Antibody against TDP-43

Abstract

The present invention relates to an antibody or antigen-binding fragment thereof, which binds to TDP-43, i.e. TAR DNA-binding protein 43, and to therapeutic and diagnostic uses thereof.

Description

[0001] ANTIBODY AGAINST TDP-43

[0002] FIELD

[0003] The present invention relates to an antibody or antigen-binding fragment thereof, which binds to TDP-43, i.e. TAR DNA-binding protein 43, and to therapeutic and diagnostic uses thereof.

[0004] BACKGROUND

[0005] Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that results in the progressive loss of motor neurons that control voluntary muscles. ALS is the most common form of motor neuron diseases and exists in both sporadic form (about 90-95% of cases) as well as familial form (about 5-10% of cases). ALS is characterized by a rapid progression, for which no effective treatments are currently available. During disease progression, motor neurons degenerate and die, which causes the muscles to weaken, start to twitch (fasciculations), and waste away (atrophy). Eventually, the brain loses its ability to initiate and control voluntary movements, and finally the ability to breathe is lost.

[0006] 97% of all ALS cases have TAR DNA-binding protein 43 (TDP-43) inclusions as a pathological hallmark. Mutations in the highly conserved TARDBP gene encoding TDP43 are linked to familial cases of ALS. Thus, TDP-43 is considered to have a significant role in ALS pathology and consequently also represents a key therapeutic target. TDP-43 is a ubiquitously expressed nuclear protein mainly involved in RNA metabolism. Cytoplasmatic accumulation of phosphorylated and ubiquitinated TDP-43 in diseased neurons is a pathological hallmark of ALS.

[0007] TDP-43 is also present in many cases of frontotemporal lobar degeneration (FTLD) such as frontotemporal dementia (FTD), in limbic-predominant age-related TDP-43 encephalopathy (LATE), and in approximately 50% of Alzheimer's disease patients. If and when biomarkers are available, these diseases too can be considered for TDP-43 directed therapy.

[0008] Therapeutic antibodies against TDP-43 have been proposed, e.g. in W02009008529, WO2012174666, W02015117088, WO2016205522, WO2019134981, WO2018218352, W02013061163, W02016053610, W02016086320, WO2019177138, WO2020234473, WO2021217267, WO2022034228, WO2023156549 and WO2024184494.

[0009] Despite the existence of candidate antibodies within the field, no product has yet been granted regulatory approval, and there remains a need in the art for novel therapeutic, prophylactic, diagnostic and prognostic tools for detecting and treating ALS and other neurodegenerative diseases associated with TDP-43 proteopathy.

[0010] DESCRIPTION OF THE INVENTION

[0011] One object of the invention is to provide antibodies, or antigen-binding fragments thereof, having a novel and useful binding specificity.

[0012] Another object of the invention is to provide novel candidate antibodies, or antigen-binding fragments thereof, for the treatment of neurodegenerative diseases via targeting TDP-43 with a beneficial and unique binding profile.

[0013] Another object of the invention is to enable the diagnosis of ALS and other neurodegenerative disorders associated with TDP-43 proteopathy via detection of TDP-43 implicated in disease formation and / or progression.

[0014] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that bind to TDP-43 with high affinity.

[0015] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that bind to TDP-43 with high specificity.

[0016] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that bind to pathological TDP-43 variants with high affinity.

[0017] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that bind to pathological TDP-43 variants with high specificity.

[0018] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that bind to pathological TDP-43 variants with high selectivity with respect to physiological TDP-43 variants.

[0019] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that induce the clearance of pathological TDP-43 variants in vivo.

[0020] Another object of the invention is to provide antibodies, or antigen-binding fragments thereof, that do not interfere with the physiological function of TDP-43 in vivo. Another object of the invention is to provide a TDP-43-binding antibody or antigen-binding fragment thereof that combines desirable properties for development into a biopharmaceutical product.

[0021] Another object of the invention is to provide TDP-43-binding antibodies, or antigen-binding fragments thereof, that exhibit little or no immunogenicity upon administration in human subjects.

[0022] Another object of the invention is to provide TDP-43-binding antibodies, or antigen-binding fragments thereof, that show a beneficial pharmacokinetic profile upon administration in human subjects, for example evidenced by one or more of a long half-life, a high total exposure and a low clearance.

[0023] One or more of these objects, and other objects that are apparent to the skilled person from reading the entire disclosure, are met by the various aspects disclosed.

[0024] Thus, in a first aspect, the present disclosure provides an antibody or antigen-binding fragment thereof, comprising an immunoglobulin heavy chain variable (VH) region and an immunoglobulin light chain variable (VL) region, said VH and VL regions forming a VH / VL pair comprising an antigen-binding surface, wherein said antigen-binding surface is composed of three complementarity-determining regions (CDRs) from said VH region and three CDRs from said VL region, and wherein said CDRs consist of the following amino acid sequences:

[0025] VH-CDR1: X1YX2X3X4 (SEQ ID NO:1), wherein

[0026] XI is selected from E, D and N,

[0027] X2 is selected from N and Y,

[0028] X3 is selected from M and I,

[0029] X4 is selected from S and H;

[0030] VH-CDR2: X5IX6X7X8X9X10X11TX12YX13X14X15X16X17X18 (SEQ ID NO:2), wherein

[0031] X5 is selected from V and A,

[0032] X6 is selected from N and S,

[0033] X7 is selected from P, T and N,

[0034] X8 is selected from Y, E and N, such as from Y and E, X9 is selected from S and G,

[0035] X10 is selected from E, G and D,

[0036] Xll is selected from N and S,

[0037] X12 is selected from N and H,

[0038] X13 is selected from N and P,

[0039] X14 is selected from P, Q and D,

[0040] X15 is selected from K and T,

[0041] X16 is selected from F and V,

[0042] X17 is selected from K and T,

[0043] X18 is selected from D and G;

[0044] VH-CDR3: X19X20GX21X22X23X24 (SEQ ID NO:3), wherein

[0045] X19 is A or missing,

[0046] X20 is selected from D, E and G, such as from D and E, X21 is selected from Y and S,

[0047] X22 is selected from F and L,

[0048] X23 is selected from D and A,

[0049] X24 is selected from F and Y;

[0050] VL-CDR1: X25SSQX26X27X28X29X30NX31X32X33X34YLX35 (SEQ ID NO:4), wherein

[0051] X25 is selected from R and K,

[0052] X26 is selected from I, Y and S,

[0053] X27 is selected from I and L,

[0054] X28 is selected from V and L,

[0055] X29 is selected from H and Y,

[0056] X30 is selected from S and T,

[0057] X31 is selected from G, N and Q, such as from G and N, X32 is Qor missing,

[0058] X33 is selected from A, D and K,

[0059] X34 is selected from N and T,

[0060] X35 is selected from E and A;

[0061] VL-CDR2: X36X37SX38RX39S (SEQ ID NO:5), wherein X36 is selected from K and W,

[0062] X37 is selected from V and A,

[0063] X38 is selected from K, N and T,

[0064] X39 is selected from F and E;

[0065] VL-CDR3: X40QX41X42X43X44PX45T (SEQ ID NO:6), wherein

[0066] X40 is selected from Q and F,

[0067] X41 is selected from G and Y,

[0068] X42 is selected from Y and S,

[0069] X43 is selected from H and T,

[0070] X44 is selected from V and Y,

[0071] X45 is selected from L and P.

[0072] As evident according to the first aspect of the disclosure, the VH and VL regions may for example be provided in a full-length traditional antibody, or in an antibody fragment selected from the group consisting of Fab fragments, Fab' fragments, Ffab' fragments, Fv fragments, single chain Fv fragments, (scFv and domain antibodies. In a particular embodiment of the antibody or antigen-binding fragment thereof it is thus selected from the group consisting of Fab fragments, Fab' fragments, Ffab' fragments, Fv fragments, single chain Fv (scFv) fragments, (scFv and domain antibodies. In one embodiment, said antibody or antigen-binding fragment thereof is selected from full-length antibodies, Fab fragments and scFv fragments. In one embodiment, the antibody or antigen-binding fragment thereof is selected from full-length antibodies, Ffab' fragments and (scFv fragments. In some embodiments, the antigen-binding fragments of the disclosure are bivalent or multivalent. In one particular embodiment, the antibody is a full-length antibody. In one embodiment, said antibody or antigen-binding fragment thereof is monoclonal.

[0073] As used herein, the term "antibody or antigen-binding fragment thereof" encompasses not only full-length or intact polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof, such as Fab, Fab', F(ab')2, Fab3, Fv and variants thereof, fusion proteins comprising one or more antibody portions, humanized antibodies, chimeric antibodies, minibodies, diabodies, triabodies, tetrabodies, linear antibodies, single chain antibodies, multispecific antibodies (e.g. bispecific antibodies) and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site of the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies and covalently modified antibodies. Further examples of modified antibodies and antigen-binding fragments thereof include nanobodies, AlbudAbs, DARTs (dual affinity re-targeting), BiTEs (bispecific T-cell engager), TandAbs (tandem diabodies), DAFs (dual acting Fab), two-in-one antibodies, SMIPs (small modular immunopharmaceuticals), FynomAbs (fynomers fused to antibodies), DVD-lgs (dual variable domain immunoglobulin), CovX-bodies (peptide modified antibodies), duobodies and triomAbs. This listing of variants of antibodies and antigen-binding fragments thereof is not to be seen as limiting, and the skilled person is aware of other suitable variants.

[0074] A full-length antibody comprises two heavy chains and two light chains. Each heavy chain contains a heavy chain variable region (VH) and first, second and third constant regions (CHI, CH2 and CH3). Each light chain contains a light chain variable region (VL) and a light chain constant region (CL). Depending on the amino acid sequence of the constant domain of its heavy chains, antibodies are assigned to different classes. There are six major classes of antibodies: IgA, IgD, IgE, IgG, IgM and IgY, and several of these may be further divided into subclasses, e.g., IgGl, lgG2, lgG3, lgG4, IgAl and lgA2. The term "full-length antibody" as used herein, refers to an antibody of any class, such as IgD, IgE, IgG, IgA, IgM or IgY (or any sub-class thereof). The subunit structures and three-dimensional configurations of different classes of antibodies are well known.

[0075] The term "antigen-binding fragment" refers to a portion or region of an antibody molecule, or a derivative thereof, that retains all or a significant part of the antigen binding of the corresponding full-length antibody. An antigen-binding fragment may comprise the heavy chain variable region (VH), the light chain variable region (VL), or both. Each of the VH and VL regions or domains typically contains three CDRs, i.e. CDR1, CDR2 and CDR3, denoted VH-CDR1, VH-CDR2 and VH-CDR3 for the CDRs from the VH domain and VL-CDR1, VL-CDR2 and VL-CDR3 for the CDRs from the VL domain. The three CDRs in VH or VL are flanked by framework regions (FR1, FR2, FR3 and FR4). As briefly listed above, examples of antigen-binding fragments include, but are not limited to: (1 ) a Fab fragment, which is a monovalent fragment having a VL-CL chain and a VH-CH1 chain; (2) a Fab' fragment, which is a Fab fragment with the heavy chain hinge region, (3) a F(ab')2 fragment, which is a dimer of Fab' fragments joined by the heavy chain hinge region, for example linked by a disulfide bridge at the hinge region; (4) an Fc fragment; (5) an Fv fragment, which is the minimum antibody fragment having the VL and VH domains of a single arm of an antibody; (6) a single chain Fv (scFv) fragment, which is a single polypeptide chain in which the VH and VL domains of an scFv are linked by a peptide linker; (7) an (scFv)2, which comprises two VH domains and two VL domains, which are associated through the two VH domains via disulfide bridges and (8) a domain antibody, which may be an antibody single variable domain (VH or VL) polypeptide that specifically binds antigen. In some embodiments, the antigen-binding fragments are selected from bivalent and mutivalent constructs, such as Ffab' fragments and (SCFV)2 fragments. Antigen-binding fragments can be prepared via routine methods. For example, F(ab')2 fragments can be produced by pepsin digestion of a full-length antibody molecule, and Fab fragments can be generated by reducing the disulfide bridges of F(ab')2 fragments. Alternatively, portions can be prepared via recombinant technology by expressing the heavy and light chain portions in suitable host cells (e.g., E. coli, yeast, mammalian, plant or insect cells) and having them assembled to form the desired antigen-binding fragments either in vivo or in vitro. A single-chain antibody can be prepared via recombinant technology by linking a nucleotide sequence coding for a heavy chain variable region and a nucleotide sequence coding for a light chain variable region. For example, a flexible linker may be incorporated between the two variable regions.

[0076] Furthermore, the skilled person is aware of the meaning of the terms "polyclonal" and "monoclonal" antibodies. Polyclonal antibodies are normally generated by administering an antigen to an animal. Said antigen will evoke an immune response giving rise to polyclonal antibodies. Monoclonal antibodies are made by immunizing an animal, usually a mouse, with an antigen and the subsequent isolation of the spleen from said animal. Isolated spleen cells are immortalized by fusion with myeloma cells to give rise to hybridoma cells. Each hybridoma cell produces a unique monoclonal antibody.

[0077] The term "human antibody" as used herein, refers to antibodies having variable and constant regions corresponding to, or derived from, antibodies obtained from human subjects. The term "chimeric antibodies" as used herein, refers to recombinant or genetically engineered antibodies, such as for example antibodies with variable regions (VH and VL) of mouse origin and human constant region (Fc), to reduce the antibodies' immunogenicity. The term "humanized antibodies" refers to antibodies from non-human species whose protein sequences have been modified to increase their similarity to antibody variants produced naturally in humans, in order to reduce immunogenicity of the full antibody itself. In one embodiment, the antibody or antigen-binding fragment thereof is thus selected from the group consisting of recombinant human antibodies and fragments thereof; humanized antibodies and fragments thereof; and antibodies and fragments thereof that have been mutated to reduce antigenicity thereof in humans. In one embodiment, said antibody or antigen-binding fragment thereof is a recombinant human antibody or fragment thereof. In one embodiment, said antibody or antigenbinding fragment thereof is a humanized antibody or fragment thereof. In one embodiment, said antibody or antigen-binding fragment thereof has been mutated to reduce antigenicity thereof in humans. In one embodiment, said antibody or antigen-binding fragment thereof is of IgG class. In one embodiment, said IgG class is selected from the group consisting of IgG1 and IgG4. In one embodiment said IgG class is IgG1. In one embodiment, said IgG class is IgG4.

[0078] The designation of " VH / VL" as used in relation to a VH / VL pair does not limit the construct to any particular order of the VH and VL regions in the polypeptide chain, but is only used to convey that both the VH and VL regions are present, and that they are capable of pairwise association to form an Ig domain with an antigenbinding surface, as e.g. found in naturally occurring antibodies. As such, the term " VH / VL pair" encompasses, for example, constructs in which the VL and VH regions associate as part of a Fab fragment or full-length antibody, constructs in which the VL region precedes the VH region in a single chain Fv, constructs in which the VH region precedes the VL region in a single chain Fv, and constructs in which the VH and VL regions are non-covalently associated with each other. With respect to the antigen-binding surface of the VH / VL pair, it may suitably be composed of said three VL-CDRs and said three VH-CDRs as defined in any one of the embodiments discussed herein. Thus, said three VL-CDRs and said three VH-CDRs forming the antigen-binding surface of the VH / VL pair consist of the amino acid sequences as defined in any one of the embodiments discussed herein. In one embodiment, the amino acid sequence of said VH-CDR1 is selected from the group consisting of SEQ ID NO:18-20. 16. In some embodiments, the amino acid sequence of said VH-CDR1 is selected from the group consisting of SEQ ID NO:18 and SEQ ID NO:19. In some embodiments, the amino acid sequence of said VH-CDR1 is selected from the group consisting of SEQ ID NO:19 and SEQ ID NO:20. In one embodiment, the amino acid sequence of said VH-CDR1 is SEQ ID NO:18. In another embodiment, the amino acid sequence of said VH-CDR1 is SEQ ID NO:19. In yet another embodiment, the amino acid sequence of said VH-CDR1 is SEQ ID NO:20.

[0079] In one embodiment, the amino acid sequence of said VH-CDR2 is selected from the group consisting of SEQ ID NO:21-23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:21-22 and SEQ ID NO:55. In some embodiments, the amino acid sequence of said VH-CDR2 is selected from the group consisting of SEQ ID NO:21 and SEQ ID NO:22. In some embodiments, the amino acid sequence of said VH-CDR2 is selected from the group consisting of SEQ ID NO:22, SEQ ID NO:23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:22 and SEQ ID NO:55. In one embodiment, the amino acid sequence of said VH-CDR2 is SEQ ID NO:21. In another embodiment, the amino acid sequence of said VH-CDR2 is SEQ ID NO:22. In yet another embodiment, the amino acid sequence of said VH-CDR2 is SEQ ID NO:23. In yet another embodiment, the amino acid sequence of said VH-CDR2 is SEQ ID NO:55.

[0080] In one embodiment, the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:24-26 and 60. In one embodiment, the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:24-26. In some embodiments, the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:24, 25 and 60. In some embodiments, the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:24 and SEQ ID NO:25. In some embodiments, the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:25 and SEQ ID NO:26. In some embodiments, the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:25 and SEQ ID NO:60. In one embodiment, the amino acid sequence of said VH-CDR3 is SEQ ID NO:24. In another embodiment, the amino acid sequence of said VH-CDR3 is SEQ ID NO:25. In yet another embodiment, the amino acid sequence of said VH-CDR3 is SEQ ID NO:26. In still another embodiment, the amino acid sequence of said VH-CDR3 is SEQ ID NO:60.

[0081] In one embodiment, the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:27-29 and 62. In one embodiment, the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:27-29. In one embodiment, the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:27, 28 and 62. In some embodiments, the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:27 and SEQ ID NO:28. In some embodiments, the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:28 and SEQ ID NO:29. In some embodiments, the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:28 and SEQ ID NO:62. In one embodiment, the amino acid sequence of said VL-CDR1 is SEQ ID NO:27. In another embodiment, the amino acid sequence of said VL-CDR1 is SEQ ID NO:28. In yet another embodiment, the amino acid sequence of said VL-CDR1 is SEQ ID NO:29. In still another embodiment, the amino acid sequence of said VL-CDR1 is SEQ ID NO:62.

[0082] In one embodiment, the amino acid sequence of said VL-CDR2 is selected from the group consisting of SEQ ID NO:30-32. In some embodiments, the amino acid sequence of said VL-CDR2 is selected from the group consisting of SEQ ID NO:30 and SEQ ID NO:31. In some embodiments, the amino acid sequence of said VL-CDR2 is selected from the group consisting of SEQ ID NO:31 and SEQ ID NO:32. In one embodiment, the amino acid sequence of said VL-CDR2 is SEQ ID NO:30. In another embodiment, the amino acid sequence of said VL-CDR2 is SEQ ID NO:31. In yet another embodiment, the amino acid sequence of said VL-CDR2 is SEQ ID NO:32.

[0083] In one embodiment, the amino acid sequence of said VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33. In one embodiment, the amino acid sequence of said VL-CDR3 is SEQ ID NO:17. In another embodiment, the amino acid sequence of said VL-CDR3 is SEQ ID NO:33.

[0084] In some embodiments, the CDR sequences are selected from all possible combinations, subgroups and individual sequence pairs of the options listed above, as listed in the Itemized Listing of Embodiments section below. In some embodiments, the amino acid sequences of the 6 CDRs are the following: the VH-CDR1 is selected from the group consisting of SEQ ID NO:18-20; the VH-CDR2 is selected from the group consisting of SEQ ID NO:21-23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:21-22 and SEQ ID NO:55; the VH-CDR3 is selected from the group consisting of SEQ ID NO:24-26 and 60, such as the group consisting of SEQ ID NO:24-26 or the group consisting of SEQ ID NO:25 and SEQ ID NO:60; the VL-CDR1 is selected from the group consisting of SEQ ID NO:27-29 and 62, such as the group consisting of SEQ ID NO:27-29 or the group consisting of SEQ ID NO:28 and SEQ ID NO:62; the VL-CDR2 is selected from the group consisting of SEQ ID NO:30-32; and the VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33.

[0085] In more specific embodiments, the amino acid sequences of the 6 CDRs are the following: the VH-CDR1 is selected from the group consisting of SEQ ID NO:18 and SEQ ID NO:19, the VH-CDR2 is selected from the group consisting of SEQ ID NO:21 and SEQ ID NO:22, theVH-CDR3 is selected from the group consisting of SEQ ID NO:24 and SEQ ID NO:25, the VL-CDR1 is selected from the group consisting of SEQ ID NO:27 and SEQ ID NO:28, the VL-CDR2 is selected from the group consisting of SEQ ID NO:30 and SEQ ID NO:31, and the VL-CDR3 is SEQ ID NO:17.

[0086] In other embodiments, the amino acid sequences of the 6 CDRs are the following: the VH-CDR1 is selected from the group consisting of SEQ ID NO:19 and SEQ ID NO:20; the VH-CDR2 is selected from the group consisting of SEQ ID NO:22 SEQ ID NO:23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:22 and SEQ ID NO:55; the VH-CDR3 is selected from the group consisting of SEQ ID NO:25 and SEQ ID NO:26; the VL-CDR1 is selected from the group consisting of SEQ ID NO:28 and SEQ ID NO:29; the VL-CDR2 is selected from the group consisting of SEQ ID NO:31 and SEQ ID NO:32; and the VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33.

[0087] As defined herein, embodiments of the antibody or antigen-binding fragment of the first aspect of the disclosure that comprise a VH / VL pair have specific amino acid sequences in the regions determining its binding capability, such as the CDRs of the heavy and light chain variable domain, or indeed the entire VL and / or VH domains or regions. In certain embodiments, the VL-CDR and VH-CDR sequences, when present in the antibody or antigen-binding fragment thereof, are selected from any one of the listed sequences. Non-limiting examples of such specific amino acid sequences are provided herein for the specific antibodies generated and characterized as described in Examples 1-15. It is contemplated that the specific sequence information provided for the generated antibodies enables the skilled person to define combinations and variations of these sequences within the scope of the invention, such as including the combinations and variations afforded by the variation in the general CDR sequences provided as SEQ ID NO:1-16.

