Anti-tdp-43 antibodies and uses thereof

Abstract

The present disclosure provides antibodies that specifically bind to human TDP-43 and methods of using these antibodies to treat patients having TDP-43 associated diseases, including amyotrophic lateral sclerosis (ALS).

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 538,595, filed September 15, 2023, the contents of which are hereby incorporated by reference in their entirety.

[0003] sequence list This application contains a sequence list submitted electronically in the form of an XML file named “50887-0006WO1.XML”. The XML file was created on August 27, 2024, and is 106,496 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety. Technical Field

[0004] This disclosure relates to the technical fields of immunology and medicine. Background Technology

[0005] Trans-reactive DNA-binding protein 43 (“TDP-43”) is a nuclear protein primarily involved in RNA splicing, transport, stabilization, and ultimately, regulation of gene expression. TDP-43 is a nucleic acid-binding protein containing two highly conserved nucleic acid recognition motifs and has been shown to form dimers and oligomers. While TDP-43 is universally expressed in all cell types, it has been shown to be highly expressed in neuroepithelial cells, which contain all CNS precursor cells, including neurons and glial cells. Furthermore, TDP-43 has been shown to specifically bind to numerous RNAs in neuronal cells.

[0006] TDP-43 cytoplasmic aggregates (also known as inclusion bodies) are associated with several neurodegenerative diseases, conditions, or disorders. Specifically, TDP-43 cytoplasmic aggregates and / or misfolding of TDP-43 are associated with neurodegenerative diseases including amyotrophic lateral sclerosis (“ALS”), frontotemporal dementia (“FTD” or “FTLD-TDP-43”), limbic-predominant age-related TDP-43 encephalopathy (“LATE”), Alzheimer’s disease, multisystemic proteinopathy, and chronic traumatic encephalopathy.

[0007] ALS is the most common motor neuron disease in adults, ultimately leading to the loss of motor neurons that control voluntary muscle movement. The annual incidence of ALS is approximately 1–2 per 100,000 cases, with a prevalence of approximately 4–6 per 100,000 cases. ALS causes progressive degeneration of both upper and lower motor neurons, typically resulting in death primarily attributable to respiratory failure within three to five years of diagnosis. Typical symptoms include muscle stiffness, muscle twitching, progressive muscle weakness, and muscle atrophy. About half of all people with ALS experience difficulty thinking and / or behavioral symptoms, and approximately 15% develop frontotemporal dementia. Pathologically, abnormal aggregation of the TDP-43 protein is observed in up to 97% of ALS patients (Nguyen, HP et al., ALS Genes in the GenomicEra and their Implications for FTD, Trends in Genetics , 34(6): 404+423(2018)).

[0008] Currently, there is no known cure for ALS. Therefore, there is an unmet need for therapies to improve the health of patients with TDP-43-related diseases (including ALS), including mortality risk and / or quality of life. Summary of the Invention

[0009] This disclosure relates to antibodies and antigen-binding antibody fragments that specifically bind to human TDP-43, compositions comprising such antibodies or antigen-binding antibody fragments, and methods of using these antibodies and antigen-binding antibody fragments to treat TDP-43-related diseases, including amyotrophic lateral sclerosis (ALS).

[0010] Therefore, this document provides isolated monoclonal antibodies that competitively bind to TDP-43 (e.g., human TDP-43) with: antibodies comprising the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; antibodies comprising the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; antibodies comprising the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; antibodies comprising the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; antibodies comprising the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or antibodies comprising the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO: 81.

[0011] In some embodiments, the antibody binds to the same epitope on TDP-43 (e.g., human TDP-43) with: an antibody containing the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; an antibody containing the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; an antibody containing the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; an antibody containing the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; an antibody containing the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or an antibody containing the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO: 81.

[0012] This article also provides antibodies that specifically bind to TDP-43 (e.g., human TDP-43), comprising three light chain CDRs and three heavy chain CDRs of a mouse antibody, characterized by including a heavy chain variable domain of SEQ ID NO: 1 and a light chain variable domain of SEQ ID NO: 24.

[0013] In some implementations, the antibody is a humanized antibody, a chimeric antibody, or a decorative antibody.

[0014] In some implementations, the CDR has a definition selected from the group consisting of Kabat, Chothia, Kabat / Chothia, Composite, AbM, and Contact.

[0015] In some embodiments, the antibody includes a humanized mature heavy chain variable domain, including: heavy chain CDR1 as defined by the Kabat / Chothia Composite, including SEQ ID NO: 49; heavy chain CDR2 as defined by Kabat, including SEQ ID NO: 51; and heavy chain CDR3 as defined by Kabat or Chothia, including SEQ ID NO: 52; and a humanized mature light chain variable domain, including the three Kabat light chain CDRs of SEQ ID NO: 53-55.

[0016] In some embodiments, the humanized mature heavy chain variable domain contains a sequence that is at least 80% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain contains a sequence that is at least 80% identical to any of SEQ ID NO: 27-48. In some embodiments, the humanized mature heavy chain variable domain contains a sequence that is at least 85% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain contains a sequence that is at least 85% identical to any of SEQ ID NO: 27-48. In some embodiments, the humanized mature heavy chain variable domain contains a sequence that is at least 90% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain contains a sequence that is at least 90% identical to any of SEQ ID NO: 27-48. In some implementations, the humanized mature heavy chain variable domain contains a sequence that is at least 95% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain contains a sequence that is at least 95% identical to any of SEQ ID NO: 27-48.

[0017] In some implementations, at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: K19 is occupied by R; S35 is occupied by G; T40 is occupied by A; E42 is occupied by G; A49 is occupied by S; K43 is occupied by E; R44 is occupied by G or D; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; L80 is occupied by A or G; L82c is occupied by G; M83 is occupied by R; S84 is occupied by A; M89 is occupied by V; or F91 is occupied by Y.

[0018] In some implementations, at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: K43 is occupied by E; R44 is occupied by G or D; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; or F91 is occupied by Y.

[0019] In some implementations, at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: S35 is occupied by G; L78 is occupied by A or G; L80 is occupied by A or G; or L82c is occupied by G.

[0020] In some embodiments, F91 of the humanized heavy chain variable domain is occupied by Y; and at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: R44 is occupied by G; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; or M83 is occupied by R.

[0021] In some embodiments, at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: V3 is occupied by Q; L9 is occupied by S; D17 is occupied by Q; Q18 is occupied by P; K39 is occupied by R; K45 is occupied by R; T80 is occupied by A or S; L83 is occupied by V; L92 is occupied by G or A; V94 is occupied by I or A; A100 is occupied by G, D or R; or L104 is occupied by V.

[0022] In some implementations, at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: V3 is occupied by Q or A100 is occupied by D or R.

[0023] In some implementations, at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: L9 is occupied by S; T80 is occupied by A or S; L92 is occupied by G or A; or V94 is occupied by I or A.

[0024] In some implementations, V3 is occupied by Q; Q18 is occupied by P; A100 is occupied by D; and at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: T80 is occupied by A or L92 is occupied by A.

[0025] In some embodiments, at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: L5 is occupied by V; G44 is occupied by R; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; M89 is occupied by V, or F91 is occupied by Y; and at least one of the following positions in the humanized light chain variable domain is occupied by a specified amino acid: V3 is occupied by Q; D17 is occupied by Q; Q18 is occupied by P; K39 is occupied by R; K45 is occupied by R; T80 is occupied by A; L83 is occupied by V; L92 is occupied by A; A100 is occupied by D; or L104 is occupied by V.

[0026] In some implementations, the humanized antibody exhibits improved thermal stability compared to a reference antibody that includes a heavy chain variable domain having SEQ ID NO: 1 and a light chain variable domain having SEQ ID NO: 24.

[0027] In some implementations, the humanized antibody has a melting temperature of 55°C or higher.

[0028] This document also provides antibodies that specifically bind to TDP-43, said antibodies comprising heavy chain variable domains including: heavy chain CDR1 as defined by the Kabat / Chothia Composite, including SEQ ID NO: 49 or SEQ ID NO: 50; heavy chain CDR2 as defined by Kabat, including SEQ ID NO: 51; heavy chain CDR3 as defined by Kabat or Chothia, including SEQ ID NO: 52; light chain CDR1 as defined by Kabat, including SEQ ID NO: 53; light chain CDR2 as defined by Kabat, including SEQ ID NO: 54; and light chain CDR3 as defined by Kabat, including one of SEQ ID NO: 55-61.

[0029] In some implementations, heavy chain CDR1 as defined by the Kabat / Chothia Composite includes SEQ ID NO: 49; heavy chain CDR2 as defined by Kabat includes SEQ ID NO: 51; heavy chain CDR3 as defined by Kabat or Chothia includes SEQ ID NO: 52; light chain CDR1 as defined by Kabat includes SEQ ID NO: 53; light chain CDR2 as defined by Kabat includes SEQ ID NO: 54; and light chain CDR3 as defined by Kabat includes SEQ ID NO: 55 or SEQ ID NO: 61.

[0030] In some embodiments, the heavy chain variable domain comprises a sequence that is at least 95% identical to any of SEQ ID NO: 4-23. In some embodiments, the heavy chain variable domain comprises a sequence that is at least 95% identical to any of SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 23. In some embodiments, the heavy chain variable domain comprises a sequence that is at least 98% identical to any of SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 23. In some embodiments, the heavy chain variable domain comprises the sequence of SEQ ID NO: 20. In some embodiments, the heavy chain variable domain comprises the sequence of SEQ ID NO: 21. In some embodiments, the heavy chain variable domain comprises the sequence of SEQ ID NO: 23.

[0031] In some embodiments, the light chain variable domain comprises a sequence that is at least 95% identical to any of SEQ ID NO: 27-48. In some embodiments, the light chain variable domain comprises a sequence that is at least 95% identical to either SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the light chain variable domain comprises a sequence that is at least 98% identical to either SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the light chain variable domain comprises SEQ ID NO: 47. In some embodiments, the light chain variable domain comprises SEQ ID NO: 48.

[0032] This article also provides antibodies that specifically bind to TDP-43, said antibodies comprising a heavy chain variable domain and a light chain variable domain, including: heavy chain CDR1, including SEQ ID NO: 84; heavy chain CDR2, including SEQ ID NO: 85; heavy chain CDR3, including SEQ ID NO: 86; light chain CDR1, including SEQ ID NO: 87; light chain CDR2, including SEQ ID NO: 88; and light chain CDR3, including SEQ ID NO: 89.

[0033] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 63. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 63. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 63.

[0034] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 65. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 65. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 65.

[0035] This article also provides an antibody that specifically binds to TDP-43, the antibody comprising a heavy chain variable domain and a light chain variable domain, comprising: heavy chain CDR1, including SEQ ID NO: 90; heavy chain CDR2, including SEQ ID NO: 91; heavy chain CDR3, including SEQ ID NO: 92; light chain CDR1, including SEQ ID NO: 93; light chain CDR2, including SEQ ID NO: 94; and light chain CDR3, including SEQ ID NO: 95.

[0036] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 67. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 67. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 67.

[0037] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 69. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 69. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 69.

[0038] This article also provides antibodies that specifically bind to TDP-43, said antibodies comprising a heavy chain variable domain and a light chain variable domain, comprising: heavy chain CDR1, including SEQ ID NO: 96; heavy chain CDR2, including SEQ ID NO: 97; heavy chain CDR3, including SEQ ID NO: 98; light chain CDR1, including SEQ ID NO: 99; light chain CDR2, including SEQ ID NO: 100; and light chain CDR3, including SEQ ID NO: 101.

[0039] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 71. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 71. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 71.

[0040] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 73. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 73. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 73.

[0041] This article also provides antibodies that specifically bind to TDP-43, said antibodies comprising a heavy chain variable domain and a light chain variable domain, comprising: heavy chain CDR1, including SEQ ID NO: 102; heavy chain CDR2, including SEQ ID NO: 103; heavy chain CDR3, including SEQ ID NO: 104; light chain CDR1, including SEQ ID NO: 105; light chain CDR2, including SEQ ID NO: 106; and light chain CDR3, including SEQ ID NO: 107.

[0042] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 75. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 75. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 75.

[0043] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 77. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 77. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 77.

[0044] This article also provides antibodies that specifically bind to TDP-43, said antibodies comprising a heavy chain variable domain and a light chain variable domain, comprising: heavy chain CDR1, including SEQ ID NO: 108; heavy chain CDR2, including SEQ ID NO: 109; heavy chain CDR3, including SEQ ID NO: 110; light chain CDR1, including SEQ ID NO: 111; light chain CDR2, including SEQ ID NO: 112; and light chain CDR3, including SEQ ID NO: 113.

[0045] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 79. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 79. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 79.

[0046] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 81. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 81. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 81.

[0047] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, or a veneered antibody. In some embodiments, the antibody is an antigen-binding antibody fragment. In some embodiments, the antigen-binding antibody fragment is a Fab fragment, a Fab'2 fragment, or a single-chain Fv.

[0048] In some implementations, the antibody is a complete antibody. In some implementations, the antibody has a human IgG1 isotype.

[0049] In some implementations, the variable structural domains of the heavy chain are fused into the constant region of the heavy chain, and the variable structural domains of the light chain are fused into the constant region of the light chain.

[0050] In some embodiments, the heavy chain constant region is a mutant form of the natural human heavy chain constant region, which binds less to the Fcγ receptor compared to the natural heavy chain constant region. In some embodiments, the heavy chain constant region has an IgG1 isotype. In some embodiments, the antibody has at least one mutation in the constant region. In some embodiments, at least one mutation reduces complement fixation or activation in the constant region. In some embodiments, at least one mutation is located at one or more of the following positions according to EU numbers: 241, 264, 265, 270, 296, 297, 318, 320, 322, 329, and 331. In some embodiments, the antibody contains alanine at positions 318, 320, and 322 according to EU numbers.

[0051] In some embodiments, the antibody selectively binds to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43). In some embodiments, the antibody selectively binds to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) compared to unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43). In some embodiments, the antibody binds to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) with at least 100-fold greater affinity compared to unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43). In some embodiments, the antibody binds to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) with at least 1000-fold greater affinity compared to unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43). In some embodiments, phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) comprises phosphorylation of at least one amino acid residue selected from S409 and S410. In some implementations, phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) includes phosphorylation of both S409 and S410.

[0052] In some embodiments, the antibody selectively binds to cytoplasmic aggregates of TDP-43 (e.g., cytoplasmic aggregates of human TDP-43). In some embodiments, the antibody selectively binds to cytoplasmic aggregates of TDP-43 (e.g., cytoplasmic aggregates of human TDP-43) compared to nuclear TDP-43 (e.g., nuclear human TDP-43). In some embodiments, the cytoplasmic aggregates of TDP-43 comprise phosphorylated aggregates of TDP-43 (e.g., phosphorylated aggregates of human TDP-43). In some embodiments, the antibody substantially does not bind to unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43).

[0053] In some implementations, the antibody is conjugated to a therapeutic agent, cytotoxic agent, cell growth inhibitor, immunomodulator, neurotrophic agent, or neuroprotective agent.

[0054] This article also provides pharmaceutical compositions comprising any of the antibodies described herein and a pharmaceutically acceptable carrier.

[0055] This article also provides nucleic acids encoding the heavy chain variable domain and / or light chain variable domain of any of the antibodies described herein.

[0056] This article also provides vectors containing nucleic acids encoding mature heavy chain variable domains and light chain variable domains, said nucleic acids being operatively linked to one or more regulatory sequences to achieve expression of any of the antibodies described herein in mammalian cells.

[0057] In some embodiments, one or more regulatory sequences include one or more of an enhancer, a ribosome binding site, a transcription termination signal, and a promoter, optionally wherein the promoter is a eukaryotic promoter. In some embodiments, the nucleic acid is codon-optimized for expression in host cells.

[0058] This article also provides host cells transformed using any of the vectors described herein.

[0059] This article also provides host cells that include any of the nucleic acids described herein.

[0060] This article also provides a method for inhibiting or reducing the accumulation of TDP-43 (e.g., human TDP-43) in subjects who have or are at risk of developing TDP-43-related diseases, comprising administering an effective amount of any of the antibodies described herein to the subject, thereby inhibiting or reducing the accumulation of TDP-43 (e.g., human TDP-43) in the subject.

[0061] This article also provides a method for treating or achieving prevention of TDP-43-related disease in subjects, which includes administering a therapeutically effective amount of any of the antibodies described herein, thereby treating or achieving prevention of TDP-43-related disease.

[0062] In some implementations, TDP-43-related diseases include amyotrophic lateral sclerosis (ALS), frontotemporal degeneration (FTLD-TDP), primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease. In some implementations, TDP-43-related diseases include ALS.

[0063] This article also provides a method for detecting TDP-43 deposits (e.g., human TDP-43 deposits) in subjects who have TDP-43-related diseases or are at risk of developing such diseases, comprising administering any of the antibodies described herein to the subject and detecting antibodies in the subject that bind to TDP-43.

[0064] In some implementations, antibodies are administered by intravenous injection into the subject's body.

[0065] This article also provides a method for detecting TDP-43 in samples obtained from patients with TDP-43-related diseases or at risk of developing the aforementioned diseases, comprising detecting any of the antibodies described herein that bind to TDP-43 in the sample.

[0066] In some embodiments, the antibody is labeled. In some embodiments, the antibody is labeled with fluorescent, paramagnetic, or radioactive labels. In some embodiments, positron emission tomography (PET) or single-photon emission computed tomography (SPECT) is used to detect the radioactive label.

[0067] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference as if each individual publication, patent, patent application, or information item expressly and individually indicated to be incorporated by reference. To the extent that any incorporated publication, patent, patent application, or information item contradicts the disclosure contained in this specification, this specification is intended to supersede and / or give precedence to any such contradictory material.

[0068] When using ranges to describe values, it should be understood that the description includes disclosing all possible subranges within those ranges, as well as specific values ​​falling within those ranges, regardless of whether a specific value or a specific subrange is explicitly stated. Attached Figure Description

[0069] The following figures illustrate certain embodiments of the features and advantages of this disclosure. These embodiments are not intended to limit the scope of the appended claims in any way. Like reference numerals in the figures indicate like elements.

[0070] Figure 1 The annotation form of the variable domain of the 13D3 heavy chain of the mouse antibody is shown.

[0071] Figure 2 The annotation form of the variable domain of the light chain of the mouse antibody 13D3 is shown.

[0072] Figure 3 A graph showing the binding data of chimeric 13D3 antibody and its humanized form to phosphorylated human TDP-43 peptide.

[0073] Figure 4 A graph showing the binding data of chimeric 13D3 antibody and its humanized form to phosphorylated human TDP-43 peptide.

[0074] Figure 5 A graph showing the binding data of the humanized form of the 13D3 antibody hu13D2Hd5Ld2 with phosphorylated human TDP-43 peptide.

