred embodiment, the treatment method typically comprises the TDP-43 binding molecule of the present invention in the form of a pharmaceutical composition described herein.

[0073] The TDP-43 binding molecules of the present invention may also be used for the classification of diseases into specific types or subtypes. Thus, a method for classifying diseases, disorders and / or abnormalities associated with TDP-43 (particularly, TDP-43 aggregates), or for classifying TDP-43 proteinopathies, comprising: a. performing the method of the present invention in which the TDP-43 level is quantified as compared to an appropriate control; b. optionally, identifying mutations in a sample from a subject, including but not limited to mutations in progranulin (GRN), C9orf72, TARDBP, valosin-containing protein (VCP), TARDBP, angiogenin (ANG), valosin-containing protein (VCP), mutations in the myotilin (MYOT) gene, or mutations in the gene encoding desmin (DES), and then c. classifying diseases, disorders and / or abnormalities associated with TDP-43 (particularly, TDP-43 aggregates), or TDP-43 proteinopathies is provided.

[0074] Similarly, a method for classifying a disease, disorder and / or abnormality associated with TDP-43 (especially, TDP-43 aggregates), or for classifying TDP-43 proteinopathy, wherein the TDP-43 level is quantified in a sample obtained from a patient with a disease, disorder and / or abnormality associated with TDP-43, or TDP-43 proteinopathy, and said level is compared with a control sample taken from a subject of a different type or subtype of a disease, disorder and / or abnormality associated with TDP-43 (especially, TDP-43 aggregates), or TDP-43 proteinopathy (i.e., a representative set of control levels is examined for the type or subtype of interest); then performing the method of the present invention based on said comparison; and then classifying a disease, disorder and / or abnormality associated with TDP-43 (especially, TDP-43 aggregates), or TDP-43 proteinopathy, is provided. Thus, said classification is based on examining the closest correspondence between the test sample and one or more control samples. These methods further include identifying mutations in the sample, including but not limited to mutations in progranulin (GRN), C9orf72, TARDBP, valosin-containing protein (VCP), TARDBP, angiogenin (ANG), valosin-containing protein (VCP), mutations in the myotilin (MYOT) gene, or mutations in the gene encoding desmin (DES), and said identified mutations may also be used to classify a disease, disorder and / or abnormality associated with TDP-43 (especially, TDP-43 aggregates), or TDP-43 proteinopathy. To avoid misidentification, the identification of mutations in the sample can be performed by any suitable method based on, for example, nucleic acid sequencing of nucleic acid molecules within the sample. The sample may be a different one from the sample in which the TDP-43 level is determined, but is derived from the same subject.

[0075] In other embodiments, the present invention provides methods for preventing, reducing, and / or treating diseases, disorders, and / or abnormalities associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathies. According to one embodiment, the method of the present invention comprises administering to a subject an effective concentration of a binding molecule of the present invention specific for TDP-43 as described herein, particularly an antibody (e.g., a full-length antibody or a TDP-43 binding fragment or derivative of an antibody). In another embodiment, the present invention provides a method for preventing, reducing, and / or treating TDP-43 proteinopathy. According to certain embodiments, the binding molecule of the present invention described herein specific for TDP-43, particularly an antibody or an antigen-binding fragment thereof, is administered for treating, reducing, and / or preventing frontotemporal dementia (FTD) or amyotrophic lateral sclerosis (ALS). In another embodiment, the binding molecule of the present invention described herein specific for TDP-43, particularly an antibody or an antigen-binding fragment thereof, is administered for preventing, reducing, and / or treating a neurodegenerative disease selected from frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (including sporadic and familial AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), limbic-predominant age-related TDP-43 encephalopathy (LATE).

[0076] In another embodiment, the binding molecules of the invention described herein that are specific for TDP-43, particularly antibodies or antigen-binding fragments thereof, are used for preventing, alleviating, and / or treating a disease selected from frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), having a progranulin (GRN) mutation, having a C9orf72 mutation, having a TARDBP mutation, having a valosin-containing protein (VCP) mutation, related to chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, having a TARDBP mutation, having an 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 AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with a mutation in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia)), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy having a mutation in the myotilin (MYOT) gene or a gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB) or Parkinson's disease (PD).

Brief Description of the Drawings

[0077]

Figure 1

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Figure 7

Mode for Carrying Out the Invention

[0078] Detailed Description of Embodiments of the Invention X. Definitions As used herein, an "antigen-binding molecule" is a molecule that can specifically or selectively bind to an antigen (particularly, TDP-43). The binding molecule may comprise an antibody or a fragment thereof, or may be an antibody or a fragment thereof. An anti-TDP-43 binding molecule is a molecule that binds to the TDP-43 protein at an epitope-specific recognition site, for example, an anti-TDP-43 antibody or a fragment thereof. That is, the antigen-binding molecule of the present invention binds to an epitope within the amino acid sequence of SEQ ID NO: 1. The antigen-binding molecules provided herein, particularly antibodies or antigen-binding fragments thereof, recognize full-length TDP-43. Other anti-TDP-43 binding molecules may also include multivalent molecules, multispecific molecules (e.g., bispecific antibodies), fusion molecules, aptamers, Avimers, or other naturally occurring or recombinantly produced molecules. Exemplary antigen-binding molecules useful in the present invention include antibody-like molecules. The antibody-like molecule is a molecule that can exhibit a function by binding to a target molecule (see, for example, 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 examples thereof include DARPin (WO2002 / 020565), affibody (WO1995 / 001937), Avimer (WO2004 / 044011; WO2005 / 040229), Adnectin (WO2002 / 032925), and fynomer (WO2013 / 135588).

[0079] As used herein, the terms "anti-TDP-43 antibody" and "antibody that binds to TDP-43" or simply "antibody" mean an antibody that can bind to TDP-43 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent that targets TDP-43. In general, the term "antibody" is used herein in the broadest sense and includes, but is not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), fully human antibodies, and antibody fragments (which are limited to those that exhibit the desired antigen-binding activity). Antibodies within the present invention can also be chimeric antibodies, recombinant antibodies, antigen-binding fragments of recombinant antibodies, humanized antibodies, or antibodies presented on the surface of phage or on the surface of chimeric antigen receptor (CAR) T cells.

[0080] An "antigen-binding fragment" of an antibody includes a portion of an intact antibody and means a molecule other than an intact antibody that binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab’, Fab’-SH, F(ab’)2 formed from antibody fragments; bispecific antibodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies.

[0081] 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 the region for binding to the protein.

[0082] In certain embodiments, an "antibody that binds to an epitope" within a defined region of a protein is identified by mutagenesis analysis in which mutations are introduced into the amino acids of the protein and the binding of the antibody to the resulting mutated protein (e.g., the mutated protein that contains the epitope) is determined until it is at least 20% of the binding to the unmutated protein. In certain embodiments, an "antibody that binds to an epitope" within a defined region of a protein is identified by mutagenesis analysis in which mutations are introduced into the amino acids of the protein and the binding of the antibody to the resulting mutated protein (e.g., the mutated protein that contains the epitope) is determined until it is 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 the unmutated protein. In certain embodiments, the binding of the antibody is determined by FACS, WB, or by a suitable binding assay (e.g., ELISA).

[0083] As used herein, the term "bind to" defines the binding (interaction) of at least two "antigen interaction sites" with each other. The term "antigen interaction site" defines, according to the present invention, a motif of a polypeptide, i.e., a part of an antibody or antigen-binding fragment of the present invention that exhibits the ability to specifically interact with a specific antigen or specific group of antigens of TDP-43. Said binding / interaction is also understood to define "specific recognition". The term "specifically recognize" means, according to the present invention, that said antibody can specifically interact with and / or bind to at least two amino acids of TDP-43 as defined herein, in particular, at least two amino acids within amino acid residues 181-195, 199-213, 307-321, 352-366, 389-411, 397-411, and 140-200 of human TDP-43 (SEQ ID NO: 1), and more specifically, at least two amino acids within amino acid residues 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406, or 400-412 of human TDP-43 (SEQ ID NO: 1).

[0084] The term "pan-TDP-43 antibody" means an antibody that binds to misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, and includes monomeric TDP-43, oligomeric TDP-43, post-translationally modified TDP-43 (e.g., phosphorylation, ubiquitination, acetylation, SUMOylation, and / or methylation, etc.), aggregated TDP-43, and truncated TDP-43.

[0085] As used herein, the term "specific interaction" means that the antibody or antigen-binding fragment thereof of the present invention does not cross-react, or substantially does not cross-react, with polypeptides of similar structure. Thus, the antibody or antigen-binding fragment thereof of the present invention specifically binds / interacts with the structure of TDP-43 formed by specific amino acid sequences within amino acid residues 181-195, 199-213, 307-321, 352-366, 389-411, 397-411, and 140-200 of human TDP-43 (SEQ ID NO: 1), and more specifically, with the structure of TDP-43 formed by specific amino acid sequences within amino acid residues 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406, or 400-412 of human TDP-43 (SEQ ID NO: 1).

[0086] The cross-reactivity of a population of antigen-binding molecules, particularly antibodies or antigen-binding fragments thereof under investigation, can be tested by assessing binding of said population of antibodies or antigen-binding fragments thereof under general 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 (poly)peptides as well as to (poly)peptides that are more or less closely related (structurally and / or functionally). Only those constructs (i.e., antibodies, these antigen-binding fragments, etc.) that bind to a specific structure of TDP-43 as defined herein, e.g., a specific epitope of TDP-43 described herein or a (poly)peptide / protein, but do not bind or substantially bind to any other epitope or (poly)peptide of the same TDP-43 are considered specific for the epitope or (poly)peptide / protein of interest and are selected for further experiments by the methods provided herein. These methods can include, inter alia, binding experiments with molecules that are more or less closely related structurally and / or functionally, blocking and competition experiments. These binding experiments can also include FACS analysis, surface plasmon resonance (SPR, e.g., BIACORE®), analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy, or radiolabeled ligand binding assays.

[0087] Thus, specificity can be investigated experimentally by methods known in the art and by the methods described herein. Such methods include, but are not limited to, Western blot, ELISA-, RIA-, ECL-, IRMA tests, and peptide scan.

[0088] As used herein, the term "monoclonal antibody" means an antibody obtained from a substantially homogeneous population of antibodies, i.e., each antibody in the population is identical except for possible naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific for a single antigenic site. Monoclonal antibodies are advantageous in that they can be synthesized by hybridoma culture in which other immunoglobulins are substantially absent. The "monoclonal" prepared as described above exhibits the characteristics of an antibody as a substantially homogeneous population of antibodies and should not be construed as requiring the production of antibodies by any particular method. As described above, the monoclonal antibodies used in the present invention may be prepared by the hybridoma method described in Kohler, Nature 256 (1975), 495.

[0089] As used herein, the term "polyclonal antibody" means an antibody produced between or in the presence of one or more other non-identical antibodies. Generally, polyclonal antibodies are produced from B lymphocytes in the presence of several other B lymphocytes that have produced non-identical antibodies. Usually, polyclonal antibodies are obtained directly from immunized animals.

[0090] As used herein, the term "fully human antibody" means an antibody that contains only human immunoglobulin protein sequences. A fully human antibody may contain mouse sugar chains when produced in a mouse, mouse cells, or a hybridoma derived from mouse cells. Similarly, a "mouse antibody" or "murine antibody" means an antibody that contains only mouse / murine immunoglobulin protein sequences. Alternatively, a "fully human antibody" may contain rat sugar chains when produced in a rat, rat cells, or a hybridoma derived from rat cells. Similarly, the term "rat antibody" means an antibody that contains only rat immunoglobulin sequences. A fully human antibody can also be produced, for example, by phage display, a widely used screening technique that enables the production and screening of fully human antibodies. Phage antibodies can also be used in the present invention. The phage display method is described, for example, in US5403484, US5969108, and US5885793. Another technique that enables the development of fully human antibodies relates to a modified method of mouse hybridoma technology. Mice are introduced to contain human immunoglobulin loci instead of their own mouse genes (see, for example, US5877397).

[0091] The term "chimeric antibody" means an antibody that contains the variable region of the present invention that is fused or chimerized to an antibody region (e.g., a constant region) derived from another human or non-human species (e.g., mouse, horse, rabbit, dog, cow, chicken).

[0092] The term "antibody" also relates to recombinant human antibodies, xenogeneic antibodies, and hetero-hybrid antibodies. The term "recombinant (human) antibody" includes all human sequence antibodies prepared, expressed, made, or isolated by recombinant methods, e.g., antibodies isolated from transgenic animals (e.g., mice) of human immunoglobulin genes; antibodies expressed using recombinant expression vectors transfected into host cells, antibodies isolated from recombinant combinatorial human antibody libraries, or any other method related to splicing of human immunoglobulin gene sequences to other DNA sequences that are prepared, expressed, made, or isolated. Such recombinant human antibodies have variable and constant regions derived from (if present) human germline immunoglobulin sequences. However, such antibodies can be subjected to in vitro mutagenesis (or in vivo somatic mutagenesis if using animal transgenic of human Ig sequences), whereby the amino acid sequences of the VH and VL regions of the recombinant antibody are derived from and related to human germline VH and VL sequences but are sequences that could not naturally occur within the human antibody germline repertoire in vivo.

[0093] "Xenogeneic antibody" is defined with respect to transgenic non-human organisms that generate such antibodies. This term means an antibody having an amino acid sequence or corresponding nucleic acid sequence found in an organism that is not a transgenic non-human animal, generally a species other than the species of the transgenic non-human animal.

[0094] The term "hetero-hybrid antibody" means an antibody having light and heavy chains from different organisms. For example, an antibody having a human heavy chain bound to a mouse light chain is a hetero-hybrid antibody. Examples of hetero-hybrid antibodies include chimeric and humanized antibodies.

[0095] The term "antibody" also relates to humanized antibodies. A "humanized" form of a non-human (e.g., mouse or rabbit) antibody is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., Fv, Fab, Fab’, F(ab’)2, or other antigen-binding subsequence of an antibody) that contains minimal sequences derived from the non-human immunoglobulin. In many cases, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues derived from the recipient's complementarity determining regions (CDRs) have been replaced by residues from the CDRs (donor antibody) of a non-human species such as mouse, rat or rabbit having the desired specificity, affinity, and capacity. In one example, the Fv framework residues of the human immunoglobulin are replaced by the corresponding non-human residues. Additionally, a humanized antibody may contain residues not found in the CDRs or framework sequences introduced into the recipient antibody. These modifications are made to further improve and optimize the function of the antibody. Generally, the humanized antibody includes substantially all of at least one, typically two variable regions, wherein all or substantially all of the CDR regions correspond to those of the non-human immunoglobulin and all or substantially all of the FR regions correspond to those of the human immunoglobulin consensus sequence. The humanized antibody may also include at least a portion of the 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.

