Anti-TDP-43 binding molecules and uses thereof

Abstract

The invention relates to anti-TDP-43 binding molecules and uses thereof. The present invention is in the field of trans-activation responsive DNA binding proteins (TARDB or also referred to as TDP-43) having a molecular weight of 43 kDa. The present invention relates to TDP-43 specific binding molecules, in particular to anti-TDP-43 antibodies or antigen binding fragments or derivatives thereof, and uses thereof. The present invention provides means and methods for diagnosing, preventing, ameliorating and / or treating diseases, disorders and / or abnormalities associated with TDP-43 aggregates, including, but not limited to, frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), and edge-dominant age-related TDP-43 encephalopathy (LATE).

Description

[0001] This application is a divisional application of Chinese patent application filed on May 22, 2020, with application number “202080038441.4” and invention title “Anti-TDP-43 binding molecule and its use”. The original application was the Chinese national phase application of international application PCT / EP2020 / 064335. Technical Field

[0002] This invention pertains to the field of trans-activating response DNA-binding proteins (TARDB or TDP-43) with a molecular weight of 43 kDa. The invention relates to TDP-43-specific binding molecules, particularly anti-TDP-43 antibodies or their antigen-binding fragments or derivatives, and their uses. This invention provides means and methods for diagnosing, preventing, mitigating, and / or treating diseases, disorders, and / or abnormalities, or TDP-43 proteinopathy related to 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 limbic-predominant age-related TDP-43 encephalopathy (LATE). Background Technology

[0003] Age-related encephalopathy (ANE), characterized by the pathological aggregation of proteins in the central nervous system (CNS) and peripheral organs, is one of the leading causes of disability and mortality worldwide. The protein best characterized by aggregate formation is β-amyloid, found in Alzheimer's disease and related disorders. Other disease-related proteins that tend to aggregate, contributing to neurodegeneration, include, but are not limited to, Tau, α-synuclein (aSyn), huntingtin, fused in sarcoma (FUS), dipeptide repeat protein (DPR) generated through unconventional translation via C9orf72 repeat amplification, superoxide dismutase 1 (SOD1), and TDP-43. Diseases involving TDP-43 aggregates are generally classified as TDP-43 proteopathies, including but not limited to ALS and FTD.

[0004] I.TDP-43 Introduction

[0005] The transactive 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 through 6 are protein-coding). TDP-43 belongs to the heteroribonucleoprotein (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 (Warraich et al., The International Journal of Biochemistry & Cell Biology 42 (2010) 1606–1609). Figure 1TDP-43 consists of two RNA recognition motifs (RRM1 and RRM2) with two highly conserved hexaribonucleoprotein 2 (RNP2) and octaribonucleoprotein 1 (RNP1) regions; a nuclear export signal (NES) and a nuclear localization signal (NLS) that enable it to shuttle between the nucleus and cytoplasm to transport bound mRNA; and a C-terminal glycine-rich domain that 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). TDP-43 is a highly conserved, ubiquitous, and strictly self-regulated protein that constantly shuttles between the nucleus and cytoplasm, but is primarily localized in the nucleus. In 2006, TDP-43 was identified as a protein that accumulates in the vast majority of cases of frontotemporal lobar degeneration (FTLD) with tau-negative, ubiquitin-positive inclusions (hereinafter referred to as FTLD-TDP) and in most cases of amyotrophic lateral sclerosis (ALS) (Arai et al., Biochemical and Biophysical Research Communications 351 (2006) 602–611; Neumann et al., Science 314, (2006), 130–133).

[0006] Thirty-eight negative dominant TDP-43 mutations have been identified in patients with sporadic and familial ALS, as well as those with inherited FTD. These mutations are primarily located in glycine-rich domains (Lagier-Tourenne and Cleveland, Cell 136, 2009, 1001-1004). Figure 1 TDP-43 is inherently prone to aggregation, as shown by sedimentation assays, and this tendency is further enhanced by some ALS-associated TARDBP mutations (Ticozzi et al., CNS Neurol. Disord. Drug Targets. 2010, 9(3), 285-296.), linking TDP-43 aggregation to clinical disease presentation. II. TDP-43 in Neurodegeneration

[0007] TDP-43 aggregates have been identified in an increasing number of neurodegenerative diseases (Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol.).19. Review Issues 1R46-R64), including but not limited to: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valosine-containing protein (VCP) mutations, linked to chromosome 9p, cortical-basal degeneration, frontotemporal degeneration (FTLD) with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic grainy disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), and nonfluent variant primary progressive aphasia (Nonfluent Variant Primary Aphasia). Progressive Aphasia (NFVPPA), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy (CTE), Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado Joseph disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis; inclusion body myopathy with valine-containing protein mutation (VCP); and Paget's disease). Myositis (including bone disease) and frontotemporal dementia; oculopharyngeal dystrophy with rimmed vacuoles; myofibril disease with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES); traumatic brain injury (TBI); dementia with Lewy body (DLB); or Parkinson's disease (PD).

[0008] Accumulated TDP-43 from patient brains exhibits numerous aberrant modifications, including hyperphosphorylation, ubiquitination, acetylation, and C-terminal fragments cleaved by proteolytic processes (Arai et al., Biochemical and Biophysical Research Communications 351(2006)602–611; Neumann et al., Science 314,(2006),130–133; Neumann et al., Acta Neuropathol.(2009)117:137–149; Hasegawa et al.,(2008) Annals of Neurology Vol 64 No 1,60–70; Cohen et al., Nat Commun.6:5845,2015). Another characteristic feature of TDP-43 pathology is the redistribution and accumulation of TDP-43 from the nucleus to the cytoplasm. The hallmark lesions of FTLD-TDP are neuronal cytoplasmic inclusions and glial cytoplasmic inclusions (NCI and GCI, respectively) and dystrophic neurites (DN), which are immunoreactive to TDP-43, ubiquitin, and p62, but negative for other neurodegenerative disease-related proteins. Differences in inclusion body morphology and histological distribution are associated with specific mutations and / or clinical presentations. To date, four types of TDP-43 pathological conditions have been described histologically (Mackenzie and Neumann, J. Neurochem. (2016) 138(Supplement 1), 54-70). FTLD-TDP type A cases are characterized by numerous short, dystrophic neuritis (DN) and compact, oval or crescent-shaped NCIs, primarily in the neocortex II (Mackenzie et al., 2016 J. Neurochem. 138(Supplement 1), 54–70). Figure 2f). This pathological condition typically presents clinically in behavioral variant frontotemporal dementia (bvFTD) or non-fluent / grammatical variant primary progressive aphasia (nfvPPA) and is associated with mutations in granular protein precursor (GRN). Type B cases show a moderate number of compact or granular NCIs in both the superficial and deep cortex, with relatively few DNs and NIIs (neuronal intranuclear inclusions); Mackenzie et al., 2016 J. Neurochem. 138 (Supplement 1), 54–70. Figure 2 g). Most cases presenting with both FTD and ALS symptoms had an FTLD-TDP type B pathology. Type C cases had numerous long, tortuous neurites, primarily in the superficial cortex, with little or no NCI (Mackenzie et al., 2016 J. Neurochem. 138(Supplement 1), 54–70). Figure 2 This pathology is particularly observed in cases of semantically variant primary progressive aphasia (svPPA). FTLD-TDP type D shows abundant intranuclear inclusions (NIIs) and short dendritic cells (DNs) in the neocortex, with only sparse NCIs (Mackenzie et al., 2016 J. Neurochem. 138(Supplement 1), 54–70). Figure 2 Type E is characterized by the presence of granulofilamentous neuronal inclusions (GFNIs) and very fine punctate neuropil aggregates in addition to curved oligodendrocyte inclusions in the white matter, affecting all layers of the neocortex (Edward B. Lee et al., Acta Neuropathol. 2017 July; 134(1):65–78.). This pathological pattern is found only in cases of VCP associated with inclusion body myositis.

[0009] III. TDP-43 in FTD

[0010] Frontotemporal dementia (FTD) is a clinical term that encompasses a broad spectrum of disorders based on degeneration of the frontal and temporal lobes—a pathological feature known as frontotemporal degeneration (FTLD). FTD is the second leading cause of early-onset dementia in the age group under 65 (Le Ber, Revue Neurologique 169 (2013) 811-819). FTD manifests as several syndromes, including bvFTD characterized by personality and behavioral changes; semantic dementia (SD) and progressive nonfluent 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 syndromes is complex, and definitive conclusions can only be reached through post-mortem histopathological analysis to detect aggregated proteins and identify affected brain regions. Regarding pathological protein inclusion bodies, approximately 45% of cases showed pathological accumulation of misfolded Tau, 45% of cases had pathological TDP-43, and a smaller subgroup had aggregates of FUS and other proteins.

[0011] IV. TDP-43 in ALS

[0012] Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the premature loss of upper and lower motor neurons. The progression of ALS is characterized by fatal paralysis and respiratory failure, with a course of 1 to 5 years from diagnosis to death. In most sporadic cases of ALS, the neuropathology is characterized by abnormal cytoplasmic accumulation of TDP-43 in neurons and glial cells of the primary motor cortex, brainstem motor nuclei, spinal cord, and related white matter tracts. ALS with dementia involves the accumulation of TDP-43 in the extramotor neocortex and hippocampus. The role of TDP-43 phosphorylation in ALS patients has been explored using antibodies that specifically bind to phosphorylated TDP-43 in the nucleus and cytoplasmic inclusion bodies, with amino acids S379, S403, S404, S409, and S410 being the major sites of TDP-43 phosphorylation (Hasegawa et al., Ann Neurol 2008; 64:60–70; Neumann et al., Acta Neuropathol (2009) 117:137-149).

[0013] TDP-43 in V.AD and other diseases

[0014] TDP-43 pathology occurs in the brains of up to 57% of patients with Alzheimer's disease (Josephs KA et al., Acta Neuropathol. 2014; 127(6):811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3):441–450; McAleese et al., Brain Pathol. 2017 Jul; 27(4):472-479). TDP-43 aggregation is age-related and associated with cognitive decline, memory loss, and medial temporal atrophy in AD. Clearly, in AD, TDP-43 represents a second or independent pathology that shares an overlapping brain distribution with β-amyloid and tau pathology in the medial temporal lobe. Pathological TDP-43 follows a conventional progressive deposition pattern, which has been described by the so-called TDP-43 (TAD) staging scheme in AD: TDP-43 is first deposited in the amygdala (stage I), then in the hippocampus, limbic system, temporal system, and finally the frontostriatum (stage V) (Josephs KA et al., Acta Neuropathol. 2014; 127(6):811-824; Josephs KA et al., Acta Neuropathol. 2014; 127(3):441–450).

[0015] VI. TDP-43 diffusion

[0016] Although the onset and initial symptoms of ALS and FTD differ significantly among patients, a common feature of disease progression is the spread of pathology from the initial lesion area to most neurons. The persistent worsening of symptoms can be explained by the progressive spread of TDP-43 pathology. TDP-43 pathology in the brains of ALS patients has been shown to spread in a four-stage process and is thought to be transmitted via synaptic propagation through transcortical axonal transmission using anterograde axonal transport (Brettschneider et al., Ann Neurol. 2013 July; 74(1):20–38.). Recent experimental evidence supports the hypothesis that β-amyloid, Tau, α-synuclein, and TDP-43 spread through a prion-like mechanism in neuronal tissue (Hasegawa et al., 2017), where the origin and topographical spread patterns differ for the four proteins (Brettschneider J et al., Nature Rev. Neuroscience, 2015, 109). A common unified mechanism of the disease is considered to be the intercellular spread of pathological protein aggregates. The mechanism consists of the following: aggregates are released from diseased cells, taken up by immature cells, and seeded with pathological protein conformations through template conformational changes of endogenous proteins.

[0017] Intercellular diffusion of TDP-43 has been studied at the molecular level in a few in vitro models, where insoluble TDP-43 preparations derived from patient brains have induced the formation of intracellular aggregates in recipient somatic cells (Nonaka et al., Cell Reports 4 (2013), 124–134; Feiler et al., 2015; Porta et al., Nat. Comm., 2018). Furthermore, intracellular TDP-43 aggregates have been observed to be released in conjunction with efflux bodies before diffusing to the next cell (Nonaka et al., Cell Reports 4 (2013, 124–134)). Similarly, adenovirus-transduced TDP-43 expression leads to phosphorylated, ubiquitinated, and, more importantly, cytoplasmic aggregates that act as seeds initiating intercellular diffusion (Ishii et al., PLoS ONE 12(6):e0179375, 2017). Patient-derived pathological TDP-43 can lead to widespread deposition of endogenous TDP-43 after intracranial inoculation into transgenic and wild-type mice (Porta et al., Nat. Comm., 2018).

[0018] VII. Prevention and Treatment of TDP-43 Protein Disorders

[0019] TDP-43 aggregation and pathological spread are key hallmarks of ALS and FTD—currently incurable and fatal diseases. Mutations in TDP-43 are associated with familial cases of ALS and FTD, providing a causal link between TDP-43 misfolding and disease progression.

[0020] VIII. Diagnosis of TDP-43 protein disorders

[0021] A diagnosis of FTD based on clinical presentation is inadequate because clinical presentations can overlap with other diseases, especially in the early stages.

[0022] Many approaches aim to develop biochemical biomarkers to differentiate between different types of FTD pathology. The development of antibodies targeting different conformations of TDP-43 could allow for the creation of more sensitive and specific diagnostic tools. In parallel with biochemical biomarkers, the development of imaging biomarkers enables early and specific detection of pathology in TDP-43 protein diseases. The ability to image TDP-43 deposition in the brain could be a significant achievement for the diagnosis and drug development of TDP-43 protein diseases. Such detection can be achieved using cell-permeable antibody fragments.

[0023] The earliest event in neurodegenerative diseases based on the misfolding of different proteins was the acquisition of alternative conformations that made the proteins toxic. Furthermore, these misfolded conformations can self-propagate by recruiting endogenous normal proteins into them, serving as the mechanistic basis for the observed diffusion through affected tissues.

[0024] To develop antibodies targeting different conformational states of a given protein, supramolecular antigen constructs were designed in which the conformation of the presented antigen is controlled to generate conformation-specific antibodies against a given target in a specific conformational state (WO2012 / 055933 and WO2012 / 020124). Conformation-specific antibodies offer many advantages because they can distinguish between disease-associated conformations and functional endogenous conformations of these proteins. This approach offers numerous advantages in therapeutic applications because such antibodies are less likely to be attracted to the normal conformation of a protein while targeting its misfolded, disease-associated isoform. Similarly, in diagnostic applications, such antibodies recognize only the disease-associated structural state of a protein, which is crucial for the development of sensitive and specific diagnostics.

[0025] The application of TDP-43-based biomarkers in TDP-43 proteopathies remains to be established. Such evaluation is hampered, in part by the lack of high-affinity antibodies available for use in appropriate immunoassays to quantify pathological TDP-43 in biological fluids (Feneberg et al., Molecular Neurobiology, 2018).

[0026] Therefore, there is a clear need for biomarkers capable of detecting, particularly in human samples, misfolded aggregated TDP-43 and non-aggregated physiological TDP-43 for the diagnosis of different types of TDP-43 proteopathies and / or for monitoring the efficacy of therapeutics for TDP-43-related diseases, disorders and abnormalities, particularly those associated with TDP-43 aggregates, or TDP-43 proteopathies.

[0027] TDP-43 protein disorders are defined as a group of neurodegenerative diseases characterized by pathological TDP-43.

[0028] IX. Existing Technology

[0029] Patent application WO 2008 / 151055 discloses a method and materials for determining whether a mammal has a neurodegenerative disease by using the levels of TDP-43 peptides and / or TDP-43 peptide cleavage products (e.g., 25kD and 35kD TDP-43 peptide cleavage products) in biological fluids.

[0030] Patent application WO 2013 / 061163 discloses TDP-43 specific binding molecules, including polypeptides such as human antibodies and their fragments, derivatives and variants. Summary of the Invention

[0031] Given the foregoing, there is a need to combine misfolded aggregated TDP-43 with non-aggregated physiological TDP-43, particularly anti-TDP-43 binding molecules of human TDP-43. Furthermore, developing sensitive and specific biomarkers that allow differentiation of pathological types within the FTD spectrum is an urgent task.

[0032] This technical problem is solved by the implementation scheme provided in this article.

[0033] Therefore, this 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. In this invention, misfolded TDP-43 includes misfolded monomeric TDP-43 and / or misfolded oligomeric TDP-43 and / or misfolded aggregates and / or post-translational modified TDP-43 and / or misfolded truncated TDP-43. Post-translational modified TDP-43 comprises phosphorylated, ubiquitinated, acetylated, ubiquitin-like, and / or methylated TDP-43. Physiological TDP-43 includes soluble nuclear TDP-43. It is shown herein that the binding molecules of this invention are capable of binding pathological TDP-43, including TDP-43 aggregates and phosphorylated TDP-43 (see Example 13). Therefore, this 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 illustrated herein, the TDP-43 binding molecules of the present invention can bind equally to misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, or preferentially bind to one type of TDP-43 when specifically binding to both. 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 abnormalities associated with TDP-43, particularly with TDP-43 aggregates, or TDP-43 proteases. The present invention also provides binding molecules, particularly antibodies or antigen-binding fragments thereof, for the detection and / or understanding (i.e., identification) of specific pathological types causing neurodegeneration. Uses are envisioned as diagnostic biomarkers, enabling more effective and precise subject selection for longitudinal monitoring in clinical studies, and supporting the development of new treatments for TDP-43 proteases.

[0034] The present invention also provides TDP-43 binding molecules, particularly antibodies or antigen-binding fragments thereof, as drugs (therapeutic agents).

[0035] Unwilling to be bound by theory, this invention is developed based on the following assumptions: modified conformation-specific antigenic peptides and peptide fragments or the whole TDP-43 protein derived from TDP-43 protein, and antibodies obtainable through or via said peptides or fragments or the whole TDP-43 protein, block the intercellular spread of TDP-43, and / or depolymerize TDP-43 aggregates and / or block TDP-43 seeding and / or inhibit the aggregation of TDP-43 protein or fragments thereof. The binding molecules of this invention, particularly polypeptides, and more particularly antibodies or antigen-binding fragments thereof, bind to misfolded aggregated TDP-43, particularly to cytoplasmic and extracellular misfolded TDP-43. The binding molecules of this invention, particularly polypeptides, and 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 this invention, particularly polypeptides, and more particularly antibodies or antigen-binding fragments thereof, specifically bind to cytoplasmic misfolded TDP-43.

[0036] Misfolded aggregates of TDP-43, or pathologically associated TDP-43, consist of TDP-43 proteins that have lost their normal folding (i.e., are misfolded) and localized. Misfolded aggregates of TDP-43 can be found in: preinclusions, as well as neuronal cytoplasmic inclusions and glial cell cytoplasmic inclusions (NCI and GCI, respectively), intranuclear inclusions (NII) of neurons, and dystrophic neurites (DN), which are immunoreactive to TDP-43.

[0037] Non-aggregated physiological TDP-43 is a physiologically functional TDP-43 protein that is mainly located in the nucleus and shuttles into the cytoplasm, and is in a state that can perform its desired function in the in vivo cellular environment.

[0038] The binding molecules of the present invention, particularly antibodies or their antigen-binding fragments, unexpectedly possess at least one, preferably two, more preferably three, and even more preferably all four of the following features:

[0039] -Block the intercellular transmission of TDP-43;

[0040] -Depolymerizes TDP-43 aggregates;

[0041] -Inhibits the aggregation of TDP-43 protein or its fragments;

[0042] -Block TDP-43 seeding.

[0043] Independent of one, two, three or four of the above-listed features, the binding molecules of the present invention, preferably antibodies or antigen-binding fragments thereof, can improve / inhibit / reduce the formation of TDP-43 pathology in in vivo models of TDP-43 protein pathology and, more importantly, in patients with TDP-43 pathology.

[0044] The TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can recruit and / or activate microglia. More particularly, this is shown herein (see Example 10 and...). Figure 5 The TDP-43 binding molecule of the present invention can affect microglial morphology in terms of cell size and activation state. This may contribute to a reduction in TDP-43 pathological conditions indicated by the TDP-43 binding molecule of the present invention.

[0045] In this invention, binding molecules, particularly antibodies or their antigen-binding fragments, specifically recognize TDP-43. The binding molecules of this invention include peptides and / or antibodies and / or their antigen-binding fragments that are specific to the TDP-43 protein. "Specifically recognizing TDP-43" means that, compared to other epitopes, the binding molecules of this invention specifically, generally, and collectively bind with greater affinity to TDP-43, particularly certain epitopes of TDP-43, especially epitopes of the TDP-43 protein exposed / accessible in one or more pathological conformations. The binding molecules of this invention that specifically bind to TDP-43, particularly peptides, and more particularly antibodies or their antigen-binding fragments, specifically recognize misfolded aggregated TDP-43 and non-aggregated physiological TDP-43. In a preferred embodiment, full-length human TDP-43 is included, preferably having the sequence of SEQ ID NO:1. In another preferred embodiment of the invention, the binding molecule, particularly an antibody or its antigen-binding fragment, specifically binds to a binding region defined in full-length and / or truncated TDP-43, wherein the binding region is preferably contained in 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, more preferably, the binding region is contained in amino acids 183 to 188, 203 to 213, 204 to 208, 204 to 211, 205 to 210, 316 to 323, 358 to 361, 400 to 405, 400 to 406, or 400 to 412. Therefore, the binding molecule, particularly the antibody or its antigen-binding fragment, preferably binds specifically to a peptide containing, and preferably consisting of, a binding region, said binding region being composed 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 the full-length human TDP-43 having the sequence SEQ ID NO:1. In another preferred embodiment of the invention, the binding molecule, particularly an antibody or its antigen-binding fragment, preferably binds specifically to a peptide comprising a binding region, preferably composed of the binding region, said binding region being composed of amino acids at positions 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). In some embodiments, the TDP-43 binding molecule, particularly an antibody or its antigen-binding fragment, binds to the C-terminal region of TDP-43.This can be advantageous, for example, because the C-terminal fragment of TDP-43 is found in the insoluble fraction and is therefore likely pathologically relevant. More specifically, TDP-43 binding molecules, particularly antibodies or their antigen-binding fragments, can bind to epitopes within amino acid residues 400-405, 400-406, or 400-412 of human TDP-43 (SEQ ID NO:1). In some embodiments of the invention, the antibody is a monoclonal antibody. In some embodiments, the antibody is a mouse antibody, a murine-derived antibody, a human antibody, a humanized antibody, or a chimeric antibody. It should be understood that equivalent binding regions exist in non-human TDP-43. Thus, for example, the length of the mouse TDP-43 amino acid sequence (see Uniprot accession number Q921F2) is also 414 amino acids and has 96% (398 / 414 residues) identity with the human sequence. This invention covers binding molecules, particularly antibodies or antigen-binding fragments thereof, that bind to non-human TDP-43, particularly mouse TDP-43, to those equivalent regions / peptides specified in the above reference SEQ ID NO:1.

[0046] In particular, the invention is summarized in the following embodiments:

[0047] 1. TDP-43 binding molecules, which bind misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, particularly human TDP-43.

[0048] 2. The TDP-43 binding molecule described in the aforementioned embodiments binds to epitopes of human TDP-43 (SEQ ID NO:1) at positions 181 to 195, 199 to 213, 307 to 321, 352 to 366, 389 to 411, 397 to 411, or 140 to 200.

[0049] 3. The TDP-43 binding molecule described in the aforementioned embodiments binds to epitopes of amino acid residues at positions 183 to 188, 203 to 213, 204 to 208, 204 to 211, 205 to 210, 316 to 323, 358 to 361, 400 to 405, 400 to 406, or 400 to 412 of human TDP-43 (SEQ ID NO:1).