[0075] Figures 6A to 6C The image shows brain tissue stained with 13D3 antibody in frontotemporal dementia (“FTD”). Figure 6A and Figure 6B ) and healthy brain tissue ( Figure 6C Immunohistochemical images of ). Figure 6B for Figure 6A The illustration shows the co-localization of the 13D3 antibody with neuronal cytoplasmic aggregates associated with phosphorylated human TDP-43 FTD.

[0076] Figures 7A to 7C Immunohistochemical images of brain tissue from a TDP-43 proteopathic model stained with 13D3 antibody and inhibited by rNLS8 dox. The data show that 13D3 antibody binding to rNLS8 dox in the TDP-43 proteopathic model inhibits cytoplasmic aggregates.

[0077] Figures 8A to 8C Results are shown for HEK cells transfected with GFP-2a-TDP43 or GFP alone. Figure 8A The staining of pTDP-43, GFP, and the cell nucleus in transfected cells is shown. Figure 8B A graph showing cell counts. Figure 8C Immunohistochemical images of HEK cells are shown, demonstrating the ability of antibodies 13D3, 13C13, and 2D4 to detect mislocalized human TDP-43 overexpressed in HEK cells. Detailed Implementation

[0078] This disclosure provides antibodies and antigen-binding antibody fragments that specifically bind to TDP-43 (e.g., human TDP-43) or aggregates of TDP-43 (e.g., aggregates of human TDP-43). Pharmaceutical compositions are also provided comprising these antibodies or antigen-binding antibody fragments and a pharmaceutically acceptable carrier, a nucleic acid and / or vector encoding these antibodies or antigen-binding antibody fragments, and a host cell expressing the aforementioned nucleic acid and / or vector.

[0079] Therefore, this document provides isolated monoclonal antibodies that competitively bind to TDP-43 (e.g., human TDP-43) with: antibodies comprising the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; antibodies comprising the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; antibodies comprising the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; antibodies comprising the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; antibodies comprising the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or antibodies comprising the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO: 81. In some embodiments, the monoclonal antibody is a humanized antibody. Therefore, this article also provides antibodies comprising a humanized mature heavy chain variable domain containing any of the sequences in SEQ ID NO: 4-23 and a humanized mature light chain variable domain containing any of the sequences in SEQ ID NO: 27-48.

[0080] Furthermore, any of the antibodies, antigen-binding antibody fragments, and compositions described herein can be used to treat TDP-43-related diseases. Therefore, this document provides a method for inhibiting or reducing TDP-43 accumulation in subjects suffering from or at risk of developing TDP-43-related diseases, comprising administering an effective amount of any of the antibodies described herein to the subject, thereby inhibiting or reducing TDP-43 accumulation in the subject. This document also provides a method for treating or achieving prevention of TDP-43-related diseases in subjects, comprising administering a therapeutically effective amount of any of the antibodies described herein, thereby treating or achieving prevention of TDP-43-related diseases.

[0081] I. Definition The term "antibody" includes both intact antibodies and their antigen-binding fragments. Typically, fragments compete with the intact antibody from which they are derived for specific binding to a target, including individual heavy chains, light chains Fab, Fab', F(ab')2, F(ab)c, Dab, nanobodies, and Fv. Fragments can be generated via recombinant DNA technology or through enzymatic or chemical separation of intact immunoglobulins. The term "antibody" also includes bispecific or multispecific antibodies and / or humanized antibodies. Bispecific or bifunctional or multifunctional antibodies are artificial hybrid antibodies having two or more distinct heavy / light chain pairs and two or more distinct binding sites (see, for example, Songsivilai and Lachmann). Clin. Exp. Immunol ., 79:315-321 (1990); Kostelny et al. , J. Immunol ., 148:1547-53 (1992)).

[0082] The term "epitope" refers to the site on an antigen where an antibody binds. Epitopes can be formed from consecutive or non-consecutive amino acids juxtaposed through the ternary folding of one or more proteins. Epitopes formed from consecutive amino acids (also known as linear epitopes) are generally retained upon exposure to denaturing solvents, while epitopes formed through ternary folding (also known as conformational epitopes) are generally lost upon treatment with denaturing solvents. Epitopes typically consist of at least three, and more usually at least five, or eight to ten amino acids, exhibiting a unique spatial conformation. Methods for determining the spatial conformation of epitopes include, for example, X-ray crystallography and two-dimensional nuclear magnetic resonance (NMR). See also, For example, Epitope Mapping Protocols, Methods in Molecular Biology, Vol. 66, edited by Glenn E. Morris (1996).

[0083] Antibodies that recognize the same or overlapping epitopes can be identified in a simple immunoassay that shows the ability of one antibody to compete with another for binding to a target antigen. An antibody epitope can also be defined by X-ray crystallography of the antibody binding to its antigen to identify contact residues. Alternatively, if all amino acid mutations in the antigen that reduce or eliminate the binding of one antibody also reduce or eliminate the binding of another antibody, then the two antibodies have the same epitope. If some amino acid mutations that reduce or eliminate the binding of one antibody also reduce or eliminate the binding of another antibody, then the two antibodies have overlapping epitopes.

[0084] The terms "humanized immunoglobulin" or "humanized antibody" refer to a chain that includes at least one humanized immunoglobulin or antibody chain. Right now At least one humanized light or heavy chain of immunoglobulin or antibody. The terms "humanized immunoglobulin chain" or "humanized antibody chain" (…) Right now"Humanized immunoglobulin light chain" or "humanized immunoglobulin heavy chain" refers to an immunoglobulin or antibody chain with a variable region. Right now The variable region (either a light chain or a heavy chain) comprises a variable framework region substantially derived from human immunoglobulins or antibodies and a complementation-determining region (CDR) substantially derived from non-human immunoglobulins or antibodies (e.g., at least one CDR, preferably two CDRs, more preferably three CDRs), and further comprises a constant region (e.g., at least one constant region or a portion thereof in the case of the light chain, and preferably three constant regions in the case of the heavy chain). The term "humanized variable region" (e.g., "humanized light chain variable region" or "humanized heavy chain variable region") refers to a variable region comprising a variable framework region substantially derived from human immunoglobulins or antibodies and a complementation-determining region (CDR) substantially derived from non-human immunoglobulins or antibodies.

[0085] Competition between antibodies is determined by measuring the specific binding of the test antibody to the reference antibody and the common antigen. (See also, For example Junghans et al., Cancer Res. 50: 1495, 1990). If an excessive amount of test antibody is used... ( For example, at least 2x, 5x, 10x, 20x, or 100x) inhibits the binding of the reference antibody by at least 50%, as measured in a competitive binding assay, where the test antibody competes with the reference antibody. Some test antibodies inhibit the binding of the reference antibody by at least 75%, 90%, or 99%. Antibodies identified by competitive assays (competitive antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to adjacent epitopes that are sufficiently close to the epitope bound by the reference antibody to cause steric hindrance.

[0086] The term "pharmaceutically acceptable" means that the carrier, diluent, excipient, or adjuvant is compatible with the other components of the formulation and is substantially harmless to the recipient.

[0087] The term "TDP-43-related disease" refers to a disease or condition that is at least partly directly or indirectly characterized and / or mediated by the formation of TDP-43 aggregates and / or the mislocalization of TDP-43. This document describes non-limiting examples of TDP-43-related cancers.

[0088] The term "patient" includes people and other mammalian subjects (e.g., humans) who receive preventative or therapeutic treatment.

[0089] If an individual has at least one known risk factor ( For example, genetic, biochemical, family history, and situational exposure factors (such as genetic, biochemical, family history, and situational exposure) give individuals with said risk factors a statistically significantly greater risk of disease than individuals without said risk factors, thus increasing the individual's risk of disease.

[0090] The term "biological sample" refers to a sample of biological material that is within or obtainable from a biological source (such as a human or mammalian individual). Such samples can be organs, organelles, tissues, tissue sections, body fluids, peripheral blood, plasma, serum, cells, molecules (such as proteins and peptides), and any part or combination thereof. The term "biological sample" may also cover any material derived from the processing of the sample. Derived material may include cells or their progeny. Processing of biological samples may involve one or more of the following: filtration, distillation, extraction, concentration, fixation, deactivation of interfering components, etc.

[0091] The term "control sample" refers to a biological sample that is unknown or suspected of including a TDP-43-affected region, or at least unknown or suspected of including a diseased region of a given type. Control samples may be obtained from individuals who do not have TDP-43-related disease. Alternatively, control samples may be obtained from patients who have TDP-43-related disease. Such samples may be obtained simultaneously with or on different occasions from biological samples believed to contain TDP-43-related disease. The biological sample and control sample may be obtained from the same tissue. Preferably, the control sample consists substantially or entirely of normal healthy regions and can be used for comparison with biological samples believed to contain regions affected by TDP-43-related disease. Preferably, the tissue in the control sample is of the same type as the tissue in the biological sample. Preferably, the TDP-43-related disease-affected cells believed to be in the biological sample are derived from the same cell type (e.g., neurons or glial cells) as the cell type in the control sample.

[0092] For the purpose of classifying amino acid substitutions as conserved or non-conserved, amino acids are grouped as follows: Group I (hydrophobic side chains): Met, Ala, Val, Leu, Ile; Group II (neutral hydrophilic side chains): Cys, Ser, Thr; Group III (acidic side chains): Asp, Glu; Group IV (basic side chains): Asn, Gln, His, Lys, Arg; Group V (residues affecting chain orientation): Gly, Pro; and Group VI (aromatic side chains): Trp, Tyr, Phe. Conservative substitution involves substitution between amino acids within the same category. Non-conservative substitution involves exchanging members of one category for members of another.

[0093] The percentage of sequence identity is determined by aligning antibody sequences to the maximum extent possible using the Kabat numbering convention. After alignment, if the antibody region of the present invention is... ( For example ，The sequence identity percentage between the antibody region of the present invention and the reference antibody region is calculated by comparing the entire mature variable region of the heavy chain or light chain with the same region of the reference antibody. The percentage is calculated by dividing the number of positions occupied by the same amino acids in the regions of the present invention and the reference antibody by the total number of alignment positions in the two regions (excluding vacancies) and multiplying by 100 to convert it into a percentage.

[0094] Unless otherwise apparent from the context, the term "about" covers non-substantial changes, such as those within the standard measurement error limit of a specified value. ( For example ， The value within SEM).

[0095] The phrase "substantially derived from human immunoglobulins or antibodies" means that, when compared with the amino sequence of human immunoglobulins or antibodies for comparative purposes, the region shares at least 80-90%, preferably 90-95%, and more preferably 95-99% identity with the human framework or constant region sequence. Right now This allows for, for example, conserved substitutions, shared sequence substitutions, germline substitutions, and reversion mutations. The introduction of conserved substitutions, shared sequence substitutions, germline substitutions, and reversion mutations is often referred to as the "optimization" of humanized antibodies or chains. The phrase "substantially derived from non-human immunoglobulins or antibodies" or "substantially non-human" means having a similar composition to non-human organisms (e.g., [missing information]). ， The immunoglobulin or antibody sequence is at least 80-95%, preferably 90-95%, more preferably 96%, 97%, 98% or 99% identical to that of a non-human mammal.

[0096] Therefore, all regions or residues (possibly excluding CDRs) of a humanized immunoglobulin or antibody, or a humanized immunoglobulin or antibody chain, are substantially identical to the corresponding regions or residues of one or more natural human immunoglobulin sequences. The terms "corresponding region" or "corresponding residue" refer to a region or residue on a second amino acid or nucleotide sequence that occupies the same position as on a first amino acid or nucleotide sequence when the first and second sequences are optimally aligned for comparative purposes. Right now (equivalent) regions or residues.

[0097] II. TDP-43 As described in this article, TDP-43 is a nuclear protein primarily involved in RNA splicing, transport, stabilization, and ultimately, regulation of gene expression. More specifically, TDP-43 is a multi-domain heterogeneous ribonucleoprotein (hnRNP). Proper function of TDP-43 is crucial for regulating the hundreds of mRNA transcripts it binds to. One of TDP-43's primary functions is regulating spliced ​​mRNA transcripts; however, TDP-43 also participates in various mechanisms of RNA processing and transport. For example, TDP-43 is an inhibitor of cryptic exon inclusion bodies and regulates alternative polyadenylation in >1,000 genes. TDP-43 can form ribonucleoprotein particles in various cell types, including Cajal bodies and parafoils in the nucleus, and is recruited to mRNA transport particles in neurons. As described in this article, various neurodegenerative diseases are associated with cytoplasmic aggregates of TDP-43 (e.g., TDP-43-related diseases).

[0098] Unless otherwise obvious from the context, references to TDP-43 refer to the natural human form of TDP-43, including any isoforms and / or post-translational modifications (e.g., phosphorylation, glycosylation, and / or acetylation). The amino acid sequence of human TDP-43 is shown below (SEQ ID NO: 82): MSEYIRVTEDENDEPIEIPSEDDGTVLLSTVTAQFPGACGLRYRNPVSQCMRGVRLVEGILHAPDAGWGNLVYVVNYPKDNKRKMDETDASSAVKVKRAVQKT SDLIVLGLPWKTTEQDLKEYFSTFGEVLMVQVKKDLKTGHSKGFGFVRFTEYETQVKVMSQRHMIDGRWCDCKLPNSKQSQDEPLRSRKVFVGRCTEDMTEDEL REFFSQYGDVMDVFIPKPFRAFAFVTFADDQIAQSLCGEDLIIKGISVHISNAEPKHNSNRQLERSGRFGGNPGGFGNQGGFGNSRGGGAGLGNNQGSNMGGGMNFGAFSINPAMMAAAQAALQSSWGMMGMLASQQNQSGPSGNNQNQGNMQREPNQAFGSGNNSYSGSNSGAAIGWGSASNAGSGSGFNGGFGSSMDSKSSGWGM TDP-43 can be phosphorylated at one or more amino acids, including serines at positions 409 and 410. In some embodiments, TDP-43 can be phosphorylated at one or more positions, including 373, 375, 379, 387, 389, 393, 395, 403, 404, 407, 409, and 410 (see, for example, Gruijs da Silva, LA, et al., Disease-linked TDP-43 hyperphosphorylation suppresses TDP-43 condensation and aggregation). The EMBO Journal , 41: e108443 (2022)).

[0099] Unless otherwise obvious from the context, references to TDP-43 or fragments thereof include natural human amino acid sequences, including isoforms, mutants, and allele variants. The ability of an antibody or antigen-binding antibody fragment to bind to TDP-43 can be determined using, for example, surface plasmon resonance.

[0100] III. Antibodies This disclosure provides antibodies or antigen-binding antibody fragments that specifically bind to TDP-43 (e.g., human TDP-43). In some embodiments, the antibodies or antigen-binding antibody fragments described herein specifically bind to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43). In some embodiments, the antibodies or antigen-binding antibody fragments described herein specifically bind to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) wherein one or both of the serine residues at positions 409 and / or 410 of SEQ ID NO: 82 are phosphorylated. In some embodiments, the antibodies or antigen-binding antibody fragments bind to a 23-amino acid peptide comprising amino acids at positions 392 to 414 of SEQ ID NO: 82, wherein the serine residues at positions 409 and 410 are phosphorylated (“TDP-43 (pS409 / pS410)”).

[0101] Therefore, this document provides isolated monoclonal antibodies and antigen-binding antibody fragments that competitively bind to TDP-43 (e.g., human TDP-43) with: antibodies containing the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; antibodies containing the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; antibodies containing the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; antibodies containing the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; antibodies containing the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or antibodies containing the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO: 81.

[0102] In some embodiments, the antibody binds to the same epitope on TDP-43 (e.g., human TDP-43) with: an antibody containing the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; an antibody containing the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; an antibody containing the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; an antibody containing the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; an antibody containing the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or an antibody containing the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO: 81.

[0103] This article also provides antibodies that specifically bind to TDP-43, including heavy chain variable domains and light chain variable domains, comprising: heavy chain CDR1, including SEQ ID NO: 84; heavy chain CDR2, including SEQ ID NO: 85; heavy chain CDR3, including SEQ ID NO: 86; light chain CDR1, including SEQ ID NO: 87; light chain CDR2, including SEQ ID NO: 88; and light chain CDR3, including SEQ ID NO: 89.

[0104] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 63. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 63. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 63.

[0105] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 65. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 65. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 65.

[0106] This article also provides antibodies that specifically bind to TDP-43, including heavy chain variable domains and light chain variable domains, comprising: heavy chain CDR1, including SEQ ID NO: 90; heavy chain CDR2, including SEQ ID NO: 91; heavy chain CDR3, including SEQ ID NO: 92; light chain CDR1, including SEQ ID NO: 93; light chain CDR2, including SEQ ID NO: 94; and light chain CDR3, including SEQ ID NO: 95.

[0107] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 67. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 68. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 67.

[0108] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 69. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 69. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 69.

[0109] This article also provides antibodies that specifically bind to TDP-43, including heavy chain variable domains and light chain variable domains, comprising: heavy chain CDR1, including SEQ ID NO: 96; heavy chain CDR2, including SEQ ID NO: 97; heavy chain CDR3, including SEQ ID NO: 98; light chain CDR1, including SEQ ID NO: 99; light chain CDR2, including SEQ ID NO: 100; and light chain CDR3, including SEQ ID NO: 101.

[0110] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 71. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 71. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 71.

[0111] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 73. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 73. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 73.

[0112] This article also provides antibodies that specifically bind to TDP-43, including heavy chain variable domains and light chain variable domains, comprising: heavy chain CDR1, including SEQ ID NO: 102; heavy chain CDR2, including SEQ ID NO: 103; heavy chain CDR3, including SEQ ID NO: 104; light chain CDR1, including SEQ ID NO: 105; light chain CDR2, including SEQ ID NO: 106; and light chain CDR3, including SEQ ID NO: 107.

[0113] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 75. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 75. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 75.

[0114] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 77. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 77. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 77.

[0115] This article also provides antibodies that specifically bind to TDP-43, including heavy chain variable domains and light chain variable domains, comprising: heavy chain CDR1, including SEQ ID NO: 108; heavy chain CDR2, including SEQ ID NO: 109; heavy chain CDR3, including SEQ ID NO: 110; light chain CDR1, including SEQ ID NO: 111; light chain CDR2, including SEQ ID NO: 112; and light chain CDR3, including SEQ ID NO: 113.

[0116] In some embodiments, the heavy chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 79. In some embodiments, the heavy chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 79. In some embodiments, the heavy chain variable domain contains the sequence of SEQ ID NO: 79.

[0117] In some embodiments, the light chain variable domain contains a sequence that is at least 95% identical to SEQ ID NO: 81. In some embodiments, the light chain variable domain contains a sequence that is at least 98% identical to SEQ ID NO: 81. In some embodiments, the light chain variable domain contains the sequence of SEQ ID NO: 81.

[0118] This article also provides antibodies that specifically bind to TDP-43 (e.g., human TDP-43), comprising three light chain CDRs and three heavy chain CDRs of a mouse antibody, characterized by comprising a heavy chain variable domain of SEQ ID NO: 1 and a light chain variable domain of SEQ ID NO: 24.