[0096] A common method for humanizing antibodies relates to CDR grafting, in which a functional antigen-binding site derived from an antibody of a non-human donor is transplanted into a human acceptor antibody. CDR grafting methods are known in the art and are described, for example, in US5225539, US5693761, and US6407213. Another related method is the generation of humanized antibodies from transgenic animals that have been genetically engineered to contain one or more humanized immunoglobulin loci capable of undergoing gene rearrangement and gene conversion (see, for example, US7129084).

[0097] Thus, in the present invention, the term "antibody" relates to whole immunoglobulin molecules as well as portions of such immunoglobulin molecules (i.e., "antigen-binding fragments thereof"). Further, the term relates to antibody molecules that have been modified and / or converted as described above. The term also relates to antibodies made / recombined or synthesized recombinantly or synthetically. The term also relates to intact antibodies, as well as antibody fragments thereof (e.g., isolated light and heavy chains, Fab, Fv, Fab', Fab'-SH, F(ab')2). The term antibody also includes, but is not limited to, fully human antibodies, chimeric antibodies, humanized antibodies, CDR-grafted antibodies, and antibody constructs such as single-chain Fvs (scFv) or antibody fusion proteins

[0098] A "single-chain Fv" or "scFv" antibody fragment, in the context of the present invention, is one having the VH and VL regions of an antibody, and these regions are present in a single polypeptide chain. Generally, the scFv polypeptide further includes a polypeptide linker between the VH and VL regions that enables the scFv to form the desired structure for antigen binding. Techniques for making single-chain antibodies are disclosed, for example, in Pluckthun in The Pharmacology of Monoclonal Antibodies, Rosenburg and Moore eds. Springer-Verlag, N.Y. (1994), 269-315.

[0099] As used herein, "Fab fragment" consists of one light chain and C H 1, and the variable region of one heavy chain. The heavy chain of the Fab molecule cannot form a disulfide bond with another heavy chain molecule.

[0100] The "Fc" region contains two heavy chain fragments including the C H 2 and C H 3 regions. The two heavy chain fragments are bound by two or more disulfide bonds and hydrophobic interactions of the C H 3 region.

[0101] "Fab' fragment" includes one light chain and a part of one heavy chain containing the V H region and C H 1 region, and further includes the region between C H 1 and C H 2 regions such that an interchain disulfide bond can be formed between the two heavy chains of the two Fab' fragments to form an F(ab')2 molecule.

[0102] "F(ab')2 fragment" contains two light chains and two heavy chains including a part of the constant region between C H 1 and C H 2 regions, and an interchain disulfide bond is formed between the two heavy chains. Thus, the F(ab')2 fragment is composed of two Fab' fragments bound by the disulfide bond between the two heavy chains.

[0103] The "Fv region" includes variable regions derived from both the heavy chain and the light chain, but lacks the constant region.

[0104] The antibodies, antibody constructs, antibody fragments, antibody derivatives (all derived from Ig), or their corresponding immunoglobulin chains used according to the present invention can be further modified by using conventional techniques known in the art, for example, amino acid deletions, insertions, substitutions, additions, and / or recombinations, and / or any other modifications known in the art, either alone or in combination. Methods for introducing such modifications in the DNA sequences inherent in the amino acid sequences of immunoglobulin chains are well known to those skilled in the art; for example, Sambrook et al., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, 2 nd edition (1989) and 3 rdSee edition (2001). The term "region derived from Ig" relates in particular to a (poly)peptide construct comprising at least one CDR. Fragments or derivatives of the regions derived from Ig described are defined as being part of the above antibody molecules and / or (poly)peptides that have been modified by chemical / biochemical or molecular biological methods. The corresponding methods are known in the art and are described in particular 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)).

[0105] As used herein, the term "CDR" relates to "complementary determining regions" well known in the art. CDRs are part of immunoglobulins that determine the specificity of the molecule and contact a particular ligand. CDRs are the most variable parts of the molecule and contribute to the diversity of these molecules. There are three CDR regions (CDR1, CDR2, and CDR3) in each V region. CDR-H represents the CDR regions of the variable heavy chain, and CDR-L relates to the CDR regions of the variable light chain. VH means variable heavy chain, and VL means variable light chain. The CDR regions of regions derived from Ig can 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). The CDR sequences provided herein are defined by Kabat. However, it will be understood by those skilled in the art that the present invention is intended to include binding molecules in which the CDR sequences are defined by any useful identification / numbering method.For example, the numbering methods of 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(1):206-11 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) may be used to define the CDR.

[0106] Therefore, in the present invention, the antibody molecule described in the above specification is selected from the group consisting of full-length antibodies (immunoglobulins, e.g., IgG1, IgG2, IgG2A, IgG2b, IgA1, IgGA2, IgG3, IgG4, IgA, IgM, IgD, or IgE), F(ab)-, Fab’-SH-, Fv-, Fab’-, F(ab’)2 fragments, chimeric antibodies, CDR-grafted antibodies, fully human antibodies, bivalent antibody constructs, antibody fusion proteins, synthetic antibodies, bivalent single-chain antibodies, trivalent single-chain antibodies, and multivalent single-chain antibodies.

[0107] "Humanization methods" are well known in the art and are particularly described for antibody molecules, e.g., Ig-derived molecules. The term "humanized" means a humanized form of a non-human (e.g., murine) antibody or fragment thereof (e.g., Fv, Fab, Fab’, F(ab’)₂, scFvs, or antigen-binding portion sequences of other antibodies) that contains a portion of the sequence derived from a non-human antibody. Humanized antibodies include human immunoglobulins in which residues derived from the complementarity-determining regions (CDRs) of human immunoglobulins have been replaced by residues derived from the CDRs of a non-human species (e.g., mouse, rat, or rabbit) having the desired binding specificity, affinity, and capacity. In general, the humanized antibody substantially comprises at least one, and generally two, variable regions, wherein all or substantially all of the CDR regions correspond to those of the non-human immunoglobulin and all or substantially all of the framework (FR) regions are of human immunoglobulin consensus sequences. The humanized antibody may optionally contain at least a portion of the immunoglobulin constant region (Fc), typically that of a human immunoglobulin; see, e.g., 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. In general, humanized antibodies have one or more amino acids introduced therein from a non-human source that still maintains the original binding activity of the antibody. Methods for humanization of antibodies / antibody molecules are described in further detail 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.Humanized antibodies, for example, specific examples of antibodies against EpCAM, are known in the art (see, for example, 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).

[0108] Thus, in the present invention, there is provided an antibody molecule or an antigen-binding fragment thereof that is humanized and can further be used in a pharmaceutical composition.

[0109] The specificity of the antibody or antigen-binding fragment of the present invention is expressed not only by the amino acid sequence of the antibody or antigen-binding fragment defined above, but also by the epitope to which the antibody can bind. Thus, in one embodiment, the present invention relates to an anti-misfolded TDP-43 antibody or an antigen-binding fragment thereof that recognizes the same epitope as the antibody of the present invention.

[0110] Those skilled in the art can understand that the epitope may be contained in the TDP-43 protein, may be contained in the degradation products, or may be a chemically synthesized peptide. The amino acid positions are shown only to indicate the positions of the corresponding amino acid sequences in the TDP-43 protein sequence. The present invention encompasses all peptides containing the epitope. The peptide may be part of a polypeptide having a length of 100 or more amino acids, or may be a peptide of 100 or less, preferably 50 or less, more preferably 25 or less, and even more preferably 16 or less amino acids. The amino acids of such peptides may be natural amino acids or unnatural amino acids (e.g., beta amino acids, gamma amino acids, D-amino acids) or combinations thereof. Furthermore, the present invention may encompass each retro-inverso peptide of the epitope. The peptide may or may not bind. It may be bound, for example, to a small molecule (e.g., a drug or a fluorophore), a high molecular weight polymer (e.g., polyethylene glycol (PEG), polyethyleneimine (PEI), hydroxypropyl methacrylate (HPMA), etc.), or a protein, fatty acid, sugar moiety, or may be inserted into the membrane.

[0111] To test whether the antibody in question and the antibody of the present invention recognize the same epitope, the following competition experiment can be performed: Vero cells infected at 3 MOI (multiplicity of infection) are incubated for 1 hour with various concentrations of the antibody in question as a competing factor 20 hours later. In the second incubation step, the antibody of the present invention is applied at a defined concentration of 100 nM, and its binding is detected by flow cytometry using a fluorescently labeled antibody against the constant region of the antibody of the present invention. Binding that occurs inversely proportional to the concentration of the antibody in question is an indicator that both antibodies recognize the same epitope. However, many other assays that can be used are known in the art.

[0112] The present invention also relates to the generation of specific antibodies against endogenous and recombinant polypeptides of TDP-43. This generation is based, for example, on the immunization of an animal (e.g., a mouse). However, other animals for the generation of antibodies / antisera are also included within the present invention. For example, monoclonal and polyclonal antibodies can be generated by rabbits, mice, goats, donkeys, etc. A polynucleotide encoding a polypeptide selected corresponding to TDP-43 can be subcloned into a suitable vector, and the recombinant polypeptide is expressed in an organism capable of expression, such as bacteria. Thus, the expressed recombinant protein can be injected intraperitoneally into a mouse, and the resulting specific antibodies can be obtained from mouse serum provided, for example, by intracardiac blood puncture. The present invention also contemplates the generation of specific antibodies against endogenous and recombinant polypeptides by using the DNA vaccination method as exemplified in the described examples. The DNA vaccination method is well known in the art and includes gene gun or jet injection and liposome-mediated delivery by intramuscular or subcutaneous injection. Thus, the polypeptide or protein or epitope of TDP-43, particularly the epitope of the antibodies provided herein, is the desired polypeptide or protein or epitope of TDP-43, particularly the epitope of the antibodies of the present invention present within amino acid residues 181-195, 199-213, 307-321, 352-366, 389-411, 397-411, and 140-200 of SEQ ID NO: 1, more specifically, the epitope of the antibodies of the present invention present within amino acid residues 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406, 400-412, and can be obtained by directly immunizing an animal by directly injecting a vector expressing the epitope of the antibodies of the present invention intramuscularly. The amount of the resulting specific antibodies can be quantified using ELISA, which is also described in the following specification.Additional methods for antibody generation are well known in the art, see, for example, Harlow and Lane, "Antibodies, A Laboratory Manual", CSH Press, Cold Spring Harbor, 1988.

[0113] Thus, under certain assay conditions, a particular antibody to TDP-43 and the corresponding epitope bind to each other and do not bind to a significant amount of other components present in the sample. Specific binding to the target analyte under such conditions may require a binding moiety selected for its properties with respect to the particular target analyte. Various immunoassay methods may be used to select antibodies that specifically react with a particular antigen. For example, solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies having specific immunoreactivity with an analyte. For descriptions of immunoassay methods and conditions that can be used to examine specific immunoreactivity, see Shepherd and Dean (2000), Monoclonal Antibodies: A Practical Approach, Oxford University Press and / or Howard and Bethell. Typically, a specific or selective reaction is at least 2-fold, more typically 10 - 100-fold or more above the background signal due to noise. One of ordinary skill in the art is in a position to provide and generate binding molecules specific for a novel polypeptide. For a particular binding assay, it can be readily used to avoid unwanted cross-reactivity. For example, polyclonal antibodies can be readily purified and selected by known methods (see Shepherd and Dean, loc. cit).

[0114] The "class" of an antibody refers to the type of constant region or the constant regions that its heavy chain has. There are five main antibody classes (IgA, IgD, IgE, IgG, and IgM), and some of these can be further divided into subclasses (isotypes), for example, IgG1, IgG2, IgG2a, IgG2b, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant regions corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0115] In certain embodiments, amino acid sequence variants of the antibodies provided herein are included. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions from residues within the amino acid sequence of the antibody and / or insertions into such residues and / or substitutions of such residues. Any and all combinations of deletions, insertions, and substitutions can be made to the final construct as long as the final construct has the desired characteristics, such as antigen binding.

[0116] In certain embodiments, antibody variants having one or more amino acid substitutions are provided. Sites of interest for substitution mutagenesis include CDRs and FRs. Conservative substitutions are shown in Table 1 under the item "Preferred substitutions". More changes are provided in Table 2 under the item "Typical substitutions" and are further described below with respect to amino acid side chain classes. Amino acid substitutions can be introduced into the antibody of interest, and the product can be screened for the desired activity, such as maintenance / improvement of antigen binding, reduction of immunogenicity, or improvement of ADCC or CDC. Table 1

Table 1

[0117] Amino acids can be grouped according to common side chain properties: (1) Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile; (2) Neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) Acidic: Asp, Glu; (4) Basic: His, Lys, Arg; (5) Residues affecting chain orientation: Gly, Pro; (6) Aromatic: Trp, Tyr, Phe. Non-conservative substitutions involve changing one member of these classes to another.

[0118] One type of substitution variant relates to substituting one or more hypervariable region residues of a parent antibody (e.g., humanized or human antibodies). Generally, the resulting variant selected for further study has a modification (e.g., improvement) in certain biological properties (e.g., high affinity, low immunogenicity) compared to the parent antibody and / or has certain biological properties that are substantially maintained from the parent antibody. A typical substitution variant is, for example, an affinity matured antibody that can be conveniently generated using affinity maturation techniques based on phage display (e.g., the techniques described herein). Briefly, mutations are introduced into one or more CDR residues, the variant antibody is displayed on phage, and screened for a particular biological activity (e.g., binding affinity).

[0119] Conversion (e.g., substitution) may be made in the CDR, for example, to improve the affinity of an antibody. Such a conversion may be made at CDR “hot spots”, i.e., residues encoded by codons that mutate frequently during the process of somatic maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and / or in the SDR (a-CDR), and at the same time, the resulting mutant VH or VL may be tested for binding affinity. Affinity maturation by constructing and reselecting from a secondary library is described, for example, in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001).). In certain embodiments of affinity maturation, diversity is introduced into the variable genes selected 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. Subsequently, the library is screened to identify any antibody mutants having the desired affinity. Another method for introducing diversity relates to a CDR-directed method in which several CDR residues (e.g., 4 to 6 residues at a time) are randomized. CDR residues involved in antigen binding can be specifically identified, for example, using alanine-scanning mutagenesis or modeling. CDR-H3 and CDR-L3 are particularly often targeted.

[0120] In certain embodiments, a substitution, insertion, or deletion may occur within one or its CDRs as long as such a conversion does not substantially reduce the ability of the antibody to bind to the antigen. For example, conservative conversions that do not substantially reduce binding affinity (e.g., conservative substitutions provided herein) may be made in the CDR. Such a conversion may also be outside of the CDR “hot spots” or SDR. In certain embodiments of the mutant VH and VL sequences provided above, each CDR may be unconverted or may contain one, two, or three or fewer amino acid substitutions.