[0050] 4. The binding molecule in any one of the foregoing embodiments is an antibody or its antigen-binding fragment.

[0051] 5. The binding molecule or TDP-43 binding molecule described in any of the foregoing embodiments, comprising:

[0052] 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, and 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; or

[0053] 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, and 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; or

[0054] 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, and 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; or

[0055] 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, and 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; or

[0056] 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, and 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; or

[0057] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO: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; or

[0058] 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, and 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; or

[0059] 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, and 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; or

[0060] 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, and 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; or

[0061] 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, and 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

[0062] 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, 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.

[0063] 6. The binding molecule or TDP-43 binding molecule described in any of the foregoing embodiments comprises an antibody or its antigen-binding fragment:

[0064] a. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:10 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:14; or

[0065] b. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:20 or a heavy chain variable region (VH) having at least 98% or 99% sequence identity with 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 a light chain variable region (VL) having at least 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:24; or

[0066] c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:30 or a heavy chain variable region (VH) having at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 98% or 99% sequence identity with the amino acid sequence of SEQ ID NO:34; or

[0067] d. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:40, and a light chain variable region (VL) containing the sequence of SEQ ID NO:44; or

[0068] e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:60 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:60, and a light chain variable region (VL) comprising the sequence of SEQ ID NO:64 or a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:64; or

[0069] f. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:70 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:74; or

[0070] g. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:80 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:84; or

[0071] h. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:100 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:104; or

[0072] i. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:120 or a heavy chain variable region (VH) having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:124; or

[0073] j. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:140 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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

[0074] k. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 or a heavy chain variable region (VH) having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:154.

[0075] 7. The binding molecule or TDP-43 binding molecule described in any of the foregoing embodiments, comprising an antibody or its antigen-binding fragment:

[0076] a. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:10 and the light chain variable region (VL) containing the sequence of SEQ ID NO:14; or

[0077] c. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:20 and the light chain variable region (VL) containing the sequence of SEQ ID NO:24; or

[0078] d. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:30 and the light chain variable region (VL) containing the sequence of SEQ ID NO:34; or

[0079] e. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 and the light chain variable region (VL) containing the sequence of SEQ ID NO:44; or

[0080] f. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 60 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 64; or

[0081] g. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:70 and the light chain variable region (VL) containing the sequence of SEQ ID NO:74; or

[0082] h. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 80 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 84; or

[0083] i. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:100 and the light chain variable region (VL) containing the sequence of SEQ ID NO:104; or

[0084] j. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:120 and the light chain variable region (VL) containing the sequence of SEQ ID NO:124; or

[0085] k. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:140 and the light chain variable region (VL) containing the sequence of SEQ ID NO:144; or

[0086] l. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 and the light chain variable region (VL) containing the sequence of SEQ ID NO:154.

[0087] In some embodiments, the antibody comprises:

[0088] 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, and 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; or

[0089] 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, and 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; or

[0090] 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, and 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; or

[0091] 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, and 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; or

[0092] 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, and 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; or

[0093] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO: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; or

[0094] 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, and 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; or

[0095] 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, and 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; or

[0096] 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, and 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; or

[0097] 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, and 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

[0098] 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, 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.

[0099] In some embodiments, the antibody comprises:

[0100] 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, and VH-CDR3 containing the amino acid sequence ES (Glu-Ser); or

[0101] 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, and VH-CDR3 containing the amino acid sequence ES (Glu-Ser); or

[0102] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:33; or

[0103] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:43; or

[0104] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:63; or

[0105] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:73; or

[0106] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:83; or

[0107] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:103; or

[0108] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:123; or

[0109] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:143; or

[0110] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO:153.

[0111] In some embodiments, the antibody comprises:

[0112] a) 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; or

[0113] b) 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; or

[0114] c) 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; or

[0115] d) 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; or

[0116] e) 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; or

[0117] f) 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; or

[0118] g) 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; or

[0119] h) 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; or

[0120] i) 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.

[0121] In some embodiments, the antibody comprises:

[0122] a) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:11; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0123] b) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:21; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0124] c) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:31; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:32; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0125] d) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:41; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:42; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0126] e) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:61; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:62; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0127] f) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:71; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:72; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO: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; or

[0128] g) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:81; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:82; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0129] h) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:101; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:102; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0130] i) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:121; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:122; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0131] j) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:141; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:142; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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

[0132] k) VH-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:151; VH-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:152; VH-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with 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; or

[0133] In some embodiments, the antibody comprises:

[0134] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:15, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:16, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:17; or

[0135] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:25, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:16, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:27; or

[0136] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:35, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:36, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:37; or

[0137] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:45, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:46, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:47; or

[0138] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:65, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:66, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:67; or

[0139] 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 of SEQ ID NO:73, VL-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:75, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:16, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:77; or

[0140] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:85, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:86, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:87; or

[0141] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:105, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:106, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:107; or

[0142] 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 an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:125, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:16, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:127; or

[0143] 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, VL-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:145, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:146, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:147; or

[0144] 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, VL-CDR1 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:155, VL-CDR2 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:156, and VL-CDR3 containing an amino acid sequence having at least 80%, 90%, 95%, or 100% sequence identity with SEQ ID NO:157.

[0145] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six CDRs selected from the following: (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.

[0146] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six CDRs selected from the following: (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.

[0147] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0148] In some embodiments, the TDP-43 antibody comprises at least one, two, or three 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; and (c) VH-CDR3 comprising the amino acid sequence of SEQ ID NO:43.

[0149] In some embodiments, the TDP-43 antibody comprises at least four, five, or six 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.

[0150] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0151] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0152] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0153] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0154] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0155] In some embodiments, the TDP-43 antibody comprises at least one, two, or three 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.

[0156] In some embodiments, the TDP-43 antibody comprises at least four, five, or six 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.

[0157] In some embodiments, the TDP-43 antibody comprises at least one, two, three, four, five, or six 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.

[0158] In another embodiment, the TDP-43 antibody comprises a heavy chain variable domain (VH) selected from the following: SEQ ID NO: 10, 20, 30, 40, 60, 70, 80, 100, 120, 140, 150, including post-translational modifications of the sequence. In one specific embodiment, the heavy chain variable domain (VH) comprises at least one, two, or three CDRs selected from: (a) VH-CDR1 comprising an amino acid sequence selected from SEQ ID NO: 11, 21, 31, 41, 61, 71, 81, 101, 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; and (c) VH-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 33, 43, 63, 73, 89, 103, 123, 143, 153, and ES (Glu-Ser).

[0159] In another embodiment, the TDP-43 antibody comprises a light chain variable domain (VL) selected from SEQ ID NO: 14, 24, 34, 64, 74, 84, 104, 124, 154, including post-translational modifications of said sequences. In one specific embodiment, the light chain variable domain (VL) comprises at least one, two, or three 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; and (b) VL-CDR2 comprising an amino acid sequence selected from SEQ ID NO: 16, 36, 66, 86, 106, 156; and (c) VL-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 17, 27, 37, 67, 77, 87, 107, 127, 157, and ES (Glu-Ser).

[0160] In some embodiments, the TDP-43 antibody comprises at least one, two, or three 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; and (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).

[0161] In some embodiments, the TDP-43 antibody comprises at least one, two, or three 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; and (c) VL-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 17, 27, 37, 67, 77, 87, 107, 127, 157.

[0162] In some embodiments, the light chain variable domain (VL) comprises at least one, two, or three 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; and (b) VL-CDR2 comprising an amino acid sequence selected from SEQ ID NO: 16, 36, 66, 86, 106, 156; and (c) VL-CDR3 comprising an amino acid sequence selected from SEQ ID NO: 17, 27, 37, 67, 77, 87, 107, 127, 157.

[0163] In some embodiments, the present invention relates to antibodies derived from hybridoma clones 631B2A2, 633B12C8, 634H10H7, 636E5B8, 641H1E7, 642A10B11, 642D12B4, 646B7F7, 712A6B10, 809D9C2, or 809F12D8.

[0164] In some embodiments, the present invention relates to antibodies selected from the following: 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.

[0165] In some embodiments, the dissociation constant (KD) of the binding molecule or antibody provided herein is ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or ≤0.001 nM (e.g., 10).-8 M or smaller, such as 10 -8 M to 10 -13 M, for example, 10 -9 M to 10 -13 M), particularly relating to binding TDP-43, especially soluble TDP-43, aggregated TDP-43, and / or oligomeric TDP-43. In some embodiments, 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 compared to soluble TDP-43. For example, the KD of the TDP-43 binding molecules of the present invention for aggregated TDP-43 may be 30 nM or less, in some specific embodiments 1 nM or less, and for soluble TDP-43 500 nM or less. This is shown in Example 8A with reference to Table 8 for the TDP-43 binding molecules of the present invention.

[0166] In one embodiment, the binding affinity to soluble or aggregated FL TDP-43 can be evaluated by determining the dissociation constant (KD) using surface plasmon resonance (SPR; Biacore T200, GE Healthcare Life Sciences). A detailed description of suitable SPR methods that can be used is provided in Examples 8A and 8B.

[0167] The TDP-43 binding molecules of the present invention, particularly antibodies or their antigen-binding fragments, typically bind TDP-43 with high affinity. For example, they may exhibit EC50 values ​​of 200 pM or less, more preferably 20 pM or less, and even more preferably 10 pM or less, as determined by Luminex assay. Further details of suitable assays can be found in Example 3. Similarly, they may exhibit EC50 values ​​of 1600 ng / ml or less, more preferably 120 ng / ml or less, and even more preferably 60 ng / ml or less, as determined by indirect ELISA. Further details of suitable assays can be found in Example 4.

[0168] 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. Therefore, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can bind well to both soluble TDP-43 and aggregated TDP-43 in approximately equal quantities. The TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can bind to aggregated TDP-43 in approximately equal quantities compared to non-aggregated TDP-43. More particularly, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can bind to aggregated TDP-43 in approximately equal quantities compared to non-aggregated TDP-43 in the cell nucleus. In other embodiments, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, can preferentially bind to aggregated TDP-43 compared to non-aggregated TDP-43 when binding to both substances. More specifically, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, preferentially bind to aggregated TDP-43 in the cytoplasm compared to non-aggregated TDP-43 in the cell nucleus when bound to both substances. Alternatively, in other embodiments, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, preferentially bind to non-aggregated TDP-43 in the cell nucleus compared to aggregated TDP-43 when bound to both substances. More specifically, the TDP-43 binding molecules of the present invention, particularly antibodies or antigen-binding fragments thereof, preferentially bind to non-aggregated TDP-43 in the cell nucleus compared to aggregated TDP-43 in the cytoplasm when bound to both substances. These binding properties can be demonstrated, for example, using immunohistochemistry. Suitable methods are described herein with reference to Example 6, in which relevant controls are provided. Results are shown in Table 7.

[0169] The present invention also relates to compositions comprising the binding molecules described herein, particularly antibodies of the present invention or antigen-binding fragments thereof (including TDP-43 binding antibody fragments and derivatives). The present invention further relates to immunotherapeutic and / or immunodiagnostic methods using such compositions for the prevention, diagnosis, and / or treatment of TDP-43 protein disorders, wherein an effective amount of the composition is administered to a subject in need.

[0170] In some embodiments, the present invention encompasses the binding molecules described herein that specifically bind to TDP-43, particularly the antibodies and antigen-binding fragments of the present invention, and the use of these binding molecules in diagnosing, preventing, mitigating, and / or treating diseases, disorders, and / or abnormalities, or TDP-43 protein disorders associated with TDP-43, particularly with TDP-43 aggregates, including but not limited to: frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), and limbic-dominant age-related TDP-43 encephalopathy (LATE). The methods and compositions disclosed herein can be used to diagnose, prevent, mitigate, and / or treat diseases, disorders, and / or abnormalities, or TDP-43 protein disorders associated with TDP-43, particularly with TDP-43 aggregates, including but not limited to frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). Preferably, these uses, which combine molecular diagnostic, preventative, mitigating, and / or therapeutic applications for diseases, disorders, and / or abnormalities associated with TDP-43, particularly those related to TDP-43 aggregates, or TDP-43 protein disorders, are targeted at amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), or frontotemporal dementia (FTD). More preferably, the use is targeted at amyotrophic lateral sclerosis (ALS). More preferably, the use is targeted at Alzheimer's disease (AD). More preferably, the use is targeted at frontotemporal dementia (FTD).

[0171] In another embodiment, a TDP-43-specific binding molecule described herein, particularly an antibody or antigen-binding fragment of the present invention, is contacted with a sample to detect, diagnose, and / or monitor diseases, disorders, and / or abnormalities associated with TDP-43, particularly with TDP-43 aggregates, or TDP-43 protein disorders selected from frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), aeropoietic granulopathy, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), and non-fluent variant primary progressive aphasia (nf (vPPA), etc.), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine tyrosinase (VCP) mutation; and Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with myoconstrictor protein (MYOT) gene mutation or mutation in the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB) or Parkinson's disease (PD).

[0172] In one embodiment, the invention covers binding molecules described herein that specifically bind to TDP-43, particularly the antibodies of the present invention or their antigen-binding fragments, and the use of these molecules, particularly these antibodies, to detect the presence of TDP-43 in a sample. Therefore, the TDP-43 binding molecules of the present invention, such as the anti-TDP43 antibodies described herein, can be particularly used to screen clinical samples for the presence of TDP-43 in a sample, particularly human blood, CSF, interstitial fluid (ISF), and / or urine, for example, by using ELISA-based assays or surface adaptation assays. In some cases, tissue samples, such as brain tissue samples, can be used. The methods and compositions of the present invention can also be used to diagnose presymptomatic diseases and / or monitor disease progression and / or treatment efficacy. According to some implementation schemes, antibodies specific to TDP-43 (e.g., full-length antibodies or TDP-43-binding fragments or derivatives of antibodies) are contacted with samples (e.g., blood, cerebrospinal fluid (CSF), interstitial fluid (ISF), or brain tissue) to detect, diagnose, and / or monitor frontotemporal dementia (FTD, such as sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), aurophilic granulomatosis, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), muscular atrophy Amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Mayo's disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; and Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).The TDP-43 binding molecule of this invention can be used to quantify TDP-43 in suitable samples, particularly clinical samples such as blood, CSF, ISF, or urine, where a relatively high TDP-43 level compared to a suitable control indicates disease and / or a later stage of disease. Many suitable forms of immunoassay are known. Therefore, this method can be performed for diagnostic purposes (e.g., ELISA, MSD (Meso Scale Discovery), HTRF (Homogeneous Time-Resolved Fluorescence), and AlphaLISA), where high TDP-43 levels indicate disease. Alternatively, this method can be performed for monitoring purposes. Levels increasing over time may indicate disease progression. Levels decreasing over time may indicate disease regression. This method can also be used to monitor treatment, particularly the efficacy of a specific treatment. Successful treatment can be measured by referring to stable or declining TDP-43 levels after treatment. This document demonstrates (Example 12) that, when measured using the antibody of the present invention, the TDP-43 level in CSF samples from patients with TDP-43 proteinopathy was higher than that in control samples taken from healthy subjects (healthy controls). Control samples may or may not be run in parallel with the test samples. In some embodiments, the control level is determined by a series of control samples taken from healthy subjects under similar or identical experimental conditions and used as a comparison level for the level determined in the test samples. The method of quantifying TDP-43 in suitable samples using the binding molecule of the present invention can also be used to select treatments (for further treatment of the subject). Therefore, personalized treatment methods are envisioned. Sampling is performed before and after treatment. If the treatment results in a stable or preferably reduced TDP-43 level after treatment, the treatment is selected for the subject. If the treatment does not result in a stable or preferably reduced TDP-43 level after treatment, the treatment is not selected for the subject. The treatment can be any suitable candidate therapeutic agent for treating TDP-43 proteinopathy. In some preferred embodiments, the treatment comprises the TDP-43 binding molecule of the present invention, typically in the form of a pharmaceutical composition as described herein.

[0173] The TDP-43 binding molecule of the present invention can also be used to classify diseases into specific types or subtypes. Therefore, methods are provided for classifying diseases, disorders, and / or abnormalities associated with TDP-43, particularly with TDP-43 aggregates, or for classifying TDP-43 protein disorders, comprising:

[0174] a. The method of the present invention, wherein the level of TDP-43 is quantified in comparison with a suitable control;

[0175] b. Optionally identify mutations in samples from the subject, including but not limited to mutations in granular protein precursor (GRN), C9orf72, TARDBP, valproic acid protein (VCP), angiotensin (ANG), myoconstrictor protein (MYOT) gene mutations, or genes encoding desmin (DES); and

[0176] c. Diseases, disorders, and / or conditions associated with TDP-43, particularly those related to TDP-43 aggregates.

[0177] They can be classified as either abnormal or TDP-43 protein disorders.

[0178] Similarly, methods are provided for classifying diseases, disorders, and / or abnormalities associated with TDP-43, particularly with TDP-43 aggregates, or for classifying TDP-43 protein disorders, comprising: performing the method of the invention, wherein the level of TDP-43 in a sample obtained from a subject suffering from a TDP-43-related disease, disorder, and / or abnormality, or TDP-43 protein disorder, is quantified, wherein the level is compared with control samples taken from subjects suffering from different types or subtypes of TDP-43-related diseases, disorders, and / or abnormalities, or TDP-43 protein disorders (i.e., determining a set of representative control levels for the target type or subtype); and classifying the TDP-43-related disease, disorder, and / or abnormality, or TDP-43 protein disorder based on the comparison results. Thus, the classification is based on determining the closest match between the test sample and one or more control samples. These methods may also include identifying mutations in the sample, including but not limited to mutations in granular protein precursor (GRN), C9orf72, TADBP, valproic acid protein (VCP), TARDBP, angiogenic protein (ANG), myoconstrictor protein (MYOT) gene, or genes encoding desmin (DES). The identified mutations are also used to classify diseases, disorders, and / or abnormalities associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 protein disorders. For the avoidance of doubt, identification of mutations in the sample can be performed by any suitable method; for example, nucleic acid sequencing based on nucleic acid molecules within the sample. The sample may be separate and different from samples in which TDP-43 levels are determined, but may originate from the same subject.

[0179] In other embodiments, the present invention provides methods for preventing, alleviating, and / or treating diseases, disorders, and / or abnormalities associated with TDP-43, particularly with TDP-43 aggregates, or TDP-43 protein disorders. According to one embodiment, the method of the present invention includes administering to a subject an effective concentration of the binding molecules described herein, particularly antibodies of the present invention specific to TDP-43 (e.g., full-length antibodies or TDP-43-binding fragments or derivatives of antibodies). In another embodiment, the present invention provides methods for preventing, alleviating, and / or treating TDP-43 protein disorders. According to some embodiments, administration of the binding molecules described herein specific to TDP-43, particularly antibodies of the present invention or their antigen-binding fragments, is used to treat, alleviate, and / or prevent frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS). In another embodiment, administration of a binding molecule described herein that is specific to TDP-43, particularly an antibody of the present invention or an antigen-binding fragment thereof, is used to prevent, alleviate, and / or treat neurodegenerative diseases selected from: frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD, including sporadic and familial forms of AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), and limbic dominant age-related TDP-43 encephalopathy (LATE).

[0180] In another embodiment, administration of a binding molecule described herein that is specific to TDP-43, particularly an antibody or antigen-binding fragment thereof of the present invention, is used to prevent, alleviate, and / or treat diseases selected from: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), aurophilic granulomatosis, 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 or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, with cortical-basal degeneration, with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), aurophilic granulomatosis, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc.), and amyotrophic lateral sclerosis (ALS, e.g., sporadic or familial, with MND mutations ... Sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation, Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuolar lesions, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD). Attached Figure Description

[0181] Figure 1 Detection of TDP-43 in tissue sections from subjects with frontotemporal dementia (FTD) with type A pathology. Immunohistochemistry was performed on 10 μm thick frozen sections of the frontal cortex from subjects with FTD and type A pathology using a fluorescently labeled secondary antibody for detection. The following antibodies were used as controls: rabbit polyclonal pan-TDP-43 antibody (Proteintech, 10782-2-AP) for detecting pathological inclusion bodies and physiological nuclear TDP-43; and rabbit monoclonal phosphorylated TDP-43p409 / 410 antibody (Cosmobio, TIP-PTD-P02) for detecting pathological aggregation and phosphorylated TDP-43. Arrows indicate TDP-43 aggregates; thick arrows indicate physiological TDP-43 in the nucleus (nuclei visualized by DAPI staining). Hybridoma name or source of commercial antibody is indicated in the upper left corner of each image.

[0182] Figure 2 Detection of TDP-43 in sarkosyl soluble and insoluble fractions obtained from FTD type A post-mortem brain tissue (frontal cortex). Immunoblotting with commercial antibodies binding to either the N-terminal region (A, B) or the C-terminal region (C) showed the presence of TDP-43 in the sarkosyl soluble (lane 1) and insoluble (lane 2) fractions. Immunoblotting of mAbs against TDP-43 generated in this study with epitopes at the N-terminal region of TDP-43 (D to I). Immunoblotting of mAbs against TDP-43 binding to the C-terminal region of TDP-43 (J to N). All mAbs against TDP-43 specifically recognized full-length TDP-43. Additionally, some mAbs (K, M, N) recognized pathological features of lesion states, such as the C-terminal fragment in the insoluble fraction.

[0183] Figure 3 (c) The density of pTDP-43 immunoreactive material measured in two brain regions of mice treated with the carrier (n=30, gray bars) and ACI-7069-633B12-Ab1 (IgG2a variant) (n=25, dotted gray bars) is shown. (d) The insoluble fraction of total TDP-43 obtained from the cortex of the left hemisphere was quantified in the groups treated with the carrier (n=30) and ACI-7069-633B12-Ab1 (IgG2a variant) (n=25) (*p<0.05, **p<0.01, ****p<0.0001).

[0184] Figure 4 TDP-43 aggregation induced by TEV cleavage was measured at 600 nm after 30 h by turbidity measurement in the presence of ACI-7069-633B12-Ab1 (IgG2a variant) or allotype control. The endpoint after 30 h was normalized relative to the allotype control (grey bar), and the aggregated TDP-43 percentage was calculated for ACI-7069-633B12-Ab1 (dotted gray bar). Mean ± SD of the three independent experiments are shown, and statistical differences between the allotype control and ACI-7069-633B12-Ab1 (IgG2a variant) were analyzed by Welch t-test (***p < 0.001).

[0185] Figure 5(A) shows the area of ​​Iba1-positive immunoreactivity in the cerebral cortex of mice treated with the carrier (n=16, gray bars) and those treated with ACI-7069-633B12-Ab1 (IgG2a variant) (n=16, dotted gray bars). Error bars represent the standard error (SEM) of the mean. (B to C) show the mean microglia size in the cerebral cortex of mice treated with the carrier (n=16, gray bars) and those treated with ACI-7069-633B12-Ab1 (IgG2a variant) (n=16, dotted gray bars). Microglia were classified into three types based on their morphology: (B) large hypertrophic, (C) small ramifying, and (D) ramified resting. Statistical differences between the carrier control and ACI-7069-633B12-Ab1 (IgG2a variant) were analyzed by t-test (*p<0.05).

[0186] Figure 6 TDP-43 levels in CSF from multiple FTLD-TDP patients and healthy controls were quantified using AlphaLISA assays with ACI-7069-633B12-Ab1 (IgG2a variant) and ACI-7071-809F12-Ab1 (IgG2a variant). Raw AlphaLISA counts of total TDP-43 were obtained from multiple CSF samples (x-axis) (y-axis). Data from three independent experiments were statistically analyzed using a linear mixed model with group, experiment, sex, and age as fixed factors and individuals as random factors (**p<0.01).