[0088] In a specific embodiment of the antibody or antigen-binding fragment thereof of the disclosure, the amino acid sequences of the 6 CDRs are the following: VH-CDR1: EYNIH (SEQ ID NO:18), VH-CDR2: VINPYSENTNYNPKFTD (SEQ ID NO:21), VH-CDR3: DGYFDF (SEQ ID NO:24), VL-CDR1: RSSQIIVHTNGATYLE (SEQ ID NO:27), VL-CDR2: KVSKRFS (SEQ ID NO:30), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0089] In another particular embodiment, the amino acid sequences of the 6 CDRs are the following:

[0090] VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: DGYFDY (SEQ ID NO:25), VL-CDR1: RSSQYIVHSNGDTYLE (SEQ ID NO:28), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0091] In yet another particular embodiment, the amino acid sequences of the 6 CDRs are the following:

[0092] VH-CDR1: NYYMS (SEQ ID NO:20), VH-CDR2: AINNNGDSTHYPDTVKG (SEQ ID NO:23), VH-CDR3: AEGSLAY (SEQ ID NO:26), VL-CDR1: KSSQSLLYSNNQKNYLA (SEQ ID NO:29), VL-CDR2: WASTRES (SEQ ID NO:32), VL-CDR3: QQYYTYPLT (SEQ ID NO:33).

[0093] In yet another particular embodiment, the amino acid sequences of the 6 CDRs are the following: VH-CDR1: DYNMH (SEQ ID N0:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: GGYFDY (SEQ ID NO:60), VL-CDR1: RSSQYIVHSNGDTYLE (SEQ ID NO:28), VL-CDR2: KVSNRFS (SEQ ID N0:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0094] In yet another particular embodiment, the amino acid sequences of the 6 CDRs are the following:

[0095] VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: DGYFDY (SEQ ID NO:25), VL-CDR1: RSSQYIVHSNQDTYLE (SEQ ID NO:62), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0096] In yet another particular embodiment, the amino acid sequences of the 6 CDRs are the following:

[0097] VH-CDR1: NYYMS (SEQ ID NO:20), VH-CDR2: AINNEGDSTHYPDTVKG (SEQ ID NO:55), VH-CDR3: AEGSLAY (SEQ ID NO:26), VL-CDR1: KSSQSLLYSNNQKNYLA (SEQ ID NO:29), VL-CDR2: WASTRES (SEQ ID NO:32), VL-CDR3: QQYYTYPLT (SEQ ID NO:33).

[0098] In one embodiment, CDR sequences in an antigen-binding interface comprised in the antibody or antigen-binding fragment thereof of the disclosure are as defined using the Kabat convention, which is well known to a person of skill in the art of antibody technology (see e.g. Kabat (1991), Sequences of Proteins of Immunological Interest, 5thedition, NIH Publication no 91-3242 from the US Department of Health and Human Services).

[0099] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:40-44 and 61, such as consisting of SEQ ID NO:40-44; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0100] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:40, 41 and 61, such as consisting of SEQ ID NO:40 and SEQ ID NO:41; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0101] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:42-44; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0102] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:43 and SEQ ID NO:44; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0103] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:40; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:40, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:40. In a specific embodiment, the VH region comprises or consists of SEQ ID NO:40.

[0104] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:41; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:41, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:41. In a specific embodiment, the VH region comprises or consists of SEQ ID NO:41.

[0105] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:61; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:61, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:61. In a specific embodiment, the VH region comprises or consists of SEQ ID NO:61.

[0106] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:42; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:42, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:42. In a specific embodiment, the VH region comprises or consists of SEQ ID NO:42.

[0107] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:43; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43. In a specific embodiment, the VH region comprises or consists of SEQ ID NO:43.

[0108] In one embodiment, said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:44; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44. In a specific embodiment, the VH region comprises or consists of SEQ ID NO:44.

[0109] In more specific embodiments, the heavy chain variable region comprises a sequence selected from all possible combinations, subgroups and individual sequences inherent in the options listed above, as listed in the Itemized Listing of Embodiments section below.

[0110] In one embodiment said VL region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:45-48 and 63, such as consisting of SEQ ID NO:45-48; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i).

[0111] In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:45, 46 and 63, such as consisting of SEQ ID NO:45 and SEQ ID NO:46; and

[0112] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i). In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:47 and SEQ ID NO:48; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of the embodiments above, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i).

[0113] In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:45; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:45, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:45. In a specific embodiment, the VL region comprises or consists of SEQ ID NO:45.

[0114] In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:46; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:46, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:46. In a specific embodiment, the VL region comprises or consists of SEQ ID NO:46.

[0115] In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:63; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:63, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:63. In a specific embodiment, the VL region comprises or consists of SEQ ID NO:63. In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:47; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47. In a specific embodiment, the VL region comprises or consists of SEQ ID NO:47.

[0116] In one embodiment, said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:48; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48. In a specific embodiment, the VL region comprises or consists of SEQ ID NO:48.

[0117] In more specific embodiments, the light chain variable region comprises a sequence selected from all possible combinations, subgroups and individual sequences inherent in the options listed above, as listed in the Itemized Listing of Embodiments section below.

[0118] In some embodiments, the antibody or antigen-binding fragment thereof of the disclosure comprises a VH / VL pair, wherein the VL and VH sequences can be freely combined among the options listed above. Thus, in some embodiments of the antibody or antigen-binding fragment thereof, it comprises a VL region and a VH region as defined in any one of the embodiments above. In more specific embodiments, the light and heavy chain variable regions comprise sequence pairs selected from all possible combinations, subgroups and individual sequence pairs inherent in the options listed above, as listed in the Itemized Listing of Embodiments section below.

[0119] In one particular embodiment of the antibody or antigen-binding fragment thereof of the disclosure, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:40; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:40, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:40; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:45; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:45, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:45; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:40; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:40, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:40; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:63; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:63, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:63; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:61; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:61, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:61; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:45; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:45, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:45; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:61; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:61, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:61; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:63; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:63, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:63; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:41; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:41, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:41; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:46; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:46, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:46; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:42; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:42, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:42; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48.

[0120] In specific embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:40 and the VL region comprising or consisting of SEQ ID NO:45; or the VH region comprising or consisting of SEQ ID NO:61 and the VL region comprising or consisting of SEQ ID NO:45; or the VH region comprising or consisting of SEQ ID NO:40 and the VL region comprising or consisting of SEQ ID NO:63; or the VH region comprising or consisting of SEQ ID NO:61 and the VL region comprising or consisting of SEQ ID NO:63; or the VH region comprising or consisting of SEQ ID NO:41 and the VL region comprising or consisting of SEQ ID NO:46; or the VH region comprising or consisting of SEQ ID NO:42 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:48; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:48.

[0121] In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:40; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:40, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:40; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:45; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:45, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:45; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:41; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:41, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:41; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:46; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:46, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:46.

[0122] In specific embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:40 and the VL region comprising or consisting of SEQ ID NO:45; or the VH region comprising or consisting of SEQ ID NO:61 and the VL region comprising or consisting of SEQ ID NO:45; or the VH region comprising or consisting of SEQ ID NO:40 and the VL region comprising or consisting of SEQ ID NO:63; or the VH region comprising or consisting of SEQ ID NO:61 and the VL region comprising or consisting of SEQ ID NO:63; or the VH region comprising or consisting of SEQ ID NO:41 and the VL region comprising or consisting of SEQ ID NO:46.

[0123] In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:42; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:42, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:42; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48.

[0124] In particular embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:42 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:48; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:48.

[0125] In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48.

[0126] In particular embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:48; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:48.

[0127] In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47.

[0128] In particular embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:47.

[0129] In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48. In particular embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:48; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:48.

[0130] In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48.

[0131] In particular embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:48. In one embodiment, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48.

[0132] In particular embodiments, the antibody or antigen-binding fragment thereof comprises the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:48.

[0133] As described above, the VH and VL sequences, when present in the antibody or antigen-binding fragment thereof, are, in certain embodiments, selected from any one of the listed sequences and sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, such as 100% identity thereto. In some embodiments, the VH and VL sequences, when present in the antibody or antigen-binding fragment thereof, are selected from any one of the listed sequences and sequences having at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, such as 100% identity thereto. Non-limiting examples of such specific amino acid sequences are provided herein for the specific antibodies generated and characterized as described in Examples 1-15. It is contemplated that the specific sequence information provided for the generated antibodies enables the skilled person to define combinations and variations of these sequences within the scope of the invention, such as including the combinations and variations afforded by the variation in the general VH and VL sequences provided as SEQ ID NO:34-48. For embodiments wherein the variable domains of the antibodies or antigen-binding fragments are defined by a particular percentage sequence identity to a reference sequence, the VH and / or VL domains retain identical CDR sequences to those present in the reference sequence such that the variation is present only within the framework regions. In specific embodiments, the combinations of VH / VL are those present in the antibodies exemplified in Examples 1-21 (see Table 3, 10 and 11 in particular).

[0134] In one particular embodiment of the antibody or antigen-binding fragment thereof of the disclosure, the antibody or antigen-binding fragment thereof further comprises a heavy chain constant (CH) region. In one embodiment, said CH region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:49 and SEQ ID NO:50; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i). In a specific embodiment, the CH region comprises or consists of SEQ ID NO:49 or SEQ ID NO:50.

[0135] In one embodiment, said CH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:49; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:49. In a specific embodiment, the CH region comprises or consists of SEQ ID NO:49.

[0136] In one embodiment, said CH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:50; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:50. In a specific embodiment, the CH region comprises or consists of SEQ ID NO:50.

[0137] In one particular embodiment of the antibody or antigen-binding fragment thereof of the disclosure, the antibody or antigen-binding fragment thereof further comprises a light chain constant (CL) region. In one embodiment, said CL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:51; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:51. In a specific embodiment, the CL region comprises or consists of SEQ ID NO:51.

[0138] In some embodiments, the antibody or antigen-binding fragment thereof of the disclosure comprises the CH and CL regions, wherein the CH and CL sequences can be freely combined among the options listed above. Thus, in some embodiments of the antibody or antigen-binding fragment thereof, it comprises a CH region and a CL region as defined in any one of the embodiments above. In more specific embodiments, the light and heavy chain constant regions comprise sequence pairs selected from all possible combinations, subgroups and individual sequence pairs inherent in the options listed above, as listed in the Itemized Listing of Embodiments section below.

[0139] In one embodiment, the antibody or antigen-binding fragment thereof that comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:40; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:40, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:40; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:45; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:45, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:45; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:41; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:41, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:41; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:46; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:46, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:46;

[0140] further comprises the CH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:49; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:49; and the CL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:51; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:51.

[0141] In one embodiment, the antibody or antigen-binding fragment thereof that comprises the VH region comprising or consisting of SEQ ID NO:40 and the VL region comprising or consisting of SEQ ID NO:45; or the VH region comprising or consisting of SEQ ID NO:61 and the VL region comprising or consisting of SEQ ID NO:45; or the VH region comprising or consisting of SEQ ID NO:40 and the VL region comprising or consisting of SEQ ID NO:63; or the VH region comprising or consisting of SEQ ID NO:61 and the VL region comprising or consisting of SEQ ID NO:63; or the VH region comprising or consisting of SEQ ID NO:41 and the VL region comprising or consisting of SEQ ID NO:46;

[0142] further comprises the CH region comprising or consisting of SEQ ID NO:49 and the CL region comprising or consisting of SEQ ID NO:51.

[0143] In one embodiment, the antibody or antigen-binding fragment thereof that comprises the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:42; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:42, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:42; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:43; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:47; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47; or the VH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:44; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44; and the VL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:48; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48;

[0144] further comprises the CH region comprising or consisting of an amino acid sequence selected from SEQ ID NO:50; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:50; and the CL region comprising or consisting of an amino acid sequence selected from SEQ ID NO:51; and an amino acid sequence having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:51.

[0145] In one embodiment, the antibody or antigen-binding fragment thereof that comprises the VH region comprising or consisting of SEQ ID NO:42 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:43 and the VL region comprising or consisting of SEQ ID NO:48; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:47; or the VH region comprising or consisting of SEQ ID NO:44 and the VL region comprising or consisting of SEQ ID NO:48;

[0146] further comprises the CH region comprising or consisting of SEQ ID NO:50 and the CL region comprising or consisting of SEQ ID NO:51.

[0147] As demonstrated in the appended Examples, the antigen-binding surface formed by the listed CDRs, in one embodiment, provides the antibody or antigenbinding fragment thereof with the capacity to bind selectively to an epitope of TDP-43. The epitope, in some embodiments, comprises or consists of SEQ ID NO:52. Without wishing to be bound by theory, the binding of said antibody or antigenbinding fragment thereof to said epitope is contemplated to offer advantages in terms of avoiding the drawbacks associated with known binders to TDP-43.

[0148] As known to a person skilled in the art, an epitope (or "antigenic determinant") is a group of amino acids or other chemical groups exposed on the surface of a molecule, frequently a protein, here TDP-43, which can generate an antigenic response and bind antibody. An epitope is a localized region on the surface of an antigen that is recognized by the immune system, specifically by antibodies. An epitope for a given antibody may be determined by epitope mapping, for example as described in Example 4 for antibodies according to the present disclosure. During epitope mapping, the shortest amino acid sequence region recognized by an antibody within the amino acid sequence of the target protein may be revealed. The recognized amino acid region may be part of linear epitope, which consists of consecutive amino acids of the target protein. The number of consecutive amino acids in such linear epitopes is typically within the range of from 3 to 15 amino acids, such as the range of from 5 to 12 amino acids. The recognized amino acid region revealed by epitope mapping may be part of a conformational epitope, also known as a non-linear epitope or discontinuous epitope, which is composed of neighboring amino acid residues located on an antigenic protein surface structure. Epitopes are bound by their complementary paratopes on B-cell receptors and / or on antibodies.

[0149] Antibodies which "recognize" an epitope have an affinity for the recognized epitope. " Affinity", or "binding affinity" as also referred to herein, corresponds to the strength with which the antigen-binding site of an antibody (also known as paratope) binds an epitope of a protein. As used herein, the terms "binding to X", "specific binding to X", "selective binding to X" and "affinity for X", wherein X is a target (e.g. an antigen or an epitope, such as TDP-43 or an epitope of TDP-43 bound by the antibody or an antigen-binding fragment thereof as defined herein), refer to a property of a binding molecule, such as a property of an antibody or antigen-binding fragment thereof or of a bi- or multispecific construct incorporating such an antibody or antigen-binding fragment thereof, which may be tested for example by ELISA, by surface plasmon resonance (SPR) or by bio-layer interferometry (BLI), as further discussed below. The skilled person is aware of these methods and others.

[0150] The present disclosure is based on detailed insights into the pathophysiology of diseases characterized by TDP-43 deposition, and the identification of particular forms of TDP-43 in brain tissue from patients suffering from such diseases. As a nonlimiting example, aggregated forms of TDP-43 were found to be a hallmark of TDP-43 proteopathies, further highlighting the importance of obtaining antibodies that bind such species in a specific and / or selective manner. However, these insights also point to the potential benefits of having antibodies that bind TDP-43 in all forms thereof that are present in connection with disease. These insights have enabled the generation of inventive antibodies, or antigen-binding fragments thereof, that are specific and / or selective for TDP-43 in its various forms. In one embodiment, an antibody according to the disclosure is selective for pathological TDP-43 over physiological TDP-43, as discussed extensively below.

[0151] Without wishing to be bound by theory, it is contemplated that such novel antibodies, or antigen-binding fragments thereof, are useful in the diagnosis, prognosis and / or preventive or curative treatment of neurodegenerative diseases such as disorders associated with TDP-43 proteopathies, through specific binding to the putatively disease-causing TDP-43 variants.

[0152] In one embodiment of the antibody or antigen-binding fragment thereof of the disclosure, said TDP-43 comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:53; and (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% identity to SEQ ID NO:53, provided that said amino acid sequence comprises SEQ ID NO:52. In a particular embodiment, said TDP-43 comprises or consists of SEQ ID NO:53.

[0153] As a person of skill in the art is aware, the TDP-43 protein exists in various biophysical forms when present in the body. As appreciated by those skilled in the art, these various biophysical forms of TDP-43 typically comprise or consist of the same amino acid sequence, such as the amino acid sequence defined immediately above. These forms are instead differentiated by for example their biophysical characteristics, oligomerization or aggregation states, phosphorylation states and / or (sub)cellu la r localization. Of particular relevance to the present disclosure are pathological TDP-43 variant forms, in that some embodiments of the antibody or antigen-binding fragment thereof herein bind to said pathological TDP-43 variant forms. Thus, in certain embodiments, said TDP-43 is pathological TDP-43, i.e. TDP-43 present in the body in a pathological form. As used herein, the term "pathological TDP-43" refers to a TPD-43 variant form which is associated with a TDP-43 proteopathy.

[0154] Thus, in certain embodiments, said pathological TDP-43 is a TDP-43 aggregate, a TDP-43 oligomer and / or phosphorylated TDP-43 or any combination, subgroup or individual selection thereof as listed in the Itemized Listing of Embodiments section below.

[0155] The skilled person is aware that a TDP-43 aggregate may be a reversible or a non-reversible (i.e. irreversible) TDP-43 aggregate. While a reversible TDP-43 aggregate is typically considered to have a physiological function, a non-reversible TDP-43 aggregate is commonly associated with a proteopathy, and is thus considered pathological. Thus, in a particular embodiment, said TDP-43 aggregate is a non-reversible TDP-43 aggregate. Non-reversible TDP-43 aggregates can typically be enriched in insoluble protein fractions, such as in cell or tissue homogenates, as demonstrated in Example 10. Thus, said TDP-43 aggregate may be an insoluble TDP-43 aggregate. As for example presented in Examples 1 and 6, whether TDP-43 is present in an aggregated form or non-aggregated form (such as a monomeric form and / or an oligomeric state) can be determined by conventional biochemical methods, which provide information on the molecular weight and / or size of said TDP-43 form. In one embodiment, the molecular weight of said TDP-43 aggregate is higher than the molecular weight of a TDP-43 monomer, wherein said molecular weight of said TDP-43 aggregate is at least about 2x higher, such as at least about 3x higher, such as at least about 4x higher, such as at least about 5x higher, such as at least about 6x higher, such as at least about 7x higher, such as at least about 8x higher, such as at least about 9x higher, such as at least about 10x higher, such as at least about 20x higher, such as at least about 30x higher, such as at least about 40x higher, such as at least about 50x higher, such as at least about 100x higher when compared to the molecular weight of said TPD-43 monomer, optionally as determined by SDS-PAGE and Western blot, wherein said TPD-43 monomer has a molecular weight of about 49 kDa. In one embodiment, the size of said TDP-43 aggregate is greater than the size of a TDP-43 monomer, wherein said size of said TDP-43 aggregate is at least about 2x greater, such as at least about 3x greater, such as at least about 4x greater, such as at least about 5x greater, such as at least about 6x greater, such as at least about 7x greater, such as at least about 8x greater, such as at least about 9x greater, such as at least about 10x greater when compared to the size of said TPD-43 monomer, optionally as determined by HPLC and / or transmission electron microscopy (TEM).

[0156] One skilled in the art appreciates that the accumulation of pathological TDP-43 in disease affected neurons typically manifests as cytoplasmic deposition of C-terminally phosphorylated TDP-43 (see e.g. Eck et al (2021), GeroScience 43(4):1605-1614). Thus, in one embodiment, said phosphorylated TDP-43 is a phosphorylated TDP-43 aggregate. In addition to being a robust and consistent pathological marker forTDP-43-positive inclusions, phosphorylated TDP-43 manifests in other neurodegenerative disease and aging. As known to a person skilled in the art, TDP-43 has 64 potential phosphorylation sites of which a subset, such as phosphorylation at S369, S379, S403 / 404, and S409 / 410, consistently occurs in context of the disease, suggesting a potential pathological role. The best characterized phosphorylation event occurs at S409 and S410 (S409 / 410). Thus, in a particular embodiment said phosphorylated TDP-43 is TDP-43 phosphorylated at S409 and S410 (S409 / 410). As appreciated by a person skilled in the field of characterizing phosphorylation events, due to a lack of phosphorylation site-specific antibodies, it is often difficult to determine the pathological role of phosphorylation events at certain phosphorylation sites. Thus, it is envisioned that pathological TDP-43 may be any phosphorylated TDP-43 variant, provided that the given phosphorylation event or events is / are associated with a TDP-43 proteopathy, for example by leading to the accumulation of cytoplasmic TDP-43 deposits.

[0157] It is appreciated that TDP-43 undergoes self-assembly into oligomeric complexes upon misfolding that act as templates for large aggregates. A person of skill in the art is also aware that TDP-43 may exist in various forms along the progressive aggregation thereof from monomers to insoluble deposits. This process may be altered in disease conditions, such as in the presence of patient-linked mutations. Thus, as discussed above, said pathological TDP-43 may be a TDP-43 oligomer. In one specific embodiment, said TDP-43 oligomer comprises mutant TDP-43 protein species (see e.g. French et al (2019) J Biol Chem 294:6696-6709).

[0158] Furthermore, as ALS-linked mutants potently affect aggregation by increasing the rate of assembly, in certain embodiments mutant TDP-43 form TDP-43 aggregates. Thus, in another embodiment, said TDP-43 aggregate comprises mutant TDP-43 protein species.

[0159] Thus, as described above, in certain embodiments, an antibody or antigenbinding fragment thereof of the first aspect has an affinity for a pathological form of TDP-43. Alternatively, the antibody or antigen-binding fragment thereof may exhibit a preference or selectivity for one form of TDP-43 over another. In one such embodiment of the antibody or antigen-binding fragment thereof as disclosed herein, it has a higher binding affinity for pathological TDP-43 than for physiological TDP-43. It is appreciated that physiological TDP-43 may be a TDP-43 variant form which is not associated with any TDP-43 proteopathy. Thus, in certain embodiments, said physiological TDP-43 is a TDP-43 monomer, a non-disease associated TDP-43 oligomer and / or a non-disease associated reversible TDP-43 aggregate. Without wishing to be bound by theory, such higher affinity for pathological TDP-43 in embodiments of the antibody or fragment thereof may be due to avidity effects, insofar as pathological forms of TDP-43 are thought to present a plurality of epitopes for the antibody to bind, in comparison to the monomeric form. Alternatively, such higher affinity may be due to the antibody or fragment thereof having affinity for at least one epitope of TDP-43 as described herein. As such, the affinity of the antibody for pathological forms of TDP-43 may be measured in a manner known to the skilled person.

[0160] In certain embodiments, the antibodies and antigen-binding fragments thereof of the first aspect bind selectively to pathological TDP-43. As used herein, the term "bind selectively" refers to the preferential binding of the antibody or antigen-binding fragment thereof to the pathological TDP-43 target. In certain embodiments, the antibodies and antigen-binding fragments thereof of the first aspect do not bind to any appreciable extent to physiological TDP-43 variants. Put slightly differently, in one such embodiment, the antibody or antigen-binding fragment thereof has a higher binding affinity for pathological forms of TDP-43 than for physiological forms of TDP-43.