[0121] A method useful for identifying residues or regions of an antibody that can be targeted for mutagenesis is called "alanine scanning mutagenesis" as described in Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, residues or groups of target residues (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) are identified and substituted with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction of the antibody with its antigen is affected. Further substitutions can be introduced at amino acid positions that show functional sensitivity to the first substitution. Alternatively, or in addition, the crystal structure of the antigen-antibody complex is used to identify the contact points between the antibody and the antigen. Residues in such contact and adjacent residues can be targeted as candidates for substitution or excluded. Mutant forms may be screened to examine whether they have the desired properties.

[0122] Insertions of amino acid sequences include amino-terminal and / or carboxyl-terminal fusions ranging in length from 1 residue to polypeptides containing 100 or more residues, as well as in-sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antibodies having an N-terminal methionine residue. Other insertion mutant forms of the antibody molecule include fusions of enzymes (e.g., ADEPT) or polypeptides that increase the serum half-life of the antibody to the N or C terminus of the antibody.

[0123] In certain embodiments, the antibodies provided herein are engineered to increase or decrease the degree to which the antibody is glycosylated. Addition or deletion of glycosylation sites to the antibody can be conveniently done by engineering the amino acid sequence to create or remove one or more glycosylation sites.

[0124] If the antibody contains an Fc region, the sugar chains bound thereto may be modified. Endogenous antibodies produced by mammalian cells typically contain a bi - branched oligosaccharide bound by an N - terminal linkage to Asn297 in the CH2 region of the Fc region. See, for example, Wright et al., TIBTECH 15:26 - 32 (1997). The oligosaccharide may include various sugar chains such as mannose, N - acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose that binds to GlcNAc in the "stem" of the bi - branched oligosaccharide structure. In certain embodiments, the modification of the oligosaccharide in the antibodies of the present invention can be made to create antibody variants with certain improved properties.

[0125] In one embodiment, an antibody variant is provided that has a glycan structure lacking fucose (either directly or indirectly) linked to the Fc region. For example, the amount of fucose in such an antibody can be 1% - 80%, 1% - 65%, 5% - 65%, or 20% - 40%. The amount of fucose is determined, for example, as described in WO2008 / 077546, by calculating the average amount of fucose within the glycan at Asn297 relative to the total of all glycan structures (e.g., complex, hybrid, and high-mannose structures) linked to Asn297. Asn297 refers to the asparagine residue located at approximately 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)), although Asn297 can also be located within approximately ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to small sequence variations in the antibody. Such fucosylation variants can have enhanced ADCC function. See, for example, US Patent Publication No. US2003 / 0157108 (Presta, L.); US2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of literature regarding "defucosylated" or "fucose-lacking" antibody variants include US2003 / 0157108; WO2000 / 61739; WO2001 / 29246; US2003 / 0115614; US2002 / 0164328; US2004 / 0093621; US2004 / 0132140; US2004 / 0110704; US2004 / 0110282; US2004 / 0109865; WO2003 / 085119; WO2003 / 084570; WO2005 / 035586; WO2005 / 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 generating afucosylated antibodies include Lec13 CHO cells lacking protein fucosylation (Ripka et al., Arch. Biochem. Biophys. 249:533-545 (1986); US Patent Application No. 2003 / 0157108A1 (Presta, L); and WO2004 / 056312A1 (Adams et al.), particularly Example 11), and knockout cell lines, such as the alpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, for example, Yamane-Ohnuki et al., Biotechnol. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107). For example, antibody variants having a bisected oligosaccharide in which the bisected oligosaccharide bound to the Fc region of the antibody is bisected by GlcNAc are further provided. Such antibody variants may exhibit reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, for example, in WO2003 / 011878 (Jean-Mairet et al.); US Patent No. 6602684 (Umana et al.); and US2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide bound to the Fc region are also provided. Such antibody variants may exhibit improved CDC function. Such antibody variants are described, for example, in WO1997 / 30087 (Patel et al.); WO1998 / 58964 (Raju, S.); and WO1999 / 22764 (Raju, S.).

[0126] In certain embodiments, the modification of one or more amino acids may be introduced into the Fc region of the antibodies provided herein, thereby creating a variant of the Fc region. The variant of the Fc region may comprise a human Fc region sequence (e.g., a human, IgG1, IgG2, IgG3, or IgG4 Fc region) comprising a modification (e.g., substitution) of an amino acid at one or more amino acid positions.

[0127] In certain embodiments, the invention contemplates antibody variants that have effector function to some, but not all, extent, which would make them desirable candidates for applications where the in vivo half-life of the antibody is important, but where some effector function (e.g., complement activation and ADCC) is unnecessary or harmful. In vitro and / or in vivo cytotoxicity assays can be performed to confirm the reduction / loss of CDC and / or ADCC activities. For example, an Fc receptor (FcR) binding assay can be performed to confirm that the antibody lacks FcγR binding (which may result in the loss of ADCC activity), but retains the ability to bind to FcRn. NK cells, which are primary cells for mediating ADCC, express only FcγRIII, while monocytes and microglia express FcγRI, FcγRII, and FcγRIII. FcR expression in 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 for evaluating the ADCC activity of a target molecule are described in U.S. Patent No. 5,500,362 (see, e.g., Hellstrom, I. et al. Proc. Nat'l 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 (Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)).

[0128] Alternatively, a non-radioactive assay method may be used (e.g., ACTI® Non-Radioactive Cytotoxicity Assay for Flow Cytometry (Cell Technology, 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.

[0129] Alternatively, or in addition, the ADCC activity of the molecule of interest can be evaluated in vivo, for example, in model animals such as those disclosed in Clynes et al., Proc. Nat'l Acad. sci. USA 95:652-656 (1998).

[0130] A C1q binding assay may also be performed to confirm that the antibody cannot bind to C1q and thus lacks CDC activity. See, for example, the Clq and C3c binding ELISAs in WO2006 / 029879 and WO2005 / 100402. A CDC assay may be performed to evaluate complement activation (see, for example, Gazzano-Santoro et al., 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)). Measurement of FcRn binding and in vivo clearance / half-life can also be performed using methods known in the art (see, for example, Petkova, S.B. et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

[0131] Antibodies having reduced effector function include antibodies having one or more substitutions of residues 238, 265, 269, 270, 297, 327, and 329 in the Fc region (U.S. Patent No. 6,737,056). Such Fc mutations include Fc mutations having substitutions at two or more of amino acid positions 265, 269, 270, 297, and 327 (including so-called "DANA" Fc mutations having substitutions of residues 265 and 297 to alanine) (U.S. Patent No. 7,332,581). Alternatively, antibodies having reduced effector function include antibodies having one or more substitutions of residues 234, 235, and 329 in the Fc region (so-called "PG-LALA" Fc mutations having substitutions of residues 234 and 235 to alanine and 329 to glycine) (Lo, M. et al., Journal of Biochemistry, 292, 3900-3908).

[0132] Certain antibody variants having improved or reduced binding to FcR have been described (see, e.g., U.S. Patent No. 6,737,056; WO2004 / 056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)).

[0133] In certain embodiments, the antibody variant includes an Fc region having one or more amino acid substitutions that improve ADCC, such as substitutions at positions 298, 333, and / or 334 in the Fc region (EU numbering of residues).

[0134] In certain embodiments, the modification is made in the Fc region that results in altered (i.e., improved or reduced) C1q binding and / or complement-dependent cytotoxicity (CDC) as described, for example, in U.S. Patent No. 6,194,551, WO99 / 51642, and Idusogie et al., J. Immunol. 164: 4178-4184 (2000).

[0135] Antibodies having an increased half-life and improved binding to the neonatal Fc receptor (FcRn) (associated with the transfer of maternal IgG to the fetus) (Guyer et al., J. Immunol. 117:587 (1976) and Kirn et al., J. Immunol. 24:249 (1994)) are described in US2005 / 0014934A1 (Hinton et al.). These antibodies include an Fc region having one or more substitutions that improve the binding of the Fc region to FcRn. Such Fc variants include substitutions of one or more of the residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424, or 434 of the hFc region, for example, those having a substitution of residue 434 of the Fc region (U.S. Patent No. 7371826). For other examples of variants of the Fc region, see also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No. 5648260; U.S. Patent No. 5624821; and WO94 / 29351.

[0136] In certain embodiments, it may be desirable to create antibodies modified with cysteine, e.g., “thioMAbs” in which one or more residues of the antibody are substituted with cysteine residues. In certain embodiments, the residue substitutions occur at acceptable sites of the antibody. By substituting these residues with cysteine, reactive thiol groups are placed at acceptable sites of the antibody, as further described herein, and may be used to conjugate the antibody to other moieties (e.g., a drug moiety or a linker-drug moiety) to create an immunoconjugate. In certain embodiments, any one or more of the following residues may be substituted with cysteine: V205 of the light chain (Kabat numbering); A118 of the heavy chain (EU numbering); and S400 of the heavy chain Fc region (EU numbering). Antibodies modified with cysteine may be created, for example, as described in U.S. Patent No. 7521541.

[0137] In certain embodiments, the antibodies provided herein may be further modified to include additional non-protein moieties that are known in the art and are readily available. Moieties suitable for derivatization of antibodies 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), ethylene glycol / propylene glycol copolymers, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly 1,3-dioxolane, poly 1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acids (homopolymers or random copolymers), and dextran or poly(n-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may be effective during production due to its stability in water. The polymers may be of any molecular weight and may be branched or unbranched. The number of polymers conjugated to the antibody may vary, and if more than one polymer is conjugated, they may be the same or different molecules. Generally, 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 is to be used therapeutically under certain conditions, and the like.

[0138] In another embodiment, conjugates of antibodies and non-protein moieties that can be selectively heated by exposure to radiation are provided. In one embodiment, the non-protein moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation can be of any wavelength and includes, but is not limited to, wavelengths that do not harm normal cells but heat the non-protein moiety to a temperature at which cells proximal to the antibody-non-protein moiety are killed.

[0139] Antibodies can be produced using recombinant methods and compositions, such as those described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding the misfolded TDP-43 antibody described herein is provided. Such a nucleic acid can encode an amino acid sequence comprising the VL of the antibody and / or an amino acid sequence comprising the VH (e.g., the light chain and / or heavy chain of the antibody). In a further embodiment, one or more vectors (e.g., expression vectors) comprising such a nucleic acid are provided. In a further embodiment, a host cell comprising such a nucleic acid is provided. In such an embodiment, the host cell comprises (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH, or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and a second vector comprising a nucleic acid encoding an amino acid sequence comprising the VH of the antibody (e.g., is transformed). In one embodiment, the host cell is a eukaryotic cell, such as a Chinese hamster ovary (CHO) cell or a lymphoid cell (e.g., YO, NSO, Sp20). In one embodiment, a method for producing a misfolded TDP-43 antibody, comprising 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) is provided.

[0140] For the recombinant production of anti-misfolded TDP-43 antibodies, for example, nucleic acids encoding antibodies as described above are isolated and inserted into one or more vectors for further cloning and / or expression in host cells or cell-free expression systems. Such nucleic acids can be readily isolated and sequenced using conventional methods (e.g., by using oligonucleotide probes that can specifically bind to the genes encoding the heavy and light chains of the antibody).

[0141] Host cells suitable for the cloning or expression of vectors encoding antibodies include the prokaryotic or eukaryotic cells described herein. For example, antibodies may be produced in bacteria, particularly if glycosylation and Fc effector functions are not required. For the expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, which discloses the expression of antibody fragments in E. coli). After expression, the antibodies can be isolated from the bacterial cell paste in the soluble fraction and further purified.

[0142] In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeast are suitable cloning or expression hosts for vectors encoding antibodies, including fungal and yeast strains whose glycosylation pathways are "humanized" to produce antibodies having partial or complete human glycosylation patterns. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).

[0143] Host cells suitable for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. In particular, many baculovirus strains that can be used in combination with insect cells have been identified for the transfection of Spodoptera frugiperda cells.

[0144] Cultures of plant cells can be used as hosts. See, for example, U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429, which describe the PL antibody (registered trademark) technology for generating antibodies in transgenic plants.

[0145] Vertebrate cells may also be used as hosts. For example, mammalian cell lines used for suspension growth may be useful. Other examples of useful mammalian host cell lines include the CVl macaque kidney cell line transformed by SV40 (COS-7); the human embryonic kidney line 293 or 293 cells (described in Graham et al., J. Gen Viral. 36:59 (1977)); baby hamster kidney cells (BHK); mouse Sertoli cells (e.g., TM4 cells described in Mather, Biol. Reprod. 23:243-251 (1980)); macaque kidney cells (CVl); African green monkey kidney cells (VER0-76); human cervical carcinoma cells (HeLa); dog kidney cells (MDCK); buffalo rat liver cells (BRL3A); human lung cells (Wl38); human liver cells (HepG2); mouse mammary tumor (MMT060562); TRI cells (described in Mather et al., Annals N. Y Aead. Sei. 383:44-68 (1982)), for example; MRC5 cells; and FS4 cells. Other useful mammalian host cell details 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, for example, Yazaki and Wu, Methods in Molecular Biology, Val. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

[0146] The method for generating the TDP-43 binding molecule of the present invention, particularly an antibody, is a. culturing a suitable host cell or cell-free expression system under conditions suitable for generating the binding molecule, particularly an antibody; and then b. isolating the binding molecule, particularly an antibody It may include the steps of engineering. Appropriate culturing and isolation techniques can be utilized by those skilled in the art.

[0147] The anti-TDP-43 antibodies provided herein may be identified, screened, or characterized for their physical / chemical properties and / or biological activities by various assays known in the art.

[0148] In one embodiment, the antibodies of the invention are tested for their antigen-binding activity by known methods such as, for example, ELISA, BIACORE®, FACS, immunofluorescence, or immunohistochemistry.

[0149] In another embodiment, a competition assay can be used to identify antibodies that compete with any of the antibodies described herein for binding to misfolded TDP-43. In certain embodiments, such competing antibodies bind to the same epitope (e.g., linear or conformational epitope) as the epitope bound by the antibodies described herein. Detailed and exemplary methods for mapping the epitope to which an antibody binds are provided in Morris (1996) "Epitope Mapping Protocols," in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ).

[0150] In a typical competitive assay, immobilized TDP-43 is incubated in a solution containing a first labeled antibody that binds to TDP-43 (e.g., any of the antibodies described herein) and a second unlabeled antibody, and tested for its ability to compete with the first antibody for binding to TDP-43. As a control, immobilized TDP-43 is incubated in a solution that contains the first labeled antibody but does not contain the second unlabeled antibody. After incubation under conditions that permit binding of the first antibody to TDP-43, excess unbound antibody is removed and the amount of label bound to the immobilized TDP-43 is measured. If the amount of label bound to the immobilized TDP-43 is substantially decreased in the test sample as compared to the control sample, this indicates that the second antibody competes with the first antibody for binding to TDP-43. See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

[0151] The invention also provides an immunoconjugate comprising an anti-TDP-43 antibody provided herein conjugated to one or more therapeutic agents, such as chemotherapeutic agents or drugs, growth inhibitors, toxins (e.g., protein toxins, enzymatically active toxins (derived from bacteria, fungi, plants, or animals), or fragments thereof), radioisotopes (i.e., radioactive conjugates), blood-brain barrier penetrating moieties, or detectable labels.