[0187] Figure 7 Immunodepletion of TDP-43 and pTDP-43 in detergent (sarkosyl) insoluble fractions obtained from FTD type A postmortem brain tissue, using antibodies ACI-7069-633B12-Ab1 (IgG2a variant) (1), ACI-7069-642D12-Ab1 (IgG2a variant) (2), and a mouse IgG2a control (3). Immunodepletion fractions 1 through 3 were analyzed by Western blotting using TDP-43 or pTDP-43 specific detection antibodies. IN refers to input material (before immunodepletion). Detailed Implementation

[0188] X. Definition

[0189] As used herein, "antigen-binding molecule" refers to any molecule that can specifically or selectively bind to an antigen, particularly TDP-43. Binding molecules may include or may be antibodies or fragments thereof. Anti-TDP-43 binding molecules are molecules that bind to the TDP-43 protein at a specific recognition site (epitope), such as anti-TDP-43 antibodies or fragments thereof. That is, the antigen-binding molecules of the present invention bind to the epitope in 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., diabody), fusion molecules, aptamers, avimers, or other naturally occurring or recombinantly generated molecules. Illustrative antigen-binding molecules that may be used in the present invention include antibody-like molecules. Antibody-like molecules are molecules that function by binding to target molecules (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 include, for example, DARPin (WO 2002 / 020565), affibody (WO 1995 / 001937), affixosome (WO 2004 / 044011; WO 2005 / 040229), adnectin (WO 2002 / 032925), and fynomer (WO2013 / 135588).

[0190] As used herein, the terms "anti-TDP-43 antibody" and "antibody that binds to TDP-43," or simply "antibody," refer to antibodies that bind to TDP-43 with sufficient affinity, making them suitable for use as diagnostic and / or therapeutic agents targeting TDP-43. Generally, the term "antibody" is used in the broadest sense herein and encompasses a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific or bicomplementary antibodies), fully human antibodies, and antibody fragments, provided they exhibit the desired antigen-binding activity. Antibodies within the scope of this invention may also be chimeric antibodies, recombinant antibodies, antigen-binding fragments of recombinant antibodies, humanized antibodies, or antibodies displayed on the surface of bacteriophages or chimeric antigen receptor (CAR) T cells.

[0191] An antibody “antigen-binding fragment” refers to a molecule that contains a portion of the complete antibody and binds to an antigen that binds to the complete antibody, distinct from the complete antibody. Some examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; biantibodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

[0192] "Antibodies that bind to epitopes in protein-defined regions" are antibodies that require the presence of one or more amino acids in that region to bind to the protein.

[0193] In some embodiments, the "antibody that binds to an epitope in a protein-defined region" is identified by mutation analysis, wherein an amino acid of the protein is mutated, and the binding of the antibody to the resulting modified protein (e.g., a modified protein containing the epitope) is determined to be at least 20% of the binding to the unmodified protein. In some embodiments, the "antibody that binds to an epitope in a protein-defined region" is identified by mutation analysis, wherein an amino acid of the protein is mutated, and the binding of the antibody to the resulting modified protein (e.g., a modified protein containing the epitope) is determined to be at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the binding to the unmodified protein. In some embodiments, antibody binding is determined by FACS, WB, or by a suitable binding assay such as ELISA.

[0194] In the context of this invention, the term "binding with" defines the binding (interaction) of at least two "antigen interaction sites" to each other. According to the invention, the term "antigen interaction site" defines a motif of a polypeptide, i.e., a portion of the antibody or antigen-binding fragment of the invention, which exhibits the ability to specifically interact with a specific antigen or group in the TDP-43 antigen. The binding / interaction should also be understood to define "specific recognition." According to the invention, the term "specific recognition" means that the antibody is capable of specifically interacting with and / or binding to at least two amino acids of TDP-43 as defined herein, particularly interacting / binding to at least two amino acid residues of human TDP-43 (SEQ ID NO:1) at positions 181-195, 199-213, 307-321, 352-366, 389-411, 397-411, and 140-200, even more particularly with human TDP-43 (SEQ ID NO:1). At least two amino acids in positions 183 to 188, 203 to 213, 204 to 208, 204 to 211, 205 to 210, 316 to 323, 358 to 361, 400 to 405, 400 to 406, or 400 to 412 of NO:1 interact / bind.

[0195] The term "pan-TDP-43 antibody" refers to an antibody that binds to misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, including monomeric TDP-43, oligomeric TDP-43, post-translational modified TDP-43 (e.g., phosphorylated, ubiquitinated, acetylated, ubiquitinated-like, and / or methylated), aggregated TDP-43, and truncated TDP-43.

[0196] The term "specific interaction" as used in this invention means that the antibody or its antigen-binding fragment of this invention does not cross-react with, or substantially does not cross-react with, peptides having similar structures. Therefore, the antibody or its antigen-binding fragment of the present invention specifically binds to / interacts with the TDP-43 structure formed by a specific amino acid sequence in amino acid residues 181 to 195, 199 to 213, 307 to 321, 352 to 366, 389 to 411, 397 to 411, and 140 to 200 of human TDP-43 (SEQ ID NO:1), and more particularly, binds to / interacts with the TDP-43 structure formed by a specific amino acid sequence in amino acid residues 183 to 188, 203 to 213, 204 to 208, 204 to 211, 205 to 210, 316 to 323, 358 to 361, 400 to 405, 400 to 406, or 400 to 412 of human TDP-43 (SEQ ID NO:1).

[0197] The cross-reactivity of the group of antigen-binding molecules under investigation, particularly antibodies or their antigen-binding fragments, can be tested, for example, by assessing the binding of the group of antibodies or their antigen-binding fragments to the target (poly)peptide and to many (poly)peptides that are more or less (structurally and / or functionally) closely related, under normal conditions (see, for example, 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)). Only those constructs (i.e., antibodies, their antigen-binding fragments, etc.) that bind to certain TDP-43 structures as defined herein, such as specific epitopes or (poly)peptides / proteins of TDP-43 as defined herein, but not to or substantially not to any other epitopes or (poly)peptides of the same TDP-43, are considered specific to the target epitope or (poly)peptide / protein and are selected for further investigation according to the methods provided herein. These methods may in particular include binding studies, blocking, and competition studies of molecules that are structurally and / or functionally closely related. These binding studies also include FACS analysis, surface plasmon resonance (SPR), and, for example, BIACORE. TM Methods include analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, fluorescence spectroscopy, or determination by binding of radiolabeled ligands.

[0198] Therefore, specificity can be determined experimentally using methods known in the art and those described herein. Such methods include, but are not limited to, Western blotting, ELISA, RIA, ECL, IRMA assays, and peptide scanning.

[0199] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies constituting that population are identical except for the possibility of naturally occurring mutations present in small amounts. Monoclonal antibodies are highly specific, targeting a single antigenic site. An advantage of monoclonal antibodies is that they can be synthesized from hybridoma cultures and are substantially unaffected by contamination from other immunoglobulins. The modifier "monoclonal" indicates the antibody's characteristics within a substantially homogeneous population of antibodies and should not be construed as requiring the antibody to be produced by any particular method. As described above, the monoclonal antibody used according to the invention can be prepared by the hybridoma method described in Kohler, Nature 256 (1975), 495.

[0200] As used in this article, the term "polyclonal antibody" refers to an antibody produced in the presence of one or more other different antibodies. Generally, polyclonal antibodies are produced by B lymphocytes in the presence of several other B lymphocytes that produce different antibodies. Typically, polyclonal antibodies are obtained directly from immunized animals.

[0201] As used herein, the term "fully human antibody" refers to an antibody containing only the sequence of human immunoglobulin proteins. If generated in mice, mouse cells, or hybridomas derived from mouse cells, a fully human antibody may contain mouse glycans. Similarly, "mouse antibody" or "mouse antibody" refers to an antibody containing only the sequence of mouse / mouse immunoglobulin proteins. Alternatively, if generated in rats, rat cells, or hybridomas derived from rat cells, a "fully human antibody" may contain rat glycans. Similarly, the term "rat antibody" refers to an antibody containing only the sequence of rat immunoglobulins. Fully human antibodies can also be generated, for example, by phage display, a widely used screening technique capable of generating and screening fully human antibodies. Phage antibodies can also be used in the context of this invention. Phage display methods are described, for example, in US 5,403,484, US 5,969,108, and US 5,885,793. Another technique enabling the development of fully human antibodies involves improvements to mouse hybridoma technology. Mice are genetically modified to include human immunoglobulin loci in exchange for their own mouse genes (see, for example, US 5,877,397).

[0202] The term "chimeric antibody" refers to an antibody of the present invention that comprises a variable region fused or chimeric with an antibody region (e.g., a constant region) from another, human or non-human species (e.g., mouse, horse, rabbit, dog, cow, chicken).

[0203] The term antibody also includes recombinant human antibodies, heterologous antibodies, and heterohybrid antibodies. The term "recombinant (human) antibody" includes antibodies against all human sequences prepared, expressed, produced, or isolated by recombinant means, such as antibodies isolated from animals (e.g., mice) transgenic to target human immunoglobulin genes; antibodies expressed using recombinant expression vectors transfected into host cells; antibodies isolated from recombinant, combined human antibody libraries; or antibodies prepared, expressed, produced, or isolated by any other means involving splicing human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions (if present) derived from human germline immunoglobulin sequences. However, such antibodies can be mutagenized in vitro (or, when using animals transgenic to target human Ig sequences, in vivo somatic cell mutagenization), and therefore the amino acid sequences of the VH and VL regions of recombinant antibodies are sequences that, although derived from and associated with human germline VH and VL sequences, may not be naturally present in an in vivo human antibody germline library.

[0204] "Heterologous antibodies" are defined in relation to the genetically modified non-human organism that produces such antibodies. The term refers to antibodies that have an amino acid sequence or coding nucleic acid sequence that corresponds to an amino acid sequence or coding nucleic acid sequence present in an organism not composed of genetically modified non-human animals, and that the organism is typically derived from a species other than the genetically modified non-human animal species.

[0205] The term "heterogeneous antibody" refers to an antibody containing light and heavy chains from different biological sources. For example, an antibody with a human heavy chain associated with a mouse light chain is a heterogeneous antibody. Some examples of heterogeneous antibodies include chimeric antibodies and humanized antibodies.

[0206] The term antibody also refers to humanized antibodies. A “humanized” form of 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 sequence of the antibody) containing a minimal sequence derived from a non-human immunoglobulin. Typically, a humanized antibody is a human immunoglobulin (receptor antibody) in which residues from the complementary determining region (CDR) of the receptor are replaced with residues from the CDR of a non-human species (donor antibody) (e.g., mouse, rat, or rabbit) having the desired specificity, affinity, and ability. In some cases, Fv framework residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, humanized antibodies may contain residues not found in the receptor antibody or in the introduced CDR or framework sequence. These modifications are made to further refine and optimize antibody performance. Generally, humanized antibodies will contain at least one, and typically substantially all, of the two variable domains, wherein all or substantially all of the CDR regions correspond to those of non-human immunoglobulins, and all or substantially all of the FR regions are those of the common sequences of human immunoglobulins. Humanized antibodies may also contain at least a portion of the immunoglobulin constant region (Fc) (which is typically the constant region of human immunoglobulins). 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.

[0207] Popular methods for antibody humanization involve CDR grafting, in which a functional antigen-binding site from a non-human "donor" antibody is grafted onto a human "recipient" antibody. CDR grafting methods are known in the art and are described, for example, in US 5,225,539, US 5,693,761, and US 6,407,213. Another related method is the production of humanized antibodies from transgenic animals genetically modified to contain one or more humanized immunoglobulin loci capable of gene rearrangement and conversion (see, for example, US 7,129,084).

[0208] Therefore, in the context of this invention, the term "antibody" refers to the complete immunoglobulin molecule and portions of such immunoglobulin molecules (i.e., "the antigen-binding fragment thereof"). Furthermore, as mentioned above, the term refers to modified and / or altered antibody molecules. The term also refers to antibodies produced / synthesized through recombinant or synthetic processes. The term further refers to complete antibodies and antibody fragments thereof, such as 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 Fv (scFv) or antibody fusion proteins.

[0209] In the context of this invention, a "single-chain Fv" or "scFv" antibody fragment has the antibody's V. H and V L Domains, which are present within a single polypeptide chain. Typically, scFv polypeptides also contain V... H With V L The domains contain peptide linkers that enable scFv to form the desired antigen-binding structure. Techniques for generating single-chain antibodies are described, for example, in Plückthun, The Pharmacology of Monoclonal Antibodies, Rosenburg and Mooreeds, Springer-Verlag, NY (1994), 269-315.

[0210] The “Fab fragment” used in this article consists of a light chain and a heavy chain C H 1. Variable region. The heavy chain of the Fab molecule cannot form disulfide bonds with another heavy chain molecule.

[0211] The “Fc” region contains two C-cells containing antibodies. H 2 and C H 3. Heavy chain segments of structural domains. Two heavy chain segments are connected by two or more disulfide bonds and by C... H The hydrophobic interactions of the three structural domains remain together.

[0212] The “Fab” fragment comprises a light chain and a portion of a heavy chain, the portion of which contains V H Domain and C H 1. Structural domain and also has in C H 1 and C H The region between the two structural domains allows for the formation of interchain disulfide bonds between the two heavy chains of the two Fab' segments to form the F(ab')2 molecule.

[0213] The “F(ab')2 segment” contains two light chains and two heavy chains, the heavy chains being contained in C H 1 and C H Part of the constant region between the two structural domains allows for the formation of interchain disulfide bonds between the two heavy chains. Therefore, the F(ab')2 segment consists of two Fab' segments held together by disulfide bonds between the two heavy chains.

[0214] The “Fv region” contains variable regions from both the heavy and light chains, but lacks constant regions.

[0215] The antibodies, antibody constructs, antibody fragments, antibody derivatives (all of which are Ig-derived), or their corresponding immunoglobulin chains used according to the present invention may be further modified using conventional techniques known in the art, such as by using amino acid deletions, insertions, substitutions, additions, and / or recombination alone or in combination, and / or any other modifications known in the art. Methods for introducing such modifications into DNA sequences based on the amino acid sequence of the immunoglobulin chain are well known to those skilled in the art; see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, 2nd edition (1989) and 3rd edition (2001). The term “Ig-derived domain” specifically refers to a (poly)peptide construct containing at least one CDR. Fragments or derivatives of the enumerated Ig-derived domains define the following (polypeptide) peptides that are part of the above antibody molecules and / or 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)).

[0216] As used herein, the term "CDR" refers to a "complementarity-determining region," which is well known in the art. A CDR is a part of an immunoglobulin that determines the specificity of the molecule and its contact with a particular ligand. The CDR is the most variable part of the molecule and contributes to the diversity of these molecules. Three CDR regions are present in each V domain: CDR1, CDR2, and CDR3. CDR-H indicates a CDR region of a variable heavy chain, while CDR-L refers to a CDR region of a variable light chain. VH signifies a variable heavy chain, and VL signifies a variable light chain. CDR regions of Ig-derived regions can be identified 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 according to Kabat. However, those skilled in the art will understand that the present invention is intended to cover binding molecules in which CDR sequences are defined according to any useful identification / numbering scheme.For example, the following numbering scheme can be used to define CDRs: Chothia (Canonical structures for the hypervariable regions of immunoglobulins. Chothia C, Lesk AM. J Mol Biol. 1987 Aug 20; 196 (4): 901-17); S,Bontrop R,Marc L,Malik A,Lefranc MP.Nucleic Acids Res.1997Jan 1;25(1):206-11 and Uniquedatabase numbering system for immunogenetic analysis.Lefranc MP.ImmunolToday.1997Nov;18(11):509);MacCallum(MacCallum RM,Martin AC,Thornton JM,J MolBiol.1996Oct 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).

[0217] Therefore, in the context of this invention, the antibody molecules described above herein are selected from intact antibodies (immunoglobulins, such as 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.

[0218] The term "humanization" is well known in the art and is specifically described for antibody molecules, such as those derived from Ig. The term "humanization" refers to a humanized form of a non-human (e.g., mouse) antibody or fragment thereof (e.g., Fv, Fab, Fab', F(ab'), scFv, or other antigen-binding portion sequences of an antibody) that contains portions of a sequence derived from a non-human antibody. Humanized antibodies include human immunoglobulins in which residues from the complementarity-determining region (CDR) of a human immunoglobulin are replaced by residues from the CDR of a non-human species (e.g., mouse, rat, or rabbit) having the desired binding specificity, affinity, and ability. Generally, humanized antibodies will contain at least one, and typically substantially all, of two variable domains, where all or substantially all of the CDR regions correspond to those of non-human immunoglobulins, and all or substantially all of the FR regions are those of human immunoglobulin common sequences. Ideally, the humanized antibody will also contain at least a portion of the immunoglobulin constant region (Fc) (typically the constant region of human immunoglobulins); see in particular: Jones et al., Nature 321 (1986), 522-525; Presta, Curr. Op. Struct. Biol. 2 (1992), 593-596. Methods for humanizing nonhuman antibodies are well known in the art. Typically, a humanized antibody has one or more amino acids introduced from a nonhuman source while retaining the antibody's original binding activity. Methods for humanizing antibody / antibody molecules are also detailed in Jones et al., Nature 321 (1986), 522-525; Reichmann et al., Nature 332 (1988), 323-327; and Verhoeyen et al., Science 239 (1988), 1534-1536. Specific examples of humanized antibodies, such as 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).

[0219] Therefore, in the context of this invention, antibody molecules or antigen-binding fragments thereof are provided that are humanized and can be successfully used in pharmaceutical compositions.

[0220] The specificity of the antibody or antigen-binding fragment of the present invention can be represented not only by the properties of the amino acid sequence of the antibody or antigen-binding fragment as defined above, but also by the epitopes that the antibody can bind to. Therefore, in one embodiment, the present invention relates to an anti-misfolded TDP-43 antibody or its antigen-binding fragment that recognizes the same epitopes as the antibody of the present invention.

[0221] Those skilled in the art will understand that epitopes may be contained within the TDP-43 protein, but may also be contained in its degradation products or may be chemically synthesized peptides. Only amino acid positions are indicated to show the location of the corresponding amino acid sequence within the TDP-43 protein sequence. This invention covers all peptides containing epitopes. The peptide may be part of a polypeptide longer than 100 amino acids, or may be a small peptide with fewer than 100, preferably fewer than 50, more preferably fewer than 25, and even more preferably fewer than 16 amino acids. The amino acids of such peptides may be natural or non-natural amino acids (e.g., β-amino acids, γ-amino acids, D-amino acids) or combinations thereof. Furthermore, this invention may cover the corresponding retro-inverso peptide of the epitope. The peptide may be unbound or bound. It may be bound to, for example, small molecules (e.g., drugs or fluorophores), high molecular weight polymers (e.g., polyethylene glycol (PEG), polyethyleneimine (PEI), hydroxypropylmethacrylate (HPMA), etc.), or proteins, fatty acids, sugar moieties, or may be intercalated into a membrane.

[0222] To test whether the antibody in question and the antibody of the present invention recognize the same epitope, the following competitive study can be performed: Vero cells infected with three MOIs (multiples of infection) are incubated for 1 hour for different concentrations of the antibody in question as a competitor after 20 hours. In a second incubation step, the antibody of the present invention is applied at a constant concentration of 100 nM, and its binding is detected by flow cytometry using a fluorescently labeled antibody targeting a constant domain of the antibody of the present invention. Binding inversely proportional to the concentration of the antibody in question indicates that the two antibodies recognize the same epitope. However, many other assays known in the art can be used.

[0223] This invention also relates to the generation of specific antibodies against natural and recombinant peptides of TDP-43. This generation is, for example, based on immunization of animals such as mice. However, other animals are also contemplated for antibody / antiserum generation in this invention. For example, monoclonal and polyclonal antibodies can be generated from rabbits, mice, goats, donkeys, etc. A polynucleotide subcloning of a selected peptide encoding TDP-43 can be performed into a suitable vector, wherein the recombinant peptide is expressed in an organism capable of expression, such as bacteria. Thus, the expressed recombinant protein can be injected intraperitoneally into mice, and the resulting specific antibodies can be obtained, for example, from mouse serum provided via intracardiac bloodletting. This invention also contemplates the generation of specific antibodies against natural and recombinant peptides using DNA vaccine strategies as illustrated in the appended examples. DNA vaccine strategies are well known in the art and encompass liposome-mediated delivery, via gene gun or jet injection, and intramuscular or intradermal injection. Therefore, antibodies against TDP-43 peptides, proteins, or epitopes, particularly those of the antibody epitopes provided herein, can be obtained by direct intramuscular injection of a vector expressing the desired TDP-43 peptide, protein, or epitope into animals. The epitopes are particularly the following antibody epitopes of the present invention, located at amino acid residues 181-195, 199-213, 307-321, 352-366, 389-411, 397-411, and 140-200 of SEQ ID NO:1; more particularly, the following antibody epitopes of the present invention are located in SEQ ID NO:1. The specific antibody obtained can be quantified using an ELISA, which is also described below. Other methods for generating antibodies are well known in the art; see, for example, Harlow and Lane, “Antibodies, A Laboratory Manual”, CSH Press, Cold Spring Harbor, 1988.

[0224] Therefore, under specified assay conditions, a specific antibody binds to the corresponding epitope of TDP-43, without binding significantly to other components present in the sample. Specific binding to the target analyte under such conditions may require a binding moiety that is selective for its specificity to the particular target analyte. Various immunoassays can be used to select antibodies that specifically react with a particular antigen. For example, solid-phase ELISA is routinely used to select monoclonal antibodies that specifically react with the analyte. See Shepherd and Dean (2000), Monoclonal Antibodies: A Practical Approach, Oxford University Press and / or Howard and Bethell, for a description of immunoassays and conditions that can be used to determine specific immunoreactivity. Typically, a specific or selective reaction will be at least twice the background signal-to-noise ratio, and more often, more than 10 to 100 times larger. Those skilled in the art can provide and generate specific binding molecules against novel peptides. For specific binding assays, they can be readily used to avoid unwanted cross-reactivity, for example, polyclonal antibodies can be easily purified and selected by known methods (see Shepherd and Dean, loc. cit.).

[0225] An antibody's "class" refers to the type of constant domain or constant region possessed by its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these can be further subdivided into subclasses (isotypes), such as IgG1, IgG2, IgG2a, IgG2b, IgG3, IgG4, IgA1, and IgA2. The constant domains of the heavy chain corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0226] In some embodiments, amino acid sequence variants of the antibodies provided herein are considered. For example, improved binding affinity and / or other biological properties of the antibody may be desired. Amino acid sequence variants of the antibody can be prepared by introducing suitable modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions and / or insertions and / or substitutions of residues in the antibody amino acid sequence. Any combination of deletions, insertions, and substitutions can be made to obtain the final construct, provided that the final construct possesses the desired characteristics, such as antigen binding.

[0227] In some embodiments, antibody variants with one or more amino acid substitutions are provided. Target sites for substitutional mutagenesis include CDR and FR. Conserved substitutions are shown under the heading “Preferred Substitutions” in Table 1. Further variations are provided under the heading “Exemplary Substitutions” in Table 1 and are further described below with reference to the amino acid side chain categories. Amino acid substitutions can be introduced into the target antibody to target desired activities, such as retaining / improving antigen binding, reducing immunogenicity, or improving ADCC or CDC screening products.