[0119] In some implementations, the antibody is a humanized antibody, a chimeric antibody, or a decorative antibody (as described herein).

[0120] The complementarity-determining region (“CDR”) can be defined by different systems. For example, the CDR described herein can be selected from the group consisting of Kabat, Chothia, Kabat / Chothia Composite, AbM, and Contact.

[0121] In some embodiments, the antibody comprises a humanized mature heavy chain variable domain comprising: a heavy chain CDR1 as defined by the Kabat / Chothia Composite, including SEQ ID NO: 49; a heavy chain CDR2 as defined by Kabat, including SEQ ID NO: 51; and a heavy chain CDR3 as defined by Kabat or Chothia, including SEQ ID NO: 52; and a humanized mature light chain variable domain comprising the three Kabat light chain CDRs of SEQ ID NO: 53-55.

[0122] In some embodiments, the humanized mature heavy chain variable domain comprises a sequence that is at least 80% identical to any of SEQ ID NO: 4-23. In some embodiments, the humanized mature light chain variable domain comprises a sequence that is at least 80% identical to any of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises a humanized mature heavy chain variable domain comprising a sequence that is at least 80% identical to any of SEQ ID NO: 4-23; and a humanized mature light chain variable domain comprising a sequence that is at least 80% identical to any of SEQ ID NO: 27-48.

[0123] In some embodiments, the humanized mature heavy chain variable domain comprises a sequence that is at least 85% identical to any of SEQ ID NO: 4-23. In some embodiments, the humanized mature light chain variable domain comprises a sequence that is at least 85% identical to any of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises a humanized mature heavy chain variable domain comprising a sequence that is at least 85% identical to any of SEQ ID NO: 4-23; and a humanized mature light chain variable domain comprising a sequence that is at least 85% identical to any of SEQ ID NO: 27-48.

[0124] In some embodiments, the humanized mature heavy chain variable domain comprises a sequence that is at least 90% identical to any of SEQ ID NO: 4-23. In some embodiments, the humanized mature light chain variable domain comprises a sequence that is at least 90% identical to any of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises a humanized mature heavy chain variable domain comprising a sequence that is at least 90% identical to any of SEQ ID NO: 4-23; and a humanized mature light chain variable domain comprising a sequence that is at least 90% identical to any of SEQ ID NO: 27-48.

[0125] In some embodiments, the humanized mature heavy chain variable domain comprises a sequence that is at least 95% identical to any of SEQ ID NO: 4-23. In some embodiments, the humanized mature light chain variable domain comprises a sequence that is at least 95% identical to any of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises a humanized mature heavy chain variable domain comprising a sequence that is at least 95% identical to any of SEQ ID NO: 4-23; and a humanized mature light chain variable domain comprising a sequence that is at least 95% identical to any of SEQ ID NO: 27-48.

[0126] In some embodiments, the humanized mature heavy chain variable domain includes a sequence of one of SEQ ID NO: 4-23. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 4. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 5. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 6. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 7. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 8. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 9. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 10. In some embodiments, the humanized mature heavy chain variable domain includes a sequence including SEQ ID NO: 11. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 12. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 13. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 14. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 15. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 16. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 17. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 18. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 19. In some embodiments, the humanized mature heavy chain variable domain includes the sequence of SEQ ID NO: 21. In some embodiments, the humanized mature heavy chain variable domain includes the sequence comprising SEQ ID NO: 22. In some embodiments, the humanized mature heavy chain variable domain includes the sequence comprising SEQ ID NO: 23.

[0127] In some embodiments, the humanized mature light chain variable domain includes a sequence of one of SEQ ID NO: 27-48. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 27. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 28. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 29. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 30. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 31. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 32. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 33. In some embodiments, the humanized mature light chain variable domain includes a sequence including SEQ ID NO: 34. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 35. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 36. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 37. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 38. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 39. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 40. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 41. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 42. In some embodiments, the humanized mature light chain variable structural domain includes the sequence of SEQ ID NO: 43. In some embodiments, the humanized mature light chain variable domain includes the sequence comprising SEQ ID NO: 44. In some embodiments, the humanized mature light chain variable domain includes the sequence comprising SEQ ID NO: 45. In some embodiments, the humanized mature light chain variable domain includes the sequence comprising SEQ ID NO: 46. In some embodiments, the humanized mature light chain variable domain includes the sequence comprising SEQ ID NO: 47. In some embodiments, the humanized mature light chain variable domain includes the sequence comprising SEQ ID NO: 48.

[0128] The humanized mature light chain variable domains described herein contain a lysine residue at their C-terminus. However, in some embodiments, the humanized mature light chain variable domains (e.g., any of the mature light chain variable domains described herein) do not contain a C-terminal lysine residue.

[0129] In some embodiments, the antibody or its antigen-binding fragment comprises a humanized mature heavy chain variable domain, comprising the sequence of one of SEQ ID NO: 4-23; and a humanized mature light chain variable domain, comprising the sequence of one of SEQ ID NO: 27-48.

[0130] In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 4 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 5 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 6 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 7 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 8 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 9 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 10 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 11 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 12 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 13 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 14 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 15 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48.In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 16 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 17 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 18 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 19 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48.

[0131] In some embodiments, the antibody or antigen-binding fragment thereof comprises the humanized mature heavy chain variable domain of SEQ ID NO: 20 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or antigen-binding fragment thereof comprises the humanized mature heavy chain variable domain of SEQ ID NO: 21 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or antigen-binding fragment thereof comprises the humanized mature heavy chain variable domain of SEQ ID NO: 22 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48. In some embodiments, the antibody or antigen-binding fragment thereof comprises the humanized mature heavy chain variable domain of SEQ ID NO: 23 and the humanized mature light chain variable domain of any one of SEQ ID NO: 27-48.

[0132] In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 20 and the humanized mature light chain variable domain of SEQ ID NO: 47. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 20 and the humanized mature light chain variable domain of SEQ ID NO: 48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 21 and the humanized mature light chain variable domain of SEQ ID NO: 47. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 21 and the humanized mature light chain variable domain of SEQ ID NO: 48. In some embodiments, the antibody or its antigen-binding fragment comprises the humanized mature heavy chain variable domain of SEQ ID NO: 23 and the humanized mature light chain variable domain of SEQ ID NO: 47. In some embodiments, the antibody or its antigen-binding fragment comprises a humanized mature heavy chain variable domain of SEQ ID NO: 23 and a humanized mature light chain variable domain of SEQ ID NO: 48.

[0133] In some implementations, at least one of the following positions (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17) in the humanized heavy chain variable domain is occupied by a specified amino acid: K19 is occupied by R; S35 is occupied by G; T40 is occupied by A; E42 is occupied by G; A49 is occupied by S; K43 is occupied by E; R44 is occupied by G or D; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; L80 is occupied by A or G; L82c is occupied by G; M83 is occupied by R; S84 is occupied by A; M89 is occupied by V; or F91 is occupied by Y.

[0134] In some implementations, at least one of the following positions (e.g., 2, 3, 4, 5, or 6) in the humanized heavy chain variable domain is occupied by a specified amino acid: K43 is occupied by E; R44 is occupied by G or D; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; or F91 is occupied by Y.

[0135] In some implementations, at least one (e.g., 2, 3, or 4) of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: S35 is occupied by G; L78 is occupied by A or G; L80 is occupied by A or G; or L82c is occupied by G.

[0136] In some implementations, F91 of the humanized heavy chain variable domain is occupied by Y; and at least one of the following positions (e.g., 2, 3, 4, 5 or 6) in the humanized heavy chain variable domain is occupied by the specified amino acid: R44 is occupied by G; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; or M83 is occupied by R.

[0137] In some implementations, at least one of the following positions (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13) in the variable domain of the humanized light chain is occupied by a specified amino acid: V3 is occupied by Q; L9 is occupied by S; D17 is occupied by Q; Q18 is occupied by P; K39 is occupied by R; K45 is occupied by R; T80 is occupied by A or S; T46 is occupied by R; L83 is occupied by V; L92 is occupied by G or A; V94 is occupied by I or A; A100 is occupied by G, D, or R; or L104 is occupied by V.

[0138] In some implementations, at least one (e.g., two) of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: V3 is occupied by Q or A100 is occupied by D or R.

[0139] In some implementations, at least one (e.g., 2, 3, or 4) of the following positions in the humanized light chain variable domain is occupied by a specified amino acid: L9 is occupied by S; T80 is occupied by A or S; L92 is occupied by G or A; or V94 is occupied by I or A.

[0140] In some implementations, V3 is occupied by Q; Q18 is occupied by P; A100 is occupied by D; and at least one (e.g., two) of the following positions in the humanized light chain variable domain is occupied by the specified amino acid: T80 is occupied by A or L92 is occupied by A.

[0141] In some embodiments, at least one of the following positions (e.g., 2, 3, 4, 5, 6, 7, or 8) in the humanized heavy chain variable domain is occupied by a specified amino acid: L5 is occupied by V; G44 is occupied by R; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; M89 is occupied by V, or F91 is occupied by Y; and at least one of the following positions (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) in the humanized light chain variable domain is occupied by a specified amino acid: V3 is occupied by Q; D17 is occupied by Q; Q18 is occupied by P; K39 is occupied by R; K45 is occupied by R; T80 is occupied by A; L83 is occupied by V; L92 is occupied by A; A100 is occupied by D; or L104 is occupied by V.

[0142] The aforementioned substitutions may confer reduced (e.g., at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% reduction) immunogenicity to parental antibodies and / or increased (e.g., at least 1%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% increase) thermal stability to parental antibodies.

[0143] In some embodiments, the humanized antibody exhibits improved thermostability compared to a reference antibody containing the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24. In some embodiments, the humanized antibody exhibits improved thermostability compared to a reference antibody containing the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65. In some embodiments, the humanized antibody exhibits improved thermostability compared to a reference antibody containing the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69. In some embodiments, the humanized antibody exhibits improved thermostability compared to a reference antibody containing the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73. In some embodiments, the humanized antibody exhibits improved thermostability compared to a reference antibody containing the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77. In some implementations, the humanized antibody exhibits improved thermal stability compared to a reference antibody containing the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO: 81.

[0144] In some embodiments, the humanized antibody has a melting temperature of about 55°C or higher. In some embodiments, the humanized antibody has the following melting temperatures: about 56°C or higher, about 57°C or higher, about 58°C or higher, about 59°C or higher, about 60°C or higher, about 61°C or higher, about 62°C or higher, about 63°C or higher, about 64°C or higher, about 65°C or higher, about 66°C or higher, about 67°C or higher, about 68°C or higher, about 69°C or higher, about 70°C or higher, about 71°C or higher, about 72°C or higher, about 73°C or higher, about 74°C or higher, about 75°C or higher, about 76°C or higher, about 77°C or higher, about 78°C or higher, about 79°C or higher, about 80°C or higher, about 81°C or higher, about 82°C or higher, about 83°C or higher, about 84°C or higher, or about 85°C or higher. In some implementations, the humanized antibody has the following melting temperatures: at least about 55°C, at least about 56°C, at least about 57°C, at least about 58°C, at least about 59°C, at least about 60°C, at least about 61°C, at least about 62°C, at least about 63°C, at least about 64°C, at least about 65°C, at least about 66°C, at least about 67°C, at least about 68°C, at least about 69°C, at least about 70°C, at least about 71°C, at least about 72°C, at least about 73°C, at least about 74°C, at least about 75°C, at least about 76°C, at least about 77°C, at least about 78°C, at least about 79°C, at least about 80°C, at least about 81°C, at least about 82°C, at least about 83°C, at least about 84°C, or at least about 85°C.In some embodiments, the humanized antibody has the following melting temperatures: about 55°C to about 85°C, about 55°C to about 80°C, about 55°C to about 75°C, about 55°C to about 70°C, about 55°C to about 65°C, about 55°C to about 63°C, about 55°C to about 61°C, about 55°C to about 59°C, about 55°C to about 57°C, about 57°C to about 85°C, about 57°C to about 80°C, about 57°C to about 75°C, about 57°C to about 70°C, about 57°C to about 65°C, about 57°C to about 63°C, about 57°C to about 61°C, about 57°C to about 59°C, about 59°C to about 85°C, about 59°C to about 80°C, about 59°C to about 75°C, about 59°C to about 70°C, about 59°C Approximately 65°C, approximately 59°C to approximately 63°C, approximately 59°C to approximately 61°C, approximately 61°C to approximately 85°C, approximately 61°C to approximately 80°C, approximately 61°C to approximately 75°C, approximately 61°C to approximately 70°C, approximately 61°C to approximately 65°C, approximately 61°C to approximately 63°C, approximately 63°C to approximately 85°C, approximately 63°C to approximately 80°C, approximately 63°C to approximately 75°C, approximately 63°C to approximately 70°C, approximately 63°C to approximately 65°C, approximately 65°C to approximately 85°C, approximately 65°C to approximately 80°C, approximately 65°C to approximately 75°C, approximately 65°C to approximately 70°C, approximately 70°C to approximately 85°C, approximately 70°C to approximately 80°C, approximately 70°C to approximately 75°C, approximately 75°C to approximately 85°C, approximately 75°C to approximately 80°C, or approximately 80°C to approximately 85°C.

[0145] This article also provides antibodies that specifically bind to TDP-43 (e.g., human TDP-43) and contain heavy chain variable domains, including: heavy chain CDR1 as defined by the Kabat / Chothia Composite, including SEQ ID NO: 49 or SEQ ID NO: 50; heavy chain CDR2 as defined by Kabat, including SEQ ID NO: 51; heavy chain CDR3 as defined by Kabat or Chothia, including SEQ ID NO: 52; light chain CDR1 as defined by Kabat, including SEQ ID NO: 53; light chain CDR2 as defined by Kabat, including SEQ ID NO: 54; and light chain CDR3 as defined by Kabat, including one of SEQ ID NO: 55-61.

[0146] In some implementations, heavy chain CDR1 as defined by the Kabat / Chothia Composite includes SEQ ID NO: 49; heavy chain CDR2 as defined by Kabat includes SEQ ID NO: 51; heavy chain CDR3 as defined by Kabat or Chothia includes SEQ ID NO: 52; light chain CDR1 as defined by Kabat includes SEQ ID NO: 53; light chain CDR2 as defined by Kabat includes SEQ ID NO: 54; and light chain CDR3 as defined by Kabat includes SEQ ID NO: 55 or SEQ ID NO: 61.

[0147] In some embodiments, the heavy chain variable domain comprises a sequence that is at least 95% (e.g., at least 96%, at least 97%, at least 98%, at least 99%) identical to any of SEQ ID NO: 4-23. In some embodiments, the heavy chain variable domain comprises a sequence that is at least 95% (e.g., at least 96%, at least 97%, at least 98%, at least 99%) identical to any of SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 23. In some embodiments, the heavy chain variable domain comprises a sequence that is at least 98% identical to any of SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 23. In some embodiments, the heavy chain variable domain comprises the sequence of SEQ ID NO: 20. In some embodiments, the heavy chain variable domain comprises the sequence of SEQ ID NO: 21. In some embodiments, the heavy chain variable domain comprises the sequence of SEQ ID NO: 23.

[0148] In some embodiments, the light chain variable domain contains at least 95% (e.g., at least 96%, at least 97%, at least 98%, at least 99%) of the sequence identical to any of SEQ ID NO: 27-48. In some embodiments, the light chain variable domain contains at least 95% (e.g., at least 96%, at least 97%, at least 98%, at least 99%) of the sequence identical to SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the light chain variable domain contains at least 98% of the sequence identical to SEQ ID NO: 47 or SEQ ID NO: 48. In some embodiments, the light chain variable domain contains SEQ ID NO: 47. In some embodiments, the light chain variable domain contains SEQ ID NO: 48.

[0149] As previously stated, the humanized mature light chain variable domains described herein include a lysine residue at their C-terminus. However, in some embodiments, the humanized mature light chain variable domains (e.g., any of the mature light chain variable domains described herein) do not include a C-terminal lysine residue. For example, any of the mature light chain variable domains of SEQ ID NO: 24-48, 65, 69, 73, 77, or 81 may lack a C-terminal lysine residue.

[0150] In some embodiments, the antibody is a humanized antibody, a chimeric antibody, or a veneered antibody. In some embodiments, the antibody is an antigen-binding antibody fragment. In some embodiments, the antigen-binding antibody fragment is a Fab fragment, a Fab'2 fragment, or a single-chain Fv.

[0151] In some implementations, the antibody is a complete antibody. In some implementations, the antibody has a human IgG1 isotype.

[0152] In some implementations, the heavy chain variable domain is fused to the heavy chain constant region (e.g., any heavy chain constant region described herein) and the light chain variable domain is fused to the light chain constant region (e.g., any light chain constant region described herein).

[0153] In some embodiments, the heavy chain constant region is a mutant form of the natural human heavy chain constant region, which binds less to the Fcγ receptor compared to the natural heavy chain constant region. In some embodiments, the heavy chain constant region has an IgG1 isotype. In some embodiments, the antibody has at least one mutation in the constant region. In some embodiments, at least one mutation reduces complement fixation or activation in the constant region. In some embodiments, at least one mutation is located at one or more of the following positions (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) according to EU numbers: 241, 264, 265, 270, 296, 297, 318, 320, 322, 329, and 331. In some embodiments, the antibody contains alanine at positions 318, 320, and 322 according to EU numbers.

[0154] In some embodiments, the antibody or antigen-binding antibody fragment selectively binds to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43). In some embodiments, the antibody or antigen-binding antibody fragment selectively binds to phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) compared to unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43).