[0152] In another aspect of the present invention, there is provided a product comprising a material useful for the treatment, prevention, and / or diagnosis of a disease, disorder, abnormality, or TDP-43 proteinopathy associated with the above-described TDP-43, particularly a TDP-43 aggregate. The product includes a container and a label or package insert on or attached to the container. Suitable containers include, for example, bottles, vials, syringes, intravenous (IV) solution bags, and the like. The container may be formed from various raw materials, such as glass or plastic. The container contains its own composition or a composition combined with another composition effective for treating, preventing, and / or diagnosing a disease, and may have a sterilizable opening and closing means (for example, the container may be an IV solution bag or vial having a stopper pierceable by a subcutaneous injection needle). At least one active agent in the composition is an antibody of the present invention. The label or package insert indicates that the composition is used for treating an appropriate disease. Further, the product may include (a) a first container having the composition contained therein (the composition includes an antibody of the present invention); and (b) a second container having the composition contained therein (the composition includes a further therapeutic agent). The product in this embodiment of the present invention may further include a package insert indicating that the composition can be used for treating a specific disease. Alternatively, or in addition, the product may further include a second (or third) container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. Other commercially and user-desired materials, including other buffers, diluents, filters, needles, and syringes, may also be included.

[0153] It is understood that any of the above products may include, instead of or in addition to the anti-TDP-43 antibody, an immunoconjugate of the present invention.

[0154] Exemplary TDP-43 specific binding molecules or antibodies In certain embodiments of the invention, the antibody is: a) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12; VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17; b) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 21; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22; VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 25; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; c) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 31; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 32; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 33; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 35; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 36; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 37; d) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 41; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 42; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 45; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 46; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; e) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 61; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 62; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 63; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 65; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 66; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 67; f) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 71; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 72; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 73; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 75; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 77; g) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 81; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 82; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 83; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 85; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 86; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 87; h) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 101; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 102; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 103; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 105; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 106; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 107; i) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 121; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 122; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 123; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 125; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 16; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 127; j) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 141; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 142; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 143; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 143; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 145; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 146; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 147; or k) VH-CDR1 containing the amino acid sequence of SEQ ID NO: 151; VH-CDR2 containing the amino acid sequence of SEQ ID NO: 152; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 153; VH-CDR3 containing the amino acid sequence of SEQ ID NO: 153; VL-CDR1 containing the amino acid sequence of SEQ ID NO: 155; VL-CDR2 containing the amino acid sequence of SEQ ID NO: 156; and VL-CDR3 containing the amino acid sequence of SEQ ID NO: 157 comprising.

[0155] In certain embodiments, the antibody is: a. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 10 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 14; b. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 20 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 24; c. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 30 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 34; d. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 40 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 44; e. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 60 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 64; f. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 70 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 74; g. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 80 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 84; h. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 100 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 104; i. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 120 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 124; j. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 140 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 144; or k. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 150 and a light chain variable region (VL) containing the sequence of SEQ ID NO: 154 is included.

[0156] In certain embodiments, the antibody is: a. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 10 or having at least 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a light chain variable region (VL) containing the sequence of SEQ ID NO: 14 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14; b. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 20 or having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 20; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 24 or having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 24; c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 30 or having at least 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 30; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 34 or having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 34; d. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 40 or having at least 90%, 91%, 92%, 93%, 94%, 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; e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 60 or having at least 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 having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 64; f. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 70 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 70; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 74 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 74; g. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 80 or having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 80; and a light chain variable region (VL) containing the sequence of SEQ ID NO: 84 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 84; h. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 100 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 100; and a light chain variable region (VL) containing the sequence of SEQ ID NO: 104 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; i. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 120 or having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 120; and a light chain variable region (VL) containing the sequence of SEQ ID NO: 124 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; j. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 140 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 140; and a light chain variable region (VL) containing the sequence of SEQ ID NO: 144; or k. A heavy chain variable region (VH) containing the sequence of SEQ ID NO: 150 or having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 150; and a light chain variable region (VL) containing the sequence of SEQ ID NO: 154 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 154 comprising.

[0157] In certain embodiments, the invention relates to an antibody derived from hybridoma clone 631B2A2, 633B12C8, 634H10H7, 636E5B8, 641H1E7, 642A10B11, 642D12B4, 646B7F7, 712A6B10, 809D9C2, or 809F12D8, as further described herein.

[0158] In certain embodiments, the invention relates to an antibody selected from ACI-7069-631B2-Ab1, ACI-7069-633B12-Ab1, ACI-7069-634H10-Ab2, ACI-7069-636E5-Ab1, ACI-7069-641H1-Ab2, ACI-7069-642A10-Ab1, ACI-7069-642D12-Ab1, ACI-7069-646B7-Ab1, ACI-7071-712A6-Ab1, ACI-7071-809D9-Ab2, and ACI-7071-809F12-Ab1, as further described herein.

[0159] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid encodes an antibody described herein.

[0160] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 18 encoding an anti-TPD-43 antibody.

[0161] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 19 encoding an anti-TPD-43 antibody.

[0162] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 28 encoding an anti-TPD-43 antibody.

[0163] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 29 encoding an anti-TPD-43 antibody.

[0164] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 38 encoding an anti-TPD-43 antibody.

[0165] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 39 encoding an anti-TPD-43 antibody.

[0166] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 48 encoding an anti-TPD-43 antibody.

[0167] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 49 encoding an anti-TPD-43 antibody.

[0168] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 68 encoding an anti-TPD-43 antibody.

[0169] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 69 encoding an anti-TPD-43 antibody.

[0170] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 78 encoding an anti-TPD-43 antibody.

[0171] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 79 encoding an anti-TPD-43 antibody.

[0172] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 88 encoding an anti-TPD-43 antibody.

[0173] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 89 encoding an anti-TPD-43 antibody.

[0174] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 108 encoding an anti-TPD-43 antibody.

[0175] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 109 encoding an anti-TPD-43 antibody.

[0176] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 128 encoding an anti-TPD-43 antibody.

[0177] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 129 encoding an anti-TPD-43 antibody.

[0178] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 148 encoding an anti-TPD-43 antibody.

[0179] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 149 encoding an anti-TPD-43 antibody.

[0180] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 158 encoding an anti-TPD-43 antibody.

[0181] In some embodiments, an (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid comprises SEQ ID NO: 159 encoding an anti-TPD-43 antibody.

[0182] XII. Compositions and Methods In certain embodiments, an immunoconjugate is provided, the immunoconjugate comprising an (isolated) antibody and a therapeutic agent described herein. In certain embodiments, a labeled antibody comprising an antibody described herein and a detectable label is provided.

[0183] In certain embodiments, a pharmaceutical composition comprising an (isolated) antibody described herein and a pharmaceutically acceptable carrier is provided.

[0184] In certain embodiments, the TDP-43 specific binding molecule of the present invention is conjugated to a detectable label.

[0185] In certain embodiments, the TDP-43 specific binding molecule is part of an immunoconjugate in which the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent.

[0186] In certain embodiments, the TDP-43 specific binding molecule or an immunoconjugate comprising the same is present as a composition comprising a TDP-43 specific binding molecule and a TDP-43 agonist and homolog, or antagonists thereof.

[0187] In certain embodiments, the TDP-43 specific binding molecule is part of a pharmaceutical composition comprising the TDP-43 specific binding molecule or an immunoconjugate (wherein the TDP-43 specific binding molecule is covalently attached to another suitable therapeutic agent), or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologs or antagonists thereof, in combination with a pharmaceutically acceptable carrier.

[0188] In certain embodiments, the TDP-43 specific binding molecule is part of a detection and / or diagnostic kit comprising the TDP-43 specific binding molecule or an immunoconjugate (wherein the TDP-43 specific binding molecule is covalently attached to another suitable therapeutic agent), or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologs, or antagonists thereof.

[0189] Kits comprising the binding molecules of the invention are also provided. In particular, such kits may be useful for performing the diagnostic methods (including classification, monitoring, and treatment selection methods) of the invention. Thus, kits for the diagnosis of diseases, disorders and / or abnormalities associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathies, or kits for use in the methods of the invention comprising the TDP-43 specific binding molecules of the invention are provided. Such kits may contain all the components necessary to perform the methods provided herein. Typically, each component is stored separately in one whole package. Suitable additional components included in the kit are, for example, buffers, detectable dyes, laboratory equipment, reaction vessels, instructions, etc. The instructions for use may be tailored to the particular method for which the kit is used. Appropriately labeled TDP-43 binding molecules of the invention are also provided and may be included in such kits.

[0190] In certain embodiments, the TDP-43 specific binding molecule is used in an immunodiagnostic method for use in the prevention, diagnosis or treatment of TDP-43 proteinopathy.

[0191] In certain embodiments, the TDP-43 specific binding molecule is an immunotherapy for the prevention or treatment of TDP-43 proteinopathy, and an effective amount of the TDP-43 specific binding molecule, or an immunoconjugate (wherein the TDP-43 specific binding molecule is covalently attached to another suitable therapeutic agent), or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologs or antagonists thereof is administered to a subject in need thereof as part of an immunotherapy.

[0192] In certain embodiments, the TDP-43 specific binding molecule or an immunoconjugate (wherein the TDP-43 specific binding molecule is covalently attached to another suitable therapeutic agent), or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologs or antagonists thereof is administered to a subject in need thereof for the diagnosis, prevention, alleviation, or treatment of a disease, disorder, and / or abnormality associated with TDP-43, particularly 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), limbic-predominant age-related TDP-43 encephalopathy (LATE), or TDP-43 proteinopathy.

[0193] In certain embodiments, the TDP-43 specific binding molecule or 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 and a TDP-43 agonist and homolog or antagonist thereof, is administered to a subject in need thereof for use in a method of diagnosing, or monitoring, TDP-43, particularly diseases, disorders and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathies, selected from frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), having a progranulin (GRN) mutation, having a C9orf72 mutation, having a TARDBP mutation, having a valosin-containing protein (VCP) mutation, related to chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, having a TARDBP mutation, having an 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 AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathies (sporadic inclusion body myositis, inclusion body myopathy with mutations in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia)), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy having a mutation in the myotilin (MYOT) gene or a gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB) or Parkinson's disease (PD).

[0194] In other embodiments, the present invention relates to a method for detecting, diagnosing, or monitoring a disease, disorder, and / or abnormality associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathy, 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).

[0195] Preferably, the disease, disorder, and / or abnormality associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathy is 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, particularly TDP-43 aggregates, or TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS). More preferably, the disease, disorder, and / or abnormality associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathy is Alzheimer's disease (AD). More preferably, the disease, disorder, and / or abnormality associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathy is frontotemporal dementia (FTD).

[0196] In certain embodiments, the TDP-43 specific binding molecule is used in a method for diagnosing a pre-onset disease, or monitoring disease progression and treatment efficacy, or predicting responsiveness, or selecting a subject likely to respond to treatment with a TDP-43 specific binding molecule. The method is preferably performed using a sample of human blood or urine. Most preferably, the method includes an ELISA-based assay or a surface plasmon assay.

[0197] In one embodiment, the TDP-43 specific binding molecule is used in a method for detecting, diagnosing, or monitoring a disease selected from frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), having a progranulin (GRN) mutation, having a C9orf72 mutation, having a TARDBP mutation, having a valosin-containing protein (VCP) mutation, associated with chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, having a TARDBP mutation, having an 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 AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with a mutation in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia)), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy having a mutation in the myotilin (MYOT) gene or a mutation in the gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB) or Parkinson's disease (PD) by contacting the TDP-43 specific binding molecule of the invention with a sample (e.g., blood, cerebrospinal fluid, or brain tissue).

[0198] In one embodiment, the TDP-43 specific binding molecule or 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 and a TDP-43 agonist and homolog or antagonist thereof, is administered to a subject in need thereof for preventing, reducing, or treating TDP-43, particularly diseases, disorders and / or abnormalities associated with TDP-43 aggregates, or frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD, including sporadic and familial AD), chronic traumatic encephalopathy, Perry syndrome, limbic-predominant age-related TDP-43 encephalopathy (LATE), and / or Parkinson's disease (PD).

[0199] In certain embodiments, the TDP-43 specific binding molecule or immunoconjugate (wherein the TDP-43 specific binding molecule is covalently attached to another suitable therapeutic agent), or a composition comprising a TDP-43 specific binding molecule and a TDP-43 agonist and homolog or antagonist thereof, is administered to a subject in need thereof for treating a disease selected from frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), having a progranulin (GRN) mutation, having a C9orf72 mutation, having a TARDBP mutation, having a valosin-containing protein (VCP) mutation, associated with chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD) (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, having a TARDBP mutation, having an 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 AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with a mutation in valosin-containing protein (VCP) (also associated with Paget's disease of bone and frontotemporal dementia)), oculopharyngeal muscular dystrophy (with rimmed vacuoles), myofibrillar myopathy having a mutation in the myotilin (MYOT) gene or a gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB) or Parkinson's disease (PD). Preferably, the disease treatment aids in maintaining or increasing mental awareness and / or reducing the level of TDP-43 aggregates in the brain.

[0200] In certain embodiments, the TDP-43 specific binding molecule or immunoconjugate (wherein the TDP-43 specific binding molecule is covalently attached to another suitable therapeutic agent), or a composition comprising a TDP-43 specific binding molecule and a TDP-43 agonist and homologs or antagonists thereof, is administered to a subject in need thereof, and is used to manufacture a therapeutic agent for TDP-43, particularly diseases, disorders and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathy, or frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (including sporadic and familial AD), chronic traumatic encephalopathy, Perry syndrome, limbic-predominant age-related TDP-43 encephalopathy (LATE), and / or Parkinson's disease (PD).

[0201] The pharmaceutical formulations of the anti-TDP-43 antibodies (preferred types of TDP-43 specific binding molecules) or immunoconjugates described herein are prepared by mixing such an antibody or immunoconjugate (having the desired purity) in the form of a lyophilized formulation or an aqueous solution with one or more optional pharmaceutically acceptable carriers (Remington’s Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Pharmaceutically acceptable carriers are generally non-toxic to the recipient at the dosages and concentrations used and include, but are not limited to, buffers (e.g., phosphoric, citric, and other organic acids); antioxidants (including ascorbic acid and methionine); preservatives (e.g., octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzetonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens (e.g., methylparaben or propylparaben); catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (about 10 residues or less) polypeptides; proteins (e.g., serum albumin, gelatin, or immunoglobulins); hydrophilic polymers (e.g., polyvinylpyrrolidone); amino acids (e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine); monosaccharides, disaccharides, and other sugars (including glucose, mannose, or dextrin); chelating agents (e.g., EDTA); sugars (e.g., sucrose, mannitol, trehalose or sorbitol); salt-forming counterions (e.g., sodium); metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants (e.g., polyethylene glycol (PEG)).Typical pharmaceutically acceptable carriers described herein further include interstitial drug dispersants, such as soluble neutral hyaluronidase glycoprotein (sHASEGP), such as human soluble PH-20 hyaluronidase glycoprotein, such as rHuPH20 (HYLENEX®, Baxter International, Inc). Certain typical HASEGs and methods of use (including rHuPH20) are described in US Patent Publications Nos. 2005 / 0260186 and 2006 / 0104968. In one aspect, sHASEGP is combined with one or more additional glycosaminoglycanases (e.g., chondroitinase).