[0161] For example, the binding affinity for a target, antigen or epitope X may be tested in an experiment in which a binding molecule to be tested is captured on ELISA plates coated with X or a molecule comprising the epitope X, and a biotinylated detector antibody is added, followed by streptavidin-conjugated horse radish peroxidase (HRP). Alternatively, said detector antibody may be directly conjugated with HRP. Tetramethylbenzidine (TMB) substrate is added and the absorbance at 450 nm is measured using an ELISA multi-well plate reader. The skilled person may then interpret the results obtained by such experiments to establish at least a qualitative measure of the binding affinity for X of the binding molecule. If a quantitative measure is desired, for example to determine the EC₅₀ value (the half maximal effective concentration) for the interaction, ELISA may also be used. The response of the binding molecule against a dilution series of X may be measured using ELISA as described above. The skilled person may then interpret the results obtained by such experiments and EC₅₀ values may be calculated from the results, using for example GraphPad Prism v.9 and non-linear regression. As used herein, the term " EC₅₀" refers to the half maximal effective concentration of binding molecule which induces a response halfway between the baseline and maximum after a specified exposure time.

[0162] Additionally or alternatively, inhibition ELISA may be used to obtain a quantitative measure of interaction by determination of the " IC₅₀" (the half maximal inhibitory concentration). In an inhibition ELISA, the concentration of target X in a fluid sample is measured by detecting interference in an expected signal output. In principle, a known target or epitope-bearing substance is used to coat a multi-well plate. In parallel, a binding molecule with putative affinity for X is added and incubated with a solution containing target at varied concentrations. Following standard blocking and washing steps, samples containing the mixture of said binding molecule and the target are added to the well. Labeled detection antibody with affinity for the binding molecule is then applied for detection using relevant substrates (for example TMB). In principle, if there is a high concentration of target in the fluid sample, a significant reduction in signal output will be observed. In contrast, if there is very little target in the fluid sample, there will be very little reduction in the expected signal output. The skilled person appreciates that the signal output is also dependent on the affinity of the binding molecule for said target.

[0163] As used herein, the term " IC₅₀" refers to the half maximal inhibitory concentration of a binding molecule which induces a response halfway between the baseline and maximum inhibition after a specified exposure time. Herein, a lower IC₅₀ value indicates that a lower concentration of target is required to interfere with the binding of the detection antibody to the known target coated on the plate, as compared to a higher IC₅₀ value. Thus, a lower IC₅₀ value typically corresponds to a higher affinity.

[0164] The binding affinity of a binding molecule may also be tested by surface plasmon resonance (SPR). For example, the affinity may be tested in an experiment in which target or epitope X is immobilized on a sensor chip of the instrument, and the sample containing the binding molecule to be tested is passed over the chip. Alternatively, the binding molecule to be tested may be immobilized on a sensor chip of the instrument, and a sample containing X is passed over the chip. The skilled person may then interpret the results obtained by such experiments to establish at least a qualitative measure of the binding affinity for X of the binding molecule. If a quantitative measure is desired, for example to determine a KD value for the interaction, SPR may also be used. Binding values may for example be defined in a Biacore (Cytiva) or ProteOn XPR 36 (Bio-Rad) instrument. The target or epitope is suitably immobilized on a sensor chip of the instrument, and samples of the binding molecule whose affinity is to be determined are prepared by serial dilution and injected. KD values may then be calculated from the results using for example the 1:1 Langmuir binding model of the Biacore Insight Evaluation Software 2.0 or other suitable software, typically provided by the instrument manufacturer.

[0165] The binding affinity may also be measured by bio-layer interferometry (BLI), a label-free technology for measuring biomolecular interactions within the interactome. It is an optical analytical technique that analyzes the interference pattern of white light reflected from two surfaces: a layer of immobilized protein on the biosensor tip, and an internal reference layer. The binding between a ligand (target or epitope X) immobilized on the biosensor tip surface and an analyte (such as a binding molecule with a putative affinity for X) in solution produces an increase in optical thickness at the biosensor tip resulting in a wavelength shift, Δλ, which is a direct measure of the change in thickness of the biological layer. Interactions are measured in real time, providing the ability to monitor binding specificity, rates of association and dissociation, or concentration, with precision and accuracy.

[0166] The skilled person is aware of the above mentioned and other methods for measuring the affinity of a binding molecule for a target or epitope X, either qualitatively or quantitatively or both.

[0167] In one embodiment, the antibody or antigen-binding fragment thereof as disclosed herein has at least 2x higher binding affinity for pathological TDP-43 than for physiological TDP-43, such as at least about 10x higher, such as at least about 20x higher, such as at least about 30x higher, such as at least about 40x higher, such as at least about 50x higher, such as at least about 60x higher, such as at least about 70x higher, such as at least about 80x higher, such as at least about 90x higher, such as at least about 100x higher, such as at least about 200x higher, such as at least about 300x higher, such as at least about 400x higher, such as at least about 500x higher, such as at least about 600x higher, such as at least about 700x higher, such as at least about 800x higher, such as at least about 900x higher, such as at least about 1000x higher, such as at least about 2000x higher, such as at least about 3000x higher, such as at least about 4000x higher, such as at least about 5000x higher, such as at least about 6000x higher, such as at least about 7000x higher, such as at least about 8000x higher, such as at least about 8500x higher binding affinity.

[0168] In a particular embodiment, the antibody or antigen-binding fragment thereof has essentially no binding affinity for physiological TDP-43. As used herein, the term "essentially no binding affinity" refers to that said antibody or antigenbinding fragment thereof has such low binding affinity to said protein that does not enable a quantitative measure of binding affinity to be obtained, as tested for example by surface plasmon resonance (SPR) and indicated in Example 9. In particular embodiments, the term "essentially no binding affinity for physiological TDP-43" refers to a property of said antibody or antigen-binding fragment thereof, wherein said property corresponds to selective binding of the antibody or antigenbinding fragment thereof to pathological TDP-43. This for example means that when physiological and pathological TDP-43 are present at the same concentrations in an assay in which binding affinity is tested, such as an ELISA, the antibody or antigenbinding fragment thereof preferably binds to pathological TDP-43, as demonstrated in e.g. Examples 7 and 8.

[0169] In a particular embodiment, the antibody or antigen-binding fragment thereof of the disclosure has a binding affinity for pathological TDP-43 that corresponds to a KDvalue of at most about 1 nM, such as at most about 900 pM, such as at most about 800 pM, such as at most about 700 pM, such as at most about 600 pM, such as at most about 500 pM, such as at most about 400 pM, such as at most about 300 pM, such as at most about 200 pM, such as at most about 190 pM, such as at most about 180 pM, such as at most about 170 pM, such as at most about 160 pM, such as at most about 150 pM, such as at most about 140 pM, such as at most about 130 pM, such as at most about 120 pM, such as at most about 110 pM, such as at most about 100, such as at most about 90 pM, such as at most about 80 pM, such as at most about 70 pM, such as at most about 63 pM; or such as a KDvalue of from about 1 pM to about 200 pM, such as from about 10 pM to about 190 pM, such as from about 20 pM to about 180 pM, such as from about 30 pM to about 170 pM, such as from about 40 pM to about 160 pM, such as from about 50 pM to about 150 pM, such as from about 60 pM to about 140 pM, such as from about 63 pM to about 140 pM. In another particular embodiment, the antibody or antigenbinding fragment thereof of the disclosure has a binding affinity for pathological TDP-43 that corresponds to a KD value of at most about 200 pM, such as at most about 190 pM, such as at most about 180 pM, such as at most about 170 pM, such as at most about 160 pM, such as at most about 150 pM; or such as a KD value of from about 100 pM to about 200 pM, such as from about 110 pM to about 200 pM, such as from about 120 pM to about 200 pM, such as from about 130 pM to about 200 pM, such as from about 140 pM to about 200 pM, such as from about 150 pM to about 200 pM, such as from about 150 pM to about 190 pM, such as from about 150 pM to about 180 pM, such as from about 150 pM to about 170 pM.

[0170] In one embodiment, the antibody or antigen-binding fragment thereof of the disclosure has a lower EC₅₀ for binding pathological TDP-43 than for binding physiological TDP-43. In one embodiment, the antibody or antigen-binding fragment thereof has at least about 1.5x lower EC₅₀ for binding pathological TDP-43 than for binding physiological TDP-43, such as at least about 1.6x lower, such as at least about 1.7x lower, such as at least about 1.8x lower, such as at least about 1.9x lower, such as at least about 2x lower, such as at least about 3x lower, such as at least about 4x lower, such as at least about 5x lower, such as at least about 6x lower, such as at least about 7x lower, such as at least about 8x lower, such as at least about 9x lower, such as at least about 10x lower, such as at least about 11x lower EC₅₀. The EC50 may be determined for example by ELISA, such as indirect ELISA, as demonstrated in Example 7. In one embodiment, the antibody or antigen-binding fragment thereof has an EC₅₀ for binding pathological TDP-43 that corresponds to an EC₅₀ value of at most about 20 ng / ml, such as at most about 19 ng / ml, such as at most about 18 ng / ml, such as at most about 17 ng / ml, such as at most about 16 ng / ml, such as at most about 15 ng / ml, such as at most about 14 ng / ml, such as at most about 13 ng / ml, such as at most about 12 ng / ml, such as at most about 11 ng / ml, such as at most about 10 ng / ml, such as at most about 9 ng / ml, such as at most about 8 ng / ml, such as at most about 7.5 ng / ml, such as at most about 7 ng / ml, such as at most about 6 ng / ml, such as at most about 5 ng / ml, such as at most about 4 ng / ml, such as at most about 3 ng / ml, optionally as determined by ELISA, such as indirect ELISA; such as an EC₅₀ value of from 1 ng / ml to about 20 ng / ml, such as from about 1 ng / ml to about 15 ng / ml, such as from about 1 ng / ml to about 14 ng / ml, such as from about 1 ng / ml to about 13 ng / ml, such as from about 1 ng / ml to about 12 ng / ml, such as from about 1 ng / ml to about 11 ng / ml, such from about 1 ng / ml to about 12 ng / ml, such as from about 1 ng / ml to about 10 ng / ml, such as from about 1 ng / ml to about 9 ng / ml, such as from about 1 ng / ml to about 8 ng / ml, such as from about 2 ng / ml to about 8 ng / ml, such as from about 2.5 ng / ml to about 8 ng / ml, such as from about 3 ng / ml to about 7.5 ng / ml, optionally as determined by ELISA, such as indirect ELISA. In one embodiment, the antibody or antigen-binding fragment thereof has an EC₅₀ for binding physiological TDP-43 that corresponds to an EC₅₀ value of at least about 4 ng / ml, such as at least about 5 ng / ml, such as at least about 6 ng / ml, such as at least about 7 ng / ml, such as at least about 8 ng / ml, such as at least about 9 ng / ml, such as at least about 10 ng / ml, such as at least about 20 ng / ml, such as at least about 25 ng / ml, such as at least about 30 ng / ml, such as at least about 35 ng / ml, such as at least about 40 ng / ml, such as at least about 45 ng / ml, such as at least about 50 ng / ml, such as at least about 55 ng / ml, such as at least about 57 ng / ml, such as at least about 60 ng / ml, optionally as determined by ELISA, such as indirect ELISA.

[0171] In a particular embodiment of the antibody or antigen-binding fragment thereof as disclosed herein has a lower IC₅₀ for binding pathological TDP-43 than for binding physiological TDP-43. In one embodiment, said antibody or antigen-binding fragment thereof has at least about 50x lower IC₅₀ for binding pathological TDP-43 than for binding physiological TDP-43, such at least about 100x lower, such as at least about 150x lower, such as at least about 200x lower, such as at least about 250x lower, such as at least about 300x lower, such as at least about 310x lower, such at least about 314x lower, such as at least about 315x lower, such at least about 350x lower, such at least about 400x lower, such at least about 450x lower, such at least about 500x lower, such at least about 550x lower, such at least about 580x lower, such at least about 583x lower, such at least about 585x lower, such at least about 600x lower, such at least about 630x lower, such at least about 635x lower, such at least about 636x lower, such at least about 640x lower, such at least about 650x lower IC₅₀. The IC₅₀ may be determined for example by ELISA, such as by inhibition ELISA, as demonstrated in Example 8. In one embodiment, the antibody or antigen-binding fragment thereof has an IC₅₀ for binding pathological TDP-43 that corresponds to an IC₅₀ value of at most about 10 ng / ml, such as at most about 9 ng / ml, such as at most about 8 ng / ml, such as at most about 7 ng / ml, such as at most about 6 ng / ml, such as at most about 5 ng / ml, such as at most about 4 ng / ml, such as at most about 3 ng / ml, such as at most about 2 ng / ml, such as at most about 1.9 ng / ml, such as at most about 1.8 ng / ml, such as at most about 1.7 ng / ml, such as at most about 1.6 ng / ml, such as at most about 1.5 ng / ml, such as at most about 1.4 ng / ml, such as at most about 1.3 ng / ml, such as at most about 1.2 ng / ml, such as at most about 1.1 ng / ml, such as at most about 1 ng / ml, such as at most about 0.9 ng / ml, optionally as determined by ELISA, such as indirect ELISA; such as an IC₅₀ value of from about 0.5 ng / ml to about 10 ng / ml, such as from about 0.5 ng / ml to about 9 ng / ml, such as from about 0.5 ng / ml to about 8 ng / ml, such as from about 0.5 ng / ml to about 7 ng / ml, such as from about 0.5 ng / ml to about 6 ng / ml, such as from about 0.5 ng / ml to about 5 ng / ml, such as from about 0.5 ng / ml to about 4 ng / ml, such as from about 0.5 ng / ml to about 3 ng / ml, such as from about 0.5 ng / ml to about 2 ng / ml, such as from about 0.5 ng / ml to about 1.9 ng / ml, such as from about 0.5 ng / ml to about 1.8 ng / ml, such as from about 0.5 ng / ml to about 1.7 ng / ml, such as from about 0.5 ng / ml to about 1.6 ng / ml, such as from about 0.5 ng / ml to about 1.5 ng / ml, such as from about 0.6 ng / ml to about 1.5 ng / ml, such as from about 0.7 ng / ml to about 1.4 ng / ml, such as from about 0.8 ng / ml to about 1.3 ng / ml, such as from about 0.9 ng / ml to about 1.2 ng / ml, optionally as determined by ELISA, such as indirect ELISA. In one embodiment, the antibody or antigen-binding fragment thereof has an IC₅₀ for binding physiological TDP-43 that corresponds to an IC₅₀ value of at least about 15 ng / ml, such as at least about 20 ng / ml, such as at least about 25 ng / ml, such as at least about 50 ng / ml, such as at least about 100 ng / ml, such as at least about 150 ng / ml, such as at least about 200 ng / ml, such as at least about 250 ng / ml, such as at least about 280 ng / ml, such as at least about 283 ng / ml, such as at least about 285 ng / ml, such as at least about 300 ng / ml, such as at least about 350 ng / ml, such as at least about 400 ng / ml, such as at least about 450 ng / ml, such as at least about 500 ng / ml, such as at least about 550 ng / ml, such as at least about 600 ng / ml, such as at least about 650 ng / ml, such as at least about 700 ng / ml, optionally as determined by ELISA, such as inhibition ELISA.

[0172] In a particular embodiment of the antibody or antigen-binding fragment thereof as disclosed herein, it has essentially no nuclear localization in living cells. The term "essentially no nuclear localization", as used herein, means that uptake or, in other words nuclear transport, of said antibody or antigen-binding fragment thereof into the nucleus of a living cell does not occur. Particularly, no nuclear localization can be observed, even when said antibody or antigen-binding fragment thereof is internalized into the cytoplasm of said living cell. Nuclear localization can for example be tested in an antibody internalization assay in vitro, such as presented in Example 12, wherein a quantum dot labeled antibody is added to cells grown in a cell culture, and the localization of said labeled antibody is analyzed by confocal microscopy.

[0173] In a particular embodiment of the antibody or antigen-binding fragment thereof as disclosed herein, it induces Fc-receptor mediated endocytosis of pathological TDP-43, such as a TDP-43 aggregate, optionally as determined by an in vitro internalization assay, such as an internalization assay using microglia cells, for example as demonstrated in Example 11.

[0174] As discussed above, in a particular embodiment of the antibody or antigenbinding fragment thereof as disclosed herein, it has essentially no binding affinity to physiological TDP-43, such as TDP-43 monomers, non-disease associated TDP-43 oligomers and / or non-disease associated reversible TDP-43 aggregates. Besides the above explained quantitative measures, a lack of binding affinity of said antibody or antigen-binding fragment thereof to physiological TDP-43 may be determined by immunostaining, such as immunocytochemistry, wherein colocalization between the antibody or antigen-binding fragment thereof and a physiological TDP-43 variant can be tested. Said colocalization is indicative of binding of said antibody or antigenbinding fragment thereof to said physiological TDP-43 variant, whereas the lack of colocalization is indicative of the lack of binding of said antibody or antigen-binding fragment thereof to said physiological TDP-43 variant. For example, stress granules in cells are known to comprise physiological TDP-43, more specifically reversible TDP-43 aggregates. Thus, binding of said antibody or antigen-binding fragment thereof to reversible TDP-43 aggregates can be tested by evaluating colocalization of a labeled, e.g. fluorescently labeled, antibody or antigen-binding fragment thereof with labeled, e.g. fluorescently labeled, stress granules in a staining, wherein stress granule formation may be induced. Thus, in one embodiment, essentially no binding affinity of the antibody or antigen-binding fragment thereof as disclosed herein to physiological TDP-43 corresponds to a lack of binding of said antibody or antigen- binding fragment thereof to intracellular stress granules expressing TDP-43 protein, such as non-disease associated, reversible TDP-43 aggregates, optionally wherein said binding is visualized using immunocytochemistry. Accordingly, in one embodiment, the antibody or antigen-binding fragment thereof has essentially no binding affinity to intracellular stress granules.

[0175] In a second aspect, the disclosure provides a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof as defined herein and a pharmaceutically acceptable carrier and / or excipient.

[0176] Techniques for formulating polypeptides such as antibodies and their derivatives for human therapeutic use are well known in the art and are reviewed, for example, in Wang et al (2007), J Pharm Sci 96:1-26, the contents of which are incorporated herein in their entirety.

[0177] Pharmaceutically acceptable excipients that may be used to formulate the compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances (for example sodium carboxymethylcellulose), polyethylene glycol, polyacrylates, waxes, polyethylenepolyoxypropylene block polymers, polyethylene glycol and wool fat.

[0178] In certain embodiments, the pharmaceutical compositions are formulated for administration to a subject via any suitable route of administration including but not limited to intramuscular, intravenous, intradermal, intraperitoneal injection, subcutaneous, epidural, nasal, oral, rectal, topical, inhalational, buccal (e.g., sublingual), and transdermal administration. In preferred embodiments, the composition is formulated for intravenous or subcutaneous administration.

[0179] The antibody or antigen-binding fragment thereof according to the present disclosure may be useful as a therapeutic, prophylactic, diagnostic and / or prognostic agent. Hence, in a further aspect of the disclosure, there is provided an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect, for use as a medicament.

[0180] In yet another aspect of the disclosure, there is provided an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect, for use as a diagnostic agent, e.g. in vivo.

[0181] In yet another aspect of the disclosure, there is provided an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect, for use as a prognostic agent, e.g. in vivo.

[0182] Also provided are methods of preventing, treating or diagnosing disease or assessing disease prognosis, wherein an antibody or antigen-binding fragment thereof as disclosed herein is administered to a subject in need thereof, typically a human subject.

[0183] Also provided is the use of the disclosed antibody or antigen-binding fragment thereof for the manufacture of compositions (such as medicaments) for use in the prevention, treatment, diagnosis and / or prognosis of any one of the listed diseases.

[0184] Thus, there is provided an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect for use in treatment, such as for use in therapeutic treatment or for use in prophylactic treatment.

[0185] In a particular embodiment, said therapy, prophylaxis, diagnosis or prognosis is with respect to a neurodegenerative disorder associated with a TDP-43 proteopathy, such as a disorder associated with non-reversible aggregation of TDP-43.

[0186] Thus, there is provided an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect, for use in diagnosis in vivo or in prognosis in vivo.

[0187] Moreover, herein is provided a method of therapeutic or prophylactic treatment of a mammal having or being at risk of developing a neurodegenerative disorder, said method comprising administering to said mammal a therapeutically effective amount of an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect. In one embodiment of said method of therapeutic or prophylactic treatment, said neurodegenerative disorder is a disorder associated with a TDP-43 proteopathy, such as a disorder associated with non-reversible aggregation of TDP-43.

[0188] In one embodiment, said neurodegenerative disorder is selected from the group consisting of amyotrophic lateral sclerosis (ALS); Alzheimer's disease (AD), optionally wherein said AD is selected from the group consisting of familial AD and sporadic AD; limbic-predominant age-related TDP-43 encephalopathy (LATE); and frontotemporal dementia (FTD). In one embodiment, said neurodegenerative disorder is selected from the group consisting of amyotrophic lateral sclerosis (ALS), limbic-predominant age-related TDP-43 encephalopathy (LATE) and frontotemporal dementia (FTD). In one embodiment, said neurodegenerative disorder is selected from the group consisting of amyotrophic lateral sclerosis (ALS) and limbic-predominant age-related TDP-43 encephalopathy (LATE). In one embodiment, said neurodegenerative disorder is selected from the group consisting of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). In a particular embodiment, said neurodegenerative disorder is amyotrophic lateral sclerosis (ALS). In another particular embodiment, said neurodegenerative disorder is limbic-predominant age-related TDP-43 encephalopathy (LATE). In yet another particular embodiment, said disorder is frontotemporal dementia (FTD). In yet another particular embodiment, said neurodegenerative disorder is Alzheimer's disease (AD), optionally wherein said AD is selected from the group consisting of familial AD and sporadic AD.

[0189] In one aspect, there is provided a method of detecting pathological TDP-43, such as non-reversible TDP-43 aggregates, said method comprising the steps of providing a sample suspected to contain pathological TDP-43, such as non-reversible TDP-43 aggregates,

[0190] contacting said sample with an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect, and detecting the binding of said antibody or the antigen-binding fragment thereof to indicate the presence of pathological TDP-43, such as non-reversible TDP-43 aggregates, in said sample.