[0228] Table 1

[0229]

[0230]

[0231] Amino acids can be grouped according to their common side chain characteristics:

[0232] (1) Hydrophobicity: Leucine, Met, Ala, Val, Leu, Ile;

[0233] (2) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln;

[0234] (3) Acidity: Asp, Glu;

[0235] (4) Alkaline: His, Lys, Arg;

[0236] (5) Residues that affect chain orientation: Gly, Pro;

[0237] (6) Aromatics: Trp, Tyr, Phe.

[0238] Non-conservative replacement would require replacing members of one of these categories with members of another category.

[0239] One type of substitution variant involves replacing one or more hypervariable residues in a parent antibody (e.g., a humanized or human antibody). Typically, the resulting variant selected for further research will have improvements (e.g., enhancements) in certain biological properties relative to the parent antibody (e.g., increased affinity, decreased immunogenicity) and / or will substantially retain certain biological properties of the parent antibody. An exemplary substitution variant is an affinity-matured antibody, which can be conveniently generated, for example, using phage display-based affinity maturation techniques, such as those described herein. In short, by mutating one or more CDR residues, the variant antibody is displayed on a phage and screened for specific biological activities (e.g., binding affinity).

[0240] Alterations (e.g., substitutions) can be made in the CDR, for example, to improve antibody affinity. Such alterations can be made in CDR “hotspots,” which are residues encoded by codons that mutate at high frequencies during somatic maturation (see, for example, Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and / or the SDR (a-CDR), and the binding affinity of the resulting variant VH or VL is tested. Affinity maturation by constructing a secondary library and reselecting from it has been described, for example, in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brienet al., ed., Human Press, Totowa, NJ, (2001)). In some embodiments of affinity maturation, diversity is introduced into the variable gene selected for maturation by any of a variety of methods (e.g., error-prone PCR, strand mixing, or oligonucleotide directed mutagenesis). A secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity. Another approach to introducing diversity involves a CDR-guided method, where 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 scan mutagenesis or modeling. In particular, CDR-H3 and CDR-L3 are often targeted.

[0241] In some embodiments, substitutions, insertions, or deletions may occur in one or more CDRs, as long as such changes do not substantially reduce the antibody's ability to bind to the antigen. For example, conserved changes that do not substantially reduce binding affinity (e.g., conserved substitutions as provided herein) may be made in the CDRs. Such changes may occur outside of CDR "hot spots" or SDRs. In some embodiments of the variant VH and VL sequences provided above, each CDR is unchanged or contains no more than one, two, or three amino acid substitutions.

[0242] A method for identifying targetable mutagenic residues or regions of an antibody is called "alanine scan mutagenesis," as described by Cunningham and Wells (1989) Science, 244:1081-1085. In this method, residues or target residue groups (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) are identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine if the antibody-antigen interaction is affected. Additional substitutions can be introduced at amino acid positions, showing functional sensitivity to the initial substitution. As an alternative or supplement, the crystal structure of the antigen-antibody complex is used to identify the contact points between the antibody and antigen. Such contact residues and adjacent residues can be targeted or eliminated as candidates for substitution. Variants can be screened to determine if they contain the desired properties.

[0243] Amino acid sequence insertions include fusion of the amino and / or carboxyl ends of peptides ranging in length from one residue to 100 or more residues, as well as intra-sequence insertions of single or multiple amino acid residues. Some examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertional variants of antibody molecules include fusions of the N- or C-terminus of the antibody with an enzyme (e.g., for ADEPT) or peptide that enhances the serum half-life of the antibody.

[0244] In some embodiments, the antibodies provided herein are modified to increase or decrease the degree of antibody glycosylation. The addition or deletion of glycosylation sites against the antibody can be conveniently achieved by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0245] When an antibody contains an Fc region, the carbohydrates linked to it can be altered. Naturally occurring antibodies produced by mammalian cells typically contain a branched, anthelmintic oligosaccharide typically linked to the Asn297 domain of the CH2 region of the Fc region via an N-bond. See, for example, Wright et al., TIBTECH 15:26-32 (1997). The oligosaccharide may include a variety of carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose linked to GlcNAc in the “stem” of the anthelmintic oligosaccharide structure. In some embodiments, the oligosaccharides in the antibodies of the present invention may be modified to produce antibody variants with certain improved properties.

[0246] In one embodiment, an antibody variant is provided having a carbohydrate structure lacking fucose (directly or indirectly) linked to the Fc region. For example, the amount of fucose in such an antibody may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose in the glycan chain of Asn297 relative to the sum of all sugar structures linked to Asn297 (e.g., complex, heterozygous, and high-mannose structures), as measured by MALDI-TOF mass spectrometry, for example, as described in WO 2008 / 077546. Asn297 refers to the asparagine residue located approximately at position 297 in the Fc region (Eu numbering of Fc region residues; see Edelman, G.M. et al., Proc. Natl. Acad. USA, 63, 78-85 (1969)); however, due to minor sequence variations in antibodies, Asn297 can also be located approximately ±3 amino acids upstream or downstream of position 297, i.e., at positions 294 and 300. Such fucosylated variants can possess improved ADCC function. See, for example, U.S. Patent Publication No. US2003 / 0157108 (Presta, L.); US2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd.). Some examples of publications related to “defucosylated” or “fucose-deficient” antibody variants include: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US2003 / 0115614; US 2002 / 0164328; US2004 / 0093621; US2004 / 0132140; US2004 / 0110704; US2004 / 0110282; US2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al., J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al., Biotech. Bioeng. 87:614 (2004).Some examples of cell lines capable of producing defucosylation antibodies include protein fucosylation-deficient Lec13 CHO cells (Ripka et al., Arch. Biochem. Biophys. 249:533-545 (1986); US Patent Application No. US2003 / 0157108A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., especially in Example 11), and knockout cell lines, such as α-1,6-fucosylation gene FUT8 knockout CHO cells (see, for example, Yamane-Ohnuki et al., Bioteeh. Bioeng. 87:614 (2004); Kanda, Y. et al., Bioteehnol. Bioeng. 94(4):680-688 (2006); and WO2003 / 085 107).

[0247] Antibody variants having bipartite oligosaccharides are also provided, for example, wherein the biantennary oligosaccharide linked to the Fc region of the antibody is bipartite by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Some examples of such antibody variants are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and US2005 / 0123546 (Umana et al.). Antibody variants having at least one galactose residue in the oligosaccharide linked to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0248] In some embodiments, one or more amino acid modifications may be introduced into the Fc region of the antibody provided herein, thereby creating an Fc region variant. The Fc region variant may contain a human Fc region sequence (e.g., human IgG1, IgG2, IgG3, or IgG Fc region) containing amino acid modifications (e.g., substitutions) at one or more amino acid positions.

[0249] In some embodiments, the present invention considers antibody variants that possess some, but not all, effector functions, making them desirable candidates for applications where the in vivo half-life of the antibody is important and certain effector functions (e.g., complement activation and ADCC) are unnecessary or detrimental. In vitro and / or in vivo cytotoxicity assays can be performed to determine the reduction / depletion of CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks FcγR binding (and therefore may lack ADCC activity), but retains FcRn binding capacity. The primary cells mediating ADCC, NK cells, express only FcγRIII, while monocytes and microglia express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Some non-limiting examples of in vitro assays for evaluating the ADCC activity of target molecules are described in U.S. Patent Nos. 5,500,362 (see, for example, 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 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)).

[0250] Alternatively, non-radioactive assays can be used (see, for example, ACTI for flow cytometry). TM Non-radioactive cytotoxicity assay (Cell Technology, Inc., Mountain View, CA); and CytoTox Non-radioactive cytotoxicity assays (Promega, Madison, WI). Available effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells.

[0251] As an alternative or supplement, the ADCC activity of the target molecule can be evaluated in vivo, for example in animal models such as those disclosed in Clynes et al., Proc. Nat'l Acad.sci. USA 95:652-656 (1998).

[0252] C1q binding assays can also be performed to determine if the antibody cannot bind C1q and therefore lacks CDC activity. See, for example, C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, CDC assays can be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half-life can also be determined using methods known in the art (see, for example, Petkova, S.B. et al., Int'l. Immunol. 18(12):1759-1769(2006)).

[0253] Antibodies with reduced effector function include those with one or more substitutions at residues 238, 265, 269, 270, 297, 327, and 329 in the Fc region (US Patent No. 6,737,056). Such Fc mutants include those with two or more substitutions at amino acids 265, 269, 270, 297, and 327, including the so-called “DANA” Fc mutant where residues 265 and 297 are replaced with alanine (US Patent No. 7,332,581). Alternatively, antibodies with reduced effector function include antibodies with one or more of the residues at positions 234, 235, and 329 of the Fc region replaced, i.e., residues at positions 234 and 235 replaced with alanine, and residue at position 329 replaced with glycine, the so-called “PG-LALA” Fc mutant (Lo, M. et al., Journal of Biochemistry, 292, 3900-3908).

[0254] Certain antibody variants with improved or weakened binding to FcR are described. (See, for example, U.S. Patent No. 6,737,056; WO 2004 / 056312, and Shields et al., J. Biol. Chem. 9(2):6591-6604(2001)).

[0255] In some embodiments, the antibody variant comprises an Fc region with one or more amino acid substitutions that improve ADCC, such as substitutions at positions 298, 333, and / or 334 of the Fc region (EU numbers of the residues).

[0256] In some embodiments, alterations are made in the Fc region resulting in altered (i.e., improved or reduced) C1q binding and / or complement-dependent cytotoxicity (CDC), for example, as described in U.S. Patent Nos. 6,194,551, WO 99 / 51642, and Idusogie et al., J. Immunol. 164:4178-4184 (2000).

[0257] Antibodies with enhanced half-life and improved binding to the neonatal Fc receptor (FcRn) responsible for transferring 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 contain one or more alternative Fc regions that enhance the binding of the Fc region to the FcRn. Such Fc variants include Fc variants in which one or more of the following Fc region residues are replaced: position 238, position 256, position 265, position 272, position 286, position 303, position 305, position 307, position 311, position 312, position 317, position 340, position 356, position 360, position 362, position 376, position 378, position 380, position 382, ​​position 413, position 424, or position 434, for example, an Fc variant in which the Fc region residue at position 434 is replaced (U.S. Patent No. 7,371,826). See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No. 5,648,260; U.S. Patent No. 5,624,821; and WO 94 / 29351, which relates to other instances of Fc region variants.

[0258] In some embodiments, it is desirable to generate cysteine-modified antibodies, such as "thioMAb," wherein one or more residues of the antibody are replaced by cysteine ​​residues. In some specific embodiments, the replaced residues are located at accessible sites on the antibody. By replacing these residues with cysteine, a reactive thiol group is thereby located at an accessible site on the antibody and can be used to conjugate the antibody with other parts, such as a pharmaceutical part or a linker-pharmaceutical part, to produce an immunoconjugate, as further described herein. In some embodiments, any one or more of the following residues may be replaced by cysteine: V205 (Kabat number) of the light chain; A118 (EU number) of the heavy chain; and S400 (EU number) of the Fc region of the heavy chain. Cysteine-modified antibodies can be generated as described, for example, in U.S. Patent No. 7,521,541.

[0259] In some embodiments, the antibodies provided herein may also be modified to include additional non-protein moieties known in the art and readily available. Suitable moieties for antibody derivatization include, but are not limited to, water-soluble polymers. Some non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol / propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, and poly-1,3,6-trimethyloxolane. Alkane, ethylene / maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers) and dextran or poly(n-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide / ethylene oxide copolymers, polyoxyethyleneized polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may be advantageous in manufacturing due to its stability in water. The polymer can have any molecular weight and can be branched or unbranched. The number of polymers linked to the antibody can vary, and if more than one polymer is linked, they can 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 specific properties or functions of the antibody to be improved, whether the antibody derivative will be used in treatment under the following defined conditions, etc.

[0260] In another embodiment, an antibody is provided as a conjugate with a non-protein portion that can be selectively heated by exposure to radiation. In one embodiment, the non-protein portion is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102:11600-11605 (2005)). The radiation can be of any wavelength, including but not limited to wavelengths that do not damage normal cells but heat the non-protein portion to a temperature that kills cells adjacent to the antibody-non-protein portion.

[0261] Antibodies can be generated using recombinant methods and compositions, for example, as described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding the anti-misfolded TDP-43 antibody described herein is provided. Such a nucleic acid may encode an amino acid sequence comprising the VL of the antibody and / or an amino acid sequence comprising the VH of the antibody (e.g., the light chain and / or heavy chain of the antibody). In another embodiment, one or more vectors (e.g., expression vectors) comprising such a nucleic acid are provided. In another embodiment, a host cell comprising such a nucleic acid is provided. In one such embodiment, the host cell comprises (e.g., having been transformed with): (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 of the antibody; or (2) a first vector and a second vector, the first vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and the second vector comprising a nucleic acid encoding an amino acid sequence comprising the VH of the antibody. In one embodiment, the host cell is eukaryotic, for example, Chinese hamster ovary (CHO) cells or lymphoid cells (e.g., YO, NSO, Sp20). In one embodiment, a method for preparing an anti-misfolded TDP-43 antibody is provided, wherein the method comprises: culturing a host cell containing a nucleic acid encoding an antibody as provided above under conditions suitable for antibody expression, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0262] For recombinant anti-misfolded TDP-43 antibodies, the nucleic acid encoding the antibody, such as as described above, is 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 routine procedures (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of the antibody).

[0263] Suitable host cells for cloning or expressing antibody-encoding vectors include prokaryotic or eukaryotic cells as described herein. Antibodies can be generated in bacteria, for example, particularly where glycosylation and Fc effector function are not required. For expression of antibody fragments and peptides in bacteria, see, for example, U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Val. 248 (BKCLo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, which describes the expression of antibody fragments in *Escherichia coli* (E. coli). Following expression, the antibody in the soluble fraction can be isolated from the bacterial cell paste and further purified.

[0264] Besides prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeasts are also suitable cloning or expression hosts for antibody-encoding vectors, including fungal and yeast strains in which the glycosylation pathway has been "humanized," resulting in the production of antibodies with partial or complete human glycosylation patterns. See Gerngross, Nat. Biotech. 22:1409-1414 (2004) and Li et al., Nat. Biotech. 24:210-215 (2006).

[0265] Suitable host cells for expressing glycosylated antibodies also originate from multicellular organisms (invertebrates and vertebrates). Some examples of invertebrate cells include plant and insect cells. Numerous baculovirus strains have been identified that can be used in combination with insect cells, particularly for transfecting fall armyworm (Spodoptera frugiperda) cells.

[0266] Plant cell cultures can also be used as hosts. See, for example, U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (which describe PLATNIBODIES for generating antibodies in transgenic plants). TM technology).

[0267] Vertebrate cells can also be used as hosts. For example, mammalian cell lines suitable for suspension culture can be used. Other examples of available mammalian host cell lines include: rhesus monkey kidney CV1 line (COS-7) transformed with SV40; human embryonic kidney cell line (293 or 293 cells, as described in, for example, Graham et al., J. Gen Viral. 36:59 (1977)); baby hamster kidney cell (BHK); mouse serratus cells (TM4 cells, as described in, for example, Mather, Biol. Reprod. 23:243-251 (1980)); rhesus monkey kidney cells (CV1); African green rhesus monkey kidney cells (VER0-76); human cervical cancer cells (HeLa); canine kidney cells (MDCK); Buffalo rat hepatocytes (BRL 3A); human lung cells (W138); human hepatocytes (Hep G2); mouse mammary tumor (MMT 060562); and TRI cells, as described in, for example, Mather et al., Annals N. Y. In Aead. Sei. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other available mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR CHO cells (Urlaub et al., Proc. Natl. Acad. Cii. USA 77:4216 (1980)); and myeloma cell lines such as YO, NSO, and Sp2 / 0. For a review of certain mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Val. 248 (BKCLo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

[0268] A method for generating the TDP-43 binding molecule, particularly an antibody, of the present invention may include the following steps:

[0269] a. Culturing suitable host cells or cell-free expression systems under conditions conducive to the production of binding molecules, particularly antibodies; and

[0270] b. Isolation of binding molecules, especially antibodies. Suitable culture and isolation techniques are available to technicians.

[0271] The anti-TDP-43 antibodies presented herein can be identified, screened, or characterized for their physical / chemical properties and / or biological activity using a variety of assays known in the art.

[0272] In one aspect, the antibody of the present invention is produced, for example, by a known method, such as ELISA. Its antigen-binding activity can be tested by FACS, immunofluorescence, or immunohistochemistry.

[0273] In another aspect, competitive assays can be used to identify antibodies that compete with any of the antibodies described herein for binding to TDP-43. In some embodiments, such competitive antibodies bind to the same epitope (e.g., linear or conformational epitope) that the antibody described herein binds to. Detailed exemplary methods for mapping epitopes bound to antibodies are provided in Morris (1996), “Epitope Mapping Protocols,” Methods in Molecular Biology vol. 66 (Humana Press, Totowa, NJ).

[0274] In an exemplary competitive assay, immobilized TDP-43 is incubated in a solution containing a first labeled antibody (e.g., any antibody described herein) that binds to TDP-43 and a second unlabeled antibody, the second unlabeled antibody being 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 containing the first labeled antibody but not the second unlabeled antibody. After incubation under conditions allowing the first antibody to bind to TDP-43, excess unbound antibody is removed, and the amount of label associated with the immobilized TDP-43 is measured. If the amount of label associated with the immobilized TDP-43 in the test sample is significantly reduced relative to the control sample, it indicates that the second antibody is competing 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).

[0275] The present 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, bacterial, fungal, plant or animal-derived enzyme-active toxins, or fragments thereof), radioisotopes (i.e., radioconjugates), blood-brain barrier penetrating portions, or detectable markers.

[0276] In another aspect of the invention, an article is provided comprising materials that can be used to treat, prevent, and / or diagnose the aforementioned diseases, disorders, abnormalities, or TDP-43 protein disorders related to TDP-43, particularly those related to TDP-43 aggregates. The article comprises a container and a label or packaging insert on or attached to the container. Suitable containers include, for example, bottles, vials, syringes, intravenous (IV) solution bags, etc. The container may be formed from a variety of materials, such as glass or plastic. The container contains a composition that, alone or in combination with another composition, is effective in treating, preventing, and / or diagnosing the condition, and may have a sterile access port (e.g., the container may be an IV solution bag or a vial with a stopper that can be punctured by a hypodermic needle). At least one active agent in the composition is an antibody of the present invention. The label or packaging insert indicates that the composition is intended to treat a selected condition. Furthermore, the article may comprise: (a) a first container containing the composition, wherein the composition contains the antibody of the present invention; and (b) a second container containing the composition, wherein the composition contains an additional therapeutic agent. The article in this embodiment of the invention may also include a packaging insert indicating that the composition is intended to treat a specific condition. As an alternative or supplement, the article may also 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 dextran solution. It may also contain other materials desired from a commercial and user perspective, including additional buffers, diluents, filters, needles, and syringes.

[0277] It should be understood that any of the above products may contain the immunoconjugate of the present invention in place of or as a supplement to anti-TDP-43 antibody.

[0278] XI. Exemplary TDP-43 specific binding molecules or antibodies

[0279] In some embodiments of the present invention, the antibody comprises:

[0280] 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, and 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; or

[0281] 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, and 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; or

[0282] 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, and 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; or

[0283] 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, and 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; or

[0284] 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, and 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; or

[0285] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO: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; or

[0286] 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, and 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; or

[0287] 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, and 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; or

[0288] 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, and 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; or

[0289] 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, and 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

[0290] 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, 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.

[0291] In some embodiments, the antibody comprises:

[0292] a. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:10 and the light chain variable region (VL) containing the sequence of SEQ ID NO:14; or

[0293] b. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:20 and the light chain variable region (VL) containing the sequence of SEQ ID NO:24; or

[0294] c. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:30 and the light chain variable region (VL) containing the sequence of SEQ ID NO:34; or

[0295] d. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 and the light chain variable region (VL) containing the sequence of SEQ ID NO:44; or

[0296] e. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 60 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 64; or

[0297] f. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:70 and the light chain variable region (VL) containing the sequence of SEQ ID NO:74; or

[0298] g. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:80 and the light chain variable region (VL) containing the sequence of SEQ ID NO:84; or

[0299] h. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 100 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 104; or

[0300] i. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:120 and the light chain variable region (VL) containing the sequence of SEQ ID NO:124; or

[0301] j. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:140 and the light chain variable region (VL) containing the sequence of SEQ ID NO:144; or

[0302] k. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 and the light chain variable region (VL) containing the sequence of SEQ ID NO:154.

[0303] In some embodiments, the antibody comprises:

[0304] a. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:10 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:14; or

[0305] b. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:20 or a heavy chain variable region (VH) having at least 98% or 99% sequence identity with 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 a light chain variable region (VL) having at least 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:24; or

[0306] c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:30 or a heavy chain variable region (VH) having at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 98% or 99% sequence identity with the amino acid sequence of SEQ ID NO:34; or

[0307] d. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:40, and a light chain variable region (VL) containing the sequence of SEQ ID NO:44; or

[0308] e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:60 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:60, and a light chain variable region (VL) comprising the sequence of SEQ ID NO:64 or a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:64; or

[0309] f. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:70 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:74; or

[0310] g. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:80 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:84; or

[0311] h. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:100 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:104; or

[0312] i. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:120 or a heavy chain variable region (VH) having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:124; or

[0313] j. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:140 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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

[0314] k. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 or a heavy chain variable region (VH) having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:154.

[0315] In some embodiments, the present invention relates to antibodies derived from hybridoma clones 631B2A2, 633B12C8, 634H10H7, 636E5B8, 641H1E7, 642A10B11, 642D12B4, 646B7F7, 712A6B10, 809D9C2, or 809F12D8, as further described herein.

[0316] In some embodiments, the present invention relates to antibodies selected from the following: 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.

[0317] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid encodes the antibody described herein.

[0318] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:18 encoding an anti-TPD-43 antibody.

[0319] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:19 encoding an anti-TPD-43 antibody.

[0320] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:28 encoding an anti-TPD-43 antibody.

[0321] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:29 encoding an anti-TPD-43 antibody.

[0322] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:38 encoding an anti-TPD-43 antibody.

[0323] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:39 encoding an anti-TPD-43 antibody.

[0324] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:48 encoding an anti-TPD-43 antibody.

[0325] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:49 encoding an anti-TPD-43 antibody.

[0326] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:68 encoding an anti-TPD-43 antibody.

[0327] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:69 encoding an anti-TPD-43 antibody.

[0328] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:78 encoding an anti-TPD-43 antibody.

[0329] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:79 encoding an anti-TPD-43 antibody.

[0330] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:88 encoding an anti-TPD-43 antibody.

[0331] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:89 encoding an anti-TPD-43 antibody.

[0332] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:108 encoding an anti-TPD-43 antibody.

[0333] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:109 encoding an anti-TPD-43 antibody.

[0334] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:128 encoding an anti-TPD-43 antibody.

[0335] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:129 encoding an anti-TPD-43 antibody.

[0336] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:148 encoding an anti-TPD-43 antibody.

[0337] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:149 encoding an anti-TPD-43 antibody.

[0338] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:158 encoding an anti-TPD-43 antibody.

[0339] In some implementations, a (isolated) nucleic acid is provided, wherein the (isolated) nucleic acid contains SEQ ID NO:159 encoding an anti-TPD-43 antibody.

[0340] XII. Compositions and Methods

[0341] In some embodiments, an immunoconjugate is provided, wherein the immunoconjugate comprises the (isolated) antibody and therapeutic agent described herein. In some embodiments, a labeled antibody is provided, comprising the antibody described herein and a detectable label.

[0342] In some embodiments, a pharmaceutical composition is provided comprising the (isolated) antibody and pharmaceutically acceptable carrier described herein.