[0202] A typical lyophilized antibody or immunoconjugate formulation is disclosed in US Patent No. 6267958. Aqueous antibody or immunoconjugate formulations include those described in US Patent No. 6171586 and W02006 / 044908, the latter of which includes a histidine acetate buffer.

[0203] The formulations described herein may also include one or more active ingredients, preferably those having complementary activities that do not produce adverse effects on each other, as required for the particular efficacy to be treated.

[0204] The active ingredient may be encapsulated in a colloidal drug delivery system (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or a macroemulsion, such as microcapsules prepared by coacervation techniques or interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

[0205] Sustained-release formulations may be prepared. Suitable examples of sustained-release formulations include semipermeable matrices of solid hydrophobic polymers containing the antibody or immunoconjugate, which matrices are in the form of shaped products such as films or microcapsules. Formulations for in vivo administration are generally sterile. Sterility can be readily achieved, for example, by filtration through a sterile filtration membrane.

[0206] Any of the antigen-binding molecules, anti-TDP-43 antibodies, or immunoconjugates provided herein may be used in a method, such as a therapeutic method.

[0207] In another aspect, an anti-TDP-43 antibody (a preferred type of TDP-43 specific binding molecule) or an immunoconjugate for use as a medicament is provided. In a further aspect, an anti-misfolded TDP-43 antibody (a preferred type of TDP-43 specific binding molecule) or an immunoconjugate for use in a therapeutic method is provided. In certain embodiments, an anti-TDP-43 antibody (a preferred type of TDP-43 specific binding molecule) or an immunoconjugate for use in the prevention, diagnosis, and / or treatment of TDP-43 proteinopathy is provided. In a preferred embodiment of the invention, the anti-TDP-43 antibody (a preferred type of TDP-43 specific binding molecule) or an immunoconjugate is provided for use in the prevention, diagnosis, and / or treatment of diseases, disorders and / or abnormalities associated with TDP-43, particularly 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 / or limbic-predominant age-related TDP-43 encephalopathy (LATE).

[0208] In a further aspect, the present invention provides for the use of an anti-TDP-43 antibody (a preferred type of TDP-43 specific binding molecule) or an immunoconjugate in the manufacture or preparation of a medicament. In such an embodiment of 1, the method further comprises administering to the individual an effective amount of at least one further therapeutic agent (such as those described below).

[0209] A "subject" or "individual" according to any of the above embodiments can be an animal, a mammal, preferably a human.

[0210] In a further aspect, the present invention provides a pharmaceutical formulation comprising any of the anti-TDP-43 antibodies (a preferred type of TDP-43 specific binding molecule) or immunoconjugates provided herein for use, for example, in any of the above treatment methods. In one embodiment, the pharmaceutical formulation comprises any of the anti-TDP-43 antibodies (a preferred type of TDP-43 specific binding molecule) or immunoconjugates provided herein and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical formulation comprises any of the anti-TDP-43 antibodies (a preferred type of TDP-43 specific binding molecule) or immunoconjugates provided herein and at least one further therapeutic agent (such as those described below).

[0211] The antibodies or immunoconjugates of the present invention can be used in therapy alone or in combination with other agents. For example, the antibodies (a preferred type of TDP-43 specific binding molecule) or immunoconjugates of the present invention may be co-administered with at least one further therapeutic agent.

[0212] Such combination therapies as described above include combined administration (where two or more therapeutic agents are included in the same or different formulations) and separate administrations. In the case of separate administrations, administration of the antibody (preferred type of TDP-43 specific binding molecule) or immunoconjugate of the present invention can be carried out before, simultaneously with, and / or after administration of additional therapeutic agents and / or adjuvants. The antibody (preferred type of TDP-43 specific binding molecule) or immunoconjugate of the present invention can also be used in combination with radiotherapy.

[0213] The antibody (preferred type of TDP-43 specific binding molecule) or immunoconjugate (and additional therapeutic agents) of the present invention can be administered by any suitable method including parenteral, intranasal, and intranasal, and, optionally, topical therapy, intralesional administration, intrauterine, or intravesical administration. Parenteral inhalation includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Dosing can be carried out by any suitable route, for example, by injection such as intravenous or subcutaneous injection, depending in part on whether the dosing is short-term or chronic. A variety of dosing schedules (including, but not limited to, single or multiple administrations, bolus administrations, and pulse infusions at various time points) are contemplated herein.

[0214] The antibodies (preferred types of TDP-43 specific binding molecules) or immunoconjugates of the present invention can be formulated, dosed, and administered in a form corresponding to appropriate medical practice. Factors considered in the present invention include the TDP-43 to be treated, particularly specific diseases, disorders, and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathy, the specific mammalian species to be treated, the clinical condition of each subject, the etiology of diseases, disorders, and / or abnormalities associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathy, the site of drug delivery, the method of administration, the dosing schedule, and other factors known to medical practitioners. The antibodies or immunoconjugates do not require one or more drugs currently used to prevent or treat the problem TDP-43, particularly diseases, disorders, and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathy, but may be appropriately formulated. The effective amount of such other drugs depends on the amount of antibody or immunoconjugate present in the formulation, TDP-43, particularly diseases, disorders, and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathy, or the type of treatment, and other factors described above. These are generally used at the same dosages and administration routes as described herein, or at about 1 to 99% of the dosages described herein, or at any dosage and any route determined to be appropriate empirically / clinically.

[0215] In the prevention or treatment of a disease, the appropriate dosage of an antibody (a preferred type of TDP-43 specific binding molecule) or immunoconjugate of the present invention (when used alone or in combination with one or more other additional therapeutic agents) depends on the type of disease to be treated, the type of antibody or immunoconjugate, the severity and course of the disease, whether the antibody or immunoconjugate is administered for prophylactic or therapeutic purposes, past treatment history, the subject's medical history and responsiveness to the antibody or immunoconjugate, and the judgment of the treating physician. The antibody (a preferred type of TDP-43 specific binding molecule) or immunoconjugate is appropriately administered to the subject either once or over a series of treatment periods. Depending on the type and severity of the disease, about 1 μg / kg to 15 mg / kg (e.g., 0.1 mg / kg to 10 mg / kg) of the antibody (a preferred type of TDP-43 specific binding molecule) or immunoconjugate can be a first candidate dosage for administration to the subject by one or more separate administrations or continuous infusion. One typical daily dosage can range from about 1 μg / kg to 100 mg / kg or more, depending on the factors described above. For repeated administrations over several days or more, depending on the condition, the treatment is generally continued until the desired suppression of the disease occurs. One typical dosage of the antibody or immunoconjugate ranges from about 0.05 mg / kg to about 10 mg / kg. Thus, one or more dosages 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 dosages may be administered intermittently, for example, once a week or once every three weeks (e.g., such that the subject receives about 2 to about 20 doses of the antibody or, for example, about 3 doses). An initial high dosage followed by one or more low dosages may be administered. However, other dosing regimens may also be useful. The course of this treatment is easily monitored by prior art and assays.

[0216] It is understood that any of the above formulations or treatment methods may be carried out using both the immunoconjugate of the present invention and an anti-TDP-43 antibody (a preferred type of TDP-43 specific binding molecule).

[0217] In another aspect of the present invention, there is provided a product comprising a material useful for the treatment, prevention, and / or diagnosis of TDP-43 as described above, particularly diseases, disorders, and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathy. The product includes a container and a label or package insert on or attached to the container. Suitable containers include, for example, bottles, vials, syringes, intravenous (IV) solution bags, and the like. The container may be formed from various raw materials, such as glass or plastic. The container contains a composition of its own or a composition combined with another composition effective for treating, preventing, and / or diagnosing TDP-43, particularly diseases, disorders, and / or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathy, and may have a sterile opening and closing (for example, the container may be an IV solution bag or vial with a stopper pierceable by a subcutaneous injection needle). At least one active agent in the composition is an antibody of the present invention. The label or package insert indicates that the composition is used for treating an appropriate disease.

[0218] Furthermore, the product may include: (a) a first container having a composition contained therein (the composition includes an antibody of the present invention (a preferred type of TDP-43 specific binding molecule) or an immunoconjugate); and (b) a second container having a composition contained therein (the composition includes a further therapeutic agent). The product in this embodiment of the present invention may further include an indication that the composition can be used to treat a specific disease. Alternatively, or in addition, the product may further include a second (or third) container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution, and dextrose solution. Other commercially and user-desired materials, including other buffers, diluents, filters, needles, and syringes, may also be included.

[0219] In a further embodiment, the present invention relates to a method of maintaining, increasing, preventing, and / or delaying the decline of cognitive memory ability, movement, and language function in a subject, the method comprising administering a binding molecule of the present invention, an immunoconjugate of the present invention, a composition of the present invention, or a pharmaceutical composition of the present invention.

[0220] In a further embodiment, the present invention relates to a method of reducing the level of TDP-43, the method comprising administering a binding molecule of the present invention, an immunoconjugate of the present invention, a composition of the present invention, or a pharmaceutical composition of the present invention.

[0221] The method of the present invention may include administering at least one further treatment, and the further treatment is selected from, but not limited to, neuropharmaceuticals, anti-A beta antibodies, anti-tau antibodies, tau aggregation inhibitors, beta amyloid aggregation inhibitors, anti-BACE1 antibodies, and BACE1 inhibitors.

[0222] The present invention further relates to a method for detecting TDP-43, which comprises contacting a sample with the binding molecule of the present invention, preferably, the sample is a brain sample, cerebrospinal fluid sample, urine sample, or blood sample.

Examples

[0223] Example 1: Preparation of TDP-43 Vaccine Composition A liposome-based vaccine was prepared according to the protocol published in WO2012 / 055933. A vaccine containing the full-length TDP-43 (FL TDP-43) protein as an antigen (Table 2, SEQ ID NO: 1) was used for antibody production. Table 2: Description of TDP-43 Protein and Peptide Antigens

Table 2

[0224] Example 2: Generation of Anti-TDP-43 Antibodies A. Immunization of Mice Female C57BL / 6J OlaHsd (C57BL / 6) and BALB / c OlaHsd (BALB / c) wild-type mice (Harlan, USA) were administered at 9 weeks of age. Vaccination was started at week 10. The mice were vaccinated with the full-length TDP-43 protein presented on the surface of liposomes in the presence of monophosphoryl hexaacyl lipid A, 3-deacyl (synthetic) (3D-(6-acyl)PHAD (registered trademark)) as an adjuvant.

[0225] The mice were vaccinated by subcutaneous injection (sc) on days 0, 4, 8, 21, 35, and 60. Blood was collected from the mice, and heparinized plasma was prepared 7 days before immunization (pre-immune plasma), as well as on days 14, 28, 42, 81, and 121 after the first immunization. The mice used for myeloma fusion were further vaccinated with three daily booster injections of the TDP-43 protein by intraperitoneal injection without an adjuvant.

[0226] The vaccine reaction was measured in mouse plasma. Binding of plasma-derived antibodies from immunized mice to immobilized recombinant full-length (FL) TDP-43 showed a high titer of antibodies against TDP-43.

[0227] B. Generation and selection of subcloning of hybridomas Mice were euthanized, and fusion with myeloma cells was performed using splenocytes from each of four mice. Screening of antibodies from the successfully fused hybridoma cell lines was carried out as follows. Diluted (1:32) cell culture supernatants were analyzed using a Luminex bead-based multiplex assay (Luminex, Netherlands). Luminex beads were bound to FL TDP-43, and IgG was captured using anti-mouse IgG-Fc antibodies (Jackson Immunoresearch, USA) specific for IgG1, IgG2a, IgG2b, IgG2c, and IgG3 subclasses. 386 hits from mice immunized with FL TDP-43 liposome vaccine were identified by binding to beads bound to FL TDP-43.

[0228] Surviving hybridomas were grown using selective medium containing serum. Clones that preferentially bind to TDP-43 inclusions in human FTD brains and clones that bind to the C-terminus of TDP-43 were selected for further subcloning. After limiting dilution, clonal hybridomas were grown in low-immunoglobulin-containing medium, and stable colonies were selected for antibody screening and selection. The antibodies shown in Table 3 were identified from this screening.

[0229] Example 3: Determination of binding efficiency (EC50) A Luminex assay with serial dilution of the antibody was performed as described above to determine the half-maximal effective concentration (EC50) of the antibody binding to FL TDP-43. All EC50 values are summarized in Table 3. In summary, all test antibodies bind to full-length TDP-43 with high affinity. Table 3: EC50 values determined by Luminex assay

Table 3

[0230] Example 4: Antibodies Binding to Human FLTDP-43 Antibodies binding to 1 μg / ml of human FL TDP-43 were determined using indirect ELISA. Coating of the ELISA plates with 1 μg / ml of human FLTDP-43 was performed overnight at 4 °C in carbonate buffer. The plates were washed with 0.05% Tween-20 / PBS and blocked with 1% bovine serum albumin (BSA) in 0.05% Tween-20 / PBS for 1 h at 37 °C. Next, antibodies purified from the hybridoma supernatants were added in 3-fold serial dilutions from 1 μg / ml, incubated for 2 h at 37 °C, and the plates were washed. AP-conjugated anti-mouse IgG secondary antibody (Jackson Immunoresearch Laboratories, UK) was added at a 1 / 1000 dilution in 0.05% Tween-20 / PBS for 1 h at 37 °C. After the final wash, the plates were incubated with pNPP (Sigma-Aldrich, Switzerland) phosphatase substrate solution and read at 405 nm using an ELISA plate reader (Tecan, Switzerland). All tested clones bound to full-length TDP-43 with various EC50 values in the range of 10 - 1567 ng / ml (Table 4). Table 4: EC50 Values by ELISA

Table 4

[0231] Example 5: Epitope Mapping by ELISA and Peptide Array Antibodies purified from serum-free hybridoma supernatants were screened by indirect ELISA assay to determine the binding regions using a library of linear peptides of 40 - 66 aa or 15mer peptides with biotinylated N-termini that cover the entire sequence of TDP-43 with a 9aa offset and 6aa overlap. The peptide sequences are shown in Table 5.