[0191] In one embodiment of said method of detecting pathological TDP-43, said sample is a biopsy and / or a body fluid, such as wherein said sample is a skin biopsy, cerebrospinal fluid, plasma, serum and / or saliva. In one embodiment, said sample is isolated from a human.

[0192] In one embodiment, said method of detecting pathological TDP-43 is an immunoassay, such as an ELISA method and / or an immunohistochemistry method.

[0193] In another aspect, there is provided a method of determining the amount of pathological TDP-43, such as non-reversible TDP-43 aggregates, present in a subject, said method comprising the steps of

[0194] contacting the subject, or a sample isolated from the subject, with an antibody or antigen-binding fragment thereof according to the first aspect, or a pharmaceutical composition according to the second aspect, and

[0195] obtaining a value corresponding to the amount of the antibody or the antigenbinding fragment thereof or the composition that has bound in said subject or to said sample, and

[0196] comparing said value to a reference value.

[0197] In one embodiment of said method of determining the amount of pathological TDP-43, said sample is a biopsy and / or a body fluid, such as wherein said sample is a skin biopsy, cerebrospinal fluid, plasma, serum and / or saliva. In one embodiment, said sample is isolated from a human. In one embodiment, said method of determining the amount of pathological TDP-43 is an immunoassay, such as an ELISA method and / or an immunohistochemistry method.

[0198] In a further aspect of the disclosure, there is provided a polynucleotide encoding an antibody or antigen-binding fragment thereof according to the first aspect of the disclosure.

[0199] In yet another aspect of the disclosure, there is provided a cloning vector comprising a polynucleotide as defined above.

[0200] In yet another aspect of the disclosure, there is provided an expression vector comprising a polynucleotide as defined above. In yet another aspect of the disclosure, there is provided a host cell comprising a cloning vector as defined above.

[0201] In yet another aspect of the disclosure, there is provided a host cell comprising an expression vector as defined above.

[0202] In yet another aspect of the disclosure, there is provided a method of producing an antibody or antigen-binding fragment thereof according to the first aspect of the disclosure, comprising

[0203] culturing a host cell comprising an expression vector as defined above under conditions permissive for the expression of said antibody or antigen-binding fragment thereof from said expression vector, and

[0204] isolating said antibody or antigen-binding fragment thereof.

[0205] Incorporation by reference

[0206] Various publications are cited in the present application, each of which is incorporated by reference herein in its entirety.

[0207] Brief description of the figures

[0208] Figure 1 shows the results of a Western blot (WB) analysis (A), the elution profile obtained via a high-performance liquid chromatography (HPLC) analysis (B) and a transmission electron microscopy (TEM) image (C) of a human TDP-43 protein preparation enriched in aggregates of human TDP-43.

[0209] Figure 2 shows the elution profile obtained via HPLC analysis of the monomeric C-terminal human TDP-43 protein preparation (SEQ ID NO:54) used in the appended Examples.

[0210] Figure 3 shows indirect ELISA results for rec20 (A), rec30 (B) and rec41 (C), tested in different concentrations on plates coated by either human TDP-43 monomers (C-terminal human TDP-43) or human TDP-43 aggregates, as indicated.

[0211] Figure 4 shows inhibition ELISA results for rec20 (A), rec30 (B) and rec41 (C), preincubated with either human TDP-43 monomers (C-terminal human TDP-43) or human TDP-43 aggregates in different concentrations prior to testing on a plate coated with human TDP-43 aggregates.

[0212] Figure 5 presents representative sensorgrams obtained by surface plasmon resonance (SPR) analysis of antibodies rec20 (A), rec30 (B) and rec41 (C) using human TDP-43 aggregate immobilized on the chip and SPR analysis of antibodies rec30 (D) and rec41 (E) captured by an anti-mouse antibody (AMC) immobilized on the chip and using human TDP-43 monomers (C-terminal human TDP-43) as analyte.

[0213] Figure 6 shows representative sensorgrams obtained by SPR analysis of Fab fragments of antibodies rec30 (A) and rec41 (B) using human TDP-43 aggregate immobilized on the chip.

[0214] Figure 7 shows results of a Meso Scale Discovery (MSD) assay of total TDP-43 following immunodepletion of human TDP-43 from sarcosyl soluble (A) and insoluble fractions (B) of human brain extracts using the indicated antibodies of the disclosure and controls (FTD: frontotemporal dementia).

[0215] Figure 8 shows results of an MSD assay of aggregated TDP-43 following immunodepletion of human TDP-43 from sarcosyl soluble (A) and insoluble fractions (B) of human brain extracts using the indicated antibodies of the disclosure and controls.

[0216] Figure 9 shows the results of an indirect MSD assay using plates coated with human brain extracts from FTD or NDE, with serial dilutions of antibody rec30 of the disclosure. Binding to PBS coated wells was included as control.

[0217] Figure 10 shows FcyR-mediated uptake of Alexa-488-labeled human TDP-43 aggregates in the presence of increasing concentrations of the indicated murine recombinant antibodies and negative control.

[0218] Figure 11 illustrates rec30 binding to pathological human TDP-43, such as human TDP-43 aggregates, in brain sections from an induced rNLS8 mouse characterized by the presence of human TDP-43 aggregates in the cytoplasm of the cells, in comparison to a wild-type control mouse (WT) without such aggregates.

[0219] Figure 12 shows a statistically significant difference in grip strength (average percent change from baseline) of female rNLS8 transgenic mice treated biweekly with rec30 for a period of 6 weeks, in comparison to vehicle (PBS) treated controls (*p=0.038, Student's T-test) three weeks after disease induction.

[0220] Figure 13 shows the result of the compound muscle action potential (CMAP) measurement described in Example 13, on male or female, 15-week old, non-transgenic control mice (tTA) or rNLS8 mice either treated with PBS or with the rec30 antibody of the disclosure as indicated (**=p<0.01, ***=p<0.001, Tukey's unequal N post-hoc test). Figure 14 shows (A, B) the IR area % of pTDP-43 labeling in the cerebral cortex (A) and in dorsal striatum (B) of the indicated groups; and (C) IOD of pTDP-43 labeling in the spinal cord of males, measured as described in Example 13. Data are shown as mean and scatter ± SEM; *=p<0.05, **=p<0.01, ****=p<0.0001.

[0221] Figure 15 shows the result of microglia branch analysis in the male cerebral cortex in the indicated groups of mice, performed as described in Example 13. Data are shown as mean and scatter ± SEM; **=p<0.01, ***=p<0.001.

[0222] Figure 16 shows the result of quantification of GFAP in the spinal cord of females in the indicated groups of mice, performed as described in Example 13. Data are shown as mean and scatter ± SEM; *=p<0.05.

[0223] Figure 17 shows the relative quantification of Uncl3a cryptic exons where each sample demonstrates the fraction of cryptic exons compared to the level of its native Uncl3a mRNA. An unpaired t-test shows a statistical difference between the two groups (**=p<0.005).

[0224] Figure 18 shows reduced plasma levels of neurofilament light (NfL) in induced rNLS8 mice upon weekly treatment with 30 mg / kg rec30 (A) as well as biweekly treatment with 30 mg / kg rec30 and 15 mg / kg rec41 (B) in comparison to vehicle-treated controls in a low DOX model.

[0225] Figure 19 shows pharmacokinetic evaluation (plasma concentration (mean±SD) over time) of murine recombinant antibodies rec20, rec30 and rec41 after a single i.v. bolus injection of the respective antibody at a dose of 30 mg / kg in C57BL / 6J mice (n=5).

[0226] Figure 20 presents representative sensorgrams obtained by SPR analysis of humanized h30 antibody variants h30-H1L4 (A) and h30-H4L3 (B) using human TDP-43 aggregate immobilized on the chip.

[0227] Figure 21 shows inhibition ELISA results for humanized h30 antibody variants h30-H1L4 (A) and h30-H4L3 (B) preincubated with either human TDP-43 monomers (C-terminal human TDP-43) or human TDP-43 aggregates in different concentrations prior to testing on a plate coated with human TDP-43 aggregates.

[0228] Figure 22 illustrates binding of humanized h30 antibody variants h30-H1L4 and h30-H4L3 to pathological human TDP-43, such as human TDP-43 aggregates, in paraffin embedded postmortem brain tissue from a patient with FTD, characterized by the presence of non-reversible human TDP-43 aggregates in the cytoplasm of the cells, in comparison to a positive control staining (using a pSer 409 / 410 antibody) and a negative control staining (using an isotype control antibody).

[0229] Figure 23 shows pharmacokinetic evaluation (plasma concentration (mean±SD) over time) of the indicated h30 humanized antibody variants using antigen coated MSD assays (A) and sandwich based MSD assays (B) after a single i.v. bolus injection of the respective antibody at a dose of 10 mg / kg in female Tg32- / -(human FcRn tg) mice (n=4 or 5; lower limit of quantification (LLOQ) was 0.018 pg / ml (LLOQ2) for H2L1 and 0.055 pg / ml (LLOQ1) for H2L2 and H3L2; values below LLOQ were set to ½ LLOQ).

[0230] Figure 24 shows FcyR-mediated uptake of Alexa-488-labeled human TDP-43 aggregates by THP-1 cells in the presence of increasing concentrations of the indicated humanized antibodies and negative control.

[0231] Figure 25 shows indirect ELISA results for h30-H1L4 after stability test at the indicated treatment conditions, as described in Example 22.

[0232] Figure 26 shows indirect ELISA results for h30(H: D95G) and h30(L: D29Q) and h30(H: D95G-L: D29Q) tested in different concentrations on plates coated by human TDP-43 aggregates, as described in Example 23.

[0233] Figure 27 presents representative sensorgrams obtained by SPR analysis of h30 antibody variants h30(H: D95G) (A), h30(L: D29Q) (B) and h30(H: D95G-L: D29Q) (C) using human TDP-43 aggregate immobilized on the chip, measured as described in Example 23.

[0234] Figure 28 illustrates binding of h30-H1L4 and indicated variants thereof to pathological human TDP-43, such as human TDP-43 aggregates, in paraffin embedded postmortem brain tissue from patients with FTD (FTD) characterized by the presence of non-reversible human TDP-43 aggregates (white arrows) in the cytoplasm of the cells or from non-demented controls (NDE), in comparison to binding using a positive control staining (pSer 409 / 410).

[0235] Figure 29 shows the results of indirect MSD assay for h30-H1L4 and h30(H: D95G) as indicated, using plates coated with human brain extracts from FTD or NDE as indicated, with serial dilutions of the indicated antibodies.

[0236] Figure 30 shows antibody-bound total TDP-43 by MSD assay (A) and pTDP-43 by AlphaLISA human pTDP-43 (Ser409 / 410) (B) after immunoprecipitation (IP) using h30-HlL4 and h30(H: D95G) in human brain extracts from FTE or ALS as indicated. IP was performed using titrated amounts of coupled antibody as described in Example 23.

[0237] Figure 31 shows pharmacokinetic evaluation (plasma concentration (mean±SD) over time) of h30-H1L4 and variants thereof as indicated, using a sandwich based MSD assays after a single i.v. bolus injection of the respective antibody at a dose of 10 mg / kg in female SCID FcRn tg mice (n= 5) (values below LLOQ were set to ½ LLOQ).

[0238] Figure 32 shows the results of indirect ELISA of h30-H1L4 and variants thereof as indicated after a stability test at 40 °C, as described in Example 24.

[0239] EXAMPLES

[0240] While the invention has been described with reference to various exemplary aspects and embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or molecule to the teachings of the invention without departing from the essential scope thereof.

[0241] Therefore, it is intended that the invention is not to be limited to any particular embodiment, but that the invention will include all embodiments falling within the scope of the appended claims.

[0242] The invention will be further illustrated by the following non-limiting Examples. They are offered for illustrative purposes only and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of non-critical parameters which can be changed or modified to yield essentially the same results. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperatures, etc.), but some experimental error and deviations may be present. Unless otherwise indicated, the practice of the invention employs conventional methods of protein chemistry, biochemistry, recombinant DNA techniques and pharmacology, within the skill of the art. Such techniques are explained fully in the existing literature. Additionally, it will be apparent to one of skill in the art that the methods for protein engineering applied herein can also be applied to other constructs described herein and contemplated by the present inventors to fall within the scope of the disclosure. Example 1

[0243] Characterization of human TDP-43 protein preparation containing aggregates A preparation of full length human TDP-43 protein (comprising SEQ ID NO:53) was analyzed by western blot (WB), high-performance liquid chromatography (HPLC) and transmission electron microscopy (TEM) to determine its purity and aggregation state.

[0244] Materials and methods

[0245] A batch of human TDP-43 protein preparation was obtained in a buffer containing 20 mM Tris-HCI pH 8.0, 0.1 M NaCI, 1 mM DTT, 1 mM EDTA, 10% glycerol.

[0246] Western blot analyses were made using both non-reduced and reduced human TDP-43. Proteins were separated on an SDS-PAGE gel (4-12% BOLT Bis-Tris gel, Invitrogen) and then blotted using a 0.2 pm nitrocellulose transfer pack with a Trans-Blot Turbo system (BioRad). A rabbit polyclonal antibody against TDP-43 (Proteintech, 10782-2-AP) was used for detection together with goat anti Rabbit IRdye 800CW (LI-COR, IRDye 800CW).

[0247] The HPLC analysis was performed using a Superdex 200 Increase 3.2 / 300 (Cytiva) column. TEM analyses were performed according to standard methods.

[0248] Results

[0249] Analysis of the obtained human TDP-43 protein preparation by WB using reducing conditions showed a band around 49 kDa, which corresponds to the approximate size of monomeric human TDP-43. Non-reducing conditions showed the presence of larger aggregates of human TDP-43 (Figure 1A).

[0250] As shown in Figure IB, the elution profile using HPLC of the analyzed human TDP-43 protein preparation has a peak at around 12 min, suggesting the presence of larger aggregates of TDP-43.

[0251] TEM analysis of the human TDP-43 protein preparation showed the presence of aggregates of various sizes (Figure 1C). Conclusions

[0252] As can be seen from the HPLC elution profile (Figure IB), the analyzed human TDP-43 protein preparation has a high degree of purity. The human TDP-43 protein preparation comprises human TDP-43 aggregates, as shown in Figure 1. In fact, these aggregates are present predominantly over human TDP-43 monomers in the protein preparation. The analyzed human TDP-43 preparation is thus considered as a protein preparation consisting of human TDP-43 aggregates, and is referred to below as human TDP-43 aggregates. Human TDP-43 aggregates were used for immunization of mice as described in Example 2. Moreover, human TDP-43 aggregates are useful for, and have been used for, testing the binding of antibodies to human TDP-43 aggregates.

[0253] Example 2

[0254] Hybridoma generation and screening

[0255] For the generation of hybridoma, Balb / c mice were immunized with the human TDP-43 aggregates described in Example 1. Following the immunization protocol described below, fusion of spleen cells from the immunized mice with SP2 / 0 cells was performed. The antibodies produced by the hybridomas were screened for binding to the human TDP-43 aggregates.

[0256] Materials and methods

[0257] Five Balb / c mice were immunized with 20 pg of human TDP-43 aggregates (described in Example 1) using an adjuvant. In brief, the mice were immunized every four weeks, receiving a total of four immunizations. Serum antibody titers were analyzed using indirect ELISA against aggregated TDP-43 (see Example 1) two weeks after the fourth immunization. The mice were given a final booster injection with aggregated TDP-43 without adjuvant, and three days later the animals were sacrificed. The spleens were removed and used for either hybridoma generation or preparation of single cell suspensions, followed by storage in liquid nitrogen for possible future hybridoma generation.

[0258] Fusion of spleen cells from immunized Balb / c mice was performed. Briefly, the spleens were prepared to single cell suspension that was mixed with SP2 / 0 cells. The fusion was made with 50% polyethylene glycol (PEG) at 37°C. The fused cells were diluted in HAT selection medium and seeded at 1x105splenocytes / well in 96-well plates. The cells were left undisturbed for one week, and thereafter the medium was changed to fresh HAT medium. 10 days after fusion, screening for antibodies binding to human TDP-43 aggregates was done using indirect ELISA.

[0259] Indirect ELISA was used for the analysis of serum antibody titers and for screening of hybridoma antibody binding to human TDP-43 aggregates. ELISA plates were coated with 0.5 pg / ml of aggregate TDP-43 (see Example 1) and incubated at +4°C O / N or at 37°C for 1 h. The plates were blocked in PBS-BSA (1%) at RT for 1 h at 900 rpm. Titrating amounts of antibodies were added to the coat. After 1-1.5 h incubation the bound antibodies were detected by goat-anti-mouse IgG-HRP (Southern Biotechnology, 1030-05). After 1 h incubation the substrate TMB was added and the reaction was stopped after 3-10 min using 0.18 M H2SO4. The absorbance at 450 nm was recorded for each well.

[0260] Results

[0261] Serum antibody titers: The serum antibody titers for reactivity against human TDP-43 aggregates were evaluated 2 weeks after the fourth immunization using an indirect ELISA. The immunization with human TDP-43 aggregates resulted in high serum levels of antibodies recognizing human TDP-43 aggregates in all five mice.

[0262] Screening of hybridomas: Hybridomas produced from mice immunized with human TDP-43 aggregates were screened using indirect ELISA for binding to human TDP-43 aggregates as described above. The results indicated that the tested hybridomas produced antibodies which bind human TDP-43 aggregates.

[0263] Conclusions

[0264] Each tested serum from mice immunized with human TDP-43 aggregates comprised antibodies which bind human TDP-43 aggregates. Such binding was detected even at high serum dilutions. These results indicate that the immunized mice produced antibodies having advantageous characteristics in binding TDP-43 aggregates.

[0265] Hybridoma supernatants positive in the indirect ELISA for binding to a plate coated with human TDP-43 aggregates comprise monoclonal antibodies which bind to human TDP-43 aggregates. Positive hybridomas, e.g. hybridomas expressing monoclonal antibodies denoted mAb20, mAb30, and mAb41, were further expanded, and monoclonal antibodies were harvested and analyzed using immunohistochemistry for binding to human TDP-43 aggregates typically present in brains sections of patients suffering from a TDP-43 proteopathy (see Example 3).

[0266] Example 3

[0267] Evaluation of murine monoclonal antibodies in brain sections of ALS patients The murine monoclonal antibodies obtained as described in Example 2 and denoted mAb20, mAb30, and mAb41 were evaluated in postmortem brain sections from patients diagnosed with amyotrophic lateral sclerosis (ALS) using immunohistochemistry (IHC) in comparison to non-demented (NDE) controls.

[0268] Materials and methods

[0269] Briefly, for single chromogenic immunohistochemical staining of human TDP-43, an automated staining robot and a detection system based on horse radish peroxidase and 3,3'-diaminobenzidine (DAB) (Discovery XT and OmniMap DAB kit, Ventana Medical Systems) were used. The analyses were primarily performed on formalin-fixed paraffin embedded tissue sections due to their superior performance regarding tissue morphology and lower background staining as compared to fresh frozen tissue. Visualization of the primary / secondary antibody complex was done by addition of hydrogen peroxide and DAB, resulting in an insoluble brown staining representing the antibody binding. Reference antibodies were an anti-TDP-43 antibody, 2E2D3 (Abeam, ab57105), and an anti-phospho-TDP-43 (pS409 / 410) antibody (Cosmo Bio, TIP-PTD-M01). Mouse IgGl was used as isotype control.

[0270] Counterstaining was done with hematoxylin. The stained slides were scanned in bright field at 40x-107x magnification, using a Pannoramic 250 FLASH II slide scanner. The resulting image files were uploaded into a viewer software (Pannoramic Viewer) and adjusted for optimal brightness and contrast for manual assessment of the staining result.

[0271] Results

[0272] Antibodies were evaluated on brain sections from ALS patients and compared to sections from NDE controls. All antibodies bound well to pathological TDP-43 in the cytoplasm, giving a similar pattern as seen for an antibody recognizing pS409 / 410 of TDP-43.

[0273] As shown in Tables 1 and 2, the tested monoclonal antibodies (mAb) stain cytoplasmic inclusions in brain sections from ALS patients tested by IHC. These inclusions are not present in NDE controls. Thus, the tested antibodies do not show staining of brain sections of NDE controls. Immunohistochemical staining of cytoplasmic inclusions / neurites using the antibodies according to the disclosure gives a similar pattern as seen for a positive control antibody recognizing pS409 / 410 of TDP-43.

[0274] Staining pattern of mAb20 in brain sections from ALS patients revealed weak nuclear staining in cortical neurons. Neurites and inclusion bodies, identified as pathological forms of TDP-43, were stained with a comparable intensity as the reference antibody pS409 / 410.

[0275] Staining pattern of mAb30 in brain sections from ALS patients revealed staining of neurites, inclusion bodies and cytoplasmic binding, and weak binding in neural nuclei. Neurites and inclusion bodies, identified as pathological forms of TDP-43, were stained with a comparable intensity as the reference antibody pS409 / 410.

[0276] Staining pattern of mAb41 in brain sections from ALS patients revealed strong staining in neuronal nuclei and weaker staining in oligodendrocytes. Staining of cytoplasmic inclusion bodies and neurites was observed. Neurites and inclusion bodies, identified as pathological forms of TDP-43, were stained with a comparable intensity as the reference antibody pS409 / 410.

[0277] Table 1. Summary of antibody binding in brain sections from NDE controls Nuclear Inclusions / Cytoplasmic Antibodies Subclass

[0278] binding Neurites binding Reference / control antibodies

[0279] Yes, TDP-43 (2E2D3) mlgGl Yes No pyramidal neurons TDP-43

[0280] mlgGl No No No (pS409 / 410)

[0281] Mouse IgGl

[0282] mlgGl No No No Isotype control

[0283]

[0284] Antibodies of the disclosure Yes, only

[0285] mAb20 lgG2a No No cortex neurons

[0286] mAb30 lgG2a No No No

[0287]

[0288] mAb41 lgG2a Yes No No

[0289] Table 2. Summary of antibody binding in brain sections from ALS patients Nuclear Inclusions / Cytoplasmic Antibodies Subclass

[0290] binding Neurites binding Reference / control antibodies

[0291] Yes, TDP-43 (2E2D3) mlgGl Yes Yes pyramidal neurons TDP-43 (pS409 / 410) mlgGl No Yes Yes Mouse IgGl Isotype

[0292] mlgGl No No No control

[0293] Antibodies of the disclosure

[0294] Yes, only

[0295] mAb20 lgG2a Yes Yes cortex neurons

[0296] mAb30 lgG2a Yes, very weak Yes Yes

[0297]

[0298] mAb41 lgG2a Yes Yes Yes

[0299] Conclusions

[0300] Tested monoclonal antibodies according to the disclosure bind pathological forms of human TDP-43, e.g. TDP-43 inclusions present in the brain. Binding can be detected in brain sections for example by IHC. These inclusions are a hallmark of a neurodegenerative disorder, such as ALS, and are present in brains of ALS patients. It is expected that binding of these antibodies to inclusions would occur in any brain section derived from a brain wherein TDP-43 inclusions are present. The inclusions comprise pathological human TDP-43, such as human TDP-43 aggregates. Thus, the tested antibodies of the disclosure bind to pathological human TDP-43, such as human TDP-43 aggregates. Pathological human TDP-43 aggregates are considered non-reversible. Example 4

[0301] Epitope mapping

[0302] Epitopes within the amino acid sequence of human TDP-43 recognized by antibodies according to the disclosure were determined.