[0343] In some embodiments, the TDP-43 specific binding molecule of the present invention is linked to a detectable label.

[0344] In some 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.

[0345] In some embodiments, the TDP-43 specific binding molecule or an immunoconjugate thereof exists as a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologous molecules, or alternatively as its antagonist.

[0346] In some embodiments, the TDP-43 specific binding molecule is part of a pharmaceutical composition comprising, in combination with a pharmaceutically acceptable carrier, the following: a TDP-43 specific binding molecule, or an immunoconjugate wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and a homologous molecule, or alternatively, its antagonist.

[0347] In some 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 linked to another suitable therapeutic agent, or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologous molecules, or alternatively, their antagonists.

[0348] Kits containing the binding molecules of the present invention are also provided. In particular, such kits can be used for performing the diagnostic methods of the present invention (including classification, monitoring, and treatment selection methods). Therefore, kits are provided for diagnosing diseases, disorders, and / or abnormalities related to TDP-43, particularly TDP-43 aggregates, or TDP-43 proteopathies, or for use in the methods of the present invention, containing the TDP-43-specific binding molecules of the present invention. Such kits may contain all the necessary components for performing the methods provided herein. Typically, each component is stored individually in a single monolithic package. Suitable additional components included in the kit are, for example, buffers, detectable dyes, laboratory equipment, reaction vessels, instructions, etc. Instructions for use may be customized for the specific method for which the kit is to be used. Appropriately labeled TDP-43 binding molecules of the present invention are also provided, which may be included in such kits.

[0349] In some implementations, TDP-43-specific binding molecules are used in immunodiagnostic methods for the prevention, diagnosis, or treatment of TDP-43 protein disorders.

[0350] In some embodiments, the TDP-43 specific binding molecule is part of an immunotherapy for the prevention or treatment of TDP-43 protein disorders, wherein an effective amount of the TDP-43 specific binding molecule is administered to a subject in need, or wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising the TDP-43 specific binding molecule and a TDP-43 agonist and homologous molecules, or alternatively, their antagonists.

[0351] In some embodiments, a TDP-43-specific binding molecule, or an immunoconjugate wherein the TDP-43-specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising the TDP-43-specific binding molecule and a TDP-43 agonist and homologous molecules, or alternatively their antagonists, is administered to a subject in need for the diagnosis, prevention, mitigation, or treatment of diseases, disorders, and / or abnormalities associated with TDP-43, particularly those associated with TDP-43 aggregates, or TDP-43 protein disorders, 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-dominant age-related TDP-43 encephalopathy (LATE).

[0352] In some embodiments, administration to a subject in need of this procedure of a TDP-43 specific binding molecule, or an immunoconjugate wherein the TDP-43 specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising a TDP-43 specific binding molecule and a TDP-43 agonist and a homologous molecule, or alternatively its antagonist, for the diagnosis or monitoring of diseases, disorders and / or abnormalities, or TDP-43 protein disorders selected from those associated with TDP-43, particularly those associated with TDP-43 aggregates, or TDP-43 protein disorders. French and Chinese: Frontotemporal dementia (FTD), such as sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, associated with chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulation disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral changes Atypical FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA, etc.), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polymorphic Glutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis; inclusion body myopathy with valine-containing protein mutations (VCP; as well as Paget's bone disease and frontotemporal dementia); oculopharyngeal muscle dystrophy with rimmed vacuoles; myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

[0353] In other embodiments, the present invention relates to any method for detecting, diagnosing, or monitoring diseases, disorders, and / or abnormalities, or TDP-43 protein disorders selected from the following TDP-43-related, particularly TDP-43 aggregate-related, or TDP-43 protein disorders: frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), and limbic-dominant age-related TDP-43 encephalopathy (LATE).

[0354] Preferably, the disease, disorder, and / or abnormality associated with TDP-43, particularly with TDP-43 aggregates, or the TDP-43 protein disease 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 with TDP-43 aggregates, or the TDP-43 protein disease is amyotrophic lateral sclerosis (ALS). More preferably, the disease, disorder, and / or abnormality associated with TDP-43, particularly with TDP-43 aggregates, or the TDP-43 protein disease is Alzheimer's disease (AD). More preferably, the disease, disorder, and / or abnormality associated with TDP-43, particularly with TDP-43 aggregates, or the TDP-43 protein disease is frontotemporal dementia (FTD).

[0355] In some embodiments, the TDP-43-specific binding molecule is used in methods for diagnosing presymptomatic diseases, monitoring disease progression and treatment efficacy, predicting responsiveness, or selecting subjects that may respond to treatment with the TDP-43-specific binding molecule. The method is preferably performed using human blood or urine samples. Most preferably, the method involves an ELISA-based assay or surface adaptation assay.

[0356] In some embodiments, the TDP-43-specific binding molecule is used in a method of contacting the TDP-43-specific binding molecule of the present invention with a sample (e.g., blood, cerebrospinal fluid, or brain tissue) to detect, diagnose, or monitor frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), chronic traumatic encephalopathy, Perry syndrome, limbic-dominant age-related TDP-43 encephalopathy (LATE), and / or Parkinson's disease (PD).

[0357] In some embodiments, the TDP-43-specific binding molecule is used in methods of contacting the TDP-43-specific binding molecule of the present invention with a sample (e.g., blood, cerebrospinal fluid, or brain tissue) to detect, diagnose, or treat diseases selected from: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, associated with chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), aurophilic granulomatosis, 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). S, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation, Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis; inclusion body myopathy with valine-containing protein mutation (VCP; and Paget's bone disease and frontotemporal dementia); oculopharyngeal dystrophy with rimmed vacuolars; myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

[0358] In some embodiments, a TDP-43-specific binding molecule, or an immunoconjugate wherein the TDP-43-specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising the TDP-43-specific binding molecule and a TDP-43 agonist and a homologous molecule, or alternatively its antagonist, is administered to a subject in need of prevention, mitigation, or treatment of diseases, disorders, and / or abnormalities associated with TDP-43, particularly those associated with TDP-43 aggregates, or TDP-43 proteinopathy, or frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD, including sporadic and familial forms of AD), chronic traumatic encephalopathy, Perry syndrome, and limbic-dominant age-related TDP-43 encephalopathy (LATE) and / or Parkinson's disease (PD).

[0359] In some embodiments, a TDP-43-specific binding molecule, or an immunoconjugate wherein the TDP-43-specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising the TDP-43-specific binding molecule and a TDP-43 agonist and a homologous molecule, or alternatively its antagonist, is administered to the subject in need for the treatment of a disease selected from: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, associated with chromosome 9p, cortical basal ganglia degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulopathy, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia, etc. (nfvPPA, etc.), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinal cord malformation) The disease includes: ataxia type 3 (SCA3; also known as Majovidia disease), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis; inclusion body myopathy with valine-containing protein mutations (VCP; and Paget's bone disease and frontotemporal dementia); oculopharyngeal muscle dystrophy with rimmed vacuoles; myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES)), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD). Preferably, treatment of the disease helps maintain or improve mental cognition and / or reduce the level of TDP-43 aggregates in the brain.

[0360] In some embodiments, administering to a subject in need a TDP-43-specific binding molecule, or an immunoconjugate wherein the TDP-43-specific binding molecule is covalently linked to another suitable therapeutic agent, or a composition comprising a TDP-43-specific binding molecule and a TDP-43 agonist and a homologous molecule, or alternatively its antagonist, to a subject in need of manufacturing a medicament for treating: diseases, disorders and / or abnormalities associated with TDP-43, particularly TDP-43 aggregates, or TDP-43 proteinopathy, or frontotemporal degeneration (FTD) or amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD, including sporadic and familial forms of AD), chronic traumatic encephalopathy, Perry syndrome and limbic-dominant age-related TDP-43 encephalopathy (LATE) and / or Parkinson's disease (PD).

[0361] Pharmaceutical formulations of anti-TDP-43 antibodies (preferred types of TDP-43-specific binding molecules) or immunoconjugates as described herein are prepared by mixing such antibodies or immunoconjugates, having the desired purity, with one or more optional pharmaceutically acceptable carriers (Remington's Pharmaceutical Sciences 16th edition, Osol, A.Ed. (1980)) in the form of lyophilized formulations or aqueous solutions. Pharmaceutically acceptable carriers are generally non-toxic to the recipient at the doses and concentrations employed and include, but are not limited to: buffers, such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (e.g., octadecyl dimethyl benzyl ammonium chloride; hexamethyl diammonium chloride; benzalkonium chloride; benzyl chloride; phenol, butanol, or benzyl alcohol; alkyl esters of p-hydroxybenzoate, such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol, and m-cresol); low molecular weight (less than about 10 residues). Polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., Zn protein complexes); and / or nonionic surfactants, such as polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers described herein also include interstitial drug dispersants, such as soluble neutral-active hyaluronidase glycoprotein (sHASEGP), such as human soluble PH-20 hyaluronidase glycoprotein, such as rHuPH20 (…). Baxter International, Inc. Certain exemplary sHASEGP and methods of use, including rHuPH20, are described in U.S. Patent Publications Nos. 2005 / 0260186 and 2006 / 0104968. In one aspect, sHASEGP is combined with one or more additional glycosaminoglycanases, such as chondroitinase.

[0362] Exemplary lyophilized antibody or immunoconjugate formulations are described in U.S. Patent No. 6,267,958. Aqueous antibody or immunoconjugate formulations include those described in U.S. Patent Nos. 6,171,586 and WO2006 / 044908, the latter of which contain a histidine-acetate buffer.

[0363] For the specific indications being treated, the formulations described herein may, if necessary, contain more than one active ingredient, preferably those with complementary activities that do not adversely affect each other.

[0364] The active ingredient can be encapsulated in microcapsules, such as those prepared by coagulation techniques or interfacial polymerization, for example, hydroxymethyl cellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively; in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules); or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A.Ed. (1980).

[0365] Sustained-release formulations can be prepared. Some suitable examples of sustained-release formulations include a semi-permeable matrix of a solid hydrophobic polymer containing an antibody or immunoconjugate, which is in the form of a molded article, such as a membrane or microcapsule. Formulations intended for in vivo administration are typically sterile. Sterility can be readily achieved, for example, by filtration via a sterile filter membrane.

[0366] Any antigen-binding molecules, anti-TDP-43 antibodies, or immunoconjugates provided herein may be used in methods, such as therapeutic applications.

[0367] In another aspect, anti-TDP-43 antibodies (preferably of the type of TDP-43-specific binding molecules) or immunoconjugates are provided for use as medicaments. In yet another aspect, anti-misfolded TDP-43 antibodies (preferably of the type of TDP-43-specific binding molecules) or immunoconjugates are provided for use in therapeutic methods. In some embodiments, anti-TDP-43 antibodies (preferably of the type of TDP-43-specific binding molecules) or immunoconjugates are provided for the prevention, diagnosis, and / or treatment of TDP-43 proteinopathy. In a preferred embodiment of the invention, an anti-TDP-43 antibody (preferably of a TDP-43-specific binding molecule) or an immunoconjugate is provided for the prevention, diagnosis, and / or treatment of diseases, disorders, and / or abnormalities associated with TDP-43, particularly with TDP-43 aggregates, or TDP-43 proteopathies, 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 dominant age-related TDP-43 encephalopathy (LATE).

[0368] In another aspect, the present invention provides the use of anti-TDP-43 antibodies (preferred types of TDP-43-specific binding molecules) or immunoconjugates in the manufacture or preparation of pharmaceuticals. In one such embodiment, the method further includes administering an effective amount of at least one additional therapeutic agent to an individual, for example, as described below.

[0369] According to any of the above implementation schemes, the "object" or "individual" can be an animal, a mammal, preferably a human.

[0370] In another aspect, the present invention provides, for example, a pharmaceutical formulation comprising any anti-TDP-43 antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate provided herein for any of the above-described treatment methods. In one embodiment, the pharmaceutical formulation comprises any anti-TDP-43 antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate provided herein, and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical formulation comprises any anti-TDP-43 antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate provided herein, and at least one additional therapeutic agent, such as those described below.

[0371] The antibodies or immunoconjugates of the present invention can be used alone or in combination with other agents in treatment. For example, the antibodies (preferably of the type of TDP-43-specific binding molecules) or immunoconjugates of the present invention can be co-administered with at least one additional therapeutic agent.

[0372] The combination therapies described above encompass both combined administration (where two or more therapeutic agents are contained in the same or separate formulations) and separate administration. In the case of separate administration, the administration of the antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate of the present invention may occur before, simultaneously with, and / or after the administration of additional therapeutic agents and / or adjuvants. The antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate of the present invention may also be used in combination with radiotherapy.

[0373] The antibodies (preferably of the type of TDP-43-specific binding molecule) or immunoconjugates (and any other therapeutic agents) of the present invention may be administered by any suitable means, including parenteral, intrapulmonary, and intranasal administration, and, if necessary, local, intralesional, intrauterine, or intravesical administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Administration may be carried out by any suitable route, such as by injection, for example, intravenous or subcutaneous injection, depending in part on whether it is a short-term or long-term administration. Various dosing regimens are considered herein, including but not limited to single administration or multiple administrations at different time points, bolus administration, and pulsatile infusion.

[0374] The antibodies (preferably of the type of TDP-43-specific binding molecules) or immunoconjugates of the present invention can be formulated, administered, and applied in a manner consistent with good medical practice. Factors to consider in this case include: the specific disease, disorder, and / or abnormality, or TDP-43 proteinopathy, related to TDP-43, particularly related to TDP-43 aggregates, being treated; the specific mammal being treated; the clinical condition of the individual subject; the cause of the disease, disorder, and / or abnormality, or TDP-43 proteinopathy, related to TDP-43, particularly related to TDP-43 aggregates; the site of delivery of the agent; the method of administration; the administration regimen; and other factors known to medical practitioners. The antibodies or immunoconjugates do not need to be formulated, but optionally may be formulated, with one or more agents currently used for the prevention or treatment of the TDP-43-related disease, disorder, and / or abnormality, or TDP-43 proteinopathy discussed, particularly related to TDP-43 aggregates. The effective amount of such other agents depends on the amount of antibody or immunoconjugate present in the formulation; the type of disease, disorder, and / or abnormality associated with TDP-43, particularly with TDP-43 aggregates, or TDP-43 proteinopathy; or the treatment, and other factors mentioned above. These are generally used at the same dosage and route of administration as described herein, or at about 1% to 99% of the dosage described herein, or at any dosage and route of administration that is empirically / clinically determined to be appropriate.

[0375] For the prevention or treatment of disease, the appropriate dose of the antibody (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) will depend on the type of disease being treated, the type of antibody or immunoconjugate, the severity and cause of the disease, whether the antibody or immunoconjugate is administered for preventive or therapeutic purposes, prior treatment, the subject's clinical history and response to the antibody or immunoconjugate, and the judgment of the attending physician. The antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate may be administered to the subject appropriately once or via a series of treatments. Depending on the type and severity of the disease, an initial candidate dose of about 1 μg / kg to 15 mg / kg (e.g., 0.1 mg / kg to 10 mg / kg) of antibody (preferred type of TDP-43-specific binding molecule) or immunoconjugate may be used for administration to the subject, whether, for example, by one or more separate administrations or by continuous infusion. Based on the above factors, a typical daily dose may be about 1 μg / kg to 100 mg / kg or higher. For repeated administration over several days or longer, treatment is typically continued until the desired suppression of disease symptoms is achieved, depending on the condition. An exemplary dose of the antibody or immunoconjugate is from about 0.05 mg / kg to about 10 mg / kg. Therefore, one or more doses (or any combination thereof) of about 0.5 mg / kg, 2.0 mg / kg, 4.0 mg / kg, or 10 mg / kg can be administered to the subject. Such doses can be administered intermittently, for example, weekly or every three weeks (e.g., administering about 2 to about 20, or for example about 6, doses of antibody to the subject). A higher initial loading dose can be administered, followed by one or more lower doses. However, other dosing regimens may be used. Progression of the treatment can be easily monitored using routine techniques and assays.

[0376] It should be understood that any of the above formulations or treatments can be performed using both the immunoconjugate of the present invention and the anti-TDP-43 antibody (a preferred type of TDP-43-specific binding molecule).

[0377] In another aspect of the invention, an article is provided comprising the above-described material that can be used to treat, prevent, and / or diagnose diseases, disorders, or abnormalities related to TDP-43, particularly those related to TDP-43 aggregates, or TDP-43 protein disorders. The article comprises a container and a label or packaging insert on or attached to the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, etc. The container can be formed from various materials, such as glass or plastic. The container contains a composition, alone or in combination with another composition, that is effective in treating, preventing, and / or diagnosing diseases, disorders, and / or abnormalities related to TDP-43, particularly those related to TDP-43 aggregates, or TDP-43 protein disorders, and may have a sterile access port (e.g., the container may be an intravenous solution bag or a vial with a stopper that can be punctured by a hypodermic needle). At least one active agent in the composition is an antibody or immunoconjugate of the present invention. The label or packaging insert indicates that the composition is intended to treat the selected condition. Furthermore, the article may comprise: (a) a first container containing a composition comprising an antibody (preferably of the type of TDP-43-specific binding molecule) or an immunoconjugate of the present invention; and (b) a second container containing a composition comprising an additional therapeutic agent. The article in this embodiment of the invention may also comprise a packaging insert indicating that the composition can be used to treat a specific condition. Alternatively or supplementally, the article may further comprise a second (or third) container containing a pharmaceutically acceptable buffer, such as water for injection (BWFI), phosphate-buffered saline, Ringer's solution, or dextran solution. It may also comprise other materials desired from a commercial and user perspective, including other buffers, diluents, filters, needles, and syringes.

[0378] In another embodiment, the present invention relates to a method for maintaining or improving cognitive memory, motor and language functions, or for preventing and / or slowing the decline of cognitive memory, motor and language functions in a subject, comprising administering the binding molecule of the present invention, the immunoconjugate of the present invention, the composition of the present invention, or the pharmaceutical composition of the present invention.

[0379] In another embodiment, the present invention relates to a method for reducing TDP-43 levels, comprising administering the binding molecule of the present invention, the immunoconjugate of the present invention, the composition of the present invention, or the pharmaceutical composition of the present invention.

[0380] The method of the present invention may include administering at least one additional treatment, preferably wherein the additional treatment is selected from, but not limited to, neuropharmaceuticals, anti-Aβ antibodies, anti-Tau antibodies, Tau aggregation inhibitors, β-amyloid aggregation inhibitors, anti-BACE1 antibodies, and BACE1 inhibitors.

[0381] The present invention also relates to a method for detecting TDP-43, comprising contacting a sample with the binding molecules of the present invention, preferably wherein the sample is a brain sample, a cerebrospinal fluid sample, a urine sample, or a blood sample.

[0382] Example

[0383] Example 1: Preparation of TDP-43 vaccine composition

[0384] The 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.

[0385] Table 2: Description of TDP-43 protein and peptide antigens

[0386]

[0387]

[0388] Example 2: Generation of anti-TDP-43 antibody A. Immunization of mice

[0389] Female C57BL / 6JOlaHsd (C57BL / 6) and BALB / c OlaHsd (BALB / c) wild-type mice (Harlan, USA) were administered vaccines at 9 weeks of age. Vaccination began at 10 weeks. The monophosphoryl hexaacyl lipid A,3-deacylated (synthesized) (3D-(6-acyl)) was used as an adjuvant. In the case of ), mice were vaccinated with the full-length TDP-43 protein present on the surface of liposomes.

[0390] Mice were vaccinated via subcutaneous injection (sc) on days 0, 4, 8, 21, 35, and 60. Heparinized plasma was prepared from blood samples collected from mice 7 days prior to immunization (pre-immunization plasma) and on days 14, 28, 42, 81, and 121 after the first immunization. Additionally, mice induced by myeloma fusion were vaccinated with three daily booster injections of TDP-43 protein via intraperitoneal injection (ip) without adjuvant.

[0391] Vaccine response was measured in mouse plasma. The binding of plasma-derived antibodies from immunized mice to immobilized recombinant full-length (FL) TDP-43 indicated the antibody titer against TDP-43. B. Hybridoma generation and selection of subclones.

[0392] Mice were euthanized, and spleen cells from four individual mice were fused with myeloma cells. Antibodies were screened from the successfully fused hybridoma cell lines as follows. Diluted (1:32) cell culture supernatants were analyzed using a Luminex bead-based multiplex assay (Luminex, The Netherlands). Luminex beads were conjugated with FLTDP-43, and IgG was captured using an anti-mouse IgG-Fc antibody specific for IgG1, IgG2a, IgG2b, IgG2c, and IgG3 subclasses (Jackson Immunoresearch, USA). Binding to the FL TDP-43-conjugated beads identified 386 hits derived from mice immunized with an FL TDP-43 liposomal vaccine.

[0393] Live hybridomas were cultured using a serum-containing selective medium. Clones preferentially binding to TDP-43 inclusion bodies in human FTD brain and clones binding to the C-terminus of TDP-43 were selected for further subcloning. After limiting dilution, the clonal hybridomas were cultured in a medium containing low immunoglobulins, and stable colonies were selected for antibody screening and selection. The antibodies shown in Table 3 were identified from this screening.

[0394] Example 3: Determination of binding efficacy (EC50)

[0395] As previously described, Luminex assays were performed with the antibodies in serial dilutions to determine the half-maximal effect concentration (EC50) at which the antibodies bind to FL TDP-43. All EC50 values ​​are summarized in Table 3. In summary, all tested antibodies bound to full-length TDP-43 with high affinity.

[0396] Table 3: EC50 values ​​determined by Luminex measurements

[0397]

[0398] Example 4: Antibody binding to human FL TDP-43

[0399] Antibodies binding to human FL TDP-43 were identified using an indirect ELISA. ELISA plates were coated overnight at 4°C with 1 μg / ml human FL TDP-43 in carbonate buffer. The plates were washed with 0.05% Tween 20 / PBS and then blocked at 37°C for 1 h with 1% bovine serum albumin (BSA) in 0.05% Tween 20 / PBS. Antibody purified from hybridoma supernatant was then added in 3-fold serial dilutions (starting at 1 μg / ml) and incubated at 37°C for 2 h, followed by washing. AP-conjugated anti-mouse IgG secondary antibody (Jackson Immunoresearch Laboratories, United Kingdom) was added at a 1 / 1000 dilution in 0.05% Tween 20 / PBS for 1 h. After the final wash, the plates were incubated with pNPP (Sigma-Aldrich, Switzerland) phosphatase substrate solution and read at 405 nm using an ELISA reader (Tecan, Switzerland). All tested clones were bound to full-length TDP-43, with EC50 values ​​ranging from 10 to 1567 ng / ml (Table 4).

[0400] Table 4: EC50 values ​​via ELISA

[0401]

[0402] Example 5: Epitope mapping using ELISA and peptide arrays

[0403] Libraries of 40-66 aa linear peptides or 15-mer peptides biotinylated at the N-terminus and covering the entire TDP-43 sequence with a 9 aa offset and a 6 aa overlap were used to screen antibodies purified from serum-free hybridoma supernatants by indirect ELISA to determine the binding region. Peptide sequences are provided in Table 5.