[0232] A 96-well plate was coated overnight at 4°C with 5 μg / ml of non-biotinylated peptide in carbonate buffer. The plate was washed with 0.05% Tween-20 / PBS and then blocked with 1% bovine serum albumin (BSA) in 0.05% Tween-20 / PBS at 37°C for 1 hour. Next, the antibody purified from the hybridoma supernatant was added at 1 μg / ml, incubated at 37°C for 2 hours, and the plate was washed. The AP-conjugated anti-mouse IgG secondary antibody (Jackson Immunoresearch Laboratories, UK) was added at a 1 / 1000 dilution in 0.05% Tween-20 / PBS at 37°C for 1 hour. After the final wash, the plate was incubated with the pNPP (Sigma-Aldrich, Switzerland) phosphatase substrate solution and read at 405 nm using an ELISA plate reader (Tecan, Switzerland).

[0233] In the case of the biotinylated peptide, a 96-well ELISA plate coated with streptavidin was incubated with a 15-mer peptide biotinylated at 5 μg / mL. The plate was washed three times with 0.05% Tween-20 / PBS and then blocked with 1% bovine serum albumin (BSA) in 0.05% Tween-20 / PBS at 37 °C for 1 hour. Next, the antibody purified from the hybridoma supernatant was added at 1 μg / ml, incubated at 37 °C for 2 hours, and the plate was washed. The AP-conjugated anti-mouse IgG secondary antibody (Jackson ImmunoResearch Laboratories, UK) was added at a 1 / 1000 dilution in 0.05% Tween-20 / PBS at 37 °C for 1 hour. After the final wash, the plate was incubated with pNPP (Sigma-Aldrich, Switzerland), an AP substrate solution, and read at 405 nm using an ELISA plate reader (Tecan). The determined binding regions are shown in Table 6. The test antibodies were found to bind to the following peptides: TP-21, TP-23, TP-35, TP-40, TP-48, TDP-6 (corresponding to regions 181-195, 199-213, 307-321, 352-366, 389-411, 140-200 of SEQ ID NO: 1, respectively).

[0234] More accurate linear epitopes were mapped using a library of 15-mer peptides that were directly synthesized on a solid support and covered the entire sequence of TDP-43 according to SEQ ID NO: 1 with 1aa offset and 14aa overlap (Pepscan, Netherlands). The peptide array was blocked with horse serum and ovalbumin and incubated overnight at 4 °C with a purified antibody solution at a concentration of 0.75 - 5 μg / ml. After washing, the peptide array was incubated at 25 °C for 1 hour with a 1 / 1000 dilution of rabbit anti-mouse IgG (H+L) HRP conjugate (Southern Biotech, USA). After washing, the peroxidase substrates 2,2'-azinodi-3-ethylbenzothiazoline sulfonate (ABTS) and 3% H2O2 at 20 μl / ml were added. After 1 hour, the color development was quantified using a charge-coupled device (CDD) (camera and image processing system). These binding regions were confirmed by epitope mapping and the following epitopes (provided in Table 6) were identified: aa183 - 188, 203 - 213, 204 - 208, 204 - 211, 205 - 210, 316 - 323, 358 - 361, 400 - 405, 400 - 406, 400 - 412 of SEQ ID NO: 1. Table 5: Peptides used for determination of binding regions by ELISA

Table 5

Table 6

[0235] Example 6: Detection of TDP-43 in brain tissue from FTD / ALS subjects by immunohistochemistry The involvement of the target was evaluated by immunohistochemical experiments on tissues from the brains of FTD subjects. Human FTD brain tissues were obtained from the Netherlands Brain Bank (open access: www.brainbank.nl) of the Netherlands Institute for Neuroscience in Amsterdam and the Queen Square Brain Bank for Neurological Disorders (UCL). All materials were collected from donors for whom written informed consent regarding autopsy of the brain for research purposes and the use of materials and clinical information had been obtained by the brain bank. Immunohistochemistry was performed on 10-μm-thick frozen sections using secondary antibodies labeled with fluorescence for detection. The following antibodies were used as controls: rabbit polyclonal pan-TDP-43 antibody (Proteintech, 10782-2-AP) for detecting pathological inclusions and physiological nuclear TDP-43; rabbit monoclonal phospho-TDP-43 p409 / 410 antibody (Cosmobio, TIP-PTD-P02) for detecting aggregated and phosphorylated TDP-43 in lesions; and secondary antibody without primary antibody (No1 Ab) for detecting non-specific background.

[0236] All antibodies of the present invention bind to nuclear, non-aggregated, and aggregated TDP-43. Some antibodies of the present invention preferentially bind to aggregated TDP-43 in the cytoplasm in type A lesions (Figure 1). The detailed evaluation of the binding characteristics is summarized in Table 7. Table 7: Detection of TDP-43 in brain tissues from FTD subjects

Table 7

[0237] Example 7: Detection of TDP-43 in brain tissues from FTD / ALS subjects by Western blot The brain tissue region (frontal cortex) was homogenized at a 1:4 (w / v) ratio in homogenization solubilization buffer (HS buffer) at 4°C using a pre-chilled CK Mix Homogenization Tube (Labgene, BER0092) with a pre-chill. The following procedure was used for homogenization: 3 cycles of 30 seconds at 5000 rpm (with a 15-second pause between each cycle). The homogenized sample was aliquoted and stored at -80°C in 1.5 ml low-protein binding tubes (Axygen MCT-175-L-C). · HS buffer - 10 mM Tris HCl pH 7.5, 150 mM NaCl, 0.1 mM EDTA, 1 mM DTT, Complete EDTA-free protease inhibitor (Roche, 32524300) and PhosSTOP phosphatase inhibitor (Roche, 4906837001).

[0238] The brain homogenate was thawed on ice and resuspended in HS buffer to obtain a final concentration of 2% sarcosyl, 1 unit / μL benzonase, and 1 mM MgCl2. The sample was then incubated at 37°C for 45 minutes with constant shaking at 600 rpm in a thermomixer. The supernatant was collected in a new tube. The precipitate was resuspended in 1000 μl of myelin suspension buffer and centrifuged at 20,000 g for 60 minutes at 4°C in a benchtop centrifuge. The supernatant was carefully removed to remove all floating lipids. If all lipids could not be removed in one step, this step was repeated. Subsequently, the precipitate was washed with PBS and centrifuged at 4°C for 30 minutes in a benchtop centrifuge. The final precipitate was resuspended in 200 μl of PBS and stored at -80°C. The sample was analyzed by immunoblotting under denaturing conditions. · HS buffer (containing sarcosyl, benzonase and MgCl2) - 10 mM Tris HCl pH 7.5, 150 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 4% sarcosyl, 1 unit / μL benzonase (Novagen 70746-4), 4 mM MgCl2, Complete EDTA-free protease inhibitor (Roche) and PhosSTOP phosphatase inhibitor (Roche). · Myelin floating buffer - HS buffer containing 1% Triton X-100 and 30% sucrose.

[0239] Western blot was performed using MES SDS running buffer (Thermofisher) on a Bolt 12% Bis-Tris Plus gel 1.0 mm (Thermofisher). Samples (30 μl / sample) were diluted with PBS and then loaded onto the gel (loading buffer (1x, Licor, 928-40004) containing 100 mM DTT). Proteins were separated at a constant voltage of 100 V for 1 hour. After electrophoresis, proteins were transferred to a nitrocellulose membrane (Thermofisher, IB23001) using iBLOT (Thermofisher, IB21001) at 20 volts for 7 minutes. After protein transfer, the membrane was blocked with Licor blocking buffer (Odyssey blocking buffer 927-40000) diluted 1:3 with PBS for 1 hour. The membrane was incubated overnight with the following primary antibodies: total TDP-43 (Proteintech, 60019-2-Ig or 10782-2-AP), pTDP-43 (Cosmobio, TIP-PTD-M01). For the primary antibodies, the blocking buffer was diluted 1:1 with PBS-T (PBS containing 0.4% Tween-20). The membrane was washed 4 times with PBS-T (PBS containing 0.1% Tween-20) and incubated with a secondary antibody conjugated to LICOR dye. The secondary antibody (goat anti-mouse (catalog number 926-68072) or donkey anti-rabbit (catalog number 926-32211)) was diluted 1:10000 with Licor blocking buffer diluted 1:1 with PBS-T (PBS containing 0.4% Tween-20) and used at room temperature for 1 hour. The membrane was washed again 4 times with PBS-T (PBS containing 0.1% Tween-20) and scanned using the LICOR system. Figure 2 shows that all mAbs specifically recognize full-length TDP-43. Furthermore, some mAbs (K, M, N) recognize pathological features of the disease state such as C-terminal fragments and high molecular weight aggregates in the insoluble fraction.

[0240] Example 8A: Measurement of Binding Activity Using SPR The binding activity to soluble or aggregated FLTDP-43 was evaluated by examining the dissociation constant (KD) using surface plasmon resonance (SPR; Biacore T200, GE Healthcare Life Sciences). Recombinant human soluble or aggregated FLTDP-43 was immobilized on a CM5 series S sensor chip (GE Healthcare Life Sciences) by amine coupling. Soluble TDP-43 was immobilized at a concentration of 5 μg / ml in 10 mM sodium acetate (pH 4.5) at a flow rate of 5 μl / min for 420 seconds to achieve an immobilization level of 150 RU. Aggregated TDP-43 was immobilized at a concentration of 50 μg / ml in 10 mM sodium acetate (pH 4.5) at a flow rate of 5 μl / min for 840 seconds to achieve an immobilization level of 110 RU. Biotinylated TP-73 peptide (aa181 - 190 of SEQ ID NO: 1) was immobilized on a series S sensor chip SA (GE Healthcare Life Sciences) in PBS-P + at a concentration of 5 μg / ml at a flow rate of 5 μl / min for 30 seconds to achieve an immobilization level of 400 RU. To evaluate the KD value, purified antibody and control antibody (2E2-D3) were used in PBS-P +It was started from 333 nM and diluted three-fold down to 0.15 nM and then injected. The antibody was injected at a flow rate of 50 μl / min with a contact time of 90 seconds and a dissociation phase of 700 seconds, and regenerated three times with 10 mM glycine-HCl (pH 1.7). In the optimized SPR protocol, the antibody was diluted three-fold from 300 nM down to 1.2 nM, injected at 30 μl / min for 300 seconds, and dissociated for 600 seconds. The surface was regenerated by injecting 10 mM glycine-HCl (pH 1.7) once. The results obtained from the binding reaction rate were double-referenced using a blank flow cell and a buffer cycle, and evaluated using a global 1:1 fitting model with RI. The binding activities of 11 antibodies and 2 Fab fragments are shown in Table 8. The antibodies of the present invention bind to aggregated TDP-43 with a KD in the range of 0.62 nM to 4.64 nM. Furthermore, some antibodies show preferential binding to aggregated TDP-43 compared to soluble TDP-43. The two Fab fragments bind to soluble TDP-43 with a KD in the range of 2.8 nM to 21.8 nM and show a similar KD for aggregated TDP-43. Two antibodies ( * marked with) were re-analyzed using an optimized SPR protocol with longer binding and dissociation phases that enable more accurate KD measurements, especially for antibodies with a delayed dissociation rate. The two antibodies bind to soluble TDP-43 with a KD in the range of 0.22 nM to 3.9 nM and to aggregated TDP-43 with a KD in the range of 0.18 nM to 0.69 nM. The antibody ACI-7069-642D12-Ab1 binds to the TP-73 peptide with a KD of 3.6 nM. Table 8: Binding characteristics by SPR

Table 8

[0241] Example 8B: Affinity measurement using SPR Binding affinity to soluble FLTDP-43 was assessed by determining the dissociation constant (KD) using surface plasmon resonance (SPR; Biacore T200, GE Healthcare Life Sciences). Goat anti-mouse capture antibodies were immobilized on a CM5 series S sensor chip (GE Healthcare Life Sciences) by amine coupling. The antibodies were incubated in PBS-P + (GE Healthcare Life Sciences) at concentrations of 2-5 μg / ml and capture at a flow rate of 10 μl / min for 120 s, resulting in capture levels of 350-1000 RU. To assess KD values, FL TDP-43 or TP-51 peptides (aa352-414 of SEQ ID NO: 1) were incubated in PBS-P + was injected at 3-fold dilutions starting from 1.2 nM to 100 nM at a flow rate of 30 μl / min with a contact time of 300 s at a kinetic rate of one cycle. Dissociation was recorded for 1 h and regenerated once with 10 mM glycine-HCl (pH 1.7). Results from binding kinetics were double-referenced using a blank flow cell and buffer cycles and evaluated with a global 1:1 fit model using RI. On-rates (ka), off-rates (kd) and affinities (KD) of the three antibodies are shown in Table 9 as the mean ± SD of 12 (ACI-7069-633B12-Ab1), 2 (ACI-7069-642D12-Ab1) or 3 (ACI-7071-809F12-Ab1) replicates. Antibodies ACI-7069-633B12-Ab1, ACI-7069-642D12-Ab1 and ACI-7071-809F12-Ab1 bind to soluble TDP-43 with affinities ranging from 15-135 pM, 226-272 pM and 389-457 pM, respectively. Antibody ACI-7069-633B12-Ab1 binds to the TP-51 peptide with an affinity ranging from 1184-1316 pM. Table 9: Affinity of soluble FL TDP-43 and TP-51 peptides by SPR [Table 9]

[0242] Example 9: Sequencing of Antibodies The cloned hybridoma cell lysate was used for gene sequencing of the variable region. Mouse hybridomas were recovered and lysed using a lysis buffer containing guanidinium salts to inactivate RNase. Genomic DNA was then removed with DNase free of RNase, and the RNA was purified by washing multiple times with a silica-based affinity column and eluted from the column using RNase-free water. Once the RNA was extracted, its purity and concentration were measured spectrophotometrically. The integrity of the RNA was evaluated on a denaturing agarose gel, and the RNA was reverse transcribed into cDNA using reverse transcriptase (RT). The RNA was heated at 70 °C for 10 minutes to disrupt its secondary structure, and the RT reaction mixture was added. The RT product was used directly for PCR amplification. For high-fidelity PCR amplification of the cDNA, each of the variable region primers corresponding to different gene families encoding the antibody was mixed separately with the constant primer for VH and VL, respectively, in a total reaction volume of 50 μl. First, the degenerate first pool was used (12 for VH and 12 for VL), and depending on the results, the second pool was used to obtain the PCR product. After the PCR reaction, the product was analyzed by gel electrophoresis on a 2% agarose gel stained with ethidium bromide. The PCR products of VL and VH were each purified on an agarose gel using Tris-acetate-EDTA (TAE). The purified fragment excised from the gel was sequenced using the dideterminator sequencing method with the same primers used for PCR. Sequencing was performed in both directions with overlap at both ends. The sequences were analyzed using multiple sequence alignment (Clustal tool) and annotated using the Kabat algorithm described in Kabat et al., Sequences of Proteins of Immunological Interest, 91-3242 (1991). The nucleotide sequences of the heavy and light chain variable domains (VH and VL) are shown in Table 10. The translated protein sequences of the selected heavy chain (VH) and light chain (VL) variable domains, and their complementarity determining regions (CDRs) are shown in Table 11. Table 10: Nucleotide sequences of the heavy and light chain variable domains (VH and VL)

Table 10-1

Table 10-2

Table 10-3

Table 10-4

Table 10-5

Table 11-1

Table 11-2

Table 11-3

Table 11-4

Table 11-5

[0243] Example 10: In vivo efficacy of ACI-7069-633B12-Ab1 (IgG2a variant) in a transgenic mouse model of TDP-43 proteinopathy To evaluate the efficacy of ACI-7069-633B12-Ab1 (IgG2a variant) in vivo, the ability of ACI-7069-633B12-Ab1 (IgG2a variant) to reduce TDP-43 lesions in NEFH-tTA x hTDP-43ΔNLS bigenic mice (rNLS8 mice, Walker et al, 2015) was tested. The rNLS8 mice were injected weekly with ACI-7069-633B12-Ab1 (IgG2a variant) (n = 30) or vehicle (n = 30), and molecular pathological markers such as phosphorylated and / or total insoluble TDP-43 were analyzed at the end of dosing.