[0303] Materials and methods

[0304] Briefly, for epitope mapping, 15-mer peptides of human TDP-43 (Q13148, SEQ ID NO:53) with 4 amino acids of offset were synthesized and covalently immobilized with a linker on a microarray glass slide. Each of the tested antibodies mAb20, mAb30, and mAb41 (1 pg / ml) was incubated on the microarray slides for 1 h at 30°C and detected with a secondary, fluorescently labeled anti-mouse IgG antibody.

[0305] The peptides giving a positive signal were also tested in ELISA and dot blot format by coating with 10 pg / ml of the peptides. Bound antibodies were detected with goat anti-mouse IgG (Southern Biotech (1030-05)) and goat anti-human IgG, respectively. TMB was used as substrate in the ELISA.

[0306] Results

[0307] Epitope mapping revealed that all tested antibodies (mAb20, mAb30 and mAb41) have affinity for an epitope of TDP-43 (Q13148, SEQ ID NO:53) comprising the amino acid sequence within positions 377-387 of SEQ ID NO:53, namely " SNSGAAIGWGS" (SEQ ID NO:52). Binding to the amino acid sequence SEQ ID NO:52 was revealed by binding of the antibodies to 15-mers comprising this amino acid sequence. The antibodies were not found to bind to any 15-mers which did not comprise sequence SEQ ID NO:52.

[0308] Conclusions

[0309] Binding to the identified epitope by the tested antibodies enables binding to human TDP-43. Based on the results of the examples below, it is expected that the epitope binding characteristics of the tested antibodies enable selective binding of said antibodies to pathological human TDP-43, such as human TDP-43 aggregates, over physiological human TDP-43, such as human TDP-43 monomers. Example 5

[0310] Construction and production of mouse recombinant antibodies Selected mouse hybridoma antibodies were sequenced, and the variable regions and complementarity determining region (CDR) sequences determined.

[0311] Materials and methods

[0312] Sequencing of hybridoma pellets was performed by whole transcriptome shotgun sequencing (RNA-Seq) at Absolute Antibody, UK.

[0313] Recombinant antibodies were constructed from the variable sequences obtained from hybridoma sequencing. Murine constant regions IGHG2A*01 and IGKC*01 were added. Antibodies were expressed in HEK cells and purified using affinity chromatography and SEC chromatography.

[0314] Results

[0315] The sequencing results are summarized in Table 3, which shows the sequences of heavy and light chain variable regions (VH and VL, respectively) and complementarity determining regions (CDRs) of the heavy and light chain variable regions (VH- and VL-CDRs, respectively) of analyzed monoclonal antibodies. Table 3. Amino acid sequences of antibodies of the disclosure SEQID

[0316] Antibody Region Amino acid sequence

[0317] NO: Heavy chain

[0318] QVQLQQSGPELVRPGVSVKISCKVSGYTFTEYNIHWVKES VH HATSLEWIGVINPYSENTNYNPKFTDKATMTVDKSSNTAY 34

[0319] MELARLTSEDSAIYYCARDGYFDFWGQGTTLTVSS VH-CDR1 EYNIH 18 VH-CDR2 VINPYSENTNYNPKFTD 21 VH-CDR3 DGYFDF 24 mAb20 Light chain

[0320] DVLMTQTPLSLTVSLGDQASISCRSSQIIVHTNGATYLEWY VL LQKPGQPPKLLIYKVSKRFSGVPDRFSGSGSGTDFTLKISRV 37

[0321] EAEDLGVYYCFQGSHVPPTFGGGTNLEIK VL-CDR1 RSSQIIVHTNGATYLE 27 VL-CDR2 KVSKRFS 30 VL-CDR3 FQGSHVPPT 17 Heavy chain

[0322] QVQLQQSGPELVRPGVSVKISCKGSGYTFTDYNMHWVK VH QSHAKSLEWIGVISTYSGNTNYNQKFKDKATMTVDKSSST 35

[0323] AYMELARLTSEDSAIYYCARDGYFDYWGQGTTLTVSS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 VH-CDR3 DGYFDY 25 mAb30

[0324] Light chain

[0325] DVLMTQTPLSLPVSLGDQASISCRSSQYIVHSNGDTYLEW VL YLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISR 38

[0326] VEAEDLGVYYCFQGSHVPPTFGGGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31 VL-CDR3 FQGSHVPPT 17 Heavy chain

[0327] DVKLVESGGGLVKLGGSLKLSCAASGFSFNNYYMSWVRQ VH TPEKRLELVAAINNNGDSTHYPDTVKGRFTISRDNAKNTL 36

[0328] YLQMNTLKSEDTALYYCARAEGSLAYWGQGTLVTVSA VH-CDR1 NYYMS 20 VH-CDR2 AINNNGDSTHYPDTVKG 23 VH-CDR3 AEGSLAY 26 mAb41 Light chain

[0329] DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLYSNNQKNYLA VL WYQQKPGQSPKLLIYWASTRESGVPDRFTGSGSGTDFTL 39

[0330] TISSVKAEDLSVYYCQQYYTYPLTFGAGTKLELK VL-CDR1 KSSQSLLYSNNQKNYLA 29 VL-CDR2 WASTRES 32

[0331]

[0332] VL-CDR3 QQYYTYPLT 33 Conclusions

[0333] Sequence information for the heavy and light chain variable regions (VH and VL, respectively) and complementarity determining regions (CDRs) of the heavy and light chain variable regions (VH- and VL-CDRs) of the monoclonal antibodies was successfully obtained. Based on this information, recombinant antibodies according to the disclosure were successfully constructed and produced. In the following, the recombinantly produced version of each of the selected monoclonal antibodies of the disclosure is denoted "rec20" for the recombinantly produced mAb20, "rec30" for recombinantly produced mAb30, and "rec41" for recombinantly produced mAb41.

[0334] Example 6

[0335] Characterization of human TDP-43 monomers and aggregates Materials and methods

[0336] Human TPD-43 aggregates: A batch of human TDP-43 protein preparation was purchased and analyzed according to the description in Example 1.

[0337] Monomeric (C-terminal) human TDP-43: C-terminal human TDP-43 (SEQ ID NO:54), corresponding to amino acids (aa) 274-414 of SEQ ID NO:53, was produced in-house to generate monomeric forms with the C-terminal intact, using standard protocols. Briefly, a C-terminal construct of human TDP-43 (SEQ ID NO:54) was cloned into a bacterial pETlOO expression vector. Expression of the C-terminal human TDP-43 construct was induced with 0.5 mM IPTG at 18°C, 100 rpm for 16-18 h. After harvesting with centrifugation (4000 rpm, 30 min), extraction in 50 mM Na phosphate, pH 8.0; 300 mM NaCI, 5 mM DTT and 5 mM imidazole (+ protease inhibitors) was made with sonication (Branson sonifier 250) followed by centrifugation at 18500 g for 75 min at 10°C. The clarified supernatant was loaded onto a 1 ml HisTrap Excel column and purified with a gradient of 5-500 mM imidazole. The C-terminal human TDP-43 (SEQ ID NO:54), corresponding to aa274-414 of SEQ ID NO:53, is representative of monomeric forms of human TDP-43.

[0338] The human TDP-43 protein preparations were analyzed using HPLC as described in Example 1. Results

[0339] The purchased human TDP-43 protein preparation exhibited a peak at around 12 minutes in the HPLC elution profile, suggesting the presence of larger aggregates of TDP-43, similarly to that observed and described in Example 1.

[0340] C-terminal human TDP-43 (SEQ ID NO:54) was produced to generate monomeric forms of TDP-43. SEC analysis (Figure 2) shows a main peak at about 24 minutes (constituting about 97-98% of the total absorbance) suggesting monomeric C-terminal species, a smaller peak at about 19 minutes (constituting about 2-3% of the total absorbance) and no peak at 12 minutes. This suggests a lack of larger aggregates and is in line with the presence of smaller molecular species of human TDP-43, predominantly SEQ ID NO:54.

[0341] Conclusions

[0342] As indicated in Example 1 above, the analyzed purchased human TDP-43 preparation is thus considered as a protein preparation consisting of human TDP-43 aggregates, and is referred to below as human TDP-43 aggregates. The analyzed inhouse produced C-terminal human TDP-43 (SEQ ID NO:54) preparation is considered as a protein preparation consisting of human TDP-43 monomers. Said human TDP-43 aggregates and C-terminal human TDP-43 (SEQ ID NO:54) are considered useful in the evaluation of antibodies according to the disclosure for selectivity for aggregated versus monomeric forms of human TDP-43. C-terminal human TDP-43 (SEQ ID NO:54) monomers are particularly useful for testing antibodies which exhibit binding to an epitope within amino acid positions 274-414 of human TDP-43 (SEQ ID NO:53), such as antibodies of the present disclosure.

[0343] Example 7

[0344] Evaluation of antibodies according to the disclosure using indirect ELISA Recombinant antibodies produced as described in Example 5 were analyzed for selectivity for human TDP-43 aggregates versus human TDP-43 monomers using recombinant human TDP-43 protein preparations according to Example 6, wherein said human TDP-43 is in either a monomeric form or in an aggregated form. Materials and methods

[0345] Indirect ELISA was used for the evaluation of recombinant antibodies according to the present disclosure, wherein either 0.5 pg / ml of human TDP-43 aggregates or 0.5 pg / ml of C-terminal human TDP-43 (SEQ ID NO:54) was used as a coat. The method was essentially performed as described in Example 2.

[0346] Results

[0347] The results are shown in Figure 3 and the approximate ECso values determined from the indirect ELISA are presented in Table 4 below.

[0348] Exemplified antibodies according to the disclosure, such as rec20, rec30 and rec41 (as shown in Figures 3A, 3B and 3C, respectively) are characterized by a higher approximate ECso value when measuring binding to C-terminal human TDP-43 (SEQ ID NO:54) (indicated as " Monomer TDP-43" in Figure 3) in comparison to the approximate ECso value measured using a plate coated with human TDP-43 aggregates (indicated as " Aggregated TDP-43" in Figure 3). The data demonstrate that maximum absorbance values were higher for aggregated human TDP-43 in each case and rec30 exhibited the greatest difference when comparing binding to aggregated human TDP-43 and C-terminal human TDP-43 (SEQ ID NO:54), (Figure 3B and Table 4). The herein presented diagrams and ECso values indicate that one or more antibodies according to the disclosure have essentially no or a very low binding affinity to monomeric human TDP-43 when compared to their binding to human TDP-43 aggregates.

[0349] Table 4. EC50 values (ng / ml) from indirect ELISA

[0350] Human TDP-43 (C-terminal) Aggregated TDP-43

[0351] Antibody monomers (SEQ ID NO:54))

[0352] EC50 (ng / ml) Abs450nm max EC50 (ng / ml) Abs450nm max rec20 7.5 3.9 60 3.0 rec30 5 3.8 57* 1.5*

[0353]

[0354] rec41 3 3.9 5 3.0 *Uncertain values since absorbance maximum was not reached. Conclusions

[0355] As seen in Table 4 and Figure 3A-C, the tested antibodies according to the disclosure exhibited selective binding to human TDP-43 aggregates in comparison to their binding characteristics to human TDP-43 monomers, as demonstrated using C-terminal human TDP-43 (SEQ ID NO:54).

[0356] Example 8

[0357] Evaluation of antibodies according to the disclosure using inhibition ELISA Inhibition ELISA was performed to determine the selectivity of antibodies according to the disclosure for human TDP-43 aggregates versus human TDP-43 monomers, using recombinant human TDP-43 protein preparations according to Example 6, wherein said human TDP-43 is in either a monomeric form (C-terminal TDP-43) or in an aggregated form.

[0358] Materials and methods

[0359] Briefly, recombinant antibodies according to the disclosure were preincubated with titrating amounts of human TDP-43 species, either human TDP-43 aggregates or C-terminal human TDP-43 (SEQ ID NO:54), at room temperature. After 1 h, mixes were transferred to an ELISA plate precoated with TDP-43 aggregates and incubated for 15 min to allow non-occupied antibodies to bind to the coat. The bound antibodies were detected with an ALP-coupled anti-mouse IgG antibody (Mabtech, 3310-4).

[0360] Results

[0361] Results are shown in Figure 4, and the approximate IC50 values determined from the inhibition ELISA are presented in Table 5. As is shown in Table 5, IC50 values could not be determined for antibodies rec30 and rec41 when preincubated with titrating amounts of monomeric forms of human TDP-43 (C-terminal human TDP-43). For rec20, a high IC50 value was obtained. Said antibodies have essentially no or low affinity for C-terminal human TDP-43 (SEQ ID NO:54) Table 5. IC50 values (ng / ml) from inhibition ELISA

[0362] IC50 (ng / ml) IC50 (ng / ml)

[0363] Antibody

[0364] Aggregated C-terminal (monomeric)

[0365] rec20 0.9 ± 0.1 283 ± 194

[0366] rec30 1.1 ± 0.3 >700

[0367]

[0368] rec41 1.2 ± 0.1 >700

[0369] Conclusions

[0370] As seen in Figure 4A-C and Table 5, antibodies according to the disclosure, such as rec20 (Figure 4A), rec30 (Figure 4B), and rec41 (Figure 4C), exhibit high selectivity for human TDP-43 aggregates versus human TDP-43 monomers, as demonstrated using C-terminal human TDP-43 (SEQ ID NO:54). This indicates that antibodies according to the disclosure have essentially no or low affinity for human TDP-43 monomers.

[0371] Example 9

[0372] SPR analysis of binding characteristics

[0373] Binding of antibodies of the disclosure to aggregated human TDP-43 was evaluated by surface plasmon resonance (SPR), yielding binding affinities of tested antibodies for human TDP-43 aggregates.

[0374] Materials and methods

[0375] Affinity measurements with SPR using a Biacore 8K (Cytiva) were used to characterize the binding of recombinant antibodies according to the disclosure to human TDP-43 aggregates. Human TDP-43 aggregates (according to Examples 1 and 6) in 1.1 pg / ml concentration were coupled to a CM5 chip using general Biacore coupling chemistry. Antibody binding to human TDP-43 aggregates was determined using single cycle kinetics (SCK) with a starting concentration of 5 nM and a 2-fold dilution series in five steps, using a 120 s injection contact time for every concentration of antibody and a 600 or 1200 s dissociation time with a flow rate of 30 pl / min. Regeneration of the surface between cycles was done by injecting 10 mM glycine-HCI, pH 1.7. Binding data was fitted to a 1:1 interaction model.

[0376] For binding to C-terminal human TDP-43 (SEQ ID NO:54), antibodies were captured using an anti-mouse antibody coupled to a CM5 chip. Monomer according to SEQ ID NO:54 was used as analyte in SCK with a starting concentration of 5 p. M and a 2-fold dilution series in five steps with 120 s injection contact time for every concentration of antibody and 600 s dissociation time with a flow rate of 30 pl / min. Regeneration of the surface between cycles was done by injecting 10 mM glycine-HCI, pH 1.7 and 3 M MgCL Binding data was fitted to a 1:1 interaction model. The highest concentration tested for C-terminal human TDP-43 (SEQ ID NO:54) was 5 pM.

[0377] To evaluate the contribution of avidity in the binding strength, Fab parts of the antibodies were prepared and binding to aggregated TDP-43 was evaluated using SPR. To assess the avidity of the antibodies, a Fab fragment was analyzed in the same way as the entire antibodies, i.e. by SCK analysis of binding to aggregated TDP-43 immobilized on a CM5 chip. The starting concentration was 200 nM for Fab fragments.

[0378] Results

[0379] SPR of rec20, rec30 and rec41: Representative sensorgrams of the SPR measurements are shown in Figure 5. KD values for the binding of tested antibodies to human TDP-43 aggregates and monomers are presented in Table 6. Notably, KD values for the binding of tested antibodies to C-terminal human TDP-43 (SEQ ID NO:54) were high, indicating that the tested antibodies have low or essentially no binding affinity for human TDP-43 monomers.

[0380] Table 6. Kp values obtained by SPR

[0381] KD (pM) KD (pM)

[0382] Antibody

[0383] Aggregate Monomer

[0384] rec20 120 n.d.

[0385] rec30 140 1.2

[0386]

[0387] rec41 63 0.5

[0388] SPR of Fab antibodies rec30 and rec41: Representative sensorgrams of the SPR measurements are shown in Figure 6. The binding profiles from the Fab antibodies show a reduced binding to aggregated TDP-43. Conclusions

[0389] The results presented in Figure 5 and Table 6 reveal selective binding of the tested antibodies to human TDP-43 aggregates in comparison to monomeric forms of human TDP-43. The measured, low picomolar KD values are indicative of a high binding affinity. Reduced binding affinity of the Fab fragments of the antibodies to aggregated TDP-43 (Figure 6) suggests that bivalent or multivalent antigen-binding (avidity) according to the present disclosure may be advantageous for achieving the hereby demonstrated high binding affinity of the tested antibodies for aggregated TDP-43.

[0390] Example 10

[0391] Evaluation of target binding in human brain extracts by immunodepletion and indirect MSP assay

[0392] To verify target binding in biologically relevant samples, immunodepletion experiments were performed, wherein human postmortem brain extracts from patients diagnosed with frontotemporal dementia (FTD) were incubated with antibody-coupled protein G beads using antibodies according to the disclosure. Following immunodepletion, human TDP-43 bound to the antibodies was evaluated using Meso Scale Discovery (MSD) assays measuring both total and aggregated TDP-43. To assess binding to disease-associated pathological TDP-43 versus non-pathological TDP-43, brain extracts from FTD patients and non-demented controls (NDE) were coated onto MSD plates. Antibody binding to the immobilized targets was evaluated by incubation with the antibodies followed by MSD detection, in order to determine whether the antibody could discriminate between disease and control samples.

[0393] Materials and methods

[0394] Brain tissue was homogenized in high salt buffer. After centrifugation and treatment with myelin buffer as described in Sampathu et al (2006) in Am J Path 169:1343-1352, the pellets were dissolved in sarcosyl buffer to obtain a sarcosyl soluble fraction. The remaining sarcosyl insoluble pellets were either dissolved in urea or resuspended in salt buffer. For the indirect MSD assay, brain homogenates were subjected to the sarkospin protocol as described in Laferriere et al (2019), Nat Neurosci. 22(l):65-77, to extract and enrich for pathological TDP-43.

[0395] Recombinant full-length monomeric human TDP-43 was produced in-house to generate monomeric forms of TDP-43 using standard protocols. Briefly, a construct of human TDP-43 (SEQ ID NO:53) was cloned into a bacterial pETlOO expression vector. Expression of the human TDP-43 construct was induced with 0.5 mM IPTG at 18°C, 100 rpm for 16-18 h. After harvesting with centrifugation (4000 rpm, 30 min), extraction in 50 mM Na phosphate, pH 8.0; 300 mM NaCI, 5 mM DTT and 20 mM imidazole (+ protease inhibitors) was made with sonication (Branson sonifier 250) followed by centrifugation at 18500 g for 60 min at 10°C. The clarified supernatant was loaded onto a 1 ml HisTrap Excel column and purified with a gradient of 20-500 mM imidazole.

[0396] Recombinant antibodies according to the disclosure were coupled to protein G beads at 100 pg / ml. Human brain material (postmortem brain material from patients diagnosed with FTD) were sonicated, mixed with the antibody-coupled beads, and incubated over night at 4°C under continuous rotation. The beads with bound material were removed and the eluates from the beads were analyzed on Western blot. The immunodepleted samples were analyzed using both total and aggregated TDP-43 assays for remains of TDP-43.

[0397] Total TDP-43 assay: The assay for total TDP-43 used two anti TDP-43 antibodies, a mouse monoclonal antibody (Abeam, ab57105 (2E2D3)) for capture and a rabbit polyclonal antibody (Proteintech, 10782-2-AP) for detection. A standard MSD plate was coated with the ab57105 antibody and free binding sites were blocked by incubation with 1% Blocker A solution (MSD). Total TDP-43 in samples was detected by adding polyclonal 10782-2-AP antibody followed by stepwise addition of MSD goat anti-rabbit SULFO-TAG and MSD Read Buffer. The plate was washed between each incubation step. When reading in an MSD sector imager a light signal was generated by electrochemiluminescence and measured. The signal strength was correlated to the amount of total TDP-43 in the sample, by interpolation from the calibrator curve. Recombinant full-length TDP-43 was used as the assay calibrator.

[0398] Aggregated TDP-43 assay: The assay for aggregated TDP-43 is a sandwichbased assay using an anti-TDP-43 mouse monoclonal antibody (produced in-house) for both capture and detection. A standard MSD plate was coated with the antibody and free binding sites were blocked by incubation with 1% Blocker A solution. TDP-43 aggregates in samples were detected by adding biotin-conjugated antibody followed by the addition of MSD Streptavidin SULFO-TAG and MSD Read Buffer. The plate was washed between each incubation step. When reading in an MSD sector imager, a light signal was generated by electrochemiluminescence and measured. The signal strength was correlated to the amount of TDP-43 aggregates in the sample, by interpolation from the calibrator curve. TDP-43 aggregates (see Examples 1 and 6) were used as assay calibrator.

[0399] Indirect MSD assay: Sarkosyl-insoluble brain extracts obtained using the sarkospin protocol from FTD and NDE were dissolved and diluted in PBS, pH 7.4, and coated onto 384-well standard MSD plates overnight at 4 °C. Free binding sites were blocked by incubation with 1% Blocker A solution. Recombinant antibody according to the disclosure was serially diluted 4-fold in 1% Blocker A solution and incubated with the coated brain extracts at room temperature with shaking (900 rpm) for 2 h. Antibodies bound to the target were detected by adding MSD goat anti-mouse SULFO-TAG and MSD Read Buffer. The plate was washed between each incubation step. The signal was plotted against antibody concentration, and the EC50 value was estimated from sigmoidal fitted curves.