[0155] In some embodiments, the antibody or antigen binds to the antibody fragment at a concentration higher than that of unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43) by at least (e.g., at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 450-fold, at least 500-fold, at least 550-fold, at least 600-fold, at least 650-fold, at least 700-fold, at least 750-fold, at least 800-fold, at least 850-fold, at least 900-fold, at least 950-fold, at least 1,000-fold, at least 1,050-fold, at least 1,100-fold, at least 1,150-fold, at least 1,200-fold, at least...). 1,250 times, at least 1,300 times, at least 1,350 times, at least 1,400 times, at least 1,450 times, or at least 1,500 times, or about 10 times to about 1,500 times, about 10 times to 1,400 times, about 10 times to about 1,300 times, about 10 times to about 1,200 times, about 10 times to about 1,100 times, about 10 times to about 1,000 times, about 10 times to about 900 times, about 10 times to about 800 times, about 10 times to about 700 times, about 10 times to about 600 times, about 10 times to about 500 times, about 10 times to about 400 times, about 10 times to about 300 times, about 10 times to about 200 times, about 10 times to Approximately 100 times, approximately 10 times to approximately 50 times, approximately 50 times to approximately 1,500 times, approximately 50 times to 1,400 times, approximately 50 times to approximately 1,300 times, approximately 50 times to approximately 1,200 times, approximately 50 times to approximately 1,100 times, approximately 50 times to approximately 1,000 times, approximately 50 times to approximately 900 times, approximately 50 times to approximately 800 times, approximately 50 times to approximately 700 times, approximately 50 times to approximately 600 times, approximately 50 times to approximately 500 times, approximately 50 times to approximately 400 times, approximately 50 times to approximately 300 times, approximately 50 times to approximately 200 times, approximately 50 times to approximately 100 times, approximately 100 times to approximately 1,500 times, approximately 100 times to 1,400 times, approximately 100 times From about 1,300 times, from about 100 times to about 1,200 times, from about 100 times to about 1,100 times, from about 100 times to about 1,000 times, from about 100 times to about 900 times, from about 100 times to about 800 times, from about 100 times to about 700 times, from about 100 times to about 600 times, from about 100 times to about 500 times, from about 100 times to about 400 times, from about 100 times to about 300 times, from about 100 times to about 200 times, from about 200 times to about 1,500 times, from about 200 times to about 1,400 times, from about 200 times to about 1,300 times, from about 200 times to about 1,200 times, from about 200 times to about 1,100 times, from about 200 times to about 1,000 times, about 200 times to about 900 times, about 200 times to about 800 times, about 200 times to about 700 times, about 200 times to about 600 times, about 200 times to about 500 times, about 200 times to about 400 times, about 200 times to about 300 times, about 500 times to about 1,500 times, about 500 times to 1,400 times, about 500 times to about 1,300 times, about 500 times to about 1,200 times, about 500 times to about 1,100 times, about 500 times to about 1,000 times, about 500 times to about 900 times, about 500 times to about 800 times, about 500 times to about 700 times It binds to phosphorylated TDP-43 (e.g., human phosphorylated TDP-43) with an affinity of 0 times, about 500 times to about 600 times, about 800 times to about 1,500 times, about 800 times to about 1,400 times, about 800 times to about 1,200 times, about 800 times to about 1,100 times, about 800 times to about 1,000 times, about 800 times to about 900 times, about 1,000 times to about 1,500 times, about 1,000 times to about 1,400 times, about 1,000 times to about 1,300 times, about 1,000 times to about 1,200 times, or about 1,000 times to about 1,100 times.

[0156] In some embodiments, phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) comprises phosphorylation of at least one amino acid residue selected from S409 and S410. In some embodiments, phosphorylated TDP-43 (e.g., phosphorylated human TDP-43) comprises phosphorylation of both S409 and S410.

[0157] In some embodiments, the antibody selectively binds to cytoplasmic aggregates of TDP-43 (e.g., cytoplasmic aggregates of human TDP-43). In some embodiments, the antibody selectively binds to cytoplasmic aggregates of TDP-43 (e.g., cytoplasmic aggregates of human TDP-43) compared to nuclear TDP-43 (e.g., nuclear human TDP-43). In some embodiments, the cytoplasmic aggregates of TDP-43 (e.g., cytoplasmic aggregates of human TDP-43) comprise phosphorylated aggregates of TDP-43. In some embodiments, the antibody substantially does not bind to unphosphorylated TDP-43 (e.g., unphosphorylated human TDP-43).

[0158] In some implementations, the antibody is conjugated to a therapeutic agent, cytotoxic agent, cell growth inhibitor, immunomodulator, neurotrophic agent, or neuroprotective agent.

[0159] The antibodies used in the disclosed formulations may be conjugated (i.e., conjugated) to a therapeutic component (such as a cytotoxic agent, radiotherapy agent, immunomodulator, or secondary antibody) (e.g., to form an antibody heteroconjugate). Representative therapeutic components include agents known to be used to treat, manage, or improve symptoms of TDP-43-related diseases.

[0160] The therapeutic portion and / or detectable substance may be directly coupled or conjugated to any of the rodent, chimeric or humanized antibodies described herein via intermediates (e.g., linkers) using techniques known in the art. See, for example, Arnon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy,” in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243–56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery,” in Controlled Drug Delivery (2nd ed.), Robinson et al. (eds.), pp. 623–53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review,” in Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475–506 (1985); “Analysis, Results, and Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy,” in Monoclonal Antibodies For Cancer Detection and Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985) and Thorpe et al., Immunol. Rev., 1982, 62: 119-58.

[0161] The antibodies used in the disclosed formulations also include modified forms of murine, chimeric, or humanized 13D3 antibodies that have an extended in vivo half-life relative to their unmodified counterparts. Such modifications can be prepared, for example, by glycosylation, acetylation, polyethylene glycolation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, or binding to cellular ligands or other proteins. As an example, a representative method for extending antibody half-life is described in PCT International Publication No. WO 02 / 060919.

[0162] The antibody or antigen-binding fragments described herein may be present in a buffer solution. The buffer solution may have a pH of about 6 to about 7. The antibody or antigen-binding fragments described herein may be present in any pharmaceutically acceptable excipient or carrier.

[0163] Typically, the formulations are sterile, for example, through aseptic filtration using 0.2 μm or 0.22 μm filters. The formulations disclosed herein are generally stable after freezing and thawing.

[0164] Selection of constant region The heavy and light chain variable regions of chimeric, veneered, or humanized antibodies may be linked to at least a portion of the human constant region. The selection of the constant region depends in part on whether antibody-dependent cell-mediated cytotoxicity, antibody-dependent phagocytosis, and / or complement-dependent cytotoxicity are required. For example, human isotypes IgG1 and IgG3 exhibit complement-dependent cytotoxicity, while human isotypes IgG2 and IgG4 do not. Human IgG1 and IgG3 also induce stronger cell-mediated effector functions than human IgG2 and IgG4. The light chain constant region may be λ or κ. The numbering convention for constant regions includes EU numbers (Edelman, GM, et al., Proc. Natl. Acad. Sci. USA 63:78-85 (1969)), Kabat number (Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1991, IMGT unique numbering (Lefranc M.-P. et al., IMGT unique numbering for immunoglobulin and T cell receptor constant domains and Ig superfamily C-likedomains, Dev. Comp. Immunol. 29:185-203 (2005) and IMGT exon number (Lefranc, ibid.).

[0165] One or more amino acids (such as C-terminal lysine in the heavy chain) at the amino or carboxyl terminus of the light and / or heavy chains may be deleted or derivatized in a certain proportion or in all of the molecule. Substitutions may be made in the constant region to reduce or enhance effector function, such as complement-mediated cytotoxicity or ADCC. ( See, for example, Winter et al., U.S. Patent No. 5,624,821; Tso et al., U.S. Patent No. 5,834,597; and Lazar et al. Proc. Natl. Acad. Sci. USA 103:4005(2006)), or prolonging the half-life of human philtrum. ( See, for example, Hinton et al. J. Biol. Chem. 279:6213 (2004)). Exemplary substitutions include Gln at position 250 and / or Leu at position 428 (EU numbers are used for the constant region in this paragraph) to prolong the antibody's half-life. Substitutions at any or all of positions 234, 235, 236, and / or 237 reduce affinity for Fey receptors, particularly FcγRI receptors. ( See, for example, U.S. Patent No. 6,624,821. Alanine substitutions at positions 234, 235, and 237 of human IgG1 can be used to reduce effector function. Some antibodies have alanine substitutions at positions 234, 235, and 237 of human IgG1 to reduce effector function. Optionally, positions 234, 236, and / or 237 of human IgG2 are substituted with alanine, and position 235 is substituted with glutamine. ( See, for example, U.S. Patent No. 5,624,821. In some antibodies, mutations are used in human IgG1 at one or more of positions 241, 264, 265, 270, 296, 297, 322, 329, and 331 (e.g., 2, 3, 4, 5, 6, 7, 8, or 9) numbered by EU. In some antibodies, mutations are used in human IgG1 at one or more of positions 318, 320, and 322 (e.g., 2 or 3) numbered by EU. In some antibodies, positions 234 and / or 235 are substituted with alanine and / or position 329 is substituted with glycine. In some antibodies, positions 234 and 235 are substituted with alanine. In some antibodies, the isotype is human IgG2 or IgG4.

[0166] Antibodies can be expressed as tetramers containing two light chains and two heavy chains, single heavy chains, light chains, Fab, Fab', F(ab')2, and Fv, or as single-chain antibodies in which the mature variable domains of the heavy and light chains are linked by spacers.

[0167] Human constant regions exhibit both allotropic and allophotropic variation among individuals; that is, a constant region can differ in different individuals at one or more polymorphic sites. Allotropic regions differ from allotropic regions in that serum recognizing the allotropic region binds to one or more other non-polymorphic regions of the same type. Thus, for example, another heavy chain constant region is IgG1 Glm3 with or without a C-terminal lysine. References to human constant regions include constant regions having any natural allotype or any arrangement of residues occupying a position in a natural allotype.

[0168] Antibodies can also be administered in the form of nucleic acids encoding antibody heavy and / or light chains. If both heavy and light chains are present, the chains are preferably linked as single-chain antibodies.

[0169] IV. Nucleic Acids, Vectors, and Host Cells This disclosure further provides nucleic acids encoding any of the above-described heavy chain variable domains and / or light chain variable domains. In some embodiments, the nucleic acid encodes the heavy chain variable domain and / or light chain variable domain of any of the antibodies described herein. For example, the nucleic acid may encode a heavy chain variable domain comprising any of SEQ ID NO: 4-23 and / or a light chain variable domain comprising any of SEQ ID NO: 27-48. Optionally, such nucleic acids further encode a signal peptide and may be expressed together with a signal peptide linked to a constant region. The coding sequence of the nucleic acid may be operatively linked to a regulatory sequence to ensure expression of the coding sequence (such as a promoter, enhancer, ribosome binding site, transcription termination signal, etc.). The nucleic acid encoding the heavy and light chains may be present in isolated form or may be cloned into one or more vectors. The nucleic acid may be synthesized, for example, by solid-state synthesis of overlapping oligonucleotides or by PCR. The nucleic acid encoding the heavy and light chains may, for example, be conjugated into a single continuous nucleic acid within an expression vector, or may be separate, for example, each cloned into its own expression vector. In some embodiments, the nucleic acid is codon-optimized for expression in a host cell.

[0170] Several cell lines using antibody expression are known. ( For example ，Methods for generating chimeric and humanized antibodies (hybridoma). For example, well-known methods can be used to clone and sequence the immunoglobulin variable region of the antibody. In one method, the heavy chain variable VH region is cloned using mRNA prepared from hybridoma cells via RT-PCR. A common primer is used as the 5' primer and a g2b constant region-specific 3' primer for the VH region leader peptide containing the translation start codon. Exemplary primers are described in U.S. Patent Publication US 2005 / 0009150 (hereinafter referred to as "Schenk") by Schenk et al. Sequences from multiple independently derived clones can be compared to ensure that no changes are introduced during amplification. The sequence of the VH region can also be determined or confirmed by sequencing the VH fragment obtained by the 5' RACE RT-PCR method and the 3' g2b-specific primer.

[0171] Light chain variable VL regions can be cloned in a similar manner. In one approach, a shared set of primers is designed to amplify the VL region using a 5' primer designed to hybridize with the VL region containing the translation start codon and a 3' primer specific to the Ck region downstream of the VJ conjugate region. In a second approach, a 5' RACE RT-PCR method is used to clone the VL-encoding cDNA. Exemplary primers are described in Schenk, ibid. The cloned sequence is then combined with a sequence encoding a human (or other non-human species) constant region.

[0172] This document also provides vectors comprising nucleic acids encoding mature heavy chain variable domains (e.g., any of the heavy chain variable domains described herein) and light chain variable domains (e.g., any of the light chain variable domains described herein), said nucleic acids being operatively linked to one or more regulatory sequences to achieve expression of any of the antibody or antigen-binding fragments described herein in mammalian cells.

[0173] In one approach, the variable regions of the heavy and light chains are reengineered to encode splicing donor sequences downstream of the corresponding VDJ or VJ cassette and cloned into mammalian expression vectors, such as pCMV-hyl for the heavy chain and pCMV-Mcl for the light chain. These vectors encode the human γl and Ck constant regions as exon fragments downstream of the inserted variable region cassette. After sequence validation, the heavy and light chain expression vectors are co-transfected into CHO cells to generate chimeric antibodies. Conditioned culture medium is collected 48 hours post-transfection, and antibody production is analyzed by Western blotting or antigen binding by ELISA. The chimeric antibodies are humanized as described above.

[0174] Chimeric, veneered, humanized, and human antibodies are typically generated through recombinant expression. Recombinant polynucleotide constructs generally include expression control sequences operatively linked to the coding sequence of the antibody chain, including naturally associated or heterologous expression control elements such as promoters. The expression control sequence may be a promoter system within a vector capable of transforming or transfecting eukaryotic or prokaryotic host cells. Once the vector has been incorporated into a suitable host, the host is maintained under conditions suitable for high levels of nucleotide sequence expression and for the collection and purification of cross-reactive antibodies.

[0175] Therefore, this document provides host cells transformed using any of the vectors described herein. This document also provides host cells containing any of the nucleic acids described herein.

[0176] Expression vectors typically replicate in a host organism either as a cell-free genome or as part of the host's chromosomal DNA. Often, expression vectors contain selection markers, such as aminopenicillin resistance or hygromycin resistance, to allow detection of cells transformed with the desired DNA sequence.

[0177] Escherichia coli is a prokaryotic host that can be used to express antibodies, especially antibody fragments. Microorganisms such as yeast can also be used for expression. yeast A suitable yeast host is required, with the vector containing, as needed, expression control sequences, origin of replication, and termination sequences. Typical promoters include 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters particularly include promoters derived from alcohol dehydrogenases, isocytochrome C, and enzymes responsible for the utilization of maltose and galactose.

[0178] Mammalian cells can be used to express nucleotide segments encoding immunoglobulins or fragments thereof. See Winnacker, From Genes to Clones, (VCH Publishers, NY, 1987). Many suitable host cell lines capable of secreting complete heterologous proteins have been developed, including CHO cell lines, various COS cell lines, HeLa cells, HEK293 cells, L cells, and non-antibody-producing myeloma cells, including Sp2 / 0 and NSO. Cells can be non-human. Expression vectors for these cells may include expression control sequences such as origin of replication, promoters, and enhancers (Queen sequences). et al., Immunol. Rev. 89:49 (1986)), and essential processing information sites, such as ribosome binding sites, RNA splicing sites, polyadenylation sites, and transcription terminator sequences. Expression control sequences may include promoters derived from endogenous genes, cytomegalovirus, SV 40, adenovirus, bovine papillomavirus, etc. See Co et al., J. Immunol. 148: 1149 (1992). In some implementations, the promoter is a eukaryotic promoter.

[0179] Alternatively, the antibody-coding sequence can be incorporated into the transgene to introduce it into the genome of the transgenic animal and subsequently expressed in its milk. ( See, for example, U.S. Patent Nos. 5,741,957, 5,304,489, and 5,849,992. Suitable transgenes include coding sequences of light and / or heavy chains operatively linked to promoters and enhancers from breast-specific genes such as casein or β-lactoglobulin.

[0180] Vectors containing the DNA segment of interest can be transferred into host cells using methods that depend on the cell host type. For example, calcium chloride transfection is commonly used for prokaryotic cells, while calcium phosphate treatment, electroporation, lipid transfection, gene gun, or virus-based transfection can be used for other cell hosts. Other methods for transforming mammalian cells include the use of polybrene, protoplast fusion, liposomes, electroporation, and microinjection. To produce transgenic animals, trans-genes can be microinjected into the genome of fertilized oocytes or embryonic stem cells, and the nuclei of such cells can be transferred into enucleated oocytes.

[0181] After introducing vectors encoding the heavy and light chains of antibodies into cell cultures, cell pools can be screened for growth productivity and product quality in serum-free media. The top-producing cell pools can then be used for FACS-based single-cell cloning to generate monoclonal lines. Specific productivity exceeding 50 pg or 100 pg / cell / day can be used, corresponding to product titers greater than 7.5 g / L of culture. The turbidity, filtration properties, PAGE, IEF, UV scan, HPSEC, carbohydrate-oligosaccharide localization, mass spectrometry, and binding assays of antibodies generated from single-cell clones can also be tested, such as by ELISA or Biacore. The selected clones can then be stored in multiple vials and cryopreserved for later use.

[0182] Once expressed, the antibody can be purified according to standard procedures in the art, including protein A capture, HPLC purification, column chromatography, gel electrophoresis, and similar methods (see Scopes for general information). Protein Purification (Springer-Verlag, NY, 1982)).

[0183] Commercially viable methods for antibody production include codon optimization, promoter selection, transcription element selection, terminator selection, blood-free cell cloning, cell banks, copy number amplification using selectable markers, CHO terminators, or improvement of protein titers (see, for example, U.S. Patent No. 5,786,464; U.S. Patent No. 6,114,148; U.S. Patent No. 6,063,598; U.S. Patent No. 7,569,339; WO2004 / 050884; WO2008 / 012142; WO2008 / 012142; WO2005 / 019442; WO2008 / 107388; WO2009 / 027471; and U.S. Patent No. 5,888,809).

[0184] DNA can be delivered in its naked form (i.e., without colloidal or encapsulating materials). Alternatively, a variety of viral vector systems may be used, including retroviral systems (see, for example, Lawrie and Tumin, Cur. Opin. Genet. Develop. 3, 102-109 (1993)); adenoviral vectors (see, for example, Bett et al., J. Virol. 67, 5911 (1993)); adeno-associated viral vectors (see, for example, Zhou et al., J. Exp. Med. 179, 1867 (1994)); viral vectors from the pox family (including poxviruses and fowlpoxviruses); viral vectors from the alphavirus genus, such as those derived from Sindbis and Semliki forest viruses (see, for example, Dubensky et al., J. Virol. 70, 508-519 (1996)); Venezuelan equine encephalitis virus (see U.S. Patent No. 5,643,576); and rod-shaped viruses, such as vesicular stomatitis virus (see WO). 96 / 34625) and papillomavirus (Ohe et al., Human Gene Therapy 6:325-333(1995); Woo et al., WO 94 / 12629 and Xiao&Brandsma, Nucleic Acids. Res. 24:2630-2622(1996)).

[0185] DNA encoding an immunogen or a vector containing said immunogen can be packaged into liposomes. Suitable lipids and related analogues are described in U.S. Patent Nos. 5,208,036, 5,264,618, 5,279,833, and 5,283,185. The vector and the DNA encoding the immunogen can also be adsorbed onto or associated with a particulate carrier, examples of which include polymethyl methacrylate polymers and polylactide and poly(lactide-co-glycolic acid) (see, for example, McGee et al.). J. Micro Encap. 1996).

[0186] V. Conjugated Antibodies Conjugated antibodies and antigen-binding antibody fragments that specifically bind to antigens such as TDP-43 (e.g., human TDP-43) can be used to detect the presence of TDP-43; monitor and evaluate the efficacy of therapeutic agents used to treat patients diagnosed with amyotrophic lateral sclerosis (ALS), frontotemporal degeneration (FTLD-TDP), primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease; inhibit or reduce TDP-43 aggregation; reduce or eliminate TDP-43 aggregates; stabilize the non-toxic conformation of TDP-43; or treat or prevent patients with ALS, FTLD-TDP, primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease.