[0244] 10.1. Animals Before the start of the study, to ensure sufficient health and minimize non-specific stress associated with experimental procedures, all animals were acclimated to the environment, examined, handled, and weighed. Mice were housed on solid feed containing doxycycline (200 mg / kg) during the breeding period and until 8 weeks of age. At 8 weeks of age, the diet was changed to solid feed without doxycycline (DOX) to allow for transgene expression. Throughout the study, the light / dark cycle (12 / 12), room temperature (20 - 23 °C), and relative humidity (approx. 50%) were kept constant. Solid feed and water were provided ad libitum during the experimental period. When the mice began to show difficulty moving, the diet was changed to moist feed and hydrogel on the floor of the cage. All behavioral tests were performed during the light cycle phase of the animals.

[0245] 10.2. Compound Administration On the day of injection, ACI-7069-633B12-Ab1 (IgG2a variant) (60 mg / kg) and vehicle were freshly prepared and administered intraperitoneally according to a weekly dosing schedule throughout the experiment.

[0246] 10.3. Brain Collection The brain was divided into two hemispheres. The left hemisphere was dissected to collect the cortical brain regions. The mouse cortex and the remaining brain tissues were snap-frozen for further biochemical analysis. The remaining right hemisphere was perfused at room temperature for 3 hours and then immersed and fixed as it was, and collected in freshly prepared 1xPBS containing 4% paraformaldehyde (PFA).

[0247] 10.4. Immunohistochemistry The immersion-fixed right brain hemisphere was cut sagittally at a thickness of 10 microns using a Leica CM1950 cryotome in a uniform and systematic random protocol. A systematic random set of sagittal sections (7 sections from brain levels 2, 3, 4, 6, 8, 10, and 11) per mouse were immunostained for TDP-43 and phosphorylated TDP-43. Iba1 staining was performed to quantify the number and morphology of microglia in the brain. Antibody binding was visualized using a fluorescently labeled secondary antibody. Standard negative controls included wild-type brain sections and sections of transgenic animals without the primary antibody.

[0248] 10.5. Imaging and Measurement of Immunoreactivity The embedded sections were imaged as a whole at 10x magnification using an Axio.Scan Z1 slide scanner operated by ZEN software, with LED (Colibri2) illumination and a high-sensitivity Orca Flash 4.0 monochromatic camera. The brain size was examined using separate images of the target regions in the cerebral cortex and the dorsal striatum. The object density (OD) (number of objects per 1 mm 2 was examined for the region of the target size in the second image excluding all markers, labeled area ratio, and tissue artifacts (such as tissue folding).

[0249] 10.6. Preparation of Protein Samples from the Cerebral Cortex: The tissue was thawed on ice and sonicated in 5X v / w radioimmunoprecipitation assay buffer (RIPA) (50 mM Tris, 150 mM NaCl, 1% IGEPAL CA630, 5 mM EDTA, 0.5% sodium deoxycholate, and 0.1% SDS, pH 8.0) containing 1 mM PMSF and protease / phosphatase inhibitor cocktail (Roche Applied Science). The sample was centrifuged at 100,000 g for 30 minutes at 4°C, and the supernatant was taken as the soluble fraction. The precipitate was washed by sonication with RIPA, and the supernatant was discarded. The RIPA-insoluble precipitate was sonicated in 2X v / w urea buffer (7 M urea, 2 M thiourea, 4% CHAPS, and 30 mM Tris, pH 8.5) and centrifuged at 100,000 g for 30 minutes at 22°C. This supernatant was taken as the RIPA-insoluble / urea-soluble fraction. The protein concentration of the RIPA-soluble fraction was examined using the BCA protein assay (Pierce).

[0250] 10.7. Quantification of insoluble TDP-43 The total TDP-43 levels in the RIPA-insoluble fraction were analyzed by a commercially available human TDP-43 AlphaLISA kit (Perkin Elmer, AL387HV).

[0251] 10.8. Statistical analysis IHC and AlphaLISA data are presented as mean ± SEM. Statistical differences between animals treated with vehicle and ACI-7069-633B12-Ab1 (IgG2a variant) were analyzed by Welch's t-test and are indicated by asterisks above each bar ( * p < 0.05, ** p < 0.01, **** p < 0.0001). Outliers in histological measurements were excluded if they were Grubbs outliers (single measurements) for the group or level or for technical reasons (image artifacts, tissue folding, etc.).

[0252] 10.9. Results Treatment with ACI-7069-633B12-Ab1 (IgG2a variant) reduces phosphorylated TDP-43 and insoluble TDP-43 in rNLS8 mice Overexpression of the DOX-repressible (hTDP-43ΔNLS) K82A / R83A / K84A mutant human TDP-43 causes marked accumulation and aggregation of TDP-43 in the cytoplasm of neurons in the rNLS8 mouse model. The pathological features of this model are the deposition of insoluble and phosphorylated TDP-43 inclusions (pTDP-43). These small spherical cytoplasmic inclusions are present only in transgenic animals and are absent in WT or single-gene transgenic tTA control mice. Furthermore, pTDP-43 is not widespread during the first week in the absence of DOX and accumulates rapidly during 3 - 4 weeks of DOX withdrawal (Walker et al., 2015). Treatment with ACI-7069-633B12-Ab1 (IgG2a variant) statistically significantly reduces the density of phosphorylated TDP-43 in both the striatum and the cerebral cortex compared to vehicle-treated mice (Figures 3A - B), indicating its functional efficacy in reducing TDP-43 lesions. The striatum and cerebral cortex were selected for quantification because the transgene is highly expressed in these regions.

[0253] 10.10. Treatment with ACI-7069-633B12-Ab1 (IgG2a variant) reduces insoluble TDP-43 in rNLS8 mice To confirm the reduction of TDP-43 lesions observed by immunohistochemical reading, after biochemical fractionation, the amount of total insoluble / aggregated TDP-43 in the brain was quantified. The RIPA insoluble fraction was prepared from the cortex of the left cerebral hemisphere containing insoluble / aggregated TDP-43. A significant decrease in the amount of insoluble TDP-43 was observed in mice treated with ACI-7069-633B12-Ab1 (IgG2a variant) compared to animals treated with vehicle (Figure 3C). This reduction in molecular TDP-43 lesions was consistent with the results observed by immunohistochemistry, confirming the efficacy of treatment with ACI-7069-633B12-Ab1 (IgG2a variant). To the inventors' knowledge, this is the first time that peripheral antibody administration has improved the formation of TDP-43 lesions in an in vivo model of TDP-43 proteinopathy.

[0254] 10.11.rNLS8 mice treated with ACI-7069-633B12-Ab1 (IgG2a variant) increase the microglial immune response area The functional recovery of rNLS8 mice after suppression of transgene expression is associated with an increase in microglial activity. The area of microglial cell bodies increases at this stage, resulting in the elimination of TDP-43 lesions and functional recovery of motor impairment, which represents an example of treatment in the rNLS8 mouse model (Spiller KJ et al., Nature Neuroscience, 2018).

[0255] To evaluate the mechanism of action of ACI-7069-633B12-Ab1 in reducing TDP-43 lesions in rNLS8 mice, its effect on microglial activation was evaluated. Iba1 staining was performed by immunohistochemistry to quantify the number and state of microglia in the mouse cerebral cortex. Microgliosis was found in rNLS8 mice at the end stage (5 weeks after Dox removal). Treatment with ACI-7069-633B12-Ab1 significantly increased the Iba1-positive immunoreactive area in the cortex compared to the vehicle-treated control (Figure 5A). This increase could have resulted from either an increase in the number of microglial cells or a change in the morphology of microglia. Therefore, first, the density of Iba1-positive cells in the cortex was evaluated. Treatment with ACI-7069-633B12-Ab1 had no effect on the microglial cell density representing the cell number compared to the vehicle-treated control.

[0256] Next, the effect of ACI-7069-633B12-Ab1 on the morphology of microglia was evaluated. To show the correlation between the increase in the Iba1 immunoreactive area and the change in the activation state of microglia representing the morphology, microglia were classified into three states based on their size and morphology (large hypertrophic, small branched, and branched resting). Compared to the control treated with vehicle, treatment with ACI-7069-633B12-Ab1 (IgG2a variant) showed a significant increase in the average cell size for large hypertrophic microglia (Figure 5B). Little difference was seen in the other two classes of microglia representing a low activation state (Figure 5C-D). This analysis indicates that the increase in the total Iba1-positive immunoreactive area observed in the ACI-7069-633B12-Ab1 treatment cohort results from morphological changes reflected by an increase in the size and activation state of microglial cells. This suggests that ACI-7069-633B12-Ab1 (IgG2a variant) reduces TDP-43 lesions in this animal model, at least in part, through mobilization and activation of microglia.

[0257] Example 11: In Vitro Function of ACI-7069-633B12-Ab1 (IgG2a Variant) in the Recombinant TDP-43 Aggregation Assay To evaluate the function of ACI-7069-633B12-Ab1 (IgG2a variant) in vitro, the ability of ACI-7069-633B12-Ab1 (IgG2a variant) to inhibit TDP-43 aggregation was tested. FL TDP-43 was cleaved by a tobacco etch virus (TEV) protease cleavage site and C-terminally fused to maltose binding protein (MBP) generated recombinantly. Aggregation of 2.5 μM TDP-43-TEV-MBP fusion protein in 30 mM Tris, 150 mM NaCl, pH 7.4 was induced by adding TEV protease (AcTEV, Invitrogen) in the presence of 2.5 μM ACI-7069-633B12-Ab1 (IgG2a variant) or an isotype control that does not bind to TDP, and the absorbance was monitored at 600 nm for 30 hours in a micro-clear 96-well plate (Greiner). For evaluation, the endpoint was normalized against the isotype control, and the percentage of aggregated TDP-43 was calculated for ACI-7069-633B12-Ab1. The antibody ACI-7069-633B12-Ab1 significantly inhibits TDP-43 aggregation up to 98% compared to the isotype control (Figure 4).

[0258] Example 12: Detection and Quantification of TDP-43 in Biological Fluids by ACI-7069-633B12-Ab1 (IgG2a Variant) and ACI-7071-809F12-Ab1 (IgG2a Variant) Method: An AlphaLISA immunoassay based on PerkinElmer beads was established using ACI-7069-633B12-Ab1 (IgG2a variant) and ACI-7071-809F12-Ab1 (IgG2a variant). For CSF samples, dilution linearity was confirmed by a spike recovery experiment. Next, the concentration of TDP-43 was measured in diluted CSF samples. Samples were prepared in white optiplate (registered trademark) 384-well microplates, and luminescence at 615 nm was measured as the raw AlphaLISA count.

[0259] Results: Total TDP-43 in cerebrospinal fluid (CSF) samples from healthy controls and FTLD-TDP (semantic dementia, C9orf72 or GRN) patients was quantified by this immunoassay (Figure 6). Relative quantification of TDP-43 by CSF samples from various patients with FTLD-TDP with GRN mutations showed significantly higher TDP-43 levels compared to healthy controls in three independent experiments (Figure 6). Relative quantification of TDP-43 by CSF samples from various patients with FTLD-TDP with C9orf72 mutations and semantic dementia also showed significantly higher TDP-43 levels compared to healthy controls in three independent experiments (Figure 6).

[0260] Example 13: Binding to pathological TDP-43 evaluated by immunodepletion in FTD brain extracts To evaluate the effectiveness of antibodies that specifically bind to TDP-43 aggregates in their native state, an immunodepletion experiment was performed on brain extracts rich in pathological TDP-43.

[0261] Method: The insoluble fraction from the postmortem brain of FTD-A was prepared as described in Example 7. Immunodepletion was performed using Dynabeads® magnetic beads, Protein G (Thermoscientific 10003D). It was resuspended in the tube and then 130 μl of beads was transferred to a 1.5 ml low-binding tube. The beads were washed twice with PBS containing 0.05% Tween-20 using a magnet to remove the supernatant. The beads were evenly distributed into three different low-binding tubes. Antibodies (ACI-7069-633B12-Ab1 (IgG2a isotype), ACI-7069-642D12-Ab1 (IgG2a isotype), mouse IgG2a control) were diluted to 100 μg / ml, and after removing the supernatant (using a magnet), 100 μl was added to each tube. The antibody-bead mixture was incubated at room temperature for 1 hour. The bead-antibody complex was washed once with 500 μl of PBS (0.05% Tween-20) and once with PBS, and then resuspended in 250 μl of PBS. The antibody-beads were aliquoted into two new tubes (120 μl per tube). The insoluble fraction was thawed on ice and sonicated for 30 seconds at an amplitude of 30 on ice. After removing the supernatant, 30 micrograms of brain material was added to each antibody-bead tube and incubated overnight at 4°C with constant rotation. The tubes were placed on a magnet and the supernatant was collected as the immunodepleted fraction. The input and immunodepleted materials were further analyzed by Western blot. The Western blot was performed as described in Example 7. 20 μl of sample was loaded per lane. Immunoblotting was performed using the following antibodies: total TDP-43 (ACI-7069-633B12-Ab1 conjugated to DyLight680), pTDP-43 (Biolegend, 829901) (used at dilutions of 1:2000 and 1:1000 respectively). A goat anti-rat secondary antibody (catalog number 925-32219) was used at a dilution of 1:10000.

[0262] Results: ACI-7069-633B12-Ab1 and ACI-7069-642D12-Ab1 specifically bound to and depleted TDP-43 and pTDP-43 from the Sarkosyl-insoluble fraction obtained from brain tissues of patients with A-type FTD, compared to isotype control antibodies (Figure 7). This data indicates that these antibodies possess the property of targeting the said targets in human patients.