[0400] Results

[0401] Total TDP-43 assay: Figure 7 presents results of the total TDP-43 assay, wherein it is revealed that immunodepletion of human TDP-43 from human brain extracts (both sarcosyl soluble (Figure 7A) and sarcosyl insoluble fractions (Figure 7B)) occurs when the indicated test antibodies are used. Such immunodepletion is not observed when the beads are coupled to a non-TDP-43-specific antibody, an IgG isotype control, or the beads are not coupled to any antibody. Thus, the observed immunodepletion is specific for the antibodies according to the disclosure. As seen in Figure 7, about 70-75% of total TDP-43 is depleted from soluble brain extracts. On the other hand, it is apparent that about 80-90% of total TDP-43 is depleted from insoluble brain extracts.

[0402] Aggregated TDP-43 assay: Figure 8 presents results of aggregated TDP-43 assay, wherein it is revealed that immunodepletion of human TDP-43 from human brain extracts (both sarcosyl soluble (Figure 8A) and sarcosyl insoluble fractions (Figure 8B)) occurs when the indicated test antibodies are used. Such immunodepletion is not observed when the beads are coupled to a non-TDP-43-specific antibody, an IgG isotype control, or the beads are not coupled to any antibody. Thus, said immunodepletion is specific for the antibodies according to the disclosure. As seen in Figure 8, about 20-40% of aggregated TDP-43 is depleted from soluble brain extracts. On the other hand, it is apparent that about 70-80% of total TDP-43 is depleted from insoluble brain extracts.

[0403] Indirect MSP assay: Figure 9 shows the results from the indirect MSD assay, demonstrating clear selectivity of rec30 for pathological versus physiological TDP-43. Rec30 exhibited strong binding to TDP-43 extracted from FTD brains, with minimal or no binding to TDP-43 from NDE brains. Rec30 showed no binding to PBS-coated wells, demonstrating that the detected signal was specific for the target.

[0404] Conclusions

[0405] As shown in Figures 7 and 8, tested antibodies according to the disclosure have affinity for TDP-43 in human brain extracts. More specifically, as apparent from an increased level of immunodepletion in sarcosyl insoluble fractions, wherein TPD-43 aggregates are enriched, the tested antibodies preferentially bind TDP-43 aggregates from brain extracts wherein TDP-43 aggregates are present, such as in brain extracts of patients diagnosed with FTD. Data in Figure 9 demonstrate that rec30 displays a distinct selectivity profile for pathological (FTD) versus physiological (NDE) TDP-43 species.

[0406] Example 11

[0407] FcyR-mediated endocytosis of human TDP-43 aggregates in microglia cells in vitro In order to evaluate the ability of murine recombinant antibodies according to the disclosure to promote Fcy-receptor-mediated endocytosis of human TDP-43 aggregates, an internalization assay was performed using microglia cells.

[0408] Materials and methods

[0409] Two vials (2x200 pl) of human TDP-43 (Novus Biologica Is, Cat.no.: NBP2-59551, Lot no.: 20351602) at 0.25 mg / ml concentration were thawed on ice, wherein said human TDP-43 consists of human TDP-43 aggregates as described in Example 1. The protein was concentrated using 10 kDa centrifugal filter units (Dynamic Biosensors, Cat. no.: CF-010-5, Lot no.: R8HA79014) by centrifugation at 4000 g for 40 min to a volume of 150 pl. The buffer was exchanged to PBS using a 7 kDa Zeba column and the concentration was measured using NanoDrop (0.941 mg / ml). The protein was labeled with Alexa-488 using the A20181 Kit (Thermo Scientific) according to the manufacturer's instructions and cleanup was performed using a 7 kDa Zeba column. Final concentration of the Alexa-488-labeled human TDP-43 aggregates was determined at 0.857 mg / ml (19.9 pM).

[0410] Non-adherent BV-2 microglia cells were discarded, and adherent cells were trypsinized. 200000 cells in 1 ml PBS were added to FACS tubes (5 ml, BD) and washed (300 g, 5 min). Cells were treated with 200 pg / ml fucoidan (Sigma) in 150 pl medium (RPMI, 10% FBS, PEST, L-Glut) for 30 min at 37°C prior to incubation with Alexa-488-labeled human TDP-43 aggregates alone or in complex with an antibody according to the disclosure.

[0411] To create said antigen / antibody complexes, 10 pg / ml of each tested antibody was mixed 1:1 with Alexa-488-labeled human TDP-43 aggregates at 140 nM concentration (both diluted in culture medium) for 30 min at room temperature, giving a final concentration of 5 pg / ml and 70 nM, respectively.

[0412] Cells were washed in medium and incubated with 300 pl Alexa-488-labeled human TDP-43 aggregates alone or in complex with test antibodies for 1 h at 37°C to allow antibody mediated endocytosis to occur. Fucoidan was included to prevent unspecific internalization of the aggregates.

[0413] The cells were washed 2x in PBS (1 ml) and uptake of Alexa-488-labeled human TDP-43 aggregates was analyzed using flow cytometry.

[0414] Results

[0415] The uptake of Alexa-488-labeled human TDP-43 aggregates in microglia in the presence of test antibodies rec30 and rec41 was evaluated in a mouse microglia cell line. A concentration-dependent internalization of aggregated human TDP-43 was observed in the presence of the antibodies with EC50 values of 1438 and 1059 ng / ml, respectively (Figure 10). Conclusions

[0416] Antibodies according to the disclosure induce Fcy-receptor-mediated endocytosis of human TDP-43 aggregates in vitro. Thus, these antibodies are capable of inducing clearance of pathological human TDP-43, such as irreversible human TDP-43 aggregates, by the immune system.

[0417] Example 12

[0418] Antibody internalization in vitro

[0419] To investigate whether antibodies according to the disclosure are capable of cellular entry and thus engage in cytoplasmatic human TDP-43 binding, antibody internalization was assessed using HEK293-T cells.

[0420] Since TDP-43 is an intracellular protein, a suitable antibody needs to be internalized into living cells. In case of successful internalization, the binding of the candidate antibody to the aggregated protein, also known as target engagement, can be studied. Since physiological TDP-43 is most prominent in the nucleus, the ability of a candidate antibody to penetrate the nucleus was investigated simultaneously.

[0421] Materials and methods

[0422] Internalization and nuclear localization of antibodies rec20, rec30 and rec41 was tested using mouse primary neurons cultured in 24-well plates prepared with poly-D-lysine coated coverslips.

[0423] Antibody internalization was studied by conjugating candidate antibodies to pHrodo™ Red dye conjugates (ThermoFisher). This pH-sensitive dye becomes fluorescent in acidic environments, such as intracellular compartments. pHrodo-conjugated antibodies were added to mouse primary neurons (1-10 pg / ml).

[0424] Nuclear localization was studied by conjugating candidate antibodies to QuantumDots 655 using a SiteClick™ labeling approach (ThermoFisher). Since conjugation can affect the antibody binding site, binding affinity of the conjugated antibodies was validated by ELISA. QuantumDots 655 conjugated antibodies (5 pg / ml) were added to living cells and fixed 24 hours later, Hoechst33342 (20 mM) was added for nuclear labeling. Results

[0425] Antibody internalization: Monitoring the change of the fluorescent signal in a time-lapse analyses (LionHeart microscope (BioTek)), revealed that all candidate antibodies were able to be internalized.

[0426] Nuclear localization: Confocal imaging with 3D reconstruction revealed no nuclear localization of the antibody candidates although all tested antibodies were readily internalized into the cytoplasm.

[0427] Conclusions

[0428] The results indicate that antibodies according to the disclosure were internalized into live human cells in vitro, in particular into the cytoplasm thereof. Here, the antibody may bind pathological forms of human TDP-43, such as human TDP-43 aggregates if present in the cytoplasm. This is beneficial for the clearance of pathological forms of human TDP-43 present in the cytoplasm of living cells.

[0429] Moreover, it is advantageous that an antibody according to the disclosure does not enter the nuclei of said living cells. Thus, said antibody does not bind physiological forms of human TDP-43 present in the nuclei, such as human TDP-43 monomers. Thus, physiological functions of human TDP-43 in the nucleus are not affected by said antibody.

[0430] Example 13

[0431] Ex vivo target engagement and in vivo therapeutic efficacy Binding of antibodies of the disclosure to pathological forms of human TDP-43, such as non-reversible human TDP-43 aggregates enriched in inclusions in the brain of a mouse model of TDP-43 proteopathy, was evaluated ex vivo. Furthermore, therapeutic effect of the disclosed antibodies was evaluated in the same mouse model by behavioral analysis and IHC for quantitative analysis of phosphorylated TDP-43 pathology, as well as other parameters. For these studies, rNLS8 mice expressing a regulatable form of human TDP-43, which lacks a nuclear localization signal under the control of the tetracycline activator protein (tTA), were used (Walker et al (2015), Acta Neuropathol 130:643-660), wherein disease induction is normally initiated at 8-10 weeks of age by removal of doxycycline (DOX) diet. tTA is a key component of this system, since its activity is regulated by DOX. The model can either be used as a "low DOX" model where a low amount of doxycycline is introduced one week after removal of DOX, thus generating a less aggressive model, or as an "off DOX" model in which full expression of the transgene is allowed. Both models have been used below. A small group of non-transgenic mice (tTA insert only) were included in the off DOX studies as a reference to physiological levels of the different read-outs.

[0432] Materials and methods

[0433] The ability of the antibodies to recognize pathological TDP-43 in this model was first evaluated ex vivo by IHC. Briefly, the antibodies were tested at three dilutions on formalin-fixed, paraffin-embedded brains from mice that had been off DOX for 1 and 3 weeks. Prior to staining, tissue sections were treated with proteinase-K to remove soluble TDP-43. Binding was visualized using AEC-la bel ling and slides were digitized and staining was qualitatively evaluated.

[0434] In vivo efficacy in low DOX model of rNLS8 tg mice: To evaluate the therapeutic efficacy, rNLS8 mice at 8-10 weeks of age were treated twice weekly with 30 mg / kg of rec30. After 1 week off DOX, the animals were switched to an intermittent, low-dose DOX diet (~46 pg DOX per g body weight per week) to extend the timing of clinical deterioration. Plasma was collected at various time points during the study (see below) to be able to measure plasma levels of NfL (a marker for neurodegeneration). In another, shorter study, female rNLS8 mice were treated twice weekly with either vehicle (PBS) or 30 mg / kg rec30 for a period of 6 weeks. Dosing started three weeks prior to disease induction (off DOX). After the disease induction, the animals were switched to an intermittent, low-dose DOX diet (~46 pg DOX per g body weight per week). Grip strength was measured at baseline (prior to first dose) and again at the study endpoint using the Bioseb Grip Strength Test (BIO-GS4, Bioseb, France). The grip strength test measures neuromuscular function by determining the maximal peak force developed by the animal. Mice were held by the tail and lowered onto a wire grid connected to a grip strength meter. The mouse was allowed to grab the metal grid and then they were pulled backwards. The force applied to the grid just before the mouse lost grip was recorded as the peak tension (recorded in grams). At each timepoint, each mouse underwent three trials with at least 5 min rest between trials. The 3-trial mean was used for the analysis. In vivo efficacy in off DOX model of rNLS8 tg mice: In a third study, non-transgenic tTA mice and one group of rNSL8 mice were treated twice weekly with PBS. In addition, one group of rNSL8 mice was treated twice weekly with 30 mg / kg rec30. The mice were kept on DOX, 200 mg / kg. After 3 weeks of treatment, DOX was completely removed to allow full transgene expression throughout the study, i.e., "off DOX". In the end of the study, compound muscle action potentials (CMAP) were measured as follows, to assess the extent of axonal degeneration and demyelination. Mice were anesthetized by isoflurane / O2and monitored for loss of reflexes in which all of the responses to external stimuli ceased. The fur from the hind limb was removed and the skin was cleaned with alcohol to remove any dirt and body oil. Recording electrodes were placed at the position where the gastrocnemius muscle has its maximum diameter. The reference electrode was placed just beneath the recording electrode. The stimulating electrodes were placed along the axis of the sciatic nerve proximal to the sciatic notch. For motor nerve conduction velocity estimates following the initial set of recordings, a second set of responses was recorded with the stimulating electrodes positioned at an intermediate site, while leaving the recording electrodes in the initial position. All leads of stimulating and recording electrodes were connected to an EMG machine (Nicolet Viking Quest, Natus Neurology, USA). For all analyses, five responses evoked by stimulation were averaged to generate a smooth compound muscle action potential (CMAP). Amplitude (mV) and the onset latency (ms) were measured based on the averaged CMAPs. Neuromuscular nerve conduction velocity (mm / ms) was calculated using response latency and the distance from stimulating electrode to recording electrode. Motor nerve conduction velocity (MNCV; mm / ms) was calculated using the differences in latencies between the proximal and intermediate stimulation sites. A maximal CMAP response was obtained by increasing stimulating intensity until the response no longer increased.

[0435] Collection of material for analysis: For analyses of plasma NfL levels, blood plasma was collected prior to dosing, at various time points during the study and at termination, and stored at -80°C. Following terminal plasma collection, mice were perfused with 20 ml cold PBS. The brains were immediately extracted and dissected along the midline. One hemisphere of the brain and the spinal cord were immersion-fixed in 10% formalin for IHC studies. The other hemisphere was further dissected into different brain regions and snap frozen. Selected snap frozen brain regions were used for preparation of RNA.

[0436] IHC staining - off DOX model: The left brain hemisphere was drop fixed in freshly prepared lx PBS containing 4% paraformaldehyde (PFA) for 24 h and then transferred to PBS until embedding. Spinal cords were extracted and post-fixed overnight in freshly prepared lx PBS containing 4% PFA. Thereafter, PFA was replaced with PBS and vials were stored in the refrigerator until embedding.

[0437] Immersion-fixed and cryo-protected samples were then embedded in cryo molds with tissue freezing medium and frozen on liquid, dry-ice cooled isopentane. All tissue blocks were stored at -80°C until sectioning. Tissue was cut in sagittal plane in a uniform systematic random protocol on an Avantik QS12 cryotome at 10 μm section thickness. Sections were stored frozen at -80°C until used. Primary antibodies against the following were used: phosphorylated TDP43, [1D3] (Sigma, MABN14); astrocytes, GFAP (Abeam, ab53554); microglia, Ibal [EPR16588] (Abeam, abl78846). Sections were stained with DAPI to visualize nuclei. Antibody binding was visualized using highly cross-absorbed, fluorescently labeled secondary antibodies. Standard negative controls included tTA genotype brain sections and sections running through the whole protocol but without the incubation with the primary antibody. Entire brain sections were imaged on a Zeiss AxioScan. Zl slide scanner microscope, equipped with LED illumination, corresponding narrow band filter sets fitting to each dye and a sensitive Orca Flash 4.0 B& W camera (mounted with lx opto-coupler thus no demagnification). Delineations and quantifications were performed on Image Pro Premier (IPP v10.0.11) from Media Cybernetics. The brain size was determined using separate delineation of the regions of interest, the cerebral cortex and the dorsal striatum. The spinal cord was delineated as a whole. Mean signal (in lumen), immunoreactive (IR) surface area (in %), and integrated optical density (IOD) (in lumen x μm2) were determined for all markers. For the lba-1 marker, measurement of single unbranched processes not attached to a soma (deriving from cells in neighboring sections) as well processes adjacent to somata as well as their branching was performed. Single process density (n / mm2) describes microglial processes that are not attached to any soma and derive from cells in neighboring sections. Lower density indicates early activation due to shortening of long processes (de-ramification). Statistical analysis, IHC: An alpha level of.05 was selected for all inferential statistics. The statistical analysis was performed by an analyst blinded to the treatment of the respective group. Histological data were analyzed by analysis of variance (one or two way-ANOVA) followed by post-hoc comparisons where appropriate. An effect was considered significant if p <.05 (*=p < 0.05; **=p < 0.01; ***=p < 0.001; ****=p < 0.0001). Data are represented as the mean, scatter and standard error of the mean (SEM). No statistical outliers were excluded.

[0438] qPCR analysis of Uncl3a cryptic exons: Dissected left striata were stored at -80°C and thawed overnight while soaked at -20°C in RNAIater-ICE (ThermoFisher Scientific, AM7030) to crosslink RNA and prevent its degradation during extraction. RNA was extracted using the SPINeasy DNA / RNA / Protein All-In-One Kit (MP Biomedicals, 6544050) following the manufacturer's protocol. Subsequent cDNA synthesis was performed using the High Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific, 4368814) and the reaction was conducted using a VeritiPro™ Thermal Cycler (ThermoFisher Scientific) as per the kit's protocol.

[0439] Quantitative PCR assays were executed on a QuantStudio 5 Real-Time PCR System (Thermo Fisher Scientific). The reactions were set up following the manufacturer's protocol of the TaqMan Fast Advanced Master Mix (ThermoFisher Scientific, A44360) with a total volume adjusted to 15 μl. Custom made TaqMan probes and primers (Table 7) were used alongside a β-actin assay (ThermoFisher Scientific, ID: Mm02619580_gl,) as endogenous reference. The custom-made assays were designed by ThermoFisher TaqMan Custom Support for the respective target sequence.

[0440] The sequence of the mouse Uncl3a cryptic exon was derived from Rosa Ma et al (2022), Nature 603:124-130 (see extended data, Figure 2 of the article). The native sequence was obtained from public databases. The native probe was designed to span the junction of exons 20 and 21, ensuring a cryptic exon insertion (breaking the junction at this position) would not be detected. Table 7. Custom made TaqMan assay sequences

[0441] Mouse Uncl3a cryptic exon Mouse Uncl3a native Forward G AAATCTGTCCCTAAGTG G AG CTT G CCCTACCATGTCC AGTAC AC primer (SEQ ID NO:64) (SEQ ID NO:67)

[0442] Probe TCGAGTCACACTCATTTGA CTGCATGAGAACCTGTTCC (SEQ ID NO:65) (SEQ ID NO:68) Reverse CTTATTCTTTATTCCTCCAGTTAGTAATCCA TG C ACGTCCGTC ACAAAGT

[0443]

[0444] primer (SEQ ID NO:66) (SEQ ID NO:69)

[0445] Thermal cycling conditions included an initial denaturation at 95°C for 20 s, followed by 40 cycles of denaturation at 95°C for 1 s and annealing / extension at 60°C for 20 s. Data analysis was performed using QuantStudio Design and Analysis Software v.1.5.3. All samples were multiplexed, including β-actin as an endogenous reference. Relative expression levels of the Uncl3a cryptic exon were quantified by normalizing to the Uncl3a native levels. All samples were analyzed in duplicate.

[0446] Plasma levels of NfL: Plasma NfL was measured using two anti-NfL mouse monoclonal antibodies, one for capture (Uman Diagnostics, UD1) and a biotinylated one for detection (Uman Diagnostics, UD3). A standard MSD plate was coated with UD1 antibody and free binding sites were blocked by incubation with 3% BSA TBS-T blocking buffer. NfL in samples was detected by adding biotinylated UD3 antibody followed by addition of MSD Streptavidin SULFO-TAG and MSD Read Buffer. The plate was washed between each incubation step. When reading in an MSD sector imager, a light signal was generated by electrochemiluminescence and measured. The signal strength was correlated to the amount of NfL in the sample, by interpolation from the calibrator curve. Lyophilized bovine spinal cord NfL (Uman Diagnostics, #27001) was used as assay calibrator.

[0447] Results

[0448] Ex vivo target engagement - IHC: Before an in vivo efficacy study in rNLS8 mice was performed, rec30 was used for IHC staining to verify that antibodies of the present disclosure recognize pathological human TDP-43 present in these mice. The tested antibody, rec30, recognized pathological human TDP-43, such as non- reversible human TDP-43 aggregates, in brains from induced rNLS8 mice. Staining with rec30 revealed binding restricted to cytoplasmatic human TDP-43 inclusions (Table 8). Example of rec30 ex vivo target engagement is illustrated in Figure 11.

[0449] Table 8. IHC conditions and results of ex vivo target engagement Antigen TDP-43 staining Antibody Dilutions Protocol

[0450] Retrieval WT rNLS8 EDTA + cytoplasmic rec30 4000 AEC no staining

[0451]

[0452] Proteinase K inclusions

[0453] In vivo therapeutic efficacy - behavioral analysis, low DOX model: Female low DOX rNLS8 mice were tested for grip strength longitudinally at baseline (prior to first dose) and at week 3. Results are shown in Figure 12. Plotting percent change in grip strength over time revealed that rec30 treated female mice showed a slightly stronger grip at week 3 compared to the baseline, while grip strength in the PBS treated control group decreased over time. The percent change from baseline grip strength reached a statistically significant difference between the groups (*p = 0.05), as determined by Student's T-test.

[0454] CMAP analysis in the off DOX model: Data was analyzed using a 2-way ANOVA with sex and treatment as independent variables, and a significant sex*treatment interaction was detected in onset latency (F2, si = 5.050, p = 0.01). The onset latency was significantly shorter for PBS treated tTA control mice compared to PBS treated rNLS8 mice in both female and male animals (p < 0.001, Tukey's post-hoc test). Additionally, the onset latency was significantly shorter for rec30 treated rNLS8 female mice compared to PBS treated rNLS8 female mice (p < 0.01, Tukey's post-hoc test; Figure 13).

[0455] In vivo therapeutic efficacy - IHC, off DOX model: Phosphorylated TDP-43 (pTDP-43) was detected using the 1D3 antibody (Sigma, MABN14 [1D3]), which stains pTDP-43 inclusions. In the cerebral cortex and dorsal striatum (Figures 13A and 13B), the % immunoreactive area was significantly increased in rNLS8 groups compared to tTA-vehicle group. However, treatment with rec30 significantly reduced the aggregate TDP-43 load in both regions compared to the vehicle treated group (-25% and -37%, respectively). Separated into sexes, data revealed a lowering of rec30 vs vehicle by 44% in the male spinal cord (Figure 14C). The microglial morphology analysis (Ibal labeling) showed a significant response to rec30 in both females and males. Especially, single process density increased significantly in the cortex of males (p<0.01, Figure 15), indicating a reduction in inflammatory activation of microglial cells, with stronger effect in males than females. The significantly reduced single process density in vehicle treated rNLS8 mice compared to the tTA group suggests de-ramification and inflammatory activation in the tg animals. The reduced process density in the rNLS8 vehicle treated mice was ameliorated in the rec30 treated group. In spinal cords, the mean GFAP signal was significantly lowered in female rNLS8 mice treated with rec30 compared to vehicle treatment suggesting a reduced astrocytosis (Figure 16).