[0404] 96-well plates were coated overnight at 4°C with 5 μg / ml non-biotinylated peptides in carbonate buffer. The plates were washed with 0.05% Tween 20 / PBS and then blocked at 37°C for 1 hour with 1% bovine serum albumin (BSA) in 0.05% Tween 20 / PBS. Antibody purified from hybridoma supernatant was then added at 1 μg / ml and incubated at 37°C for 2 hours, followed by washing. AP-conjugated anti-mouse IgG secondary antibody (Jackson Immunoresearch Laboratories, United Kingdom) was added at a 1 / 1000 dilution in 0.05% Tween 20 / PBS at 37°C for 1 hour. After a final wash, the plates were incubated with pNPP (Sigma-Aldrich, Switzerland) phosphatase substrate solution and read at 405 nm using an ELISA reader (Tecan, Switzerland).

[0405] For biotinylated peptides, 96-well ELISA plates coated with streptoacidin were incubated with 5 μg / mL of biotinylated 15-mer peptide. The plates were washed three times with 0.05% Tween 20 / PBS and then blocked at 37°C for 1 h with 1% bovine serum albumin (BSA) in 0.05% Tween 20 / PBS. Antibody purified from hybridoma supernatant was then added at 1 μg / mL and incubated at 37°C for 2 h, followed by washing. AP-conjugated anti-mouse IgG secondary antibody (Jackson ImmunoResearch Laboratories, United Kingdom) was added at a 1 / 1000 dilution in 0.05% Tween 20 / PBS at 37°C for 1 h. After a final wash, the plates were incubated with pNPP (Sigma-Aldrich, Switzerland) AP substrate solution and read at 405 nm using a Tecan ELISA reader. The identified binding regions are provided in Table 6. The test antibody was found to bind to the following peptides: TP-21, TP-23, TP-35, TP-40, TP-48, and TDP-6, corresponding to positions 181-195, 199-213, 307-321, 352-366, 389-411, and 140-200 of SEQ ID NO:1, respectively.

[0406] More precise linear epitopes were mapped using a library (Pepscan, Netherlands) of 15-mer peptides synthesized directly on a solid support and overlaid with a 1aa offset and a 14aa overlap covering the entire TDP-43 sequence according to SEQ ID NO:1. The peptide array was blocked with horse serum and ovalbumin and incubated overnight at 4°C with purified antibody solutions at concentrations from 0.75 to 5 μg / ml. After washing, the peptide array was incubated for 1 hour at 25°C with a 1 / 1000 dilution of rabbit anti-mouse IgG (H+L) HRP conjugate (Southern Biotech, USA). After washing, the peroxidase substrate 2,2'-azido-di-3-ethylbenzothiazoline sulfonate (ABTS) and 20 μl / ml of 3% H2O2 were added. After one hour, quantification was performed using a charge-coupled device (CCD) camera and image processing system. These binding regions were determined by epitope mapping, and the following epitopes were identified (provided in Table 6): amino acids at positions 183-188, 203-213, 204-208, 204-211, 205-210, 316-323, 358-361, 400-405, 400-406, and 400-412 of SEQ ID NO:1.

[0407] Table 5: Peptides used to determine the binding region by ELISA

[0408]

[0409] Table 6: Binding regions and epitopes of the tested antibodies

[0410] Hybridoma clone name Combined region, aa Epitope, aa 631B2A2 397-411 400-406 633B12C8 397-411 400-405 634H10H7 140-200 183-188 636E5B8 352-366 358-361 641H1E7 199-213 204-211 642A10B11 389-411 400-412 642D12B4 181-195 183-188 646B7F7 199-213 205-210 712A6B10 307-321 316-323 809D9C2 199-213 203-213 809F12D8 199-213 204-208

[0411] Example 6: Detection of TDP-43 in brain tissue from FTD / ALS subjects by immunohistochemistry

[0412] Target binding was evaluated in immunohistochemical experiments using tissues from the brains of FTD subjects. Human FTD brain tissues were obtained from the Netherlands Brain Bank and the Netherlands Institute for Neuroscience (Amsterdam). (Open access: [link to open access information]) www.brainbank.nlMaterials were obtained from the Queen Square Brain Bank for Neurological Disorders (UCL). All materials were collected from donors, from whom written informed consent was obtained regarding brain autopsies and the use of materials and clinical information for research purposes. Immunohistochemistry was performed on 10 μm thick frozen sections using fluorescently labeled secondary antibodies for detection. The following antibodies were used as controls: rabbit polyclonal pan-TDP-43 antibody (Proteintech, 10782-2-AP) for detecting pathological inclusion bodies and physiological nuclear TDP-43; rabbit monoclonal phosphorylated TDP-43p409 / 410 antibody (Cosmobio, TIP-PTD-PO2) for detecting pathological aggregation and phosphorylated TDP-43; and a primary antibody-free secondary antibody (without 1°Ab) for detecting nonspecific background.

[0413] All antibodies of this invention bind to nuclear TDP-43, non-aggregated TDP-43, and aggregated TDP-43. Some antibodies of this invention preferentially bind to aggregated TDP-43 in the cytoplasm of type A pathology. Figure 1 A detailed evaluation of the binding characteristics is summarized in Table 7.

[0414] Table 7: Detection of TDP-43 in brain tissue from FTD subjects

[0415] Antibody name IHC detection of TDP-43 IHC detection of non-aggregated TDP-43 in the cell nucleus ACI-7069-631B2-Ab1 +++ +++ ACI-7069-633B12-Abl +++ +++ ACI-7069-634H10-Ab2 ++ ++ ACI-7069-636E5-Abl + / - +++ ACI-7069-641H1-Ab2 +++ + ACI-7069-642A10-abl +++ + ACI-7069-642D12-Abl ++ + ACI-7069-646B7-Ab1 +++ +++ ACI-7071-712A6-Abl ++ + ACI-7071-809D9-Ab2 +++ +++ ACI-7071-809F12-Abl +++ +++

[0416] NA: Data unavailable; -: Does not exist; + / -: Unclear; +: Weak; ++: Medium; +++: Rich

[0417] Example 7: Detection of TDP-43 in brain tissue from FTD / ALS subjects by Western blot

[0418] Using Precellys CK homogenization tubes (Labgene, BER0092), brain tissue regions (frontal cortex) were homogenized at 4°C in homogenization-solubilization buffer (HS buffer) at a ratio of 1:4 (w / v). The following sequence was used for homogenization: three cycles of 5000 rpm for 30 seconds each (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-LC).

[0419] • HS buffer - 10mM Tris.HCl pH 7.5, 150mM NaCl, 0.1mM EDTA, 1mM DTT, completely free of EDTA protease inhibitor (Roche, 32524300) and PhosSTOP phosphatase inhibitor (Roche, 4906837001).

[0420] Brain homogenate was thawed on ice and resuspended in HS buffer to obtain a final concentration of 2% Sarkosyl, 1 unit / μL Benzosase, and 1 mM MgCl2. The sample was then incubated on a hot mixer at 37°C with constant shaking at 600 rpm for 45 minutes. The supernatant was collected in a new tube. The precipitate was resuspended in 1000 μL of myelin flotation buffer and centrifuged at 20,000 g for 60 minutes at 4°C. The supernatant was carefully removed to remove all floating lipids. If all lipids could not be removed in a single step, this step was repeated. The precipitate was then washed with PBS and centrifuged at 4°C for 30 minutes. The final precipitate was resuspended in 200 μL of PBS and stored at -80°C. The sample was analyzed by immunoblotting under denaturing conditions.

[0421] • HS buffer containing Sarkosyl, Benzonase and MgCl2 - 10mM Tris.HCl pH 7.5, 150mM NaCl, 0.1mM EDTA, 1mM DTT, 4% Sarkosyl, 1 unit / μL Benzonase (Novagen 70746-4), 4mM MgCl2, completely free of EDTA protease inhibitors (Roche) and PhosSTOP phosphatase inhibitors (Roche).

[0422] • Myelin flotation buffer – HS buffer containing 1% Triton X-100 and 30% sucrose

[0423] Western blotting was performed on a Bolt 12% Bis-Tris Plus gel 1.0 mm (Thermofisher) using MESSDS running buffer (Thermofisher). Once diluted in PBS, samples (30 μl / sample) were loaded onto the gel using loading buffer (1×, Licor, 928-40004) containing 100 mM DTT. Proteins were allowed to degrade at a constant voltage of 100 V for 1 hour. After electrophoresis, proteins were transferred to a nitrocellulose membrane (Thermofisher, IB23001) at 20 volts using an iBLOT (Thermofisher, IB21001) for 7 minutes. Following protein transfer, the membrane was blocked for 1 hour in Licor blocking buffer (Odyssey blocking buffer 927-40000) diluted 1:3 in PBS. Incubate the membrane overnight with the following primary antibodies: total TDP-43 (Proteintech, 60019-2-Ig or 10782-2-AP) or pTDP-43 (Cosmobio, TIP-PTD-M01). For the primary antibody, dilute the blocking buffer 1:1 in PBS-T (PBS containing 0.4% Tween 20). After washing four times with PBS-T (PBS containing 0.1% Tween 20), incubate the membrane with the secondary antibody conjugated to LICOR dye. Use the secondary antibody—donkey anti-mouse (catalog number 926-68072) or goat anti-rabbit (catalog number 926-32211)—at room temperature at a dilution of 1:10000 in Licor blocking buffer diluted 1:1 with PBS-T (PBS containing 0.4% Tween 20) for 1 hour. The membrane was washed four times again with PBS-T (PBS containing 0.1% Tween 20) and scanned using the LICOR system. Figure 2 The results showed that all mAbs specifically recognized full-length TDP-43. Additionally, some mAbs (K, M, N) identified pathological features of disease states, such as C-terminal fragments and high-molecular-weight aggregates in the insoluble fraction.

[0424] Example 8A: Affinity Measurement Using SPR

[0425] The binding affinity to soluble or aggregated FL TDP-43 was evaluated using surface plasmon resonance (SPR; Biacore T200, GE Healthcare Life Sciences) to determine the dissociation constant (KD). Recombinant human soluble or aggregated FL TDP-43 was immobilized on a CM5 Series S sensor chip (GE Healthcare Life Sciences) via 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, resulting in 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, resulting in an immobilization level of 110 RU. Biotinylated TP-73 peptide (amino acids 181 to 190 of SEQ ID NO:1) was immobilized in PBS-P + A concentration of 5 μg / ml was immobilized on a Series S Sensor Chip SA (GE Healthcare Life Sciences) at a flow rate of 5 μl / min for 30 seconds to achieve a fixation level of 400 RU. To evaluate the KD value, purified antibody and control antibody (2E2-D3) were prepared in PBS-P +The antibody was initially diluted 3-fold at 333 nM and then further diluted to 0.15 nM before injection. It was injected at a flow rate of 50 μl / min for a 90-second contact time and a 700-second dissociation phase, followed by three regenerations at 10 mM glycine-HCl pH 1.7. For the optimized SPR protocol, the antibody was 3-fold diluted from 300 nM to 1.2 nM and injected at 30 μl / min for 300 seconds, followed by a 600-second dissociation phase. Surface regeneration was achieved by a single injection of 10 mM glycine-HCl pH 1.7. Results from binding kinetics were evaluated using a double reference of blank flow cell and buffer cycling, and a 1:1 fit model with RI was used. The affinities of 11 antibodies and two Fab fragments are shown in Table 8. The antibodies of this invention bound aggregated TDP-43 at KD values ​​ranging from 0.62 nM to 4.64 nM. Additionally, some antibodies showed preferential binding to aggregated TDP-43 compared to soluble TDP-43. Both Fab fragments bound to soluble TDP-43 with KD ranging from 2.8 nM to 21.8 nM and showed similar KD for aggregated TDP-43. The two antibodies (marked *) were reanalyzed using an optimized SPR protocol (with longer association and dissociation phases) that allows for more accurate KD determination, especially for antibodies with slower dissociation rates. Both antibodies bound to soluble TDP-43 with KD ranging from 0.22 nM to 3.9 nM and to aggregated TDP-43 with KD ranging from 0.18 nM to 0.69 nM. Antibody ACI-7069-642D12-Ab1 bound to the TP-73 peptide with a KD of 3.6 nM.

[0426] Table 8: Binding characterization by SPR

[0427]

[0428]

[0429] NA, Not applicable, because there are fewer than three curves available for fitting.

[0430] *The binding of the recombinant IgG2a isotype antibody was characterized using an optimized SPR protocol.

[0431] Example 8B: Affinity Measurement Using SPR

[0432] The binding affinity to soluble FL TDP-43 was evaluated using surface plasmon resonance (SPR; Biacore T200, GE Healthcare Life Sciences), determining the dissociation constant (KD). Goat anti-mouse capture antibody was immobilized on a CM5 Series S sensor chip (GE Healthcare Life Sciences) via amine conjugation. The antibody was then subjected to PBS-P + Concentrations of 2 to 5 μg / ml in GE Healthcare Life Sciences were captured at a flow rate of 10 μl / min for 120 seconds, resulting in a capture level of 350 to 1000 RU. To evaluate the KD value, FL TDP-43 or TP-51 peptides (amino acids 352 to 414 of SEQ ID NO:1) were in PBS-P + The antibodies were injected 3-fold at a flow rate of 30 μl / min at a rate starting from 1.2 nM and continuing up to 100 nM using a single-cycle kinetic approach, with a contact time of 300 seconds. Dissociation was recorded for 1 hour, followed by a single regeneration with 10 mM glycine-HCl at pH 1.7. Results from binding kinetics were evaluated using a double reference with blank flow cell and buffer cycling, and were assessed using a 1:1 fit model with RI. The binding rate (on-rate) (ka), dissociation rate (off-rate) (kd), and affinity (KD) of the three antibodies are shown in Table 9 as mean ± SD of 12 replicates (ACI-7069-633B12-Ab1), 2 replicates (ACI-7069-642D12-Ab1), or 3 replicates (ACI-7071-809F12-Ab1). Antibodies ACI-7069-633B12-Ab1, ACI-7069-642D12-Ab1, and ACI-7071-809F12-Ab1 bound to soluble TDP-43 with affinities of 15–135 pM, 226–272 pM, and 389–457 pM, respectively. Antibody ACI-7069-633B12-Ab1 bound to TP-51 peptide with an affinity of 1184–1316 pM.

[0433] Table 9: Affinity of SPR to soluble FL TDP-43 and TP-51 peptides

[0434]

[0435] Example 9: Antibody Sequencing

[0436] Clonal hybridoma cell lysates were used for gene sequencing of variable regions. Mouse hybridomas were harvested and lysed using a lysis buffer containing guanidine salts to inactivate RNases. Genomic DNA was then eliminated with RNase-free DNase, and RNA was purified using a silica-based affinity column with multiple washes, eluting it from the column with RNase-free water. Once RNA was extracted, its purity and concentration were measured spectrophotometrically. RNA integrity was assessed on a denaturing agarose gel, and RNA was reverse transcribed into cDNA using reverse transcriptase (RT). RNA was heated to 70°C for 10 minutes to disrupt its secondary structure before adding the RT reaction mixture. The RT products were used directly for PCR amplification. For high-fidelity PCR amplification of cDNA, each of the variable region primers corresponding to different gene families encoding antibodies was individually mixed with constant primers for VH and VL in a total reaction volume of 50 μl. Initially, a degenerate primer library (VH = 12 and VL = 12) was used, and a second library was used based on the results to obtain PCR products. Following the PCR reaction, the products were analyzed by gel electrophoresis on 2% agarose gels stained with ethidium bromide. PCR products of VL and VH were purified individually on agarose gels using tris acetate EDTA (TAE). The purified fragments excised from the gel were sequenced using dye-terminated sequencing with the same primers used for PCR. Sequencing was performed in two directions to provide overlap at both ends. Sequences were analyzed using multiple sequence alignment (Clustal tool) and annotated using the Kabat algorithm, as 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.

[0437] Table 10: Nucleotide sequences of variable domains (VH and VL) in the heavy and light chains

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444] Table 11: Amino acid sequences of heavy and light chain variable domains (VH and VL) and their CDRs

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452] Example 10: In vivo potency of ACI-7069-633B12-Ab1 (IgG2a variant) in a transgenic mouse model of TDP-43 proteinopathy

[0453] To evaluate the in vivo potency of ACI-7069-633B12-Ab1 (IgG2a variant), the ability of ACI-7069-633B12-Ab1 (IgG2a variant) to reduce TDP-43 pathological status in NEFH-tTAx hTDP-43ΔNLS double transgenic mice (rNLS8 mice, Walker et al. 2015) was tested. rNLS8 mice were injected weekly with either ACI-7069-633B12-Ab1 (IgG2a variant) (n=30) or the carrier (n=30), and molecular pathological markers, such as phosphorylated TDP-43 and / or total insoluble TDP-43, were analyzed at the end of administration.

[0454] 10.1 Animals

[0455] Before the start of the study, all animals were acclimatized, examined, handled, and weighed to ensure adequate health and minimize non-specific stress associated with experimental procedures. Mice were fed a diet containing doxycycline (200 mg / kg) during rearing and until 8 weeks of age. At 8 weeks of age, the diet was changed to a doxycycline-free (DOX) diet to allow for transgene expression. Throughout the study, light / dark cycles (12 / 12), room temperature (20°C to 23°C), and relative humidity (approximately 50%) were kept constant. Food and water were provided freely during the study. When mice began to exhibit difficulty moving, the diet was changed to wet food and hydrogel on the cage floor. All behavioral tests were conducted during the animals' light cycle phase.

[0456] 10.2. Compound application

[0457] On the day of injection, ACI-7069-633B12-Ab1 (IgG2a variant) (60 mg / kg) and the loading agent were freshly prepared and administered via ip weekly throughout the study.

[0458] 10.3. Collection of the brain

[0459] The brain was divided into two hemispheres. The left hemisphere was dissected to collect the cortical brain region. The mouse cortex and remaining brain tissue were rapidly frozen for further biochemical analysis. The remaining right hemisphere was perfused at room temperature for 3 hours, then directly immersed for fixation and collected in freshly prepared 1×PBS containing 4% paraformaldehyde (PFA).

[0460] 10.4. Immunohistochemistry

[0461] The submerged and fixed right hemisphere was sagittally sectioned at a thickness of 10 micrometers using a homogeneous, systematic randomization protocol on a Leica CM1950 cryostat. A systematically randomized sagittal section set from each mouse (seven sections from brain levels 2, 3, 4, 6, 8, 10, and 11) was immunostained against 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 fluorescently labeled secondary antibodies. Standard negative controls included wild-type brain sections and sections from transgenic animals that had not received primary antibodies.

[0462] 10.5. Imaging and determination of immunoreactivity

[0463] Mounted slides were imaged holistically on an Axio.Scan Z1 slide scanner driven by ZEN software at 10× magnification using LED (Colibri2) illumination and a sensitive Orca Flash 4.0 monochrome camera. Brain size was determined using separate delineations of the target regions in the cerebral cortex and dorsal striatum. Object density (OD) (in mm²) was also determined. 2 The number of objects is determined for the following: all markers, percentage of marked area, and OD relative to the size of the target area depicted in the second drawing (excluding any tissue artifacts such as tissue folds).

[0464] 10.6. Preparation of protein samples from the cerebral cortex:

[0465] Tissues were thawed on ice and then sonicated in 5X v / w radioimmunoprecipitation assay buffer (RIPA, 50mM Tris, 150mM NaCl, 1% IGEPAL CA630, 5mM EDTA, 0.5% sodium deoxycholate, and 0.1% SDS, pH 8.0) containing a mixture of 1mM PMSF and protease- / phosphatase inhibitors (Roche Applied Science). Samples were centrifuged at 100,000g for 30 min at 4°C, and the supernatant was considered the soluble fraction. The precipitate was washed with RIPA sonication, and the supernatant was discarded. The RIPA-insoluble precipitate was sonicated in 2X v / w urea buffer (7M urea, 2M thiourea, 4% CHAPS, and 30mM Tris, pH 8.5) and centrifuged at 100,000g for 30 min at 22°C. This supernatant was considered the RIPA-insoluble / urea-soluble fraction. The protein concentration of the RIPA soluble fraction was determined using the BCA protein assay (Pierce).

[0466] 10.7. Quantification of insoluble TDP-43

[0467] The total TDP-43 level in the RIPA insoluble fraction was analyzed using a commercial human TDP-43 AlphaLISA kit (PerkinElmer, AL387HV).

[0468] 10.8. Statistical Analysis

[0469] IHC and AlphaLISA data are expressed as mean ± SEM. Statistical differences between animals treated with the loading agent and those treated with ACI-7069-633B12-Ab1 (IgG2a variant) were analyzed by Welch t-test and indicated by an asterisk above the corresponding bar (*p<0.05, **p<0.01, ****p<0.0001). Outliers in histological measurements were excluded as Grubbs outliers (single measure) at the group or level, or due to technical reasons (image artifacts, tissue folds, etc.).

[0470] 10.9. Results

[0471] Treatment with ACI-7069-633B12-Ab1 (IgG2a variant) reduced phosphorylated TDP-43 and insoluble TDP-43 in rNLS8 mice.

[0472] In the rNLS8 mouse model, overexpression of the DOX-repressible form of K82A / R83A / K84A mutant human TDP-43 (hTDP-43ΔNLS) led to significant accumulation and aggregation of TDP-43 in the neuronal cytoplasm. The pathological marker of this model was the deposition of insoluble and phosphorylated TDP-43 inclusion bodies (pTDP-43). These small, spherical cytoplasmic inclusion bodies were present only in the transgenic animals and were completely absent in WT or single-gene transgenic tTA control mice. Furthermore, pTDP-43 was generally absent during the first week of DOX absence, but accumulated rapidly during weeks 3–4, after DOX removal (Walker et al., 2015). Compared with mice treated with the loading agent, treatment with ACI-7069-633B12-Ab1 (IgG2a variant) resulted in a statistically significant reduction in the density of phosphorylated TDP-43 in both the striatum and cerebral cortex. Figure 3 (A to B) indicates its functional efficacy in reducing TDP-43 pathological conditions. The striatum and cerebral cortex were chosen for quantification due to the high expression of the transgene in these regions.

[0473] 10.10. Treatment with ACI-7069-633B12-Ab1 (IgG2a variant) reduced insoluble TDP-43 in rNLS8 mice.

[0474] To determine the pathological reduction in TDP-43 observed in immunohistochemical readouts, the amount of total insoluble / aggregated TDP-43 in the brain was quantified after biochemical grading. The RIPA insoluble fraction was prepared from the cortex of the left hemisphere containing insoluble / aggregated TDP-43. A significantly reduced amount of insoluble TDP-43 was observed in mice treated with ACI-7069-633B12-Ab1 (IgG2a variant) compared to animals treated with the carrier. Figure 3 C). This reduction in molecular TDP-43 pathological status is consistent with findings observed by immunohistochemistry, confirming the efficacy of treatment with ACI-7069-633B12-Ab1 (IgG2a variant). To our knowledge, this is the first time that administration of a peripheral antibody has improved the formation of TDP-43 pathological status in an in vivo model of TDP-43 proteopathy.

[0475] Treatment with ACI-7069-633B12-Ab1 (IgG2a variant) in 10.11.rNLS8 mice increased the surface area of ​​microglial immune responses.

[0476] Functional recovery in rNLS8 mice following suppression of transgene expression involved increased microglial activity. Microglial body area increased at this stage, leading to clearance of TDP-43 pathology and functional recovery of motor deficits, demonstrating a therapeutic paradigm in the rNLS8 mouse model (Spiller KJ et al., Nature Neuroscience, 2018).

[0477] To evaluate the mechanism of action of ACI-7069-633B12-Ab1 in reducing TDP-43 pathological status in rNLS8 mice, its effect on microglial activation was assessed. Iba1 staining was performed by immunohistochemistry to quantify the number and status of microglia in the mouse cerebral cortex. Microglial proliferation was observed in rNLS8 mice at the terminal stage (5 weeks from Dox). Compared with the carrier-treated control, ACI-7069-633B12-Ab1 treatment significantly increased the area of ​​Iba1-positive immunoreactivity in the cortex (…). Figure 5 A). This increase may be due to an increase in the number of microglia or changes in their morphology. Therefore, the density of Iba1-positive cells in the cortex was first evaluated. Compared with the carrier-treated control, ACI-7069-633B12-Ab1 treatment did not affect microglia density, representing cell number.