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Claims

1. A TDP-43 binding molecule that is an antibody or an antigen-binding fragment thereof that binds to misfolded, aggregated TDP-43 and non-aggregated physiological TDP-43, a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 11; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 12; a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 15; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 17; b) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 21; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22; a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 25; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; c) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 31; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 32; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 33; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 35; 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: 37; d) 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; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 43; 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; e) 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; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO:63; 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; f) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 71; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 72; a VH-CDR3 comprising the amino acid sequence of 73; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 75; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 77; g) 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; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; 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; h) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 101; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 102; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 103; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 105; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 106; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 107; i) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 121; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 122; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 123; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 125; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 127; j) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 141; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 142; a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 143; a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 145; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 146; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 147; or k) VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 151; VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 152; VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 153; VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 155; VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 156; and VL-CDR3 comprising the amino acid sequence of SEQ ID NO:

157. A TDP-43 binding molecule comprising:

2. Features as below: a) inhibiting aggregation of TDP-43 protein or a fragment thereof; b) blocking the cell-to-cell spread of TDP-43; c) disintegrating TDP-43 aggregates; and d) Blocking TDP-43 Propagation The TDP-43 binding molecule of claim 1, wherein the TDP-43 binding molecule exhibits one or more up to all of the following:

3. (a) reducing TDP-43 pathology in vivo; and / or (b) a TDP-43 binding molecule according to claim 1 or 2, which reduces the levels of aggregated TDP-43 and / or phosphorylated TDP-43 in vivo.

4. 4. The TDP-43 binding molecule of any one of claims 1 to 3, which binds to an epitope within amino acid residues 400-405, 183-188, 181-195, 199-213, 307-321, 352-366, 389-411, 397-411, 140-200, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-406, or 400-412 of human TDP-43 (SEQ ID NO:1), or an equivalent epitope in a non-human TDP-43.

5. a. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 10 or having at least 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 10; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 14 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 14; b. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO:20 or having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO:20; and a light chain variable region (VL) comprising the sequence of SEQ ID NO:24 or having at least 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO:24; c. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO:30 or having at least 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO:30; and a light chain variable region (VL) comprising the sequence of SEQ ID NO:34 or having at least 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO:34; d. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 40 or having at least 90%, 91%, 92%, 93%, 94%, 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; e. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO:60 or having at least 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 having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:64; f. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 70 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 70; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 74 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 74; g. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 80 or having at least 90%, 91%, 92%, 93%, 94%, 95%, 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 having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 84; h. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 100 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 100; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 104 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 104; i. a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 120 or having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 120; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 124 or having at least 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 124; j. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 140 or having at least 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 140; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 144; or k. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 150 or having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence of SEQ ID NO: 150; and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 154 or having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:

154. The TDP-43 binding molecule of any one of claims 1 to 4, comprising:

6. a. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 10 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 14; b. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 20 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 24; c. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 30 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 34; d. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 40 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 44; e. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO:60 and a light chain variable region (VL) comprising the sequence of SEQ ID NO:64; f. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 70 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 74; g. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 80 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 84; h. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 100 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 104; i. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 120 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 124; j. a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 140 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 144; or k. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 150 and a light chain variable region (VL) comprising the sequence of SEQ ID NO:

154. The TDP-43 binding molecule of any one of claims 1 to 5, comprising:

7. a) a VH-CDR1 comprising the amino acid sequence of SEQ ID NO: 21; a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 22; and a VH-CDR3 comprising the amino acid sequence ES (Glu-Ser); a VL-CDR1 comprising the amino acid sequence of SEQ ID NO: 25; a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 16; and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 27; b) a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 20 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 24; c) 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; 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; and / or d) a heavy chain variable region (VH) comprising the sequence of SEQ ID NO: 80 and a light chain variable region (VL) comprising the sequence of SEQ ID NO: 84; The TDP-43 binding molecule of any one of claims 1 to 6, comprising:

8. a) a monoclonal antibody or an antigen-binding fragment thereof; b) a murine, chimeric, or humanized antibody or an antigen-binding fragment thereof; and / or c) an IgA, IgD, IgE, IgM, IgG1, IgG2, IgG2a, IgG2b, IgG3 or IgG4 antibody or an antigen-binding fragment thereof The TDP-43 binding molecule of any one of claims 1 to 7,

9. A pharmaceutical composition comprising a TDP-43 binding molecule according to any one of claims 1 to 8 for use in human or animal therapy and / or diagnosis.

10. A pharmaceutical composition comprising a TDP-43 binding molecule according to any one of claims 1 to 8 for research use, in particular as an analytical tool or reference molecule.

11. i. TDP-43-related diseases, disorders and / or disorders, or ii. TDP-43 proteinopathy A pharmaceutical composition comprising a TDP-43 binding molecule according to any one of claims 1 to 8 for use in the prevention, alleviation, treatment and / or diagnosis of

12. 10. The pharmaceutical composition of claim 9 for use as a diagnostic tool to monitor a TDP-43 proteinopathy.

13. The TDP-43-related disease, disorder and / or abnormality or TDP-43 proteinopathy is selected from the group consisting of frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), progranulin (GRN) mutated, C9orf72 mutated, TARDBP mutated, valosin-containing protein (VCP) mutated, and associated with chromosome 9p; corticobasal degeneration; Frontotemporal lobar degeneration (FTLD) with ubiquitin-positive inclusions (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, with TARDBP mutation and angiogenin (ANG) mutation), Alexander disease (AxD), limbic system dominant senile Early stage TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy (CTE), Perry syndrome, Alzheimer's disease (AD) (including sporadic and familial AD), Down's syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with mutations in valosin-containing protein (VCP) (bone myopathy) and / or TDP-43-associated disease, disorder and / or abnormality, including, but not limited to, myofibrillar myopathy with mutations in the myotilin (MYOT) gene or mutations in the gene encoding desmin (DES), traumatic brain injury (TBI), dementia with Lewy bodies (DLB), or Parkinson's disease (PD), and optionally the TDP-43-associated disease, disorder and / or abnormality, or TDP-43 proteinopathy, is a) frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), or limbic-predominant senile TDP-43 encephalopathy (LATE); b) amyotrophic lateral sclerosis (ALS); c) Alzheimer's disease (AD), or d) Frontotemporal lobe dementia (FTD) The pharmaceutical composition of claim 11 ,

14. A pharmaceutical composition comprising the TDP-43 binding molecule of any one of claims 1 to 8 and a pharma- ceutically acceptable carrier and / or excipient.

15. i. encoding a TDP-43 binding molecule according to any one of claims 1 to 8, or ii. a. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO: 18 and a light chain variable region (VL) encoding the sequence of SEQ ID NO: 19; b. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO:28 and a light chain variable region (VL) encoding the sequence of SEQ ID NO:29; c. A heavy chain variable region (VH) encoding the sequence of SEQ ID NO:38 and a light chain variable region (VL) encoding the sequence of SEQ ID NO:39; d. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO:48 and a light chain variable region (VL) encoding the sequence of SEQ ID NO:49; e. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO:68 and a light chain variable region (VL) encoding the sequence of SEQ ID NO:69; f. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO:78 and a light chain variable region (VL) encoding the sequence of SEQ ID NO:79; g. A heavy chain variable region (VH) encoding the sequence of SEQ ID NO:88 and a light chain variable region (VL) encoding the sequence of SEQ ID NO:89; h. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO: 108 and a light chain variable region (VL) encoding the sequence of SEQ ID NO: 109; i. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO: 128 and a light chain variable region (VL) encoding the sequence of SEQ ID NO: 129; j. a heavy chain variable region (VH) encoding the sequence of SEQ ID NO: 148 and a light chain variable region (VL) encoding the sequence of SEQ ID NO: 149; or k. A heavy chain variable region (VH) encoding the sequence of SEQ ID NO: 158 and a light chain variable region (VL) encoding the sequence of SEQ ID NO: 159 A nucleic acid molecule comprising the nucleotide sequence represented by:

16. A recombinant vector or expression vector comprising the nucleic acid molecule of claim 15.

17. A host cell which (a) comprises the nucleic acid molecule of claim 15 and / or the vector of claim 16, or (b) expresses the TDP-43 binding molecule of any one of claims 1 to 8.

18. A cell-free expression system comprising the expression vector of claim 16.

19. 1. A method for producing a TDP-43 binding molecule that is an antibody or antigen-binding fragment thereof, comprising: a) culturing a host cell according to claim 17 or a cell-free expression system according to claim 18 under conditions suitable for producing said binding molecule; and b) isolating the binding molecule A method comprising the steps of:

20. 10. A method for quantifying TDP-43 in a sample obtained from a subject, comprising contacting the sample with a TDP-43 binding molecule according to any one of claims 1 to 8, and then comparing the level of TDP-43 in the sample with a control sample.

21. 21. A pharmaceutical composition comprising a TDP-43 binding molecule according to any one of claims 1 to 8 for use in a method for diagnosing a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, said method comprising carrying out the method according to claim 20, wherein an elevated level of TDP-43 in a sample compared to a control level based on healthy subjects is indicative of said disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy.

22. 21. A pharmaceutical composition comprising a TDP-43 binding molecule according to any one of claims 1 to 8 for use in a method for diagnosing a TDP-43 associated disease, disorder and / or abnormality, or a TDP-43 proteinopathy, said method comprising carrying out the method according to claim 20, wherein a similar or elevated level of TDP-43 in the sample compared to a diseased control is indicative of a TDP-43 associated disease, disorder and / or abnormality, or a TDP-43 proteinopathy.

23. A pharmaceutical composition for use in a method for classifying diseases, disorders and / or abnormalities associated with TDP-43 or for classifying TDP-43 proteinopathies, comprising a TDP-43 binding molecule according to any one of claims 1 to 8, said method comprising: a. carrying out the method according to claim 21 and / or 22, b. Classifying diseases, disorders and / or disorders associated with TDP-43 or TDP-43 proteinopathies A pharmaceutical composition comprising:

24. The method further comprises, after step a. and before step b., the steps of: Identifying in the sample a progranulin (GRN) mutation, a C9orf72 mutation, a TARDBP mutation, a valosin-containing protein (VCP) mutation, a TARDBP mutation, an angiogenin (ANG) mutation, a valosin-containing protein (VCP) mutation, a mutation in the myotilin (MYOT) gene, or a mutation in the gene encoding desmin (DES).

24. The pharmaceutical composition of claim 23, further comprising:

25. A pharmaceutical composition for use in a method for classifying diseases, disorders and / or abnormalities associated with TDP-43 or for classifying TDP-43 proteinopathies, comprising a TDP-43 binding molecule according to any one of claims 1 to 8, said method comprising: i. carrying out the method according to claim 22 on a sample obtained from a subject with a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, wherein the comparison with the diseased control is based on a plurality of control samples from subjects with different types or subtypes of a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, and then ii. classifying a disease, disorder and / or condition associated with TDP-43 or a TDP-43 proteinopathy based on said comparison. A pharmaceutical composition comprising:

26. A pharmaceutical composition for use in a method for monitoring a disease, disorder and / or abnormality associated with TDP-43 or for monitoring a TDP-43 proteinopathy at two or more time points using a sample obtained from a subject, comprising a TDP-43 binding molecule according to any one of claims 1 to 8, said method comprising contacting said sample with said TDP-43 binding molecule, (a) a higher level of TDP-43 in a subsequent sample compared to one or more earlier samples indicates the progression of a TDP-43-related disease, disorder and / or condition, or a TDP-43 proteinopathy; or (b) a lower level of TDP-43 in a subsequent sample compared to one or more earlier samples indicates regression of a TDP-43 associated disease, disorder and / or abnormality, or a TDP-43 proteinopathy.

27. 10. A pharmaceutical composition for use in a method for monitoring the treatment of a TDP-43 associated disease, disorder and / or disorder, or for monitoring the treatment of a TDP-43 proteinopathy, at two or more time points using samples obtained from a subject treated with a particular therapy, comprising a TDP-43 binding molecule according to any one of claims 1 to 8, said method comprising contacting said sample with said TDP-43 binding molecule, wherein a lower level of TDP-43 in a later sample compared to one or more earlier samples indicates successful treatment of the TDP-43 associated disease, disorder and / or disorder, or TDP-43 proteinopathy.

28. 28. The pharmaceutical composition of claim 26 or 27, wherein the first time point is prior to treatment with the therapy and the second time point is after treatment with the therapy.

29. 10. A pharmaceutical composition comprising a TDP-43 binding molecule according to any one of claims 1 to 8 for use in a method for selecting a therapy for the treatment of a disease, disorder and / or disorder associated with TDP-43 or for selecting a therapy for the treatment of a TDP-43 proteinopathy, the method comprising contacting samples taken before and after treatment with said therapy with the TDP-43 binding molecule, wherein a lower level of TDP-43 in the sample taken after treatment compared to the sample taken before treatment indicates successful treatment of the disease, disorder and / or disorder associated with TDP-43 or a TDP-43 proteinopathy, whereby said therapy is selected for treatment.

30. The pharmaceutical composition of claim 29, wherein the treatment comprises a TDP-43 binding molecule of any one of claims 1 to 8 or a pharmaceutical composition of claim 14.

31. The pharmaceutical composition of any one of claims 21 to 30, wherein the sample comprises a blood, CSF, ISF, or urine sample.

32. The TDP-43-related disease, disorder and / or abnormality or TDP-43 proteinopathy is selected from the group consisting of frontotemporal dementia (FTD) (e.g., sporadic or familial, with or without motor neuron disease (MND), progranulin (GRN) mutated, C9orf72 mutated, TARDBP mutated, valosin-containing protein (VCP) mutated, and associated with chromosome 9p; corticobasal degeneration; Frontotemporal lobar degeneration (FTLD) with ubiquitin-positive inclusions (FTLD-TDP), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS) (e.g., sporadic ALS, with TARDBP mutation and angiogenin (ANG) mutation), Alexander disease (AxD), limbic system dominant senile Early stage TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy (CTE), Perry syndrome, Alzheimer's disease (AD) (including sporadic and familial AD), Down's syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with mutations in valosin-containing protein (VCP) (bone myopathy) and / or TDP-43-associated disease, disorder and / or abnormality, including, but not limited to, myofibrillar myopathy with mutations in the myotilin (MYOT) gene or mutations in the gene encoding desmin (DES), traumatic brain injury (TBI), dementia with Lewy bodies (DLB), or Parkinson's disease (PD), and optionally the TDP-43-associated disease, disorder and / or abnormality, or TDP-43 proteinopathy, is (a) frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), or limbic-predominant senile TDP-43 encephalopathy (LATE); (b) amyotrophic lateral sclerosis (ALS); (c) Alzheimer's disease (AD); or (d) frontotemporal dementia (FTD), The pharmaceutical composition according to any one of claims 21 to 31.

33. 21. The method of claim 20, wherein the sample comprises a blood, CSF, ISF, or urine sample.

34. A kit for diagnosing a disease, disorder and / or abnormality associated with TDP-43 or a TDP-43 proteinopathy, comprising a TDP-43 binding molecule according to any one of claims 1 to 8 or a pharmaceutical composition according to any one of claims 21 to 32.

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