[0187] For example, such antibody or antigen-binding antibody fragments may be conjugated to other therapeutic portions, other proteins, other antibodies, and / or detectable markers. See WO 03 / 057838; U.S. Patent No. 8,455,622. Such therapeutic portions may be any agent that can be used to treat, counteract, improve, prevent, or modify an unwanted condition or disease in a patient, such as amyotrophic lateral sclerosis (ALS), frontotemporal degeneration (FTLD-TDP), primary lateral sclerosis, and progressive muscular atrophy and Parkinson's disease.

[0188] The conjugated therapeutic portion may include cytotoxic agents, cell growth inhibitors, neurotrophic agents, neuroprotective agents, radiotherapy agents, immunomodulators, or any bioactive agent that promotes or enhances antibody activity. Cytotoxic agents may be any agent that is toxic to cells. Cell growth inhibitors may be any agent that inhibits cell proliferation. Neurotrophic agents may be any agent that promotes the maintenance, growth, or differentiation of neurons, including chemical or protein agents. Neuroprotective agents may be agents that protect neurons from acute injury or degenerative processes, including chemical or protein agents. Immunomodulators may be any agent that stimulates or inhibits the development or maintenance of an immune response. Radiotherapy agents may be any molecule or compound that emits radiation. If such a therapeutic portion is conjugated to a TDP-43-specific antibody or antigen-binding antibody fragment, such as the antibodies and antigen-binding antibody fragments described herein, the conjugated therapeutic portion will have a specific affinity for cells affected by TDP-43-related diseases compared to normal cells.

[0189] Therefore, the application of conjugated antibodies or conjugated antigen-binding antibody fragments will directly target cells with minimal damage to surrounding normal healthy tissue. This is particularly suitable for treatment fractions that are too toxic to be administered alone. Additionally, smaller amounts of the treatment fraction can be used.

[0190] Some of these antibodies or antigen-binding antibody fragments can be linked to radioactive isotopes. Examples of radioactive isotopes include, for example, yttrium. 90 (90Y), Indium 111 (111In) 131 1. 99 mTc, radioactive silver-111, radioactive silver-199 and bismuth 213 The binding of radioactive isotopes to antibodies or antigen-binding antibody fragments can be performed using conventional bifunctional chelates. For binding of radioactive silver-111 and radioactive silver-199, sulfur-based linkers can be used. See Hazra et al. Cell Biophys. 24-25:1-7 (1994). Binding of silver radioisotopes can involve the reduction of immunoglobulins with ascorbic acid. For radioisotopes (such as 111In and 90Y), ibritumomab tiuxetan can be used and will react with such isotopes to form 111In-ibritumomab and 90Y-ibritumomab, respectively. See Witzig, Cancer Chemother. Pharmacol., 48 (Supplement 1): S91-S95 (2001).

[0191] Some of these antibodies or antigen-binding antibody fragments can be linked to other therapeutic components. These therapeutic components can be, for example, cytotoxic, cell growth-inhibiting, immunomodulatory, neurotrophic, or neuroprotective. For instance, antibodies and antigen-binding antibody fragments can be linked to toxic chemotherapeutic agents (such as maytansine and geldanamycin) or microtubule inhibitors (such as microtubule binders). ( For example, auratestatin or minor groove conjugates (such as calicheamicin). Other representative therapeutic components include agents known to be used to treat, manage, or improve amyotrophic lateral sclerosis (ALS), frontotemporal degeneration (FTLD-TDP), primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease.

[0192] Antibodies or antigen-binding antibody fragments can also be conjugated to detectable markers. Such antibodies and antigen-binding antibody fragments can be used, for example, to diagnose ALS, FTLD-TDP, primary lateral sclerosis, and progressive muscular atrophy and Parkinson's disease. Representative detectable markers that can be conjugated to or linked to antibody or antigen-binding antibody fragments include various enzymes such as horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; prosthetic groups such as streptavidin / biotin and avidin / biotin; fluorescent materials such as umbelliferone, luciferin, luciferin isothiocyanate, rhodamine, dichlorotriazine aminoluciferin, dansyl chloride, or phycoerythrin; luminescent materials such as luminol; bioluminescent materials such as luciferase, luciferin, and jellyfish luminescent protein; and radioactive materials such as radioactive silver-111, radioactive silver-199, and bismuth. 213 ,iodine( 131 I, 125 I, 123 I, 121 I), carbon ( 14 C), sulfur 5 S), tritium ( 3 H), Indium 115 In 113 In 112 In 111 In), Technetium ( 99 Tc), thallium 201 Ti, gallium 68 Ga、 67 Ga), Palladium ( 103 Pd), molybdenum ( 99 Mo), xenon ( 133 Xe), fluorine ( 18 F) 153 Sm、 177 Lu、 159 Gd, 149 Pm, 140 La、 175 Yb、 166 Ho、 90 Y、 47 Sc、 186 Re、 188 Re、 142 Pr、 105 Rh、 97 Ru、 68 Ge 57 Co、 65 ZN, 85 SR, 32 P, 153 Gd, 169 Yb、 51 CR 54 Mn,75 Se、 113 Sn and 117 Sn; positron-emitting metals obtained using various positron emission tomography (PET) scans; non-radioactive paramagnetic metal ions; and radioactively labeled molecules or molecules conjugated to specific radioactive isotopes.

[0193] Therapeutic components, other proteins, other antibodies, and / or detectable markers can be obtained directly or through intermediates. ( For example ， The linker is indirectly coupled or conjugated to the antibody or antigen-binding antibody fragment of the present invention. See, for example, Arnon et al., “Monoclonal Antibodies For Immunotargeting of Drugs in Cancer Therapy,” in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243–56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery,” in Controlled Drug Delivery (2nd ed.), Robinson et al. (eds.), pp. 623–53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers of Cytotoxic Agents in Cancer Therapy: A Review,” in Monoclonal Antibodies 84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475–506 (1985); “Analysis, Results, and Future Prospective of The Therapeutic Use of Radio-labeled Antibody in Cancer Therapy,” in Monoclonal Antibodies For Cancer Detection and Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985); and Thorpe et al. Immunol. Rev.,62:119-58 (1982). Suitable linkers include, for example, cleavable and non-cleavable linkers. Different linkers may be used to release coupled therapeutic moieties, proteins, antibodies, and / or detectable labels under acidic or reducing conditions, upon exposure to specific proteases, or under other defined conditions.

[0194] VI. Pharmaceutical compositions and products This disclosure also provides pharmaceutical compositions and products. Therefore, pharmaceutical compositions comprising any antibody or antigen-binding antibody fragment described herein and a pharmaceutically acceptable carrier are provided herein.

[0195] Pharmaceutical compositions intended for parenteral administration are preferably sterile, substantially isotonic, and manufactured under GMP conditions. The pharmaceutical composition may be provided in unit dosage forms (i.e., a single-dose dose). The pharmaceutical composition may be formulated using one or more physiologically acceptable carriers, diluents, excipients, or adjuvants. The formulation depends on the chosen route of administration. For injection, antibodies or antigen-binding antibody fragments may be formulated in an aqueous solution, preferably in a physiologically compatible buffer such as Hank's solution, Ringer's solution, or physiological saline or acetate buffer (to reduce injection site discomfort). The solution may contain formulation agents such as suspending agents, stabilizers, and / or dispersants. Alternatively, the antibody may be in lyophilized form for preparation with a suitable medium (e.g., sterile, pyrogen-free water) prior to use.

[0196] The anti-TDP-43 antibody described herein can be present in any pharmaceutically acceptable excipient or carrier. For example, the anti-TDP-43 antibody described herein can be present in a buffer solution. The buffer solution may have a pH of about 6 to about 7. Typically, the formulation is sterile, for example, by aseptic filtration using a 0.2 μm or 0.22 μm filter. The formulations disclosed herein are generally stable after freezing and thawing.

[0197] In some implementation schemes, it may be necessary to... In vitro Alternatively, a pharmaceutical composition comprising any of the antibodies described herein may be used in an in vitro method. For example, the method may be used for non-diagnostic and / or non-therapeutic purposes. In such cases, a sample (such as cells, tissues, and / or organs) removed from a patient is exposed to a pharmaceutical composition comprising any of the antibodies described herein.

[0198] In prophylactic applications, antibodies or antigen-binding antibody fragments (e.g., nucleic acids or vectors encoding any of the antibodies or antigen-binding antibody fragments described herein) or pharmaceutical compositions thereof are administered to patients suspected of or at risk of diseases such as ALS, FTLD-TDP, primary lateral sclerosis, and progressive muscular atrophy and Parkinson's disease in a regimen (dosage, frequency, and route of administration) that effectively reduces the risk of at least one sign or symptom of TDP-43-related disease, lessens its severity, or delays its onset. Specifically, the regimen preferably effectively inhibits or delays TDP-43 aggregates (e.g., human TDP-43 aggregates) in the brain, and / or inhibits or delays its toxic effects and / or inhibits / delays the development of behavioral deficits.

[0199] In therapeutic applications, antibodies or antigen-binding antibody fragments are administered to patients suspected of or already suffering from the disease in a regimen (dosage, frequency, and route) that effectively improves or at least inhibits further exacerbation of at least one sign or symptom of the disease (e.g., ALS). Specifically, the regimen preferably effectively reduces or at least inhibits further increases in cytoplasmic levels of TDP-43 (e.g., human TDP-43) and / or aggregates formed therefrom, associated toxicity, and / or behavioral deficits.

[0200] The regimen is considered effective in treatment or prevention if the outcome achieved by an individual patient is more favorable than the average outcome in a control group of comparable patients who were not treated using the methods disclosed herein.

[0201] VII. Treatment Plan As used herein, the terms “treat” and “treatment” mean to reduce or improve one or more symptoms or effects associated with a disease, to prevent, suppress or delay the onset of one or more symptoms or effects of a disease, to reduce the severity or frequency of one or more symptoms or effects of a disease, and / or to increase or tend toward the desired outcome as described herein.

[0202] The expected outcomes of the treatments disclosed herein vary depending on the TDP-43-related disease and patient profile, and are readily determined by those skilled in the art. Expected outcomes include improvements in the patient's health. Typically, expected outcomes include measurable indicators such as reductions or clearance of pathological ALS, FTLD-TDP, primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease.

[0203] Therefore, this article provides a method for inhibiting or reducing the accumulation of TDP-43 (e.g., human TDP-43) in subjects who have TDP-43-related diseases or are at risk of developing such diseases, comprising administering to the subject an effective amount of any of the antibodies or antigen-binding antibody fragments described herein, thereby inhibiting or reducing the accumulation of TDP-43 in the subject (i.e., the patient).

[0204] This article also provides methods for treating or achieving prevention of TDP-43-related disease in subjects, comprising administering a therapeutically effective amount of any of the antibodies or antigen-binding antibody fragments described herein, thereby treating or achieving prevention of TDP-43-related disease.

[0205] In some implementations, TDP-43-related diseases include ALS, FTLD-TDP, primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease. In some implementations, TDP-43-related diseases include ALS.

[0206] This article also provides a method for detecting TDP-43 deposits (e.g., human TDP-43 deposits) in individuals with or at risk of developing TDP-43-related diseases, comprising administering any of the antibodies or antigen-binding antibody fragments described herein to a subject, and detecting antibodies in the subject that bind to TDP-43.

[0207] In some embodiments, antibodies or antigen-binding antibody fragments are administered via intravenous injection into the subject's body. In some embodiments, the antibodies or antigen-binding antibody fragments are labeled. In some embodiments, the antibodies or antigen-binding antibody fragments are labeled with fluorescent, paramagnetic, or radioactive labels. In some embodiments, radioactive labeling is detected using positron emission tomography (PET) or single-photon emission computed tomography (SPECT).

[0208] Antibodies or antigen-binding antibody fragments are administered in an effective regimen, meaning the dosage, route of administration, and frequency of administration for delaying the onset of at least one sign or symptom of the disease being treated, reducing its severity, inhibiting further deterioration, and / or improving the condition. If the patient already has the disease, the regimen may be called a therapeutically effective regimen. If the patient is at an elevated risk of the disease relative to the general population but has not yet experienced symptoms, the regimen may be called a preventatively effective regimen. In some cases, therapeutic or preventative efficacy may be observed in the same patient relative to a historical control or past experience. In other cases, therapeutic or preventative efficacy may be demonstrated in a preclinical or clinical trial population relative to a control group of untreated patients.

[0209] Administration can be done parenterally, intravenously, orally, subcutaneously, intra-arterially, intracranially, intrathecally, intraperitoneally, locally, intranasally, or intramuscularly. Some antibodies can be administered into the systemic circulation via intravenous or subcutaneous administration.

[0210] Antibody or antigen-binding antibody fragment formulations can be administered intravenously or subcutaneously in a dose range of about 0.5 mg / kg to about 30 mg / kg of host body weight. For example, doses may be about 0.5 mg / kg body weight, about 1.0 mg / kg, about 1.5 mg / kg, about 2.0 mg / kg, about 4.0 mg / kg, about 5.0 mg / kg, about 8.0 mg / kg, about 10 mg / kg, about 15 mg / kg, about 16 mg / kg, about 20 mg / kg, about 24 mg / kg, about 25 mg / kg, or about 30 mg / kg body weight. Dosages may also be based on body surface area, ranging from about 0.5 mg / m². 2 Approximately 500 mg / m 2 (e.g., 0.5, 5, 10, 50, 100, 250 or 500 mg / m²) 2 Administration. For intravenous administration, an amount of antibody or antigen-binding antibody fragment formulation sufficient to achieve the individual patient's desired dose is transferred from one or more vials into one or more intravenous bags containing liquid (e.g., saline) and administered to the patient.

[0211] Antibodies or antigen-binding antibody fragments are typically administered on multiple occasions. The frequency of administration depends on factors such as the half-life of the antibody or antigen-binding antibody fragment in circulation, the patient's condition, and the route of administration. The frequency can be daily, weekly, monthly, quarterly, or at irregular intervals in response to changes in the patient's condition or the progression of the disease being treated.

[0212] Exemplary treatment regimens require administration once every two weeks, once a month, or once every 3 to 6 months. The frequency of administration depends on whether the condition is acute or chronic and the condition's response to treatment. The dosing frequency can be adjusted based on the pharmacokinetic profile of the antibody preparation in the patient. For example, the half-life of the antibody or antigen-binding antibody fragment may ensure a dosing frequency of two weeks. In some embodiments disclosed herein, the antibody or antigen-binding antibody fragment is administered to the patient for at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, 5 years, 10 years, or for the patient's lifetime.

[0213] Normal levels of TDP-43 (e.g., human TDP-43) can be represented by a sample of individuals in a general population who have not been diagnosed with a specific TDP-43-related disease (e.g., ALS) and are not considered to be at high risk of developing such a disease. ，The level can be determined in the brain of a representative sample from a disease-free individual under 50 years of age. Alternatively, if, according to the method of the invention, the PET signal in a brain region known to develop TDP-43 aggregates (e.g., human TDP-43 aggregates) is indistinguishable (within measurement accuracy) from a brain region known not to normally form such deposits, then normal levels can be identified in an individual patient. Elevated levels in an individual can be compared with normal levels. ( For example ， The TDP-43 aggregates are identified by comparing the variance of the external mean and standard deviation, or by comparing them with unknown regions associated with the deposits, using elevated signals exceeding experimental error only from brain regions associated with TDP-43 aggregates (e.g., human TDP-43 aggregates). For the purpose of comparing the levels of TDP-43 aggregates (e.g., human TDP-43 aggregates) in individuals and populations, TDP-43 aggregates should preferably be measured in the same regions of the brain, including at least one region known to form TDP-43 aggregates associated with a specific disease (e.g., ALS) (e.g., in the cytoplasm).

[0214] Patients with elevated levels of TDP-43 aggregates (e.g., human TDP-43 aggregates) are candidates for initiating immunotherapy. A decrease in TDP-43 aggregate levels (e.g., human TDP-43 aggregates) after initiation of immunotherapy can initially be considered an indication of the desired therapeutic effect. The observed decrease may, for example, be in the range of 1–100%, 1–50%, or 1–25% of baseline values. Such effects can be measured in one or more regions of the brain where deposits are known to form, or may be measured by the average of such regions. The overall therapeutic effect can be approximated by adding the percentage reduction relative to baseline to the increase in TDP-43 aggregates (e.g., human TDP-43 aggregates), which would otherwise occur in untreated patients.

[0215] Maintaining TDP-43 aggregates (e.g., human TDP-43 aggregates) at a substantially constant level or even a slight increase in TDP-43 aggregates (e.g., human TDP-43 aggregates) can also indicate a response to treatment, albeit a suboptimal one. Such responses can be compared to the time course of TDP-43 aggregate (e.g., human TDP-43 aggregate) levels in patients with an untreated specific disease (e.g., ALS) to determine whether immunotherapy has an effect in inhibiting further increases in TDP-43 aggregates (e.g., human TDP-43 aggregates).

[0216] VIII. Reagent Kit This disclosure further provides kits (e.g.) ， Containers containing any of the antibodies disclosed herein or their antigen-binding antibody fragments and related materials, such as instructions for use (e.g. ，(Packaging insert). Instructions for use may contain, for example, instructions regarding the administration of the antibody or antigen-binding antibody fragment and optionally one or more other agents. The antibody may be packaged in unit dose, bulk packaging (e.g., ...). ， (Multi-dose packaging) or subunit dose.

[0217] IX. Detection Methods In some aspects, the antibodies of this disclosure further provide a method for detecting TDP-43 in a sample. For example, in some embodiments, this disclosure provides a method for detecting TDP-43 in a sample, which includes contacting the sample with an antibody or antigen-binding fragment of this disclosure. For example, such a method may be an ex vivo or in vitro method. In some embodiments, the sample is a biological sample derived from a subject (e.g., a human subject). In some embodiments, the subject is a human. In some embodiments, the subject is a patient with a TDP-43-related disease or at risk of having said disease. In such cases, a sample (such as cells, tissues, and / or organs) removed from the patient is exposed to the antibody or antigen-binding fragment described herein. In some embodiments, the sample comprises cells derived from the patient, and the cells are lysed prior to administration of the antibody or antigen-binding fragment described herein.

[0218] Example The following embodiments are included to illustrate the patterns disclosed herein. Certain aspects of the following embodiments are described in accordance with techniques and procedures that have been discovered or anticipated by the co-inventors of the invention to work well in the practices disclosed herein. Based on this disclosure and the general level of skill of those skilled in the art, it will be understood that the following embodiments are intended to be exemplary only and that various variations, modifications, and alterations may be made without departing from the scope of this disclosure.