[0478] Next, the effect of ACI-7069-633B12-Ab1 on microglia morphology was evaluated. To correlate the increase in Iba1 immunoreactivity area with changes in the activation state of microglia, which is representative of morphology, microglia were classified into three states based on their size and morphology (mastoid, small-branched, and branched-resting). Compared with the carrier-treated control, treatment with ACI-7069-633B12-Ab1 (IgG2a variant) showed a significant increase in the mean cell size of mastoid microglia. Figure 5 B). No significant differences were found in the other two types of microglia, which represent a lower activation state. Figure 5 (C to D). This analysis indicates that the increased total Iba1-positive immunoreactivity area observed in the ACI-7069-633B12-Ab1 treatment cohort was generated by morphological changes reflecting increased microglial size and activation status. This suggests that ACI-7069-633B12-Ab1 (IgG2a variant) at least partially reduces TDP-43 pathology in this animal model by recruiting and activating microglia.

[0479] Example 11: In vitro function of ACI-7069-633B12-Ab1 (IgG2a variant) in recombinant TDP-43 aggregation assay

[0480] To evaluate the in vitro function of ACI-7069-633B12-Ab1 (IgG2a variant), its ability to inhibit TDP-43 aggregation was tested. FL TDP-43 is fused at its C-terminus to a maltose-binding protein (MBP) generated by recombination after separation from the cleavage site of Tobacco Etch Virus (TEV) protease. Aggregation of the 2.5 μM TDP-43-TEV-MBP fusion protein was induced in 30 mM Tris, 150 mM NaCl, pH 7.4 in the presence of 2.5 μM MACCI-7069-633B12-Ab1 (IgG2a variant) or an isotype control not binding to TDP-43. Absorbance was monitored at 600 nm in a μclear 96-well plate (Greiner) over 30 h. For evaluation, the endpoint was normalized relative to the isotype control, and the percentage of aggregated TDP-43 was calculated against ACI-7069-633B12-Ab1. Compared with the isotype control, antibody ACI-7069-633B12-Ab1 significantly inhibited TDP-43 aggregation, inhibiting 98% ( Figure 4 ).

[0481] Example 12: Detection and quantification of TDP-43 in biofluids using ACI-7069-633B12-Ab1 (IgG2a variant) and ACI-7071-809F12-Ab1 (IgG2a variant)

[0482] Methods: A PerkinElmer bead-based AlphaLISA immunoassay was established using ACI-7069-633B12-Ab1 (IgG2a variant) and ACI-7071-809F12-Ab1 (IgG2a variant). For CSF samples, dilution linearity was established in the incorporation recovery assay. The concentration of TDP-43 was then measured in diluted CSF samples. (The last sentence appears to be incomplete and possibly refers to a white optoplate.) TM Samples were prepared in a 384 microplate and the emission at 615 nm was measured as the original AlphaLISA count.

[0483] Results: In this immunoassay, total TDP-43 was quantified in cerebrospinal fluid (CSF) samples from healthy controls and patients with FTLD-TDP (semantic dementia, C9orf72, or GRN). Figure 6 In three independent trials, relative TDP-43 quantifications of multiple patient CSF samples from patients with GRN-mutated FTLD-TDP showed significantly higher TDP-43 levels compared to healthy controls. Figure 6 In three independent trials, relative TDP-43 quantifications of multiple patient CSF samples from FTLD-TDP patients with C9orf72 mutations and semantic dementia also showed higher TDP-43 levels compared to healthy controls. Figure 6 ).

[0484] Example 13: Binding with pathological TDP-43 in FTD brain extract as assessed by immune depletion

[0485] To evaluate the efficacy of the antibody in specifically binding to native TDP-43 aggregates, an immunodepletion assay was performed in brain extracts rich in pathological TDP-43.

[0486] Method: Insoluble fractions from post-mortem brain of FTD type A (FTD-A) were prepared as described in Example 7. Dynabeads was used. TMImmunodepletion was performed using magnetic beads and protein G (Thermoscientific 10003D). After resuspending in tubes, 130 μl of beads were transferred to 1.5 ml low-binding tubes. The beads were washed twice with PBS supplemented with 0.05% Tween 20 using a magnet to remove the supernatant. The beads were aliquoted into three separate low-binding tubes. Antibodies (ACI-7069-633B12-Ab1 (IgG2a isotype), ACI-7069-642D12-Ab1 (IgG2a isotype), and mouse IgG2a control) were diluted to 100 μg / ml, and 100 μl was added to each tube after removing the supernatant (using a magnet). The antibody-bead mixture was incubated at room temperature for 1 hour. The bead-antibody complex was washed once with 500 μl PBS-0.05% Tween 20, then washed once with PBS, and resuspended in 250 μl PBS. The antibody-beads were aliquoted into two new tubes (120 μl each). The insoluble fraction was thawed on ice and sonicated on ice at an amplitude of 30 for 30 seconds. After removing the supernatant, 30 μg of brain material was added to each antibody-bead tube and incubated overnight at 4°C with continuous 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 blotting. Western blotting was performed as described in Example 7. Each lane was loaded with 20 μl of sample. The following antibodies were used for immunoblotting: total TDP-43 (ACI-7069-633B12-Ab1 conjugated with DyLight680) and pTDP-43 (Biolegend, 829901) used at dilutions of 1:2000 and 1:1000, respectively. Use goat anti-rat secondary antibody (catalog number 925-32219) at a dilution of 1:10000.

[0487] Results: Compared with the isotype control antibody, ACI-7069-633B12-Ab1 and ACI-7069-642D12-Ab1 were able to specifically bind to and consume TDP-43 and pTDP-43 (TDP-43) from the sarkosyl insoluble fraction obtained from FTD type A brain tissue. Figure 7 This data determined the binding characteristics of these antibodies to targets in human patients.

[0488] This application also relates to the following implementation schemes:

[0489] 1. TDP-43 binding molecules, which bind misfolded aggregated TDP-43 and non-aggregated physiological TDP-43.

[0490] 2. The TDP-43 binding molecule described in Implementation Scheme 1, which binds misfolded aggregated human TDP-43 and non-aggregated physiological human TDP-43.

[0491] 3. The TDP-43 binding molecule according to embodiment 1 or 2, exhibiting one or more, up to all, of the following characteristics:

[0492] a) Inhibit the aggregation of TDP-43 protein or its fragments;

[0493] b) Block the intercellular transmission of TDP-43;

[0494] c) Depolymerizes TDP-43 aggregates; and

[0495] d) Block TDP-43 seeding.

[0496] 4. The TDP-43 binding molecule in any of the foregoing embodiments, which alleviates the TDP-43 pathological condition in vivo.

[0497] 5. The TDP-43 binding molecule in any of the foregoing embodiments, which reduces the level of accumulated TDP-43 and / or phosphorylated TDP-43 in vivo.

[0498] 6. The TDP-43 binding molecule according to any one of the preceding embodiments, which binds to an epitope in amino acid residues at positions 181 to 195, 199 to 213, 307 to 321, 352 to 366, 389 to 411, 397 to 411, or 140 to 200 of human TDP-43 (SEQ ID NO:1) or to an equivalent epitope in non-human TDP-43.

[0499] 7. The TDP-43 binding molecule according to any one of the foregoing embodiments, which binds to an epitope in amino acid residues at positions 183 to 188, 203 to 213, 204 to 208, 204 to 211, 205 to 210, 316 to 323, 358 to 361, 400 to 405, 400 to 406, or 400 to 412 of human TDP-43 or to an equivalent epitope in non-human TDP-43.

[0500] 8. The TDP-43 binding molecule according to any of the preceding embodiments, which binds to an epitope in amino acid residues at positions 400 to 405, 400 to 406, or 400 to 412 of human TDP-43 (SEQ ID NO:1) or to an equivalent epitope in non-human TDP-43.

[0501] 9. The TDP-43 binding molecule in any of the foregoing embodiments is an antibody or its antigen-binding fragment.

[0502] 10. The TDP-43 binding molecule according to any one of the foregoing embodiments, comprising:

[0503] 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, and 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; or

[0504] 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, and 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; or

[0505] 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, and 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; or

[0506] 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, and 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; or

[0507] 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, and 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; or

[0508] 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, and VH-CDR3 containing the amino acid sequence of SEQ ID NO: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; or

[0509] 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, and 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; or

[0510] 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, and 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; or

[0511] 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, and 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; or

[0512] 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, and 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

[0513] 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, 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.

[0514] 11. The TDP-43 binding molecule according to any one of the foregoing embodiments, comprising:

[0515] a. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:10 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:14; or

[0516] b. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:20 or a heavy chain variable region (VH) having at least 98% or 99% sequence identity with 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 a light chain variable region (VL) having at least 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:24; or

[0517] c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:30 or a heavy chain variable region (VH) having at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 98% or 99% sequence identity with the amino acid sequence of SEQ ID NO:34; or

[0518] d. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:40, and a light chain variable region (VL) containing the sequence of SEQ ID NO:44; or

[0519] e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:60 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:60, and a light chain variable region (VL) comprising the sequence of SEQ ID NO:64 or a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:64; or

[0520] f. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:70 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:74; or

[0521] g. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:80 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:84; or

[0522] h. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:100 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:104; or

[0523] i. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:120 or a heavy chain variable region (VH) having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:124; or

[0524] j. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:140 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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

[0525] k. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 or a heavy chain variable region (VH) having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:154.

[0526] 12. The TDP-43 binding molecule according to any one of the foregoing embodiments, comprising:

[0527] a. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:10 and the light chain variable region (VL) containing the sequence of SEQ ID NO:14; or

[0528] b. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:20 and the light chain variable region (VL) containing the sequence of SEQ ID NO:24; or

[0529] c. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:30 and the light chain variable region (VL) containing the sequence of SEQ ID NO:34; or

[0530] d. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 and the light chain variable region (VL) containing the sequence of SEQ ID NO:44; or

[0531] e. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 60 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 64; or

[0532] f. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:70 and the light chain variable region (VL) containing the sequence of SEQ ID NO:74; or

[0533] g. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:80 and the light chain variable region (VL) containing the sequence of SEQ ID NO:84; or

[0534] h. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 100 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 104; or

[0535] i. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:120 and the light chain variable region (VL) containing the sequence of SEQ ID NO:124; or

[0536] j. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:140 and the light chain variable region (VL) containing the sequence of SEQ ID NO:144; or

[0537] k. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 and the light chain variable region (VL) containing the sequence of SEQ ID NO:154.

[0538] 13. The TDP-43 binding molecule according to any one of the foregoing embodiments, comprising: 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); 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.

[0539] 14. The TDP-43 binding molecule according to any one of the foregoing embodiments, comprising: 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.

[0540] 15. The TDP-43 binding molecule according to any one of embodiments 1 to 12, comprising: 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; 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.

[0541] 16. The TDP-43 binding molecule according to any one of embodiments 1 to 12 or 15, comprising: 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.

[0542] 17. The TDP-43 binding molecule in any of the foregoing embodiments is a monoclonal antibody or its antigen-binding fragment.

[0543] 18. The TDP-43 binding molecule in any one of the preceding embodiments is a mouse antibody, a chimeric antibody, a humanized antibody or a human antibody, or an antigen-binding fragment thereof.

[0544] 19. The TDP-43 binding molecule in any of the foregoing embodiments is an antibody or antigen-binding fragment of IgA, IgD, IgE, IgM, IgG1, IgG2, IgG2a, IgG2b, IgG3 or IgG4.

[0545] 20. The TDP-43 binding molecule according to any one of the foregoing embodiments, for use in human or veterinary treatment and / or diagnosis.

[0546] 21. A TDP-43 binding molecule according to any of the foregoing embodiments for human or veterinary treatment and / or diagnosis, wherein the TDP-43 binding molecule is a diagnostic or therapeutic tool.

[0547] 22. The TDP-43 binding molecule according to any one of embodiments 1 to 19, used for research purposes, particularly as an analytical tool or reference molecule.

[0548] 23. The TDP-43 binding molecule according to any one of the foregoing embodiments, used for the prevention, mitigation, treatment and / or diagnosis of diseases, disorders and / or abnormalities associated with TDP-43.

[0549] 24. The TDP-43 binding molecule according to any one of the foregoing embodiments, used for the prevention, mitigation, treatment and / or diagnosis of TDP-43 protein disorders.

[0550] 25. The TDP-43 binding molecule described in Implementation Scheme 21, which is used as a diagnostic tool for monitoring TDP-43 protein disorders.

[0551] 26. A TDP-43 binding molecule used according to any one of embodiments 23 to 25, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valine-containing protein (VCP) mutation, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulopathy, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), nonfluent variant primary progressive aphasia (nfvPPA), etc.), amyotrophic lateral sclerosis (ALS ... MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, Sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation, Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuolar lesions, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

[0552] 27. The TDP-43 binding molecule used according to embodiment 26, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is: frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), or limbic dominant age-related TDP-43 encephalopathy (LATE).

[0553] 28. The TDP-43 binding molecule used according to embodiment 26 or 27, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is amyotrophic lateral sclerosis (ALS).

[0554] 29. The TDP-43 binding molecule used according to embodiment 26 or 27, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is Alzheimer's disease (AD).

[0555] 30. The TDP-43 binding molecule used according to embodiment 26 or 27, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is frontotemporal dementia (FTD).

[0556] 31. A pharmaceutical composition comprising the TDP-43 binding molecule as described in any of the preceding embodiments, and a pharmaceutically acceptable carrier and / or excipient.

[0557] 32. A nucleic acid molecule encoding the TDP-43 binding molecule described in any of the preceding embodiments.

[0558] 33. Nucleic acid molecules, which contain the following nucleotide sequences:

[0559] a. The heavy chain variable region (VH) coding sequence of SEQ ID NO:18 and the light chain variable region (VL) coding sequence of SEQ ID NO:19; or

[0560] b. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:28 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:29; or

[0561] c. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:38 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:39; or

[0562] d. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:48 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:49; or

[0563] e. The heavy chain variable region (VH) coding sequence of SEQ ID NO:68 and the light chain variable region (VL) coding sequence of SEQ ID NO:69; or

[0564] f. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:78 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:79; or

[0565] g. The heavy chain variable region (VH) coding sequence of SEQ ID NO:88 and the light chain variable region (VL) coding sequence of SEQ ID NO:89; or

[0566] h. The heavy chain variable region (VH) coding sequence of SEQ ID NO:108 and the light chain variable region (VL) coding sequence of SEQ ID NO:109; or

[0567] i. The heavy chain variable region (VH) coding sequence of SEQ ID NO:128 and the light chain variable region (VL) coding sequence of SEQ ID NO:129; or

[0568] j. The heavy chain variable region (VH) coding sequence of SEQ ID NO:148 and the light chain variable region (VL) coding sequence of SEQ ID NO:149; or

[0569] k. The heavy chain variable region (VH) coding sequence of SEQ ID NO:158 and the light chain variable region (VL) coding sequence of SEQ ID NO:159.

[0570] 34. A recombinant vector comprising the nucleic acid described in implementation scheme 32 or 33.

[0571] 35. A host cell comprising the nucleic acid described in embodiment 32 or 33 and / or the vector described in embodiment 34.

[0572] 36. A host cell that expresses the TDP-43 binding molecule according to any one of embodiments 1 to 30.

[0573] 37. An expression vector comprising the nucleic acid molecule described in embodiment 32 or 33.

[0574] 38. A cell-free expression system comprising the expression vector described in embodiment 37.

[0575] 39. A method for generating TDP-43 binding molecules, particularly antibodies or antigen-binding fragments thereof, comprising the following steps:

[0576] a) Culturing the host cells of embodiment 35 or 36 or the cell-free expression system of embodiment 38 under conditions suitable for producing the binding molecules, particularly the antibodies or their antigen-binding fragments; and

[0577] b) Isolate the binding molecules, particularly the antibody or its antigen-binding fragment.

[0578] 40. A method for quantifying TDP-43 in a sample obtained from an object, the method comprising: contacting the sample with a TDP-43 binding molecule according to any one of embodiments 1 to 30, and comparing the TDP-43 level in the sample with the TDP-43 level in a control sample.

[0579] 41. A method for diagnosing a disease, disorder, and / or abnormality associated with TDP-43, or a TDP-43 protein disorder, comprising the method described in embodiment 40, wherein a higher level of TDP-43 in the sample compared to a control level based on a healthy subject indicates a disease, disorder, and / or abnormality associated with TDP-43, or a TDP-43 protein disorder.

[0580] 42. A method for diagnosing a disease, disorder and / or abnormality associated with TDP-43, or a TDP-43 protein disorder, comprising the method described in embodiment 40 or 41, wherein similar or higher TDP-43 levels in the sample compared to a diseased control indicate a disease, disorder and / or abnormality associated with TDP-43, or a TDP-43 protein disorder.

[0581] 43. A method for classifying diseases, disorders, and / or abnormalities associated with TDP-43 or for classifying TDP-43 protein disorders, comprising:

[0582] a. Perform the methods described in implementation schemes 41 and / or 42;

[0583] b. Optionally identify mutations in the sample, including but not limited to mutations in granolar protein precursor (GRN), C9orf72, TARDBP, valproic acid protein (VCP), angina pectoris (ANG), myoconstrictor protein (MYOT) gene, or genes encoding desmin (DES); and

[0584] c. Classify the diseases, disorders and / or abnormalities, or TDP-43 protein disorders associated with TDP-43.

[0585] 44. A method for classifying diseases, disorders, and / or abnormalities associated with TDP-43 or for classifying TDP-43 protein disorders, comprising:

[0586] a. The method of embodiment 42 is performed in samples obtained from subjects suffering from TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders, wherein the comparison with diseased controls is based on multiple control samples from subjects suffering from different types or subtypes of TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders; and

[0587] b. Classify the TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders based on comparison.

[0588] 45. A method for monitoring TDP-43-related diseases, disorders and / or abnormalities or monitoring TDP-43 protein disorders at two or more time points using samples from a subject, the method comprising contacting the sample with a TDP-43 binding molecule according to any one of embodiments 1 to 30, wherein a higher TDP-43 level in a later sample compared to one or more earlier samples indicates the progression of a TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder.

[0589] 46. ​​A method for monitoring TDP-43-related diseases, disorders and / or abnormalities or monitoring TDP-43 proteinopathy at two or more time points using samples from a subject, the method comprising contacting the sample with a TDP-43 binding molecule according to any one of embodiments 1 to 30, wherein a lower TDP-43 level in a later sample compared to one or more earlier samples indicates remission of TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 proteinopathy.

[0590] 47. A method for monitoring the treatment of TDP-43-related diseases, disorders, and / or abnormalities, or the treatment of TDP-43 protein disorders, at two or more time points using samples from subjects treated with a specific treatment, the method comprising contacting the sample with a TDP-43 binding molecule according to any one of embodiments 1 to 30, wherein a lower TDP-43 level in a later sample compared to one or more earlier samples indicates that the TDP-43-related disease, disorder, and / or abnormality, or TDP-43 protein disorder, has been successfully treated.

[0591] 48. The method of any one of embodiments 45 to 47, wherein the first time point is before treatment with the treatment and the second time point is after treatment with the treatment.

[0592] 49. A method for selecting a treatment for a TDP-43-related disease, disorder, and / or abnormality, or for selecting a treatment for a TDP-43 protein disorder, the method comprising contacting samples collected before and after treatment with the treatment with a TDP-43 binding molecule according to any one of embodiments 1 to 30, wherein a lower TDP-43 level in the sample collected after treatment, compared to the sample collected before treatment, indicates that the TDP-43-related disease, disorder, and / or abnormality, or TDP-43 protein disorder, has been successfully treated, and thus the treatment is selected for treatment.

[0593] 50. The method of embodiment 49, wherein the treatment comprises a TDP-43 binding molecule according to any one of embodiments 1 to 30 or a pharmaceutical composition as described in embodiment 31.

[0594] 51. The method of any one of embodiments 40 to 50, wherein the sample includes blood, CSF, ISF or urine samples.

[0595] 52. The method according to any one of embodiments 40 to 51, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valine-containing protein (VCP) mutation, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulopathy, 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). The following are considered contraindications for dementia: TARDBP mutation, angiopoietin (ANG) mutation, Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

[0596] 53. The method according to embodiment 52, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease includes: frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), or limbic dominant age-related TDP-43 encephalopathy (LATE).

[0597] 54. The method according to embodiment 52, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is amyotrophic lateral sclerosis (ALS).

[0598] 55. The method according to embodiment 52, wherein the TDP-43-related disease or disorder and / or abnormality, or TDP-43 protein disease, is Alzheimer's disease (AD).

[0599] 56. The method according to embodiment 52, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is frontotemporal dementia (FTD).

[0600] 57. A kit for diagnosing diseases, disorders and / or abnormalities associated with TDP-43, or TDP-43 protein disorders, or for use in the method of any one of embodiments 40 to 56, comprising a TDP-43 binding molecule according to any one of embodiments 1 to 30.

[0601] References

[0602] Arai et al., TDP-43 is a component of ubiquitin-positive tau-negative inclusions in frontotemporal lobar degeneration and amyotrophic lateralsclerosis, Biochemical and Biophysical Research Communications 351 (2006) 602-611.

[0603] Buratti and Baralle, Nuclear factor TDP-43can affect selected microRNAlevels, FEBS Joumal 277(2010)2268-2281.

[0604] Brettschneider J et al., Spreading of pathology in neurodegenerativediseases: a focus on human studies, Nature Rev. Neuroscience, 2015, 109.

[0605] Brettschneider et al.,Stages of pTDP-43 pathology in amyotrophiclateral sclerosis,Ann Neurol.2013July;74(1):20-38.

[0606] Charlton,Methods in Molecular Biology,Val.248(B.K.C.Lo,ed.,HumanaPress,Totowa,NJ,2003),pp.245-254.

[0607] Chowdhury,Methods Mol.Biol.207:179-196(2008)

[0608] Clynes et al.,Fc receptors are required in passive and activeimmunity to melanoma Proc.Nat'l Acad.sci.USA 95:652-656(1998)

[0609] Cohen et al.,An acetvlation switch controls TDP-43 function andaggregation propensity,Nat Commun.6:5845,2015.

[0610] Cragg,M.S.et al.,Complement-mediated lysis by anti-CD20 mAbcorrelates with segregation into lipid rafts,Blood 101:1045-1052(2003)

[0611] Cragg,M.S.and M.J.Glennie,Antibody specificity controls in vivoeffector mcchanisms of anti-CD20reagents,Blood 103:2738-2743(2004)).

[0612] Cunningham and Wells,High-resolution epitope mapping of hGH-receptorinteractions by alanine-scanning mutagenesis,(1989)Science,244:1081-1085.

[0613] Duncan&Winter,The binding site for C lq on IgG,Nature 322:738-40(1988)

[0614] Edelman,G.M.et al.,The Covalent Structure of an Entire gammaGImmunoglobulin Molecule,Proc.Natl.Acad.USA,63,78-85(1969)

[0615] Feiler et al.,TDP-43 is intercellularly transmitted across axonTerminals,J.Cell Biol.Vol.211 No.4897-911.

[0616] Feneberg et al..Towards a TDP-43-Based Biomarker for ALS and FTLD,Molecular Neurobiology,2018;55(10):7789-7801.