[0456] In vivo therapeutic efficacy - Uncl3a cryptic exons, off DOX model: As shown in Figure 17, the transgenic expression of human TDP43 in rNLS8 mice leads to elevated levels of the mouse Uncl3a cryptic exon. This effect is likely due to the coaggregation of human TDP43 with endogenous mouse TDP43, which impairs its normal function as an RNA-binding protein and results in the mis-splicing of Uncl3a mRNA. Treatment with rec30, which specifically targets aggregated TDP43, prevents further co-aggregation of endogenous TDP43. This intervention leads to a significant reduction in the levels of the Uncl3a cryptic exon (p < 0.01, unpaired t-test).

[0457] In vivo therapeutic efficacy - plasma levels of neurofilament light (NfL):

[0458] Plasma levels of NfL, a marker for neurodegeneration, were analyzed, comparing the vehicle group (PBS) with the treatment groups in the low DOX model. Data shows that treatment twice a week with rec30 at 30 mg / kg results in significantly lower levels of plasma NfL in the rec30 group compared to the vehicle control group at three weeks after onset of TDP-43 tg expression (***p<0.001, Mann-Whitney U-test, Figure 18).

[0459] Conclusions

[0460] The results show that antibodies of the disclosure bind to pathological human TDP-43, such as non-reversible human TDP-43 aggregates, ex vivo in brain sections from induced rNLS8 mice, which are characterized by the presence of inclusions comprising pathological human TDP-43, such as non-reversible human TDP-43 aggregates.

[0461] Quantitative analysis of phosphorylated TDP-43 pathology revealed that an antibody according to the disclosure induces the clearance of pathological forms of human TDP-43, such as pathological phosphorylated human TDP-43, in vivo, in brain regions affected by phosphorylated TDP-43 pathology.

[0462] In addition, levels of the Uncl3a cryptic exon are decreased upon treatment with an antibody of the disclosure, showing that TDP-43 function is retained upon antibody treatment in vivo.

[0463] The results furthermore revealed that treatment of low DOX rNLS8 mice with antibodies according to the disclosure corresponds to lower plasma levels of NfL, a blood-based biomarker of neurodegeneration, which indicates decelerated disease progression in the animals.

[0464] In conclusion, antibodies as disclosed herein bind to pathological human TDP-43, such as non-reversible human TDP-43 aggregates ex vivo, and induce the clearance of pathological phosphorylated human TDP-43 in vivo, for which pathological forms are associated with TDP-43 proteopathy. The clearance of pathological forms of TDP-43 is expected to lead to improved motor function of subjects affected by TDP-43 proteopathy, as supported by the herein presented data for improved grip strength of and reaction time for rec30 treated female rNLS8 mice upon disease induction. Said antibodies thus have a therapeutic potential in the treatment of diseases associated with TDP-43 proteopathies.

[0465] Example 14

[0466] Pharmacokinetic profile of murine recombinant TDP-43 antibodies Pharmacokinetic analysis of murine recombinant antibodies rec20, rec30 and rec41 was performed in C57BL / 6J (wild type) mice. Pharmacokinetic parameters were calculated based on plasma exposure data.

[0467] Materials and methods

[0468] Administration of antibody and sample collection: Each antibody was administered intravenously (i.v.) at a dose of 30 mg / kg via the tail vein to 8 weeks old female C57BL / 6J mice (five mice per antibody). Blood was collected at 5 min, 4 h, 24 h, 72 h, 168 h (7 days), 336 h (14 days), 672 h (28 days) and 840 h (35 days) after the i.v. injection. Blood was collected into Sarstedt Microvette CB300 K2E tubes and placed on wet ice immediately after collection. The samples were centrifuged at 2400 x g for 10 min at 4°C shortly after collection (within 30 min). Plasma was collected and stored at -80°C until bioanalysis.

[0469] Determination of antibody concentrations in plasma: An MSD assay with antigen coated plates was used to determine plasma exposure. Briefly, wells of an MSD plate were coated with 2.5 pg / ml of human aggregated TDP-43 (described in Example 1) overnight at 4°C. The wells were blocked with Pierce protein free blocking buffer (Thermo Fisher) for 1-3 h and thereafter standard and plasma samples were added. After incubation for 2 h, anti-mouse IgG-SULFO (MSD, R32AC-1) was added to each well and the plates were incubated for 1 h at RT, 900 rpm after which 2x Reading buffer (MSD, R92TC-1) was added. The plate was read within 5 min using MSD software. All incubation steps were performed at RT and 900 rpm.

[0470] Pharmacokinetic analysis: Individual plasma concentration versus time data was analyzed by non-compartmental analysis (NCA) using the software Phoenix® WinNonlin®. Nominal doses and sampling timepoints were used. The maximum plasma concentration, Cmax, was directly derived from the observed concentration versus time curves. The calculation method in the NCA was set to "linear up, log down", in which the linear trapezoidal rule is used when concentrations are increasing, and the logarithmic trapezoidal rule is used when the concentrations are decreasing. Calculated parameters included the area under the concentration versus time curve (AUC) to the last observed time point (AUClast) or to infinity (AUCinf), calculated as AUClast+ Ct / Xz, where Ctis the observed concentration at the last time point and λzis the elimination rate constant estimated using log-linear regression during the terminal elimination phase. Terminal half-life (t1 / 2) was calculated as ln(2) / λzand clearance (CL) was calculated as dose / AUCinf.

[0471] Results

[0472] Pharmacokinetics for murine recombinant antibodies rec20, rec30 and rec41 were evaluated after a single i.v. bolus injection in C57BL / 6J mice based on plasma concentration over time. Mean plasma concentration (pg / ml) time profiles are shown in Figure 19 and a summary of pharmacokinetic parameters is given in Table 9. Table 9. Mean values (n=5 / group) of pharmacokinetic parameters Cmax CL AUCinf Half-life Antibody

[0473] (μg / ml) (ml / h / kg) (mg / ml*h) (days) rec20 715 0.77 40 6.5 rec30 808 0.34 89 6.5

[0474]

[0475] rec41 1000 0.11 270 14.1

[0476] Conclusions

[0477] Pharmacokinetic evaluation of the tested murine antibodies showed the lowest clearance and the longest half-life for rec41 as compared to rec20 and rec30. Rec20 and rec30 exhibited a similar half-life but rec30 showed lower clearance than rec20. The data together indicate that the tested antibodies, and in particular rec41 and rec30, show beneficial pharmacokinetic characteristics.

[0478] Example 15

[0479] Humanization of murine TDP-43 antibodies

[0480] Selected antibodies according to the present disclosure were humanized.

[0481] Materials and methods

[0482] mAb30 and mAb41 were humanized into h30 and h41, respectively; using Composite Human Antibody™ variable regions (Abzena Ltd.). Briefly, structural models of the antibody variable (V) regions were analyzed to identify amino acids, within the CDRs and framework regions, that are relevant for binding (using both Kabat and Chothia definitions). Human sequence segments were analyzed in silico for post-translational modification and immunogenicity risks using the software iTope-AI, and then selected and assembled. Human constant regions were also selected.

[0483] For h30 variants, human IgGl with a heavy chain K322A mutation to lower Clq binding and a kappa light chain were chosen. For h41, mutations were also added to remove sequence liabilities in the VH and VL sequences. For h41, a mutation in VH-CDR2 (N53E, Kabat numbering) was included to remove a deamidation liability. Human constant regions for h41 variants were human IgGl with a heavy chain K322A mutation to lower Clq binding and a YTE (M252Y / S254T / T256E) mutation to improve the half-life, and a kappa light chain. Results

[0484] Five humanized heavy chain variable regions (VH1-VH5) and five humanized light chain variable regions (VL1-VL5) were produced and analyzed in comparison to a chimeric antibody with murine variable regions and human constant regions. Following the analysis, two variants denoted h30-HlL4 and h30-H4L3 were chosen for further analysis. The amino acid sequences comprised in the complete h30 variant antibodies are given in Table 10 below, and h41 variants in Table 11.

[0485] _ Table 10. Sequences of humanized antibodies: h30-variants SEQ ID

[0486] Antibody Region Amino acid sequence

[0487] NO: Heavy chain

[0488] QVQLVQSGPELKKPGASVKISCKGSGYTFTDYNMHWVK VH QAHGKGLEWIGVISTYSGNTNYNQKFKDRATMTVDKST 40 STAYMELSSLRSEDSAIYYCARDGYFDYWGQGTTVTVSS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 VH-CDR3 DGYFDY 25 h30-HlL4

[0489] Light chain

[0490] DVVMTQSPDSLAVSLGERATINCRSSQYIVHSNGDTYLE VL WYQQKPGQPPKLLIYKVSNRFSGVPDRFSGSGSGTDFTL 45 TISSLQAEDVAVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31 VL-CDR3 FQGSHVPPT 17 Heavy chain

[0491] QVQLVQSGAEVKKPGASVKISCKGSGYTFTDYNMHWV KQASGKGLEWIGVISTYSGNTNYNQKFKDRATMTVDKS VH 56 TSTAYMELSSLRSEDTAIYYCARDGYFDYWGQGTLVTVS S VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 h30-H2Ll

[0492] VH-CDR3 DGYFDY 25 Light chain

[0493] DVLMTQTPLSLPVTLGQPASISCRSSQYIVHSNGDTYLE VL WYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTL 58 KISRVEAEDVGVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31

[0494]

[0495] VL-CDR3 FQGSHVPPT 17 Heavy chain

[0496] QVQLVQSGAEVKKPGASVKISCKGSGYTFTDYNMHWV KQASGKGLEWIGVISTYSGNTNYNQKFKDRATMTVDKS VH 56

[0497] TSTAYMELSSLRSEDTAIYYCARDGYFDYWGQGTLVTVS S VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 h30-H2L2 VH-CDR3 DGYFDY 25

[0498] Light chain

[0499] DVVMTQSPLSLPVTLGQPASISCRSSQYIVHSNGDTYLE VL WYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTL 59

[0500] KISRVEAEDVGVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31 VL-CDR3 FQGSHVPPT 17 Heavy chain

[0501] QVQLVQSGAEVKKPGASVKVSCKGSGYTFTDYNMHW VRQASGKGLEWIGVISTYSGNTNYNQKFKDRVTMTVD VH 57

[0502] KSTSTAYMELSSLRSEDTAIYYCARDGYFDYWGQGTLVT VSS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 h30-H3L2 VH-CDR3 DGYFDY 25

[0503] Light chain

[0504] DVVMTQSPLSLPVTLGQPASISCRSSQYIVHSNGDTYLE VL WYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTL 59

[0505] KISRVEAEDVGVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31 VL-CDR3 FQGSHVPPT 17 Heavy chain

[0506] VH QVQLVQSGAEVKKPGASVKISCKGSGYTFTDYNMHWV KQAPGQGLEWIGVISTYSGNTNYNQKFKDRATMTVDK

[0507] 41 STSTAYMELSSLRSEDTAIYYCARDGYFDYWGQGTLVTV SS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 h30-H4L3 VH-CDR3 DGYFDY 25

[0508] Light chain

[0509] DVLMTQSPDSLPVSLGERATINCRSSQYIVHSNGDTYLE VL WYQQKPGQPPKLLIYKVSNRFSGVPDRFSGSGSGTDFTL 46

[0510] TISSLQAEDVAVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31

[0511]

[0512] VL-CDR3 FQGSHVPPT 17 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELL

[0513] Human GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK

[0514] IgGl FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ 49 Constant (K322A) DWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYT regions LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV

[0515] Human

[0516] QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA 51 kappa

[0517]

[0518] DYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0519] Table 11. Sequences of humanized antibodies: h41 variants SEQ ID

[0520] Antibody Region Amino acid sequence

[0521] NO: Heavy chain

[0522] DVKLVESGGGLVKLGGSLKLSCAASGFSFNNYYMSWVRQ VH TPEKRLELVAAINNEGDSTHYPDTVKGRFTISRDNAKNTLY 42

[0523] LQMNSLRAEDTALYYCARAEGSLAYWGQGTLVTVSS VH-CDR1 NYYMS 20 VH-CDR2 AINNEGDSTHYPDTVKG 55 VH-CDR3 AEGSLAY 26 K41-H1L4

[0524] Light chain

[0525] DIVMTQSPDSLAVSLGERATMNCKSSQSLLYSNNQKNYL VL AWYQQKPGQAPRLLIYWASTRESGVPDRFSGSGSGTDFT 47

[0526] LTISSLQAEDVAVYYCQQYYTYPLTFGQGTKLEIK VL-CDR1 KSSQSLLYSNNQKNYLA 29 VL-CDR2 WASTRES 32 VL-CDR3 QQYYTYPLT 33 Heavy chain

[0527] DVKLVESGGGLVKPGGSLRLSCAASGFSFNNYYMSWVRQ VH TPEKRLELVAAINNEGDSTHYPDTVKGRFTISRDNAKNTLY 43

[0528] LQMNSLRAEDTALYYCARAEGSLAYWGQGTLVTVSS VH-CDR1 NYYMS 20 VH-CDR2 AINNEGDSTHYPDTVKG 55 VH-CDR3 AEGSLAY 26 h41-H3L3

[0529] Light chain

[0530] DIVMTQSPDSLAVSLGERATMNCKSSQSLLYSNNQKNYL VL AWYQQKPGQAPRLLIYWASTRESGVPDRFSGSGSGTDFT 48

[0531] LTISSLQAEDVSVYYCQQYYTYPLTFGQGTKLEIK VL-CDR1 KSSQSLLYSNNQKNYLA 29 VL-CDR2 WASTRES 32

[0532]

[0533] VL-CDR3 QQYYTYPLT 33 Heavy chain

[0534] DVKLVESGGGLVKPGGSLRLSCAASGFSFNNYYMSWVRQ VH TPEKRLELVAAINNEGDSTHYPDTVKGRFTISRDNAKNTLY 43

[0535] LQMNSLRAEDTALYYCARAEGSLAYWGQGTLVTVSS VH-CDR1 NYYMS 20 VH-CDR2 AINNEGDSTHYPDTVKG 55 VH-CDR3 AEGSLAY 26 h41-H3L4

[0536] Light chain

[0537] DIVMTQSPDSLAVSLGERATMNCKSSQSLLYSNNQKNYL VL AWYQQKPGQAPRLLIYWASTRESGVPDRFSGSGSGTDFT 47

[0538] LTISSLQAEDVAVYYCQQYYTYPLTFGQGTKLEIK VL-CDR1 KSSQSLLYSNNQKNYLA 29 VL-CDR2 WASTRES 32 VL-CDR3 QQYYTYPLT 33 Heavy chain

[0539] EVQLVESGGGLVKPGGSLRLSCAASGFSFNNYYMSWVRQ VH TPEKRLELVAAINNEGDSTHYPDTVKGRFTISRDNAKNTLY 44

[0540] LQMNSLRAEDTALYYCARAEGSLAYWGQGTLVTVSS VH-CDR1 NYYMS 20 VH-CDR2 AINNEGDSTHYPDTVKG 55 VH-CDR3 AEGSLAY 26 h41-H4L3

[0541] Light chain

[0542] DIVMTQSPDSLAVSLGERATMNCKSSQSLLYSNNQKNYL VL AWYQQKPGQAPRLLIYWASTRESGVPDRFSGSGSGTDFT 48

[0543] LTISSLQAEDVSVYYCQQYYTYPLTFGQGTKLEIK VL-CDR1 KSSQSLLYSNNQKNYLA 29 VL-CDR2 WASTRES 32 VL-CDR3 QQYYTYPLT 33 Heavy chain

[0544] EVQLVESGGGLVKPGGSLRLSCAASGFSFNNYYMSWVRQ VH TPEKRLELVAAINNEGDSTHYPDTVKGRFTISRDNAKNTLY 44

[0545] LQMNSLRAEDTALYYCARAEGSLAYWGQGTLVTVSS VH-CDR1 NYYMS 20 VH-CDR2 AINNEGDSTHYPDTVKG 55 VH-CDR3 AEGSLAY 26 h41-H4L4

[0546] Light chain

[0547] DIVMTQSPDSLAVSLGERATMNCKSSQSLLYSNNQKNYL VL AWYQQKPGQAPRLLIYWASTRESGVPDRFSGSGSGTDFT 47

[0548] LTISSLQAEDVAVYYCQQYYTYPLTFGQGTKLEIK VL-CDR1 KSSQSLLYSNNQKNYLA 29 VL-CDR2 WASTRES 32

[0549]

[0550] VL-CDR3 QQYYTYPLT 33 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ. T YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG

[0551] Human

[0552] PSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWY

[0553] IgGl

[0554] VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG 50 (K322A)

[0555] Constant KEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEL

[0556] with YTE

[0557] regions TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ

[0558] Human

[0559] WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE 51 kappa

[0560]

[0561] KHKVYACEVTHQGLSSPVTKSFNRGEC

[0562] Conclusions

[0563] Humanized versions of selected monoclonal antibodies were successfully obtained. The recombinantly produced humanized versions of each of the selected monoclonal antibodies of the disclosure were denoted "h30" for recombinantly produced mAb30, and "h41" for recombinantly produced mAb41.

[0564] Example 16

[0565] SPR analysis of binding characteristics of humanized TDP-43 antibodies Binding of humanized versions of antibodies of the disclosure to aggregated human TDP-43 was evaluated by SPR, yielding binding affinities of tested antibodies for human TDP-43 aggregates. The procedure was essentially the same as described in Example 9 above.

[0566] Results

[0567] Representative sensorgrams of the SPR measurements are shown in Figure 20 for h30-variants h30-HlL4 (Figure 20A) and h30-H4L3 (Figure 20B). KD values for the binding of tested antibodies to human TDP-43 aggregates are presented in Table 12.

[0568] Table 12. Kp values for h30-H! L4 and h30-H4L3 obtained by SPR Antibody ka (M-1s-1) kd (s-1) KD (pM) h30-HlL4 1.4 x 1062.0 x IO’4150

[0569]

[0570] h30-H4L3 1.1 x 1061.8 x IO’4170 Conclusions

[0571] The results presented in Figure 20 and Table 12 reveal that the tested antibodies have affinity for human TDP-43 aggregates. The measured, low picomolar KD values are indicative of a high binding affinity.

[0572] Example 17

[0573] Evaluation of humanized TDP-43 antibodies according to the disclosure using inhibition ELISA

[0574] Inhibition ELISA was performed to determine the selectivity of antibodies according to the disclosure for human TDP-43 aggregates versus human monomeric forms of TDP-43, using recombinant human TDP-43 protein preparations according to Example 6, wherein said human TDP-43 is in either a monomeric form or in an aggregated form. The procedure was essentially the same as described in Example 8 above.

[0575] Antibodies at a fixed concentration of 50 ng / ml were pre-incubated with titrating amounts of human TDP-43 species, either human TDP-43 aggregates or C-terminal human TDP-43 (SEQ ID NO:54), at room temperature. The antibody / antigen mix were thereafter added to TDP-43 (aggregated) coated plates for detection of free antibody.

[0576] Results

[0577] Results are shown in Figure 21 for the two h30 antibody variants h30-HlL4 (Figure 21A) and h30-H4L3 (Figure 21B).

[0578] Conclusions

[0579] As seen in Figure 21, antibodies according to the disclosure, such as h30-H1L4 (A) and h30-H4L3 (B), exhibit high selectivity for human TDP-43 aggregates versus human TDP-43 monomers, as demonstrated using C-terminal human TDP-43 (SEQ ID NO:54). This indicates that said antibodies have essentially no or low affinity for human TDP-43 monomers. Example 18

[0580] Human target binding of h30 antibody variants

[0581] The purpose of this experiment was to confirm binding of the humanized antibodies of the present disclosure to the human target, i.e. human TDP-43, in human samples.

[0582] Materials and methods

[0583] Paraffin embedded tissues from individuals with FTD were obtained from the Netherlands Brain Bank. Sections were cut at 4 pm thickness. IHC staining was performed with the recombinant antibodies h30-HlL4 and h30-H4L3, and with anti pSer 409 / 410 (Cosmo Bio, TIP-PTD-M01) as positive control. Human IgGl isotype antibody (D265A) was used as negative control. The detection system Discovery XT and kits from Ventana Medical Systems (F. Hoffmann-La Roche, Ltd) were used. The Amp HQ kit, in conjunction with a chromogenic DISCOVERY ChromoMap DAB Kit (Roche, cat. no. 760-159) enabled the enhancement of the chromogenic signal in standard IHC. The paraffin sections were incubated at 60°C for 30 min, deparaffinized in 2x xylene for 15 min, followed by gradient ethanol wash before rehydration in lx PBS. Before application of antibodies, the sections were exposed to antigen retrieval solutions during an automated heat process (standard CC1 protocol from Ventana at 95°C). Stained slides were scanned in brightfield using a Pannoramic 250 FLASH II slide scanner (3D Histech). The resulting image files were uploaded into viewer software (Pannoramic CaseViewer) and adjusted for optimal brightness and contrast, for manual assessment of the staining result.

[0584] Results

[0585] The binding to the human target was confirmed for humanized antibodies of the present disclosure. The results are shown in Figure 22. The binding pattern for humanized antibodies of the present disclosure h30-HlL4 and h30-H4L3 is similar to that for the control antibody pTDP-43409 / 410, i.e. binding to pathological TDP-43 in the cytoplasm and neurites is observed, but only weak binding to endogenous TDP-43 in the nucleus. Conclusion

[0586] Humanized antibodies of the present disclosure bind pathological TDP-43 in the cytoplasm and neurites, but only bind weakly to endogenous TDP-43 in the nucleus. Thus, antibodies as disclosed herein bind to pathological human TDP-43, such as non-reversible human TDP-43 aggregates ex vivo. Said antibodies thus have a therapeutic potential in the treatment of diseases associated with TDP-43 proteopathies.

[0587] Example 19

[0588] Pharmacokinetic profile of humanized TDP-43 antibodies Pharmacokinetic analysis of humanized antibodies according to the disclosure was performed in Tg32- / - (human FcRn transgenic) mice. This strain expresses human neonatal Fc receptor (FcRn) and therefore provides a better prediction of the pharmacokinetic profiles of antibodies in human in comparison to wild type mice. Pharmacokinetic parameters were calculated based on plasma exposure data.