[0219] Example 1. Humanized anti-TDP-43 antibody Humanized anti-TDP-43 antibody was generated from mouse monoclonal antibody 13D3. Specifically, Figure 1 Show the annotation form of the heavy-chain variable structural domain and Figure 2 The annotation format of the light chain variable domain is shown. Both the heavy chain variable domain and light chain variable domain sequences show the native signal peptide, the variable domain, and a partially constant domain. The partially heavy chain constant domain region corresponds to the IgG2a mouse constant domain. The partially light chain constant domain corresponds to the mouse κ constant domain. References to amino acid substitutions used herein refer to the Kabat numbering system (see, e.g., Kabat EA et al.). Sequences of Proteins of Immunological Interest (5th ed.). Bethesda, MD: National Institutes of Health (1991)).

[0220] Protein sequences were identified in the Protein Data Bank (PDB) database (see Deshpande et al., The RCSB Protein Data Bank: a redesigned query system and relational database based on the mmCIF schema). Nucleic Acids Research , 33:D233-D237 (2005)) to search for structures that would provide a similar structural model of 13D3. The crystal structure of the antibody Fab PDB code “5BK5”, which is a human germline antibody. Based on its overall sequence similarity with 13D3 VH and VK, resolution quality (3.0 Å), 5BK5 was selected for both VH and VK structures, and it retains the same classic structure for the complementarity-determining region (CDR) loop (Scally et al., Crystal structure of anti-cirumsporozoite protein 663 germline antibody; Directdeposit to PDB (2017)).

[0221] In addition, since the 5BK5 antibody is a human lineage-derived antibody and belongs to the same classical category as human lineage IGHV3-48'03 (SEQ ID NO: 2) for the variable heavy chain domain and the same classical category as human lineage IGKV2-30*02 (SEQ ID NO: 25) for the variable light chain domain, these sequences were used as the human receptor framework.

[0222] Therefore, the framework regions of 5BK5 VH and 5BK5 VL were selected as the receptor sequences for the 13D3 CDR. 13D3 CDR models transplanted onto the corresponding human frameworks of VH and VL were established and used as a guide for further reversal mutations to improve binding specificity and reduce immunogenicity.

[0223] More specifically, the amino acid sequence consisting of the 5BK5 VH human framework and 13D3 CDR is named hu13D3VHv1d (SEQ ID NO: 20), and the amino acid sequence consisting of the 5BK5 VL human framework and 13D3 VL CDR is named hu13D3VLv1d (SEQ ID NO: 47).

[0224] The additional forms hu13D3VH and hu13D3VL were designed to enable the assessment of the contributions of various framework residues to antigen binding, thermostability, exploitability (e.g., deamination, oxidation, N-glycosylation, proteolysis, and aggregation), and immunogenicity. The substitution positions were considered based on several factors, including the position defining the classical CDR conformation (see Martin, ACR, Protein sequence and structure analysis of antibody variable domains, In: Kontermann R and Dübel S (eds.)). Antibody Engineering Heidelberg, Germany: Springer International Publishing AG (2010); Position within the vernier area (see Foote J. and Winter, G., Antibody framework residues affecting the conformation of the hypervariable loops). J Mol Biol 224(2):487-99 (1992)); located at the VH / VL domain interface (see Léger OJP and Saldanha, J. Preparation of recombinant antibodies from immune rodent spleens and the design of their humanisation by CDRgrafting. In: Shepherd P and Dean C (eds.) Monoclonal Antibodies: A Practical Approach Oxford, UK: Oxford University Press (2000); locations readily susceptible to post-translational modifications (such as glycosylation or pyroglutamylation); locations occupied by residues predicted to interact with the CDR, based on 13D3 CDR models transplanted to the VH and VL frameworks; and / or locations occupied by residues rare in sequenced human antibodies, where parental mouse 13D3 residues or other residues are much more prevalent in the human antibody lineage.

[0225] The following is an overview of humanized 13D3 antibodies.

[0226] Heavy chain variable structural domain hu13D3VHvd1 (SEQ ID NO: 20) consists of CDR-H1, CDR-H2, and CDR-H3 loops of 13D3-VH transplanted onto a 5BK5 VH framework, and restores all framework substitutions at key locations defining the classic Chothia class. These substitutions are part of the vernier region and are located at the VH / VL domain interface or contribute to structural stability. hu13D3VHvd1 includes substitutions that are reversion mutations of germline antibodies and are most common at positions L5V and T77S.

[0227] hu13D3VHv2d (SEQ ID NO: 21) includes the following substitution: L78A. As indicated by the Immunoeptope Database (“IEDB”) analysis, leucine at position 78 is immunogenic. Therefore, deimmunization analysis predicts reduced immunogenicity with an alanine substitution at position 78.

[0228] hu13D3VHv3d (SEQ ID NO: 22) includes the following substitutions: G44R, S49A, and S74A. Arginine at position 44 undergoes a G100D (glycine to aspartic acid) substitution with the variable light chain domain to form... From the beginning Contact is established, thereby strengthening the heavy chain variable domain:light chain variable domain interface. Specifically, arginine at position 44 forms a hydrogen bond and salt bridge with G100D, and also forms a hydrogen bond with F98 in the variable light chain domain. Position 49 is a vernier residue, and the substitution from serine to alanine is a reversion mutation to assess the effect on CDR conformation and binding. Finally, the substitution from serine to alanine at position 74 is a germline substitution.

[0229] Hu13D3VHv4d (SEQ ID NO: 23) includes various substitutions made in the attempted combinations of hu1353VHv1d, hu1353VHv2d and hu1353VHv3d.

[0230] Light chain variable structural domain Hu13D3VLv1d (SEQ ID NO: 47) consists of CDR-L1, CDR-L2, and CDR-L3 rings of 13D3VL transplanted onto a 5BK5 VL frame and restores all frame substitutions at key locations defining the Chothia classical class, which are part of the vernier region and located at the VH / VL structural domain interface. Hu1353VLv1d includes the following substitutions: V3Q, P15L, E17Q, L38Q, K39R, Q100D, and L104V.

[0231] Replacing glutamine with valine at position 3 reduces the immunogenicity of the light chain variable domain. Replacing proline with leucine at position 15 is a germline substitution. Replacing glutamate with glutamine at position 17 is a rare substitution. Position 17 has significant surface exposure, resulting in negatively charged plaques on the protein surface. Substitution with glutamine reduces positive plaques on the antibody surface. Because glutamine in the mouse structural model forms interchain contacts within the light chain variable domain to maintain structural conformation, leucine is replaced with glutamine at position 38. In contrast, leucine residues do not form the same contacts; therefore, this reversion mutation improves conformational stability. Replacing lysine with arginine at position 39 improves conformational stability by forming additional contacts with adjacent residues that lysine residues cannot. Position 100 in the light chain variable domain is located at the interface between the heavy chain variable domain and the light chain variable domain; however, it does not form interchain contacts with glutamine residues. Substitution at position 44 of the heavy chain variable domain with aspartic acid and simultaneous substitution with arginine produces a stronger de novo contact and thus improves the antibody's thermal stability. Substitution of leucine at position 104 with valine is predicted to decrease immunogenicity.

[0232] hu13D3VLv2d (SEQ ID NO: 48) consists of the substitutions described above in hu13D3VLv1d, but also includes the following substitution: L92A. Since leucine at position 92 is predicted to be immunogenic, substitution with alanine at position 92 reduces the immunogenicity of the variable light chain domain.

[0233] Tables 1 and 2 below show the sequence alignments of the 13D3 variable heavy chain domain and the variable light chain domain compared to the humanized form, respectively.

[0234] Table 1.13 Sequence Alignment of D3 Humanized Variable Heavy Chain Domain

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241] Table 2. Sequence alignment of the 13D3 humanized variable light chain

[0242]

[0243]

[0244]

[0245]

[0246]

[0247] Specifically, for Vk, the human κ light chain (SEQ ID NO: 26) with NCBI accession code ABC66863 (Shringer et al., 2006) was selected. This κ light chain has the same classical category as CDR-L1 and L2, and according to IMGT convention, belongs to the hominid IGKV2-30*02 (SEQ ID NO: 25). For VH, the human Ig heavy chain AEX28899 (SEQ ID NO: 3) (GenBank: AEX28899) (Bowers et al., 2014) was selected, which also has the same classical category and belongs to the hominid IGHV3-48'03. It is a member of Kabat human heavy subgroup 3.

[0248] The AEX28899 and ABC66863 antibodies are human-derived antibodies, belonging to the same classical class and, for the heavy chain variable domain, to the human IGHV3-48'03 (SEQ ID NO: 2), and for the light chain variable domain, to the human IGHV3-48*03 (SEQ ID NO: 25). Therefore, the heavy chain variable domain sequences of AEX28899 and the light chain variable domain sequences of ABC66863 were used as the human receptor framework for the 13D3 CDR. 13D3 CDR models transplanted onto the corresponding human frameworks in VH and VL were established and used as guidance for further reversion mutations.

[0249] Humanized versions of the AEX28899 heavy chain variable domain and ABC66863 light chain variable domain sequences were used as the human receptor framework for the 13D3 CDR and were designed to allow for the assessment of the contributions of various framework residues to antigen binding, thermostability, exploitability (e.g., deamination, oxidation, N-glycosylation, proteolysis, and aggregation), and immunogenicity. The placement of substitutions was considered based on several factors, including the position defining the classical CDR conformation (see Martin, ACR, Protein sequence and structure analysis of antibody variable domains, In: Kontermann R and Dübel S (eds.)). Antibody Engineering Heidelberg, Germany: Springer International Publishing AG (2010); Position within the vernier region (see Foote J. and Winter, G., Antibody framework residues affecting the conformation of the hypervariable loops). J Mol Biol 224(2):487-99 (1992)); located at the VH / VL domain interface (see Léger OJP and Saldanha, J. Preparation of recombinant antibodies from immunerodent spleens and the design of their humanisation by CDR grafting. In: Shepherd P and Dean C (eds.) Monoclonal Antibodies: A Practical Approach Oxford, UK: Oxford University Press (2000); locations readily susceptible to post-translational modifications (such as glycosylation or pyroglutamylation); locations occupied by residues predicted to interact with the CDR, based on 13D3 CDR models transplanted to the VH and VL frameworks; and / or locations occupied by residues rare in sequenced human antibodies, where parental mouse 13D3 residues or other residues are much more prevalent in the human antibody lineage.

[0250] The following is an overview of the VH domain substitutions in SEQ ID NO: 3: R19K: K19 in the mouse antibody structure generates a π-cation interaction with W79 in SEQ ID NO: 3; G44R: Replacing Gly with Arg at this position potentially establishes a contact with the light chain, thereby improving antibody stability; S77T: Threonine reduces immunogenicity; L78A: Leucine is predicted to be immunogenic, therefore substitution with alanine reduces the predicted immunogenicity; L78G: Glycine also reduces the predicted immunogenicity; L80A: Removal of leucine from the stated position reduces immunogenicity; L80G: Similar to alanine, glycine at position 80 reduces immunogenicity; L82cG: Glycine potentially reduces heavy chain immunogenicity; and R83M: A reversion mutation that can confer antibody stability.

[0251] The following is an overview of the VL domain substitutions in SEQ ID NO: 26: I2V is a vernier domain residue, valine is retained to maintain the CDR conformation; L9S: Leucine is predicted to be immunogenic, and the substitution with serine reduces the predicted immunogenicity; P18Q: Glutamine in the mouse antibody model forms an H bond (LC) with K74, thereby stabilizing the interchain interaction; R46L: is an interface and vernier domain residue, therefore Leu is retained; A80S: Serine reduces the predicted immunogenicity; L92G: L92 is a CDR residue and is predicted to be immunogenic, and glycine residues are predicted to reduce immunogenicity; V94I: Valine at position 94 is predicted to have a low level of immunogenicity, and isoleucine is predicted to reduce immunogenicity; and V94A: Alanine is a predicted immunogenicity-reducing alternative substitution.

[0252] Example 2. Characterization of humanized 13D3 antibody variants As discussed in this paper, cytoplasmic aggregates of TDP-43 are found in many neurodegenerative diseases, particularly ALS. Typically, the individual TDP-43 proteins found in cytoplasmic aggregates are phosphorylated at serine residues at positions 409 and / or 410. A binding screening assay was established to determine whether 13D3 and its humanized variants selectively bind to the phosphorylated TDP-43 found in cytoplasmic aggregates.

[0253] Approximately 50 RU of biotin-TDP43 protein was immobilized on a CM3 chip along with NeutrAvidin. Various humanized and chimeric 13D3 antibodies were flowed through the chip, and their binding to phosphorylated TDP43 protein was analyzed. Specifically, hu13D3H9L8, hu13D3H10L6, hu13D3H10L7, hu13D3H10L8, and hu13D3H9L7 were analyzed, showing similar binding to TDP43 phosphate as the chimeric 13D3 antibody. Figure 3 Similarly, the binding of hu13D3Hd1Ld1, hu13D3Hd2Ld2, hu13D3Hd3Ld1, hu13D3HD4LD1, hu13D3HD1LD2, hu13D3HD2LD2, hu13D3HD3LD2, and hu13D3HD4LD2 to TDP43 phosphate was tested. Each variant showed similar binding to TDP43 phosphate as the chimeric 13D3 antibody. Figure 4 ).

[0254] In another experiment, the antibody was immobilized on a protein A chip (approximately 2000 RU), and a TDP-43 peptide with 23 amino acids phosphorylated at positions 409 and 410 (“TDP-43 peptide (pS409 / pS410)”) was flowed through the chip at concentrations between 1 nM and 100 nM (SEQ ID NO: 83). As a control, TDP-43 peptide with 23 amino acids not phosphorylated at positions 409 or 410 was also tested between 12 nM and 1 μM. Specifically, the humanized form hu13D3H5L2 showed a 1:1 binding to various concentrations (1.2345 nM; 3.703 nM; 11.111 nM; 33.33 nM; and 100 nM) of TDP-43 peptide (pS409 / pS410), similar to that of the 13D3 murine antibody (…). Figure 5 Furthermore, hu13D3H5L2 did not show affinity for the unphosphorylated 22 amino acid TDP-43 peptide at 12.345 nM, 37.03 nM, 111.111 nM, 333.333 nM and 1 μM (data not shown).

[0255] Thermostability of the antibody was also assessed. Thermostability analysis was performed using differential scanning calorimetry, a method characterizing the stability of proteins or other biomolecules (e.g., antibodies or their antigen-binding fragments). 2.66 mM (0.4 mg / mL) antibody was tested in 1xPBS at pH 7.4. The test temperature range was 25°C to 100°C.

[0256] Data on the binding of chimeric 13D3 and its humanized forms are summarized in Tables 3 and 4. Tables 3 and 4 also summarize thermal stability and immunogenicity data.

[0257] Table 3. Measurement of humanized anti-TDP-43

[0258] Table 4. Measurement of humanized 13D3 antibody

[0259] In summary, the data shown in Tables 3 and 4 are exemplary humanized antibodies with low predicted immunogenicity scores relative to mouse 13D3 antibodies (explained in further detail below). The data further demonstrate the high yields and melting temperatures of the humanized antibodies (e.g., similar to mouse 13D3 antibodies). See, for example, the antibodies h13D3Hd1-Ld1, h13D3Hd1-Ld2, and h13D3Hd2-Ld1 shown in Table 4. Furthermore, the data also demonstrate humanized antibodies with similar binding metrics to mouse 13D3 antibodies.

[0260] Computer-simulated immunogenicity analysis of humanized antibodies The immunogenicity scores shown in Tables 3 and 4 are predicted immunogenicity values ​​calculated by two different software programs: the Immune Epitope Database (IEDB) and EpiQuest. The IEDB immunogenicity analysis tool is sponsored by the National Institute of Allergy and Infectious Diseases.

[0261] For IEBDB, the immunogenicity prediction method used is based on the predicted potential binding of peptides within the protein sequence to major histocompatibility class (MHC) II. The program identifies potential immunogenic regions within the protein sequence. The MHC II tool uses a broad spectrum of MHC II alleles in the human population (i.e., 26 reference alleles) to predict immunogenic regions. The software generates a series of 15-residue peptides overlapping at 10 residues. The generated 15-residue peptides are predicted to bind to the 26 reference alleles.

[0262] For each peptide, a percentile rank is generated for each of three methods (combinatorial library, SMM_align, and Sturniolo) by comparing the score of each peptide with the scores of five million randomly selected 15-residue peptides from the SWISSPROT database. The adjusted percentile rank is a frequency-adjusted percentile rank based on peptide length. A low number percentile rank indicates high affinity. The median percentile rank of the three methods is then used to generate a rank for the common method. By default, the prediction results are collapsed to display only the percentile rank and the adjusted rank. The maximum median percentile rank threshold is set to 20.

[0263] The EpiQuest T-Scanner tool classifies cytotoxic T-lymphocyte (CTL) epitopes based on their predicted immunodominance. Immunodominance of an epitope is defined as its relative strength in functional assays related to the target killing or release of the corresponding cytokine. These parameters indicate the function of the T-eptope. This procedure is designed to analyze and sort CTL peptide epitopes eluted from target cells based on their immunodominance. Typically, only a few peptide epitopes binding to MHC class I possess actual functional activity. The relative strength of a CTL(T) epitope is defined by its binding strength to MHC class I and T-cell receptors (TCRs) (in the case of MHCI). The algorithm detects the structural and compositional characteristics of the peptide epitope, which enable it to elicit a high affinity for the TCR. This analysis is haplotype-specific, and the EpiQuest T-scanner has a matrix for analyzing HLA-A2 and H2kB haplotype-binding peptides.

[0264] Example 3. Binding of anti-TDP-43 antibody to phosphorylated cytoplasmic aggregates of TDP-43 in FTP brain tissue and model systems Figures 6A to 6C Displaying brain tissue in frontotemporal dementia (“FTD”) Figure 6A ) and healthy brain tissue ( Figure 6C ). Figure 6B for Figure 6A The illustration shows the co-localization of the 13D3 antibody with neuronal cytoplasmic aggregates associated with phosphorylated TDP-43 FTD. The data indicate that the 13D3 antibody specifically binds to cytoplasmic aggregates in FTD brain tissue, but not in healthy brain tissue. Similarly, Figures 7A to 7C Each rNLS8 dox, which showed TDP-43 protein disease, could inhibit 13D3-specific binding cytoplasmic aggregates in the model.

[0265] Example 4. Binding of anti-TDP-43 antibody to phosphorylated cytoplasmic aggregates and nuclear TDP-43 aggregates in transfected HEK cells. Figure 8A Confocal micrographs showing HEK cells transiently transfected with GFP-2a-TDP43 [mNLS (R82L / K83Q) DCS (C173S / C175S)] or GFP only (2a being a self-cleaving peptide that releases TDP-43 upon expression). The top image shows staining of GFP, cell nuclei (gray), and pTDP-43 (white). The bottom image shows only pTDP-43 staining, which was not present in the GFP-only control transfection. Figure 8B (Left) A graph showing cell counts in HEK cells transfected with GFP-2a-TDP43 or GFP alone. The data demonstrate that the cell counts were approximately equal between the two populations. Furthermore, Figure 8B (Right) Shows the pTDP-43 lesion count in HEK cells transfected with GFP-2a-TDP43 or GFP alone. The data indicate that TDP-43 lesions only form in HEK cells transfected with GFP-2a-TDP43. Figure 8C TDP-43 staining was performed using commercially available antibodies or antibodies disclosed herein (including 13D3, 13C13, and 2D4). Cells were treated with the antibody for 24 hours, and then incubated with 100 μg / ml antibody for another 24 hours. Cells were then washed, fixed / permeabilized, and stained with anti-pTDP-43 antibody, followed by staining with AF647-conjugated anti-mouse secondary antibody. The stained cells were imaged using a 40x water objective via high-content imaging (Operetta system). Quantitative analysis was performed using Harmony software.