[0617] Gazzano-Santoro et al.,Engineered Antibodies with Increased Activityto Recruit Complement,J.Immunol.Methods 202:163(1996)

[0618] Gerngross,Advances in the production of human therapeutic proteins inyeasts and filamentous fungi,Nat.Biotech.22:1409-1414(2004).

[0619] Gerhardt et al.,Methods for General and Molecular Bacteriology,ASMPress(1994).

[0620] Golemis,Protein-Protein Interactions:A Molecular Cloning Manual,ColdSpring Harbor Laboratory Press(2002).

[0621] Graham et al.,Characteristics of a human cell line transformed byDNAfrom human adenovirus type 5,J.Gen Viral.36:59(1977)

[0622] Guver et al.,Immunoglobulin binding by mouse intestinal epithelialcell receptors,J.Immunol.117:587(1976).

[0623] Harlow and Lane.Antibodies:A Laboratory Manual,Cold Spring HarborLaboratory Press,(1988).

[0624] Harlow and Lane.Using Antibodies:A Laboratory Manual,Cold SpringHarbor Laboratory Press,(1999).

[0625] Hasegawa et al.,Phosphorylated TDP-43 in frontotemporal lobardegeneration and amyotrophic lateral sclerosis,(2008)Annals of Neurology Vol64 No 1,60-70.

[0626] Hasegawa et al.,Prion-like mechanisms and potential therapeutictargets in neurodegenerative disorders,Pharmacol Ther.2017 Apr;172:22-33.

[0627] Hellstrom,I.et al.,Proc.Nat’l Acad.Sci.USA 83:7059-7063(1986).

[0628] Hellstrom,I et al.,Proc.Nat’l Acad.Sci.USA 82:1499-1502(1985).

[0629] Hoogenboom et al..Methods in Molecular Biology 178:1-37(O'Brien etal.,ed.,Human Press,Totowa.NJ,(2001).

[0630] Howard and Bethell.(2000)Basic Methods in Antibody Production andCharacterization,Crc.Pr.Inc.Idusogie et al.,J.Immunol.164:4178-4184(2000).

[0631] Ishii et al.,Formation and spreading of TDP-43 aggregates in culturedneuronal and glial cells demonstrated by time-lapse imaging,PLoS ONE 12(6):e0179375,2017.

[0632] Jones et al.,Replacing the complementarity-determining regions in ahuman antibody with those from a mouse,Nature 321(1986),522-525.

[0633] KA et al.,TDP-43 is a key player in the clinical features associatedwith Alzheimer's disease,Acta Neuropathol.2014;127(6):811-824.

[0634] KA et al.,Staging TDP-43 pathology in Alzheimer’s disease ActaNeuropathol.2014;127(3):441-450.Kam et al.,Proc.Natl.Acad.Sci.USA 102:11600-11605(2005).

[0635] Kanda Y.et al.,Bioteehnol.Bioeng.,94(4):680-688(2006).

[0636] Kabat et al..Sequences of Proteins of Immunological Interest.5thedit.NIH Publication no.91-3242U.S.Department of Health and Human Services(1991).

[0637] Kirn et al.,J.Immunol.24:249(1994).

[0638] Kohler.Nature 256(1975),495.

[0639] Khor,Proceedings of the American Society of Clinical OncologyAbstract(1997),847.

[0640] Lagier-Tourenne et al.,TDP-43 and FUS / TLS:emerging roles in RNAprocessing and neurodegeneration,Human Molecular Genetics,2010,Vol.19,ReviewIssue 1 R46-R64.

[0641] Lagier-Tourenne and Cleveland,Rethinking ALS:the FUS about TDP-43,Cell 136,2009,1001-1004.

[0642] Le Ber,Genetics of frontotemporal lobar degeneration:an up-date anddiagnosis algorithm,Revue Neurologique 169(2013)811-819.

[0643] Lefkovits,Immunology Methods Manual:The Comprehensive Sourcebook ofTechniques;Academic Press(1997).

[0644] LoBuglio,Proceedings of the American Society of Clinical OncologyAbstract(1997),1562.

[0645] Li et al.,Optimization of humanized IgGs in glycoengineered Pichiapastoris,Nat.Biotech.24:210-215(2006).

[0646] Mackenzie and Neumann,Molecular neuropathology of frontotemporaldementia:insights into disease mechanisms from postmortem studies,J.Neurochem(2016)138(Suppl.1),54-70.

[0647] Mather,Establishment and characterization of two distinct mousetesticular epithelial cell lines,Biol.Reprod.23:243-251(1980)

[0648] Mather et al.,Culture of testicular cells in hormone-supplementedserum-free medium,Annals N.Y Aead.Sei.383:44-68(1982)

[0649] McAleese et al.,TDP-43 pathology in Alzheimer's disease,dementia withLewy bodies and ageing,Brain Pathol.2017 Jul;27(4):472-479.

[0650] Morris,Epitope Mapping Protocols,Methods in Molecular Biology vol.66(1996)(Humana Press,Totowa,NJ).

[0651] Nonaka et al.,Prion-like Properties of Pathological TDP-43 Aggregatesfrom Diseased Brains,Cell Reports 4(2013),124-134.

[0652] Neumann et al.,Ubiquitinated TDP-43 in frontotemporal lobardegeneration and amyotrophic lateral sclerosis,Science 314,(2006),130-133.

[0653] Neumann et al..Phosphorylation of S409 / 410 of TDP-43 is a consistentfeature in all sporadic and familial forms of TDP-43 proteinopathies,ActaNeuropathol.(2009)117:137-149.

[0654] Petkova.S.B.et al.,Int'l.Immunol.18(12):1759-1769(2006).

[0655] Plückthun,The Pharmacology of Monoclonal Antibodies,Rosenburg andMoore eds.Springer-Verlag,N.Y.(1994),269-315.

[0656] Porta S.et al.,Patient-derived frontotemporal lobar degenerationbrain extracts induce formation and spreading of TDP-43 pathology in vivoNat.Comm.,2018

[0657] Okazaki et al.,Fucose depletion from human IgG1 oligosaccharideenhanccs binding enthalpy and association rate between IgG1 andFcgammaRIIIa.J.Mol.Biol.336:1239-1249(2004).

[0658] Presta LG.,Antibody engineering,Curr Op Struct Biol 2(1992),593-596.

[0659] Ravetch and Kinet,Annu.Rev.Immunol.9:457-492(1991)

[0660] Reichmann,Reshaping human antibodies for therapy,Nature 332(1998),323-327.

[0661] Remington's Pharmaceutical Sciences 16th edition,Osol,A.Ed.(1980)

[0662] Ripka et al.,Two Chinese hamster ovary glycosylation mutants affectedin the conversion of GDP-mannose to GDP-fucose.Arch.Biochem.Biophys.249:533-545(1986).

[0663] Sambrook et al.,Molecular Cloning:A Laboratory Manual;Cold SpringHarbor Laboratory Press,2nd edition(1989)and 3rd edition(2001).

[0664] Shepherd and Dean(2000),Monoclonal Antibodies:A Practical Approach,Oxford University Press Shields et al.,High Resolution Mapping of the BindingSite on Human IgG1 for FcγRI,FcγRII,FcγRIII,and FcRn and Design of IgG1Variants with Improved Binding to the FcγR,J.Biol.Chem.9(2):6591-6604(2001).

[0665] Spillcr K.J ct al.,Microglia-mcdiatcd rccovcry from ALS-rclcvantmotor ncuron dcgcncration in a mouse model of TDP-43 proteinopathy,NatureNeuroscience,2018

[0666] Ticozzi et al.,Protein Aggregation and Defective RNA Metabolism asMechanisms for Motor Neuron Damage,9(3):285-296 CNS Neurol.Disord.DrugTargets.2010,9(3),285-296.

[0667] Urlaub et al.,Isolation of Chinese hamster cell mutants deficient indihydrofolate reductase activity,Proc.Natl.Acad.cii.USA 77:4216(1980)

[0668] Verhoeyen et al.,Reshaping human antibodies:Grafting an antilysozymeactivity.,Science 239(1988),1534-1536.

[0669] Walker et al..Neuropathologically mixed Alzheimer’s and Lewy bodydisease:burden of pathological protein aggregates differs between clinicalphenotypes,Acta Neuropathol(2015)129:729-748

[0670] Wang et al.,TDP-43:an emerging new player in neurodegenerativediseases Trends in Molecular Medicine Vol.14 No.11,2008,479-485.

[0671] Warraich et al.,TDP-43:a DNA and RNA binding protein with roles inneurodegenerative diseases,The International Journal of Biochemistry&CellBiology 42(2010)1606-1609.Wright et al.,Effect of glycosylation on antibodyfunction.TIBTECH 15:26-32(1997).

[0672] Yamane-Ohnuki et al.,Establishment of FUT8 knockout Chinese hamsterovary cells:An ideal host cell line for producing completely defucosylatedantibodies with enhanced antibody-dependent cellularcytotoxicity.Biotech.Bioeng.87:614(2004).

[0673] Yazaki and Wu,Methods in Molecular Biology,Val.248(B.K.C.Lo,ed.,Humana Press,Totowa,NJ),pp.255-268(2003).

Claims

1. A TDP-43 binding molecule, which is a mouse antibody or its antigen-binding fragment, binds misfolded aggregated TDP-43 and non-aggregated physiological TDP-43, and comprises: a. VH-CDR1 containing the amino acid sequence of SEQ ID NO:151, VH-CDR2 containing the amino acid sequence of SEQ ID NO:152, and 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; or b. VH-CDR1 containing the amino acid sequence of SEQ ID NO:41, VH-CDR2 containing the amino acid sequence of SEQ ID NO:42, and 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; or c. VH-CDR1 containing the amino acid sequence of SEQ ID NO:61, VH-CDR2 containing the amino acid sequence of SEQ ID NO:62, and 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; or d. VH-CDR1 containing the amino acid sequence of SEQ ID NO:71, VH-CDR2 containing the amino acid sequence of SEQ ID NO:72, and VH-CDR3 containing the amino acid sequence of SEQ ID NO: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; or e. VH-CDR1 containing the amino acid sequence of SEQ ID NO:81, VH-CDR2 containing the amino acid sequence of SEQ ID NO:82, and 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; or f. VH-CDR1 containing the amino acid sequence of SEQ ID NO:101, VH-CDR2 containing the amino acid sequence of SEQ ID NO:102, and 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; or g. VH-CDR1 containing the amino acid sequence of SEQ ID NO:121, VH-CDR2 containing the amino acid sequence of SEQ ID NO:122, and 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; or h. VH-CDR1 containing the amino acid sequence of SEQ ID NO:141, VH-CDR2 containing the amino acid sequence of SEQ ID NO:142, and 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 i. 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; Or its chimeric or humanized versions.

2. The TDP-43 binding molecule of claim 1, which binds misfolded aggregated human TDP-43 and non-aggregated physiological human TDP-43.

3. The TDP-43 binding molecule of claim 1, which binds to epitopes of amino acid residues at positions 183 to 188, 203 to 213, 204 to 208, 204 to 211, 205 to 210, 316 to 323, 358 to 361, or 400 to 412 of human TDP-43 (SEQ ID NO:1).

4. The TDP-43-binding molecule of claim 1, comprising: a. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:30 or a heavy chain variable region (VH) having at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 98% or 99% sequence identity with the amino acid sequence of SEQ ID NO:34; or b. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:40 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:40, and a light chain variable region (VL) comprising the sequence of SEQ ID NO:44; or c. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:60 or a heavy chain variable region (VH) having at least 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:60, and a light chain variable region (VL) comprising the sequence of SEQ ID NO:64 or a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:64; or d. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:70 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:70, and a light chain variable region (VL) containing the sequence of SEQ ID NO:74 or a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:74; or e. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:80 or a heavy chain variable region (VH) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:84; or f. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:100 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:104; or g. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:120 or a heavy chain variable region (VH) having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO:124; or h. A heavy chain variable region (VH) containing the sequence of SEQ ID NO:140 or a heavy chain variable region (VH) having at least 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 i. A heavy chain variable region (VH) comprising the sequence of SEQ ID NO:150 or a heavy chain variable region (VH) having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with 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 a light chain variable region (VL) having at least 99% sequence identity with the amino acid sequence of SEQ ID NO:

154.

5. The TDP-43-binding molecule of claim 1, comprising: b. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:30 and the light chain variable region (VL) containing the sequence of SEQ ID NO:34; or c. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:40 and the light chain variable region (VL) containing the sequence of SEQ ID NO:44; or d. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:60 and the light chain variable region (VL) containing the sequence of SEQ ID NO:64; or e. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:70 and the light chain variable region (VL) containing the sequence of SEQ ID NO:74; or f. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 80 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 84; or g. The heavy chain variable region (VH) containing the sequence of SEQ ID NO: 100 and the light chain variable region (VL) containing the sequence of SEQ ID NO: 104; or h. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:120 and the light chain variable region (VL) containing the sequence of SEQ ID NO:124; or i. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:140 and the light chain variable region (VL) containing the sequence of SEQ ID NO:144; or j. The heavy chain variable region (VH) containing the sequence of SEQ ID NO:150 and the light chain variable region (VL) containing the sequence of SEQ ID NO:

154.

6. The TDP-43 binding molecule of claim 1, wherein it is a monoclonal antibody or an antigen-binding fragment thereof.

7. The TDP-43 binding molecule of claim 1, wherein it is an antibody of IgA, IgD, IgE, IgM, IgG1, IgG2, IgG2a, IgG2b, IgG3 or IgG4, or an antigen-binding fragment thereof.

8. The TDP-43 binding molecule of claim 1, used for the prevention, mitigation, treatment, and / or diagnosis of TDP-43-related diseases, disorders, and / or abnormalities, or TDP-43 protein disorders, wherein the TDP-43-related diseases, disorders, and / or abnormalities, or TDP-43 protein disorders are: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulopathy, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), and non-fluent variant primary progressive aphasia (nfvPPA). (etc.), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Mayo's disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine tyrosinase (VCP) mutation; and Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with myoconstrictor protein (MYOT) gene mutation or mutation in the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

9. The TDP-43 binding molecule of claim 8, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is: frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), or limbic dominant age-related TDP-43 encephalopathy (LATE).

10. The TDP-43 binding molecule of claim 9, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is amyotrophic lateral sclerosis (ALS).

11. The TDP-43 binding molecule of claim 9, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is Alzheimer's disease (AD).

12. The TDP-43 binding molecule of claim 9, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is frontotemporal dementia (FTD).

13. A pharmaceutical composition comprising the TDP-43 binding molecule of any one of claims 1 to 7, and a pharmaceutically acceptable carrier and / or excipient.

14. A nucleic acid molecule encoding the TDP-43 binding molecule of claim 1, optionally wherein the nucleic acid molecule comprises a nucleotide sequence as shown below: a. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:38 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:39; or b. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:48 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:49; or c. The heavy chain variable region (VH) coding sequence of SEQ ID NO:68 and the light chain variable region (VL) coding sequence of SEQ ID NO:69; or d. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:78 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:79; or e. The heavy chain variable region (VH) coding sequence of SEQ ID NO:88 and the light chain variable region (VL) coding sequence of SEQ ID NO:89; or f. The coding sequence of the heavy chain variable region (VH) of SEQ ID NO:108 and the coding sequence of the light chain variable region (VL) of SEQ ID NO:109; or g. The heavy chain variable region (VH) coding sequence of SEQ ID NO:128 and the light chain variable region (VL) coding sequence of SEQ ID NO:129; or h. The heavy chain variable region (VH) coding sequence of SEQ ID NO:148 and the light chain variable region (VL) coding sequence of SEQ ID NO:149; or i. The heavy chain variable region (VH) coding sequence of SEQ ID NO:158 and the light chain variable region (VL) coding sequence of SEQ ID NO:

159.

15. An expression vector comprising the nucleic acid molecule of claim 14.

16. A host cell comprising the nucleic acid molecule of claim 14 and / or the expression vector of claim 15.

17. A host cell that expresses the TDP-43 binding molecule according to any one of claims 1 to 7.

18. A cell-free expression system comprising the expression vector of claim 15.

19. A method for generating a TDP-43 binding molecule, said TDP-43 binding molecule being an antibody or an antigen-binding fragment thereof, said method comprising the steps of: a) Culturing the host cells of claim 16 or 17 or the cell-free expression system of claim 18 under conditions suitable for producing the binding molecule; and b) Separate the bound molecules.

20. Use of a reagent for determining the level of TDP-43 in a sample in the preparation of a kit for quantifying TDP-43 in a sample obtained from an object, wherein the kit is formulated to contact the sample with a TDP-43 binding molecule according to any one of claims 1 to 7, and to compare the level of TDP-43 in the sample with the level of TDP-43 in a control sample.

21. Use of reagents for determining TDP-43 levels in a sample in the preparation of a kit for diagnosing TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders, wherein the kit is formulated for use as claimed in claim 20, wherein higher levels of TDP-43 in the sample, compared to control levels in healthy subjects, indicate TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders, wherein the TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders are... / Or abnormal, or TDP-43 protein disorders are: frontotemporal dementia (FTD, such as sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valine-containing protein (VCP) mutation, linked to chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulopathy, 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, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British type Dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

22. Use of reagents for determining TDP-43 levels in a sample in the preparation of a kit for diagnosing TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders, wherein the kit is formulated for use as described in claim 20 or 21, wherein similar or higher TDP-43 levels in the sample compared to a diseased control indicate a TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder, wherein the TDP-43-related disease, disorder and / or abnormality indicates a TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder. Abnormalities, or TDP-43 protein disorders, include: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulation disease, Pick's disease, and semantically variant primary progressive aphasia (SPD). (vPPA), behavioral variant FTD (bvFTD), non-fluent variant primary progressive aphasia (nfvPPA), etc., amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British type Dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

23. Use of reagents for determining TDP-43 levels in samples in the preparation of kits for classifying TDP-43-related diseases, disorders, and / or abnormalities, or for classifying TDP-43 protein disorders, wherein the classification includes: a. To perform the use as described in claim 21 or 22; b. Optionally identify mutations in the sample, including but not limited to mutations in granolar protein precursor (GRN), C9orf72, TARDBP, angiogenesis protein (ANG), valine-containing protein (VCP), myoconstrictor protein (MYOT) gene, or genes encoding desmin (DES); and c. Classify the TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders, wherein the TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders are: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granuloma, 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). Examples include sporadic ALS, with TARDBP mutations, with angiogenesis protein (ANG) mutations, Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutations; and Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

24. Use of reagents for determining TDP-43 levels in a sample in the preparation of a kit for monitoring TDP-43-related diseases, disorders, and / or abnormalities or monitoring TDP-43 protein disorders at two or more time points using samples from a subject, wherein the kit is formulated to contact the sample with a TDP-43 binding molecule according to any one of claims 1 to 7, wherein a higher TDP-43 level in a later sample compared to one or more earlier samples indicates a TDP-43-related disease, disorder, and / or abnormality, or TD. Progression of P-43 protein disorders, wherein the TDP-43-related diseases, disorders and / or abnormalities, or TDP-43 protein disorders are: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilia. Granulosa 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, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, etc. The following are considered as a whole: Dementia gravidarum, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutations; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

25. Use of reagents for determining TDP-43 levels in a sample in the preparation of a kit for monitoring TDP-43-related diseases, disorders, and / or abnormalities or monitoring TDP-43 protein disorders at two or more time points using samples from a subject, wherein the kit is formulated to contact the sample with a TDP-43 binding molecule according to any one of claims 1 to 7, wherein a lower TDP-43 level in a later sample compared to one or more earlier samples indicates a TDP-43-related disease, disorder, and / or abnormality, or TD. P-43 proteinopathy regression, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 proteinopathy is: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, cortical-basal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulomas, etc. Myelopathic encephalopathy, 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, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, etc. The following are considered as a whole: Dementia gravidarum, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutations; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

26. Use of reagents for determining TDP-43 levels in a sample in the preparation of a kit for monitoring treatment or monitoring treatment of TDP-43-related diseases, disorders, and / or abnormalities at two or more time points using samples from subjects treated with a specific treatment, wherein the kit is formulated to contact the sample with a TDP-43 binding molecule according to any one of claims 1 to 7, wherein a lower TDP-43 level in a later sample compared to one or more earlier samples indicates a TDP-43-related disease, disorder, and / or abnormality. The disorder and / or abnormality, or TDP-43 proteinopathy, has been successfully treated, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 proteinopathy is: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valine-containing protein (VCP) mutations, linked to chromosome 9p, cortical-basal degeneration, or frontotemporal degeneration (FTLD) with ubiquitin-positive TDP-43 inclusion bodies. -TDP), arrhythmia granulomatosis, 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, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of A). D) Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Mayo's disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

27. The use according to claim 26, wherein the first time point is before treatment with the treatment and the second time point is after treatment with the treatment.

28. Use of reagents for determining TDP-43 levels in samples in the preparation of kits for the selective treatment of TDP-43-related diseases, disorders, and / or abnormalities, or for the selective treatment of TDP-43 protein disorders, wherein the kits are formulated to contact samples collected before and after treatment with said treatment with a TDP-43 binding molecule according to any one of claims 1 to 7, wherein a lower TDP-43 level in samples collected after treatment, compared to samples collected before treatment, indicates TDP-43-related diseases, disorders, and / or abnormalities. The TDP-43 protein disorder or abnormality has been successfully treated, and therefore the treatment is selected for treatment, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valine-containing protein (VCP) mutation, linked to chromosome 9p, cortical-basal degeneration, or frontotemporal degeneration with ubiquitin-positive TDP-43 inclusion bodies (F... TLD (FTLD-TDP), arrhythmia granulosis, 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, such as sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation), Alexander disease (AxD), borderline dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial) The following are considered as a range of conditions: AD (adolescent form), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutations; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuoles, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

29. The use of claim 28, wherein the treatment comprises the TDP-43 binding molecule according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 13.

30. The use according to any one of claims 20, 21, 24, 25, 26 or 28, wherein the sample comprises blood, CSF, ISF or urine sample.

31. The use according to any one of claims 21, 24, 25, 26 or 28, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is: frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with granulin precursor (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valine-containing protein (VCP) mutation, linked to chromosome 9p, corticobasal degeneration, frontotemporal degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic granulopathy, 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 ... MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation, with MND mutation Sporadic ALS, with TARDBP mutation, with angiogenesis protein (ANG) mutation, Alexander disease (AxD), limbic dominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Ma-Yo disease)), hippocampal sclerotic dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valine-containing protein (VCP) mutation; as well as Paget's bone disease and frontotemporal dementia), oculopharyngeal dystrophy with rimmed vacuolar lesions, myofibril myopathy with mutations in the myoconstrictor protein (MYOT) gene or the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), or Parkinson's disease (PD).

32. The use according to any one of claims 21, 24, 25, 26, or 28, wherein the TDP-43-related disease, disorder, and / or abnormality, or TDP-43 protein disorder comprises: Frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), or limbic-dominant age-related TDP-43 encephalopathy (LATE).

33. The use according to any one of claims 21, 24, 25, 26 or 28, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disease is amyotrophic lateral sclerosis (ALS).

34. The use according to any one of claims 21, 24, 25, 26 or 28, wherein the TDP-43-related disease or disorder and / or abnormality, or TDP-43 protein disease, is Alzheimer's disease (AD).

35. The use according to any one of claims 21, 24, 25, 26 or 28, wherein the TDP-43-related disease, disorder and / or abnormality, or TDP-43 protein disorder is frontotemporal dementia (FTD).

36. A kit for diagnosing diseases, disorders and / or abnormalities associated with TDP-43, or TDP-43 protein disorders, or for use in any of claims 20 to 35, comprising a TDP-43 binding molecule according to any of claims 1 to 7.

Images