[0589] Materials and methods

[0590] Administration of antibody and sample collection: Female Tg32- / - mice (human FcRn tg mice) (n=5 / group and substance) were given a single i.v. injection (via the tail vein) of 10 mg / kg of respective antibody. Blood was collected at 5 min, 4 h, 24 h, 72 h, 168 h (7 days), 336 h (14 days), 672 h (28 days) and 840 h (35 days) after injection. Blood was collected into K2 EDTA tubes. The samples were centrifuged at 2400 x g for 10 min at 4°C shortly after collection (within 30 min). Plasma was collected and stored at -80°C until bioanalysis.

[0591] Determination of antibody concentrations in plasma using antigen coat: For screening purposes an MSD assay with antigen coated plates was used to determine plasma exposure. Briefly, wells of an MSD plate were coated with 2.5 pg / ml of aggregated TDP-43 (described in Example 1) overnight at 4°C. The wells were blocked with Pierce protein free blocking buffer (Thermo Fisher) for 1-3 h and thereafter plasma samples and standard were added. After incubation for 2 h, antihuman IgG-SULFO (Mesoscale, R32A1-1) was added to each well and the plates were incubated for 1 h at RT, 900 rpm after which 2x Reading buffer (Mesoscale, R92TC-1) was added. The plate was read within 5 min using MSD software. All incubation steps were performed at RT and 900 rpm.

[0592] Determination of antibody concentrations in plasma using sandwich format: To determine the pharmacokinetic parameters of the antibodies of interest, a sandwich approach was used for a more robust method. Briefly, wells of an MSD plate were coated with goat anti-human IgG (0.5 pg / ml) (Jackson Immuno Research, 165207) overnight at 4°C. The wells were blocked with Blocker A (Mesoscale, R93BA-2) at RT, 900 rpm for 1-3 h and thereafter the plasma samples were added, diluted in diluent 100 (Mesoscale, R50AA-3). After incubation for 2 h, mouse anti-human IgG-SULFO (Mabtech, 3850-1-1000) was added to each well at a concentration of 0.5 pg / ml and the plates were incubated for 1 h at RT, 900 rpm. A detection antibody, goat anti-mouse IgG-SULFO (Mesoscale, R32AC-1) was added at a concentration of 0.5 pg / ml and after an additional hour of incubation 2x Reading buffer (MSD, R92TC-1) was added. The plate was read within 5 min using MSD software. All incubation steps were performed at RT and 900 rpm.

[0593] Pharmacokinetic analysis: Individual plasma concentration versus time profiles were subjected to pharmacokinetic evaluation as described in Example 14 using exposure data from the sandwich based MSD assay.

[0594] Results

[0595] Plasma pharmacokinetic profiles for the humanized antibody variants were evaluated after a single i.v. bolus injection in Tg32- / - (human FcRn tg) mice. Mean concentration-time profiles using the antigen coated MSD assay are shown in Figure 23A indicating that profiles for h30-HlL4 and h30-H4L3 were similar and superior to the other antibody variants, with lower plasma clearance and longer terminal halflife. Pharmacokinetic parameters for h30-HlL4 and h30-H4L3 were determined based on exposure data from a sandwich based MSD assay (Figure 23B). A summary of the pharmacokinetic parameters of h30-HlL4 and h30-H4L3 inTg32 - / - mice is given in Table 13. Similar pharmacokinetic properties were observed for h30-HlL4 and h30-H4L3. Table 13. Mean values (n=4-5 / group) of pharmacokinetic parameters CL AUCinf Half-life Antibody n

[0596] (ml / h / kg) (mg / ml*h) (days) h30-HlL4 4 0.22 47 16

[0597]

[0598] h30-H4L3 5 0.19 54 16

[0599] Conclusions

[0600] The most promising pharmacokinetic profile among the tested humanized rec30 ( h30) variants was observed for H1L4 and H4L3, characterized by low clearance and long half-life.

[0601] Example 20

[0602] FcyR-mediated endocytosis of aggregated TDP-43 in THP-1 cells in vitro by TDP-43 antibodies

[0603] An in vitro uptake assay was used to assess FcyR-mediated endocytosis of Alexa-488-labeled TDP-43 aggregates by human monocytic THP-1 cells induced by two humanized (h30) antibody variants, h30-HlL4 and h30-H4L3.

[0604] Materials and methods

[0605] THP-1 cells were purchased from Sigma / ECACC and cultured in RPMI1640 (Gibco) supplemented with 10% FBS (Hyclone), lx GlutaMax (Gibco), lx Penicillin Streptomycin (Hyclone). Alexa-488 labelling of aggregated TDP-43 was performed as described in Example 11.

[0606] h30-HlL4 or h30-H4L3 and Alexa-488-labeled TDP-43 aggregates were incubated together for 30 min at RT (at final concentrations of 10.7-0.3 nM for the antibodies and 70 nM for aggregated TDP-43).

[0607] 200,000 THP-1 cells were added to the wells of a 96-well plate (Corning) and pelleted by centrifugation at 300 x g 5 min at RT, resuspended with the antibody / TDP-43-aggregate complexes and incubated at 37°C, 5% CO2 for 60-120 min. Cells were washed in PBS before data acquisition using a BD FACS Lyric flow cytometer. Data was evaluated using FCS Express 4 Flow Research Edition software (De Novo Software). EC50 values were calculated using non-linear regression with the sigmoidal 4PL equation in GraphPad Prism. Results

[0608] The ability of H30-H1L4 and h30-H4L3 to induce uptake of Alexa-488-labeled TDP-43 aggregates by THP-1 human monocytic cells was evaluated. The results indicate that both antibodies induced uptake of Alexa-488-labeled TDP-43 aggregates by THP-1 cells in a concentration-dependent manner (Figure 24). No uptake was observed using an irrelevant control antibody (" Neg Control Ab"). The calculated EC50 values were 1 and 1.2 nM for h30-HlL4 and h30-H4L3, respectively.

[0609] Conclusions

[0610] Humanized antibodies according to the disclosure induced FcyR-mediated endocytosis of human TDP-43 aggregates in vitro by monocytes. These data indicate that these antibodies are capable of inducing clearance of pathological human TDP-43, such as irreversible human TDP-43 aggregates, by the immune system.

[0611] Example 21

[0612] Immunogenicity of h30 variant of TDP-43 antibodies This example describes the assessment of the potential immunogenicity of h30-HlL4 and h30-H4L3 antibodies generated as described in Example 15. The ability of the variant antibodies to induce CD4+ T cell responses was evaluated using the Episcreen™ time course assay.

[0613] Materials and methods

[0614] Isolation of peripheral blood mononuclear cells (PBMCs): PBMCs were isolated from healthy community donor buffy coats (from blood drawn within 24 h) obtained under consent from commercial vendors. Cells were separated by density centrifugation using a lymphocyte separation medium (StemCell Technologies Inc, London, UK) and CD8+ T cells were depleted using CD8+ RosetteSep™ (StemCell Technologies Inc). Donors were characterized by identifying HLA-DR and HLA-DQ haplotypes to 4-digit resolution by SSO HLA typing (VHBio, Gateshead, UK). T cell responses to the neo-antigen KLH (Invitrogen, Paisley, UK) were also determined. PBMCs were then frozen and stored in the vapor phase of nitrogen until required.

[0615] Preparation of samples: Endotoxin levels in the antibody samples were measured using a LAL chromogenic kinetic assay kit (Charles River, Margate, UK) according to the manufacturer's instructions and found to be within the limit acceptable for the assay (<3 EU / mg).

[0616] The antibody samples were diluted to 0.6 pM in AIM-V® culture medium (Invitrogen) prior to use (at a final assay concentration of 0.3 pM). KLH was used as a reproducibility control and stored at -20°C as a 10 mg / ml stock solution in water. For the studies, an aliquot of KLH was thawed immediately before use and diluted to 200 pg / ml in AIM-V® (at a final concentration of 100 pg / ml). A further high immunogenicity control, CEFT (pool of 13 peptides from Pepscan Ltd, Lelystad, The Netherlands) was used as a high responding control and this was stored at -20°C as a 1.538 mg / ml stock solution and diluted in AIM-V® to 2 pg / ml before use (final concentration 1 pg / ml). Herceptin® (Bionical Ltd, Willington, UK) was used as a negative clinical control and this was stored at -80°C as a 20 mg / ml stock solution (at a final assay concentration of 50 pg / ml).

[0617] Time course proliferation assay: A cohort of 50 donors was selected for the assay. PBMCs from each donor were thawed, counted and viability assessed using an acridine orange (AO) and 4',6-diamidino-2-phenylindole (DAPI) (Chemometec Ltd, Allerod, Denmark) dye exclusion. Cells were revived in room temperature AIM-V® culture medium, washed and resuspended in AIM-V® to 4-6 x 106PBMC / ml for use as the proliferation cell stock. For each donor, bulk cultures were established in which 1 ml of the proliferation cell stock was added to the appropriate wells of a 24 well plate. 1 ml of each sample was added to the PBMCs to give a final sample concentration of 0.3 pM. For each donor, a reproducibility control well (cells incubated with 100 pg / ml KLH), a further high immunogenicity control (cells incubated with 1 pg / ml CEFT peptide pool), a low immunogenicity control (cells incubated with 50 pg / ml Herceptin®) and a culture medium only well were also included. Cultures were incubated for a total of 8 d at 37°C with 5% CO2. On days 5, 6, 7 and 8, the cells in each well were gently resuspended by mixing 5x using an electronic pipette and 3 x 100 pl aliquots transferred to each well of a round bottomed 96 well plate. The cultures were pulsed with 0.75 pCi [3H]-Thymidine (Perkin Elmer, Beaconsfield, UK) in 100 pl AIM-V® culture medium and incubated for a further 18 h before harvesting onto filter mats (Perkin Elmer) using a TomTec Mach III cell harvester. CPM for each well were determined by Meltilex™ (Perkin Elmer) scintillation counting on a 1450 Microbeta Wallac Trilux Liquid Scintillation Counter (Perkin Elmer) in paralux, low background counting.

[0618] Assessment of cell viability: On day 7, bulk cultures (previously established for the proliferation assay) were gently resuspended by mixing 5x using an electronic pipette and 50 pl was removed from each well and mixed with 2.5 pl acridine orange (AO) and 4',6-diamidino-2-phenylindole (DAPI) (Chemometec Ltd) dye exclusion. Cells were then assessed for viability using a NucleoCounter® NC-250™ automated cell analyser (Chemometec Ltd).

[0619] Data analysis: For proliferation assays, an empirical threshold of a stimulation index (SI) equal to or greater than 1.9 (SI >1.90) had been previously established, whereby samples inducing responses above this threshold are deemed positive. For proliferation analysis, donors that were positive on at least one time point during the time course assay were deemed positive donors and counted towards the "% Response"parameter.

[0620] Results

[0621] The ability of H30-H1L4 and h30-H4L3 to induce CD4+ T cell responses was evaluated. EpiScreen™ analysis of the frequency and magnitude of the responses showed that h30-HlL4 and h30-H4L3 induced 0% and 4% response respectively which was below the response for the low immunogenicity control Herceptin® and were therefore considered to have a low risk for immunogenicity (Table 14).

[0622] Table 14. Summary of healthy donor proliferation Antibody Mean SI SD % Response h30-HlL4 N / A N / A 0

[0623] h30-H4L3 2.73 0.56 4

[0624] Herceptin® 2.05 0.03 6

[0625] CEFT 3.87 2.73 70

[0626]

[0627] KLH 8.84 13.39 100

[0628] Conclusions

[0629] Immunogenicity evaluation of h30 antibody variants revealed that the tested antibodies exhibit a low risk for immunogenicity. These data support therapeutic applicability of antibodies according to the present disclosure. Example 22

[0630] Stability of h30

[0631] Stability assessment was performed for h30 variants H30-H1L4 and h30-h4L3. Results were similar for both variants. Only h30-HlL4 results are shown.

[0632] Materials and methods

[0633] Stability study: h30 variants h30-HlL4 and h30-h4L3 were diluted to 1 mg / ml in PBS and incubated at 4°C, 20°C or 40°C for 2 or 4 weeks, respectively (Table 15). A reference sample (TO) and samples withdrawn after 2 and 4 weeks were frozen at - 80°C. All samples were analyzed later, at the same time point.

[0634] Indirect ELISA: Samples from the different incubation conditions were analyzed for binding to human TDP-43 aggregates using recombinant human TDP-43 protein. The procedure was essentially the same as described in Example 7 above, using 0.5 pg / ml of aggregated TDP-43 as coat.

[0635] Table 15. Conditions of stability study

[0636] Temperature TO T2W T4W -80°C X

[0637] 4°C X X 20°C X X

[0638]

[0639] 40°C X X

[0640] Results

[0641] Results from indirect ELISA showed that target binding by h30-HlL4 to aggregated TDP-43 was intact after incubation at 4°C and 20°C for 2 and 4 weeks. For incubation at 40°C, a decreased binding to target can be seen after 2 weeks, and this effect is more pronounced after 4 weeks (Figure 25). The measured EC50 values are given in Table 16. Table 16. EC50 values by ELISA for binding of h30-H1L4 to aggregated TDP-43

[0642] EC50

[0643] Sample

[0644] (ng / ml))

[0645] TO 9.2

[0646] 4°C, 2 w 7.4

[0647] 4°C, 4 w 8.9

[0648] 25°C, 2 w 7.6

[0649] 25°C, 4w 7.7

[0650] 40°C, 2 w 18

[0651]

[0652] 40°C, 4 w 53

[0653] Conclusion

[0654] Incubation at 40°C negatively affects the ability of h30-HlL4 to bind to aggregated TDP-43, showing a worsening overtime.

[0655] Example 23

[0656] Engineered h30-H1L4 variants with improved stability In forced degradation studies of h30-H1L4, the antibody exhibited deamidation and isomerization. It also lost its target binding capacity, similarly to what was seen in Example 22 after incubation at 40°C. The major sites for isomerization and deamidation were identified by peptide mapping, and found to be an isomerization site in VHCDR3 (D95) and a deamidation site in VLCDR1 (G29). To remove these motifs, mutations were made at both positions. Three mutants were chosen for further evaluation: a VHCDR3 isomerization mutant denoted h30(H: D95G), a VLCDR1 deamidation mutant denoted h30(L: G29Q), and a double mutant denoted h30(H: D95G-L: G29Q). The variable domain amino acid sequences three mutant antibodies are listed in Table 17. For the constant domains, the same heavy and light chains were used as for the h30 variants listed in Table 10, i.e. the human IgGl heavy chain with a K322A mutation of SEQ ID NO:49, and the human kappa light chain of SEQ ID NO:51.

[0657] Table 17. Seguences of h30-HlL4 mutant variable domains

[0658] SEQ ID

[0659] Antibody Region Amino acid sequence

[0660] NO: Heavy chain

[0661] h30(H: D95G)

[0662]

[0663] VH QVQLVQSGPELKKPGASVKISCKGSGYTFTDYNMHWV 61 KQAHGKGLEWIGVISTYSGNTNYNQKFKDRATMTVDK STSTAYMELSSLRSEDSAIYYCARGGYFDYWGQGTTVT VSS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 VH-CDR3 GGYFDY 60 Light chain

[0664] DVVMTQSPDSLAVSLGERATINCRSSQYIVHSNGDTYL VL EWYQQKPGQPPKLLIYKVSNRFSGVPDRFSGSGSGTDF 45

[0665] TLTISSLQAEDVAVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNGDTYLE 28 VL-CDR2 KVSNRFS 31 VL-CDR3 FQGSHVPPT 17 Heavy chain QVQLVQSGPELKKPGASVKISCKGSGYTFTDYNMHWV KQAHGKGLEWIGVISTYSGNTNYNQKFKDRATMTVDK VH 40

[0666] STSTAYMELSSLRSEDSAIYYCARDGYFDYWGQGTTVT VSS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 h30(L: G29Q) VH-CDR3 DGYFDY 25

[0667] Light chain

[0668] DVVMTQSPDSLAVSLGERATINCRSSQYIVHSNQDTYL VL EWYQQKPGQPPKLLIYKVSNRFSGVPDRFSGSGSGTDF 63

[0669] TLTISSLQAEDVAVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNQDTYLE 62 VL-CDR2 KVSNRFS 31 VL-CDR3 FQGSHVPPT 17 Heavy chain QVQLVQSGPELKKPGASVKISCKGSGYTFTDYNMHWV KQAHGKGLEWIGVISTYSGNTNYNQKFKDRATMTVDK VH 61

[0670] STSTAYMELSSLRSEDSAIYYCARGGYFDYWGQGTTVT VSS VH-CDR1 DYNMH 19 VH-CDR2 VISTYSGNTNYNQKFKD 22 h30(H: D95G- VH-CDR3 GGYFDY 60 L: G29Q)

[0671] Light chain

[0672] DVVMTQSPDSLAVSLGERATINCRSSQYIVHSNQDTYL VL EWYQQKPGQPPKLLIYKVSNRFSGVPDRFSGSGSGTDF 63

[0673] TLTISSLQAEDVAVYYCFQGSHVPPTFGQGTKLEIK VL-CDR1 RSSQYIVHSNQDTYLE 62 VL-CDR2 KVSNRFS 31

[0674]

[0675] VL-CDR3 FQGSHVPPT 17 Materials and methods

[0676] Construction of new variants of h30: Mutations were made to remove the liabilities in h30 (DG-motif in VH-CDR3 and NG-motif in VL-CDR1) without affecting in silica immunogenicity or introducing new liabilities. Standard molecular biology techniques were used.

[0677] Indirect ELISA: The new variants were analyzed for binding to human TDP-43 aggregates using recombinant human TDP-43 protein. The procedure was essentially the same as described in Example 7 above, using 0.5 pg / ml of aggregated TDP-43 as coat.

[0678] SPR: The new variants were analyzed for binding to human TDP-43 aggregates by SPR, as described in Example 9 above.

[0679] IHC on human brain sections: Selected variants were evaluated in postmortem brain sections from patients diagnosed with frontotemporal dementia (FTD) using immunohistochemistry (IHC) in comparison to non-demented (NDE) controls. The procedure was essentially the same as described in Example 18.

[0680] Indirect MSP assay: The procedures were essentially the same as described in Example 10. For detection, a goat anti-human SULFO-TAG antibody was used. Background signal obtained from binding of the anti-human SULFO-TAG antibody to the human brain extracts alone was subtracted from the signal obtained for the humanized antibody variants.

[0681] Immunoprecipitation (IP) of total TDP-43 and pTDP-43: The h 30 and h30(H: D95G) antibodies were coupled to M-280 tosyl activated magnetic beads at 15 pg antibody per mg of beads. Human brain extracts (postmortem brain material from patients diagnosed with FTD and ALS) were incubated with titrated amounts of antibody-coupled beads for 2 h with shaking, resulting in 4.5, 1.13 and 0.28 pg antibody per IP reaction. Bound target was eluted using 1% SDS and boiling at 95 °C for 5 min. The eluates were analyzed using both the total TDP-43 assay (as described in Example 10) and a commercially provided Human Phospho-TDP-43 (Ser409 / 410) specific assay (Alpha LISA, revvity).

[0682] Pharmacokinetics in SCID hFcRn tg mice (Tg32- / -): The selected variants of h30 were evaluated in a pharmacokinetic (PK) study to confirm that PK properties had not changed. The study was conducted in the same manner as in Example 19, using the sandwich based MSD assay to determine exposure levels in plasma. Results

[0683] The three h30-HlL4 mutants h30(H: D95G), h30(L: G29Q) and h30(H: D95G-L: G29Q) exhibited retained binding to aggregated TDP-43 as analyzed using indirect ELISA (Figure 26) with no significant worsening of in silica immunogenicity and no introduction of new liabilities. The measured EC50 values are given in Table 18.

[0684] Table 18. EC50 by ELISA for binding ofh30 mutants to aggregated TDP-43

[0685] EC50

[0686] Antibody

[0687] (ng / ml))

[0688] h30-HlL4 10.55

[0689] h30(L: G29Q) 12.67

[0690] h30(H: D95G) 11.64

[0691]

[0692] h30(H: D95G-L: G29Q) 17.06

[0693] The results of SPR analysis of the mutants' binding to immobilized, aggregated TDP-43 are shown in Figure 27. The measured kinetic parameters are summarized in Table 19. KD values for the two single mutants are within two-fold of the KD for h30-HlL4, while the double mutant shows a five-fold loss of binding.

[0694] Table 19. KD values for h30-HlL4 mutants as measured by SPR Antibody ka (M-1s-1) kd (s-1) KD (pM) h30(H: D95G) 8.1 x 1052.5 x IO’4320 h30(L: G29Q) 8.4 x 1051.6 x IO’4190

[0695]

[0696] h30(H: D95G-L: G29Q) 6.9 x 1055.3 x IO’4810

[0697] Further evaluation of target binding by the novel mutants revealed a similar target binding as for h30-HlL4, as determined by IHC on human brain sections (Figure 28).

[0698] Indirect MSD assay and immunoprecipitation (IP) followed by measurement of bound total TDP-43 and pTDP-43 showed that h30 and h30(H: D95G) exhibit similar binding profiles, confirming target binding to pathological TDP-43 and, to a lesser extent, to TDP-43 in NDE extracts. The results are shown in Figure 29 and Figure 30. Plasma PK profiles of the new mutants were evaluated after a single i.v. bolus injection in SCID Tg32- / - (human FcRn tg) mice. Mean concentration-time profiles are shown in Figure 31, including h30-HlL4 from Example 19, indicating that profiles for all liability variants were similar to h30-HlL4, although a direct comparison is difficult since h30-HlL4 was evaluated in a different mouse strain. A summary of the pharmacokinetic parameters of the new mutants is given in Table 20.

[0699] Table 20. Mean values (n=4 / group) of PK parameters per sandwich MSP Cmax CL AUCinf Half-life Antibody

[0700] (pg / ml) (ml / h / kg) (mg / ml*h) (days) h30(H: D95G) 342 0.12 82 15.3 h30(H: D95G-L: G29Q) 311 0.15 67 15.4

[0701]

[0702] h30(L: G29Q) 372 0.11 90 19.8

[0703] Conclusions

[0704] The three tested mutants of h30-HlL4 show retained binding to target in indirect ELISA, although the single mutants exhibit slightly higher KD values to aggregated TDP-43 by SPR and the double mutant exhibits a slightly increased KD value. The binding to pathological forms of TDP-43 on human brain sections remained, although differences among the mutants were seen, with