[0266] Antibodies 13D3, 13C13, and 2D4 were used to detect mislocalized TDP-43 overexpression in HEK cells. The top row shows transfection with GFP-2a-TDP-43, where phosphorylated TDP43 does not include the nuclear localization signal (i.e., phosphorylated TDP43 is retained in the cytoplasm), and the bottom row shows transfection with the GFP-only construct. The data indicate that TDP-43 lesions only form in HEK cells transfected with GFP-2a-TDP-43.

[0267] The assay included various control antibodies, including pTDP-43 (Cosmo)+, pTDP-43 (1D3)+, total TDP-43 (PT), and 3B12 (ED)+. The control antibodies validated TDP-43 aggregation in the cytoplasm. Similarly, antibodies 13D3, 13C13, and 2D4 were tested and showed similar binding to cytoplasmic aggregates of phosphorylated TDP-43.

[0268] References

[0269]

[0270] Sequence Appendix Variable heavy chain reference sequence SEQ ID NO: 1 m13D3-VH EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYFMSWVRQTPEKRLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTLYLQM NSL MSEDTAMYFCARQTYYSYGGFPYWGQGTLVTVSA SEQ ID NO: 2 IGHV3-48*03 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYEMNWVRQAPGKGLEWVSYIS S SGSTIYYADSVKGRFTISRDNAKNSLYLQM NSL RAEDTAVYYCARYFDYWGQGTLVTVSS SEQ ID NO: 3 AEX28899 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYEMNWVRQAPGKGLEWVSYIS S SGSTIYYADSVKGRFTISRDNAKNSLYLQM NSL RAEDTAVYYCARRNYYDS GGY GHWGQGTLVTVSS Humanized 13D3 VH design sequence SEQ ID NO: 4 h13D3VHv1 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMSWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNSLYLQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 5 h13D3VHv2 EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYFMSWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNSLYLQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 6 h13D3VHv3 EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYFMSWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNSLYAQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 7 h13D3VHv4 EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYFMSWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTLYAQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 8 h13D3VHv5 EVQLVESGGGLVQPGGSLKLSCAASGFTFSNYFMSWVRQAPGKGLEWVAYIS TGGDSANYADNVKGRFTISRDNAKNTAYAQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 9 h13D3VHv6 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMSWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTAYAQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 10 h13D3VHv7 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTAYAQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 11 h13D3VHv8 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTLYGQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 12 h13D3VHv9 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGKGLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTGYLQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 13 h13D3VHv10 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGKGLEWVAYIST GGDSANYADNVKGRFTISRDNAKNTLYLQM NSG RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 14 h13D3VHv11 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGKRLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTGYLQM NSL MAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 15 h13D3VHv12 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGERLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTGYLQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 16 h13D3VHv13 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMGWVRQAPGEDLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNTGYLQM NSL RAEDTAVYFCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 17 h13D3VHv1b ELVQSGAEVKKPGSSVKVSCKASGFTFSNYFMSWVRQAPGQGLEWVAYISTGGDSANYADNVKGRFTITRDNSTSTLYMELSSLRSEDTAVYFCARQTYYSYGGFPYWGQGTLVTVSS SEQ ID NO: 18 h13D3VHv2b ELVQSGAEVKKPGSSVKVSCKASGFTFSNYFMSWVRQAPGQGLEWVAYISTGGDSANYADNVKGRFTITKDTSTSTLYMELSSLRSEDTAVYFCARQTYYSYGGFPYWGQGTLVTVSS SEQ ID NO: 19 h13D3VHv3b ELVQSGAEVKKPGSSVKVSCKASGFTFSNYFMSWVRQAPGQGLEWVAYISTGGDSANYADNVKGRGTITKDTSTSTLYMELSSLRSEDTAVYFCARQTYYSYGGFPYWGQGTLVTVSS SEQ ID NO: 20 h13D3VHv1d EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMSWVRQAPGKGLEWVSYIS T GGDSANYADNVKGRFTISRDNSKNSLYLQM NSL RAEDTAVYYCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 21 h13D3VHv2d EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMSWVRQAPGKGLEWVSYIS T GGDSANYADNVKGRFTISRDNSKNTAYLQM NSL RAEDTAVYYCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 22 h13D3VHv3d EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMSWVRQAPGKRLEWVAYIS T GGDSANYADNVKGRFTISRDNAKNSLYLQM NSL RAEDTAVYYCARQTYYSY GGF PYWGQGTLVTVSS SEQ ID NO: 23 h13D3VHv4d EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYFMSWVRQAPGKRLEWVSYIST GGDSANYADNVKGRFTISRDNSKNTAYLQM NSL RAEDTAVYYCARQTYYSY GGF PYWGQGTLVTVSS The underlined residues are in the positions defined below.

[0271] Position 52A*# Location 82A*# Location 82B*# Location 82C*# Location 100A* Location 100B* Location 100C*.

[0272] * Applicable to SEQ ID NO: 1, 3-16 and 20-23 # Applicable to SEQ ID NO: 2 Variable light chain reference sequence SEQ ID NO: 24 m13D3-VL DVVMTQSPLSLPVSLGDQASISCRSSQ SLVHS NGKTYLHWYQQKPGQSPKLLIYKVSDRYSGVSDRFSGSGSGTDFTLKISRVETEDLGVYFCSQSLHVPLTFGAGTKLELK SEQ ID NO: 25 IGKV2-30*02 DVVMTQSPLSLPVTLGQPASISCRSSQ SLVHS DGNTYLNWFQQRPGQSPRRLIYKVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGTHWPYTFGQGTKLEIK SEQ ID NO: 26 ABC66863 DIVMTQSPLSLPVTLGQPASISCRSSQ SLVYS DGNTYLNWFQQRPGQSPRRLIYKVSNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGTHRPLTFGGGTKVEIK Humanized 13D3 VL Design Sequence SEQ ID NO: 27 h13D3VLv1 DVVMTQSPLSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 28 h13D3VLv2 DVVMTQSPLSLPVTLGQQASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 29 h13D3VLv3 DVVMTQSPSSLPVTLGQQASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 30 h13D3VLv4 DVVMTQSPSSLPVTLGQQASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSGHVPLTFGGGTKVEIK SEQ ID NO: 31 h13D3VLv5 DVVMTQSPSSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSGHVPLTFGGGTKVEIK SEQ ID NO: 32 h13D3VLv6 DVVMTQSPSSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 33 h13D3VLv7 DVVMTQSPSSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSLHIPLTFGGGTKVEIK SEQ ID NO: 34 h13D3VLv8 DVVMTQSPSSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSLHAPLTFGGGTKVEIK SEQ ID NO: 35 h13D3VLv9 DVQMTQSPSSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSGHVPLTFGDGTKVEIK SEQ ID NO: 36 h13D3VLv10 DVQMTQSPSSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVESEDVGVYFCSQSGHVPLTFGRGTKVEIK SEQ ID NO: 37 h13D3VLv1b EVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTDFTLTISSLEPEDFAVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 38 h13D3VLv2b EVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEFTLTISSLEPEDFAVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 39 h13D3VLv3b EVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEVTLTISSLEPEDFAVYFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 40 h13D3VLv4b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTDFTLTISSLEPEDFAVKFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 41 h13D3VLv5b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEFTLTISSLEPEDFAVKFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 42 h13D3VLv6b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEVTLTISSLEPEDFAVKFCSQSLHVPLTFGGGTKVEIK SEQ ID NO: 43 h13D3VLv7b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEFTLTISSLEPEDFAVYFCDQSLHVPLTFGGGTKVEIK SEQ ID NO: 44 h13D3VLv8b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHSNGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEVTLTISSLEPEDFAVYFCDQSLHVPLTFGGGTKVEIK SEQ ID NO: 45 h13D3VLv9b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEFTLTISSLEPEDFAVYFCSQQLHVPLTFGGGTKVEIK SEQ ID NO: 46 h13D3VLv10b DVVMTQSPATLSLSPGERATLSCRSSQ SLVHS NGKTYLHWYQQKPGQAPRLLIYKVSDRYSGVPARFSGSGSGTEVTLTISSLEPEDFAVYFCSQQLHVPLTFGGGTKVEIK SEQ ID NO: 47 h13D3VLv1d DVQMTQSPLSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFCSQSLHVPLTFGDGTKVEIK SEQ ID NO: 48 h13D3VLv2d DVQMTQSPLSLPVTLGQPASISCRSSQ SLVHS NGKTYLHWYQQRPGQSPRLLIYKVSDRYSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFCSQSAHVPLTFGDGTKVEIK The underlined residues are in the positions defined below.

[0273] Specifically, S represents position 27A. L represents position 27B. V is at position 27C H represents position 27D S represents position 27E.

[0274] Applicable to SEQ ID NO: 24-48.

[0275] SEQ ID NO: 49 HC CDR1 #1 GFTFSNYFMS SEQ ID NO: 50 HC CDR1 #2 GFTFSNYFMG SEQ ID NO: 51 HC CDR2 #1 YISTGGDSANYADNVKG SEQ ID NO: 52 HC CDR3 #1 QTYYSYGGFPY SEQ ID NO: 53 LC CDR1 #1 RSSQSLVHSNGKTYLH SEQ ID NO: 54 LC CDR2 #1 KVSDRYS SEQ ID NO: 55 LC CDR3 #1 SQSLHVPLT SEQ ID NO: 56 LC CDR3 #2 SQSGHVPLT SEQ ID NO: 57 LC CDR3 #3 SQSLHIPLT SEQ ID NO: 58 LC CDR3 #4 SQSLHAPLT SEQ ID NO: 59 LC CDR3 #5 DQSLHVPLT SEQ ID NO: 60 LC CDR3 #6 SQQLHVPLT SEQ ID NO: 61 LC CDR3 #7 SQSAHVPLT TDP43 mouse antibody variable domain sequence SEQ ID NO: 62 1B3_JH140 VH signal peptide MNFGLSLIFLVLVLKGVLC SEQ ID NO: 63 1B3_JH140 Variable Restructuring Domain EVKLVESGGGLVQPGGSLKLSCAASGFTFSSYTMSWVRQTPEKRLELVAEISNSGGRTNYPDTVKGRFTISRDNAKNTLYLQMSSLKSEDTAMYYCARRRYSDYYYYAMDYWGQGTSVTVSS SEQ ID NO: 64 1B3_JH140 VL signal peptide MSSAQFLGLLLLCFQGTRC SEQ ID NO: 65 1B3_JH140 Variable Lightweight Domain DIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPRTFGGGTKLEIK SEQ ID NO: 66 1C12_JH104 VH signal peptide MNFGLSLIFLVLVLKGVLC SEQ ID NO: 67 1C12_JH104 Variable Restructuring Domain EVKLVESGGGLVQPGGSLKLSCAASGFTFSSYTMSWVRQTPEKRLELVADISNSGGRTYYPDTVKGRFTISRENAKNSLYLQMSSLKSEDTAMYYCARRRYSDYYYYYAMDNWGQGTSVTVSS SEQ ID NO: 68 1C12_JH104 VL signal peptide MSSAQFLGLLLLCFQGTRC SEQ ID NO: 69 1C12_JH104 Variable Lightweight Domain DIQMTQTTSSLSASLGDRVTISCRASQDISNYLSWYQQKPDGTVKLLIYYTSRLNSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNALPRTFGGGTKLEIN SEQ ID NO: 70 11A1_LA121 VH signal peptide MEWPCIFLFLLSVTEGVHS SEQ ID NO: 71 11A1_LA121 Variable Restructuring Domain QVQLQQSGAELVRPGSSVKISCKASGYEFSRYWMNWVKQRPGQGLEWIGQIYHGDGDTNYKGKFKGKAILTADKSSSTAYMQVSSLTSEDSAVYFCVRGGYYGYAMDYWGQGTSVTVSS SEQ ID NO: 72 11A1_LA121 signal peptide MKLPVRLLVLMFWIPASSS SEQ ID NO: 73 11A1_LA121 Variable Lightweight Domain DVVMTQTPLSLPVSLGDQASISCRSSQSLLHSNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPYTFGGGTNLEIK SEQ ID NO: 74 11B1_LA121 VH signal peptide MEWPCIFLFLLSVTEGVHS SEQ ID NO: 75 11B1_LA121 Variable Restructuring Domain QVQLQQSGAELVRPGSSVKISCKASGYEFSRYWMNWVKQRPGQGLEWIGQIYHGDGDTNYKGKFKGKAILTADKSSSTAYMQVSSLTSEDSAVYFCVRGGYYGYAMDYWGQGTSVTVSS SEQ ID NO: 76 11B1_LA121 VL signal peptide MKLPVRLLVLMFWIPASSS SEQ ID NO: 77 11B1_LA121 Variable Lightweight Domain DVVMTQTPLSLPVSLGDQASISCRSSQSLLHSNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPYTFGGGTNLEIK SEQ ID NO: 78 16G5_LA121 VH signal peptide MKCSWVIFFLMAVVTGVNS SEQ ID NO: 79 16G5_LA121 Variable Restructuring Domain EVHLQQSGAELVKPGASVKLSCTGSGFNIIDTYIHWVKQRPEQGLEWIGRIDPANGNTMYASKFQDKATIIADTSSNTVYMRLGSLTSGDTAVYYCSHGDFWWGQGTTLTVSS SEQ ID NO: 80 16G5_LA121 VL signal peptide MSPAQFLFLLVLWIQETNG SEQ ID NO: 81 16G5_LA121 variable light domain DVVMTQTPLTLSIPIGQPAYISCKSSQSLLKSNGKTYLNWLFQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGIYYCVQGTHLPHTFGGGTRLEIK SEQ ID NO: 82 Human TDP-43 protein MSEYIRVTEDENDEPIEIPSEDDGTVLLSTVTAQFPGACGLRYRNPVSQCMRGVRLVEGILHAPDAGWGNLVYVVNYPKDNKRKMDETDASSAVKVKRAVQKTSDLIVLGLPWKTTEQDLKEYFSTFGEVLMVQVKKDLKTGHSKGFGFVRFTEYETQVKVMSQRHMIDGRWCDCKLPNSKQSQDEPLRSRKVFVGRCTEDMTEDELREFFSQYGDVMDVFIPKPFRAFAFVTFADDQIAQSLCGEDLIIKGISVHISNAEPKHNSNRQLERSGRFGGNPGGFGNQGGFGNSRGGGAGLGNNQGSNMGGGMNFGAFSINPAMMAAAQAALQSSWGMMGMLASQQNQSGPSGNNQNQGNMQREPNQAFGSGNNSYSGSNSGAAIGWGSASNAGSGSGFNGGFGSSMDSKSSGWGM SEQ ID NO: 83 TDP-43 peptide (pS409 / pS410) GSGSGFNGGFGSSMDSKSSGWGM SEQ ID NO: 84 1B3_JH140 variable heavy chain CDR1 SGFTFSSYTMS SEQ ID NO: 85 1B3_JH140 Variable Heavy Chain CDR2 EISNSGGRTNY SEQ ID NO: 86 1B3_JH140 Variable Heavy Chain CDR3 RRYSDYYYYAMDY SEQ ID NO: 87 1B3_JH140 Variable Light Chain CDR1 RASQDISNYLN SEQ ID NO: 88 1B3_JH140 Variable Light Chain CDR2 YTSRLHS SEQ ID NO: 89 1B3_JH140 Variable Light Chain CDR3 QQGNTLPRT SEQ ID NO: 90 1C12_JH104 Variable Heavy Chain CDR1 SGFTFSSYTMS SEQ ID NO: 91 1C12_JH104 Variable Heavy Chain CDR2 DISNSGGRTYYPDTVKG SEQ ID NO: 92 1C12_JH104 Variable Heavy Chain CDR3 RRYSDYYYYAMDN SEQ ID NO: 93 1C12_JH104 Variable Light Chain CDR1 RASQDISNYLS SEQ ID NO: 94 1C12 JH104 Variable Light Chain CDR2 YTSRLNS SEQ ID NO: 95 1C12_JH104 Variable Light Chain CDR3 QQGNALPRT SEQ ID NO: 96 11A1_LA121 Variable Heavy Chain CDR1 SGYEFSRYWMN SEQ ID NO: 97 11A1_LA121 Variable Heavy Chain CDR2 QIYHGDGDTNYKGKFKG SEQ ID NO: 98 11A1_LA121 Variable Heavy Chain CDR3 GGYYGYAMDY SEQ ID NO: 99 11A1_LA121 Variable Light Chain CDR1 RSSQSLLHSNGNTYLH SEQ ID NO: 100 11A1_LA121 Variable Light Chain CDR2 KVSNRFS SEQ ID NO: 101 11A1_LA121 Variable light chain CDR3 SQSTHVPYT SEQ ID NO: 102 11B1_LA121 Variable Heavy Chain CDR1 SGYEFSRYWMN SEQ ID NO: 103 11B1_LA121 Variable Heavy Chain CDR2 QIYHGDGDTNYKGKFKG SEQ ID NO: 104 11B1_LA121 Variable Heavy Chain CDR3 GGYYGYAMDY SEQ ID NO: 105 11B1_LA121 Variable Light Chain CDR1 RSSQSLLHSNGNTYLH SEQ ID NO: 106 11B1_LA121 Variable light chain CDR2 KVSNRFS SEQ ID NO: 107 11B1_LA121 Variable Light Chain CDR3 SQSTHVPYT SEQ ID NO: 108 16G5_LA121 Variable Heavy Chain CDR1 SGFNIIDTYIH SEQ ID NO: 109 16G5_LA121 Variable Heavy Chain CDR2 RIDPANGNTMYA SEQ ID NO: 110 16G5_LA121 Variable Heavy Chain CDR3 GDFW SEQ ID NO: 111 16G5_LA121 Variable Light Chain CDR1 KSSQSLLKSNGKTYLN SEQ ID NO: 112 16G5_LA121 Variable light chain CDR2 LVSKLDS SEQ ID NO: 113 16G5 LA121 Variable Light Chain CDR3 VQGTHLPHT

Claims

1. An isolated monoclonal antibody that competitively binds to a 43 kD trans-active reactive DNA-binding protein (TDP-43): An antibody containing the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; An antibody containing the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; An antibody containing the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; An antibody containing the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; An antibody containing the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or An antibody containing the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO:

81.

2. The antibody of claim 1, wherein the antibody binds to the same epitope on TDP-43 with the following: An antibody containing the heavy chain variable domain of SEQ ID NO: 1 and the light chain variable domain of SEQ ID NO: 24; An antibody containing the heavy chain variable domain of SEQ ID NO: 63 and the light chain variable domain of SEQ ID NO: 65; An antibody containing the heavy chain variable domain of SEQ ID NO: 67 and the light chain variable domain of SEQ ID NO: 69; An antibody containing the heavy chain variable domain of SEQ ID NO: 71 and the light chain variable domain of SEQ ID NO: 73; An antibody containing the heavy chain variable domain of SEQ ID NO: 75 and the light chain variable domain of SEQ ID NO: 77; or An antibody containing the heavy chain variable domain of SEQ ID NO: 79 and the light chain variable domain of SEQ ID NO:

81.

3. An antibody that specifically binds to TDP-43, comprising three light chain CDRs and three heavy chain CDRs of a mouse antibody, characterized in that it comprises a heavy chain variable domain of SEQ ID NO: 1 and a light chain variable domain of SEQ ID NO:

24.

4. The antibody of claim 3, wherein the antibody is a humanized antibody, a chimeric antibody, or a decorative antibody.

5. The antibody of claim 4, wherein the definition of CDR is selected from the group consisting of Kabat, Chothia, Kabat / ChothiaComposite, AbM, and Contact.

6. The antibody of claim 5, wherein the antibody comprises a humanized mature heavy chain variable domain, comprising: Heavy chain CDR1, as defined by the Kabat / Chothia Composite, includes SEQ ID NO: 49; For example, the heavy chain CDR2 as defined by Kabat contains SEQ ID NO: 51; and Heavy chain CDR3, as defined by Kabat or Chothia, includes SEQ ID NO: 52; and Humanized mature light chain variable structural domain, which contains three Kabat light chain CDRs of SEQ ID NO: 53-55.

7. The antibody of claim 6, wherein the humanized mature heavy chain variable domain comprises a sequence that is at least 80% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain comprises a sequence that is at least 80% identical to any of SEQ ID NO: 27-48.

8. The antibody of claim 7, wherein the humanized mature heavy chain variable domain comprises a sequence that is at least 85% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain comprises a sequence that is at least 85% identical to any of SEQ ID NO: 27-48.

9. The antibody of claim 7 or 8, wherein the humanized mature heavy chain variable domain comprises a sequence that is at least 90% identical to any of SEQ ID NO: 4-23, and the humanized mature light chain variable domain comprises a sequence that is at least 90% identical to any of SEQ ID NO: 27-48.

10. The antibody of any one of claims 7 to 9, wherein the humanized mature heavy chain variable domain comprises a sequence that is at least 95% identical to any one of SEQ ID NO: 4-23, and the humanized mature light chain variable domain comprises a sequence that is at least 95% identical to any one of SEQ ID NO: 27-48.

11. The antibody of any one of claims 6 to 10, wherein at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: K19 is occupied by R; S35 is occupied by G; T40 is occupied by A; E42 is occupied by G; A49 is occupied by S; K43 is occupied by E; R44 is occupied by G or D; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; L80 is occupied by A or G; L82C is occupied by G; M83 is occupied by R; S84 is occupied by A; M89 is occupied by V; or F91 is occupied by Y.

12. The antibody of any one of claims 6 to 11, wherein at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: K43 is occupied by E; R44 is occupied by G or D; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; or F91 is occupied by Y.

13. The antibody of any one of claims 6 to 12, wherein at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: S35 is occupied by G; L78 is occupied by A or G; L80 is occupied by A or G; or L82C is occupied by G.

14. The antibody of any one of claims 6 to 13, wherein F91 of the humanized heavy chain variable domain is occupied by Y; and at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: R44 is occupied by G; A49 is occupied by S; A74 is occupied by S; T77 is occupied by S; L78 is occupied by A or G; or M83 is occupied by R.

15. The antibody of any one of claims 6 to 14, wherein at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: V3 is occupied by Q; L9 is occupied by S; D17 is occupied by Q; Q18 is occupied by P; K39 is occupied by R; K45 is occupied by R; T80 is occupied by A or S; L83 is occupied by V; L92 is occupied by G or A; V94 is occupied by I or A; A100 is occupied by G, D or R; or L104 is occupied by V.

16. The antibody of any one of claims 6 to 15, wherein at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: V3 is occupied by Q or A100 is occupied by D or R.

17. The antibody of any one of claims 6 to 16, wherein at least one of the following positions in the variable domain of the humanized light chain is occupied by a specified amino acid: L9 is occupied by S; T80 is occupied by A or S; L92 is occupied by G or A; or V94 is occupied by I or A.

18. The antibody of any one of claims 6 to 17, wherein V3 is occupied by Q; Q18 is occupied by P; A100 is occupied by D; and at least one of the following positions in the variable domain of the humanized light chain is occupied by the specified amino acid: T80 is occupied by A or L92 is occupied by A.

19. The antibody of any one of claims 6 to 18, wherein at least one of the following positions in the humanized heavy chain variable domain is occupied by a specified amino acid: L5 occupied by V; G44 occupied by R; A49 occupied by S; A74 occupied by S; T77 occupied by S; L78 occupied by A or G; M89 occupied by V; or F91 occupied by Y; and At least one of the following positions in the variable domain of the humanized light chain is occupied by the specified amino acid: V3 is occupied by Q; D17 is occupied by Q; Q18 is occupied by P; K39 is occupied by R; K45 is occupied by R; T80 is occupied by A; L83 is occupied by V; L92 is occupied by A; A100 is occupied by D; or L104 is occupied by V.

20. The antibody according to any one of claims 4 to 19, wherein the humanized antibody has improved thermal stability compared to a reference antibody comprising a heavy chain variable domain comprising SEQ ID NO: 1 and a light chain variable domain comprising SEQ ID NO:

24.

21. The antibody of claim 20, wherein the humanized antibody has a melting temperature of 55°C or higher.

22. An antibody that specifically binds to TDP-43, comprising a heavy chain variable domain and a light chain variable domain, said heavy chain variable domain and light chain variable domain comprising: Heavy chain CDR1 as defined by the Kabat / Chothia Composite includes SEQ ID NO: 49 or SEQ ID NO: 50; For example, the heavy chain CDR2 as defined by Kabat contains SEQ ID NO: 51; Heavy chain CDR3, as defined by Kabat or Chothia, contains SEQ ID NO: 52; For example, the light chain CDR1 as defined by Kabat contains SEQ ID NO: 53; For example, the light chain CDR2 as defined by Kabat contains SEQ ID NO: 54; and Light chain CDR3 as defined by Kabat includes one of SEQ ID NO: 55-61.

23. The antibody of claim 22, wherein: The heavy chain CDR1 as defined by the Kabat / Chothia Composite includes SEQ ID NO: 49; The heavy chain CDR2 as defined by Kabat contains SEQ ID NO: 51; The heavy chain CDR3, as defined by Kabat or Chothia, contains SEQ ID NO: 52; The light chain CDR1 as defined by Kabat contains SEQ ID NO: 53; The light chain CDR2, as defined by Kabat, includes SEQ ID NO: 54; and The light chain CDR3 as defined by Kabat includes SEQ ID NO: 55 or SEQ ID NO:

61.

24. The antibody of claim 22 or 23, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to any of SEQ ID NO: 4-23.

25. The antibody of claim 24, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to any of the following: SEQ ID NO: 20, SEQ ID NO: 21 and SEQ ID NO:

23.

26. The antibody of claim 24 or 25, wherein the heavy chain variable domain comprises a sequence that is at least 98% identical to any of the following: SEQ ID NO: 20, SEQ ID NO: 21 and SEQ ID NO:

23.

27. The antibody of any one of claims 22 to 26, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

20.

28. The antibody of any one of claims 22 to 26, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

21.

29. The antibody of any one of claims 22 to 26, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

23.

30. The antibody of any one of claims 22 to 29, wherein the light chain variable domain comprises a sequence that is at least 95% identical to any of the following: SEQ ID NO: 27-48.

31. The antibody of any one of claims 22 to 30, wherein the light chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO: 47 or SEQ ID NO:

48.

32. The antibody of any one of claims 22 to 31, wherein the light chain variable domain comprises a sequence that is at least 98% identical to SEQ ID NO: 47 or SEQ ID NO:

48.

33. The antibody of any one of claims 30 to 32, wherein the light chain variable domain comprises SEQ ID NO:

47.

34. The antibody of any one of claims 30 to 32, wherein the light chain variable domain comprises SEQ ID NO:

48.

35. An antibody that specifically binds to TDP-43, comprising a heavy chain variable domain and a light chain variable domain, wherein the antibody comprises: Heavy chain CDR1 containing SEQ ID NO: 84; Heavy chain CDR2 containing SEQ ID NO: 85; Heavy chain CDR3 containing SEQ ID NO: 86; The light chain CDR1 containing SEQ ID NO: 87; The light chain CDR2 containing SEQ ID NO: 88; and The light chain CDR3 contains SEQ ID NO:

89.

36. The antibody of claim 35, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

63.

37. The antibody of claim 35 or 36, wherein the heavy chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

63.

38. The antibody of any one of claims 35 to 37, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

63.

39. The antibody of any one of claims 35 to 38, wherein the light chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

65.

40. The antibody of any one of claims 35 to 39, wherein the light chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

65.

41. The antibody of any one of claims 35 to 40, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

65.

42. An antibody that specifically binds to TDP-43, comprising a heavy chain variable domain and a light chain variable domain, wherein the antibody comprises: Heavy chain CDR1 containing SEQ ID NO: 90; Heavy chain CDR2 containing SEQ ID NO: 91; Heavy chain CDR3 containing SEQ ID NO: 92; The light chain CDR1 containing SEQ ID NO: 93; The light chain CDR2 containing SEQ ID NO: 94; and It contains the light chain CDR3, which is SEQ ID NO:

95.

43. The antibody of claim 42, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

67.

44. The antibody of claim 42 or 43, wherein the heavy chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

67.

45. The antibody of any one of claims 42 to 44, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

67.

46. ​​The antibody of any one of claims 42 to 45, wherein the light chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

69.

47. The antibody of any one of claims 42 to 46, wherein the light chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

69.

48. The antibody of any one of claims 42 to 47, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

69.

49. An antibody that specifically binds to TDP-43, comprising a heavy chain variable domain and a light chain variable domain, wherein the antibody comprises: Heavy chain CDR1 containing SEQ ID NO: 96; Heavy chain CDR2 containing SEQ ID NO: 97; Heavy chain CDR3 containing SEQ ID NO: 98; The light chain CDR1 containing SEQ ID NO: 99; The light chain CDR2 containing SEQ ID NO: 100; and The light chain CDR3 contains SEQ ID NO:

101.

50. The antibody of claim 49, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

71.

51. The antibody of claim 49 or 50, wherein the heavy chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

71.

52. The antibody of any one of claims 49 to 51, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

71.

53. The antibody of any one of claims 49 to 52, wherein the light chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

73.

54. The antibody of any one of claims 49 to 53, wherein the light chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

73.

55. The antibody of any one of claims 49 to 54, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

73.

56. An antibody that specifically binds to TDP-43, comprising a heavy chain variable domain and a light chain variable domain, wherein the antibody comprises: Heavy chain CDR1 containing SEQ ID NO: 102; Heavy chain CDR2 containing SEQ ID NO: 103; Heavy chain CDR3 containing SEQ ID NO: 104; The light chain CDR1 containing SEQ ID NO: 105; The light chain CDR2 containing SEQ ID NO: 106; and The light chain CDR3 contains SEQ ID NO:

107.

57. The antibody of claim 56, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

75.

58. The antibody of claim 56 or 57, wherein the heavy chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

75.

59. The antibody of any one of claims 56 to 58, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

75.

60. The antibody of any one of claims 56 to 59, wherein the light chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

77.

61. The antibody of any one of claims 56 to 60, wherein the light chain variable domain comprises a sequence that is at least 98% identical to SEQ ID NO:

77.

62. The antibody of any one of claims 56 to 61, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

77.

63. An antibody that specifically binds to TDP-43, comprising a heavy chain variable domain and a light chain variable domain, wherein the antibody comprises: Heavy chain CDR1 containing SEQ ID NO: 108; Heavy chain CDR2 containing SEQ ID NO: 109; Heavy chain CDR3 containing SEQ ID NO: 110; The light chain CDR1 containing SEQ ID NO: 111; The light chain CDR2 containing SEQ ID NO: 112; and The light chain CDR3 contains SEQ ID NO:

113.

64. The antibody of claim 63, wherein the heavy chain variable domain comprises a sequence that is at least 95% identical to that of SEQ ID NO:

79.

65. The antibody of claim 63 or 64, wherein the heavy chain variable domain comprises a sequence that is at least 98% identical to that of SEQ ID NO:

79.

66. The antibody of any one of claims 63 to 65, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

79.

67. The antibody of any one of claims 63 to 66, wherein the light chain variable domain comprises a sequence that is at least 95% identical to SEQ ID NO:

81.

68. The antibody of any one of claims 63 to 67, wherein the light chain variable domain comprises a sequence that is at least 98% identical to SEQ ID NO:

81.

69. The antibody of any one of claims 63 to 67, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

81.

70. The antibody of any one of claims 22 to 69, wherein the antibody is a humanized antibody, a chimeric antibody, or a decorative antibody.

71. The antibody according to any one of claims 1 to 71, wherein the antibody is an antigen-binding antibody fragment.

72. The antibody of claim 71, wherein the antigen-binding antibody fragment is a Fab fragment, a Fab'2 fragment, or a single-chain Fv.

73. The antibody according to any one of claims 1 to 70, wherein the antibody is a complete antibody.

74. The antibody according to any one of claims 1 to 70 and 73, wherein the antibody has a human IgG1 isotype.

75. The antibody of any one of claims 1 to 70, 73 and 74, wherein the heavy chain variable domain is fused to the heavy chain constant region and the light chain variable domain is fused to the light chain constant region.

76. The antibody of claim 75, wherein the heavy chain constant region is a mutant form of the natural human heavy chain constant region, which binds less to the Fcγ receptor relative to the natural heavy chain constant region.

77. The antibody of claim 75 or 76, wherein the heavy chain constant region has an IgG1 isotype.

78. The antibody of any one of claims 1 to 77, wherein the antibody has at least one mutation in the constant region.

79. The antibody of claim 78, wherein the at least one mutation reduces complement fixation or activation in the constant region.

80. The antibody of claim 79, wherein the at least one mutation is at one or more of the following locations numbered according to EU: 241, 264, 265, 270, 296, 297, 318, 320, 322, 329 and 331.

81. The antibody of claim 80, wherein the antibody has alanine at positions 318, 320 and 322 according to EU numbers.

82. The antibody according to any one of claims 1 to 81, wherein the antibody selectively binds to phosphorylated TDP-43.

83. The antibody according to any one of claims 1 to 82, wherein the antibody selectively binds to phosphorylated TDP-43 compared to unphosphorylated TDP-43.

84. The antibody of claim 82 or 83, wherein the antibody binds to phosphorylated TDP-43 with an affinity at least 100 times greater than that of unphosphorylated TDP-43.

85. The antibody of any one of claims 82 to 84, wherein the antibody binds to phosphorylated TDP-43 with an affinity at least 1000 times greater than that of unphosphorylated TDP-43.

86. The antibody according to any one of claims 82 to 85, wherein phosphorylated TDP-43 comprises phosphorylation of at least one amino acid residue selected from S409 and S410.

87. The antibody of claim 86, wherein the phosphorylated TDP-43 comprises phosphorylation of both S409 and S410.

88. The antibody of any one of claims 1 to 87, wherein the antibody selectively binds to cytoplasmic aggregates of TDP-43.

89. The antibody according to any one of claims 1 to 88, wherein the antibody selectively binds to cytoplasmic aggregates of TDP-43 compared to nuclear TDP-43.

90. The antibody of claim 88 or 89, wherein the cytoplasmic aggregates of TDP-43 comprise phosphorylated aggregates of TDP-43.

91. The antibody of any one of claims 1 to 90, wherein the antibody substantially does not bind to unphosphorylated TDP-43.

92. The antibody of any one of claims 1 to 55, wherein the antibody is conjugated to a therapeutic agent, a cytotoxic agent, a cell growth inhibitor, an immunomodulator, a neurotrophic agent, or a neuroprotective agent.

93. The antibody of any one of claims 1 to 92, wherein the light chain variable domain does not contain a C-terminal lysine residue.

94. A pharmaceutical composition comprising the antibody of any one of claims 1 to 93 and a pharmaceutically acceptable carrier.

95. A nucleic acid encoding the heavy chain variable domain and / or light chain variable domain of the antibody according to any one of claims 1 to 93.

96. A vector comprising a nucleic acid encoding a mature heavy chain variable domain and a light chain variable domain, said nucleic acid being operatively linked to one or more regulatory sequences to achieve expression of the antibody of any one of claims 1 to 93 in mammalian cells.

97. The vector of claim 96, wherein the one or more regulatory sequences comprise one or more of an enhancer, a ribosome binding site, a transcription termination signal, and a promoter, optionally wherein the promoter is a eukaryotic promoter.

98. The vector of claim 96 or 97, wherein the nucleic acid is codon-optimized for expression in host cells.

99. A host cell transformed with the vector according to any one of claims 96 to 98.

100. A host cell comprising the nucleic acid of claim 95.

101. A method for inhibiting or reducing TDP-43 accumulation in a subject suffering from or at risk of developing TDP-43-related disease, comprising administering to the subject an effective amount of the antibody according to any one of claims 1 to 93, thereby inhibiting or reducing TDP-43 accumulation in the subject.

102. A method of treating or achieving prevention of TDP-43-related disease in a subject, comprising administering a therapeutically effective amount of the antibody according to any one of claims 1 to 93, and thereby treating or achieving prevention of said TDP-43-related disease.

103. The method of claim 102, wherein the TDP-43-related disease is amyotrophic lateral sclerosis (ALS), frontotemporal degeneration (FTLD-TDP), primary lateral sclerosis, progressive muscular atrophy, and Parkinson's disease.

104. The method of claim 103, wherein the TDP-43-related disease is ALS.

105. A method for detecting TDP-43 deposits in a subject suffering from or at risk of developing TDP-43-related disease, comprising administering an antibody to the subject according to any one of claims 1 to 93, and detecting an antibody in the subject that binds to TDP-43.

106. The method of claim 105, wherein the antibody is administered by intravenous injection into the body of the subject.

107. A method for detecting TDP-43 in a sample obtained from a patient with or at risk of having TDP-43-related disease, comprising detecting an antibody in the sample that binds to TDP-43 as described in any one of claims 1 to 93.

108. The method of any one of claims 105 to 107, wherein the antibody is labeled.

109. The method of claim 108, wherein the antibody is labeled with fluorescent labeling, paramagnetic labeling, or radioactive labeling.

110. The method of claim 109, wherein the radioactive label is detected using positron emission tomography (PET) or single-photon emission computed tomography (SPECT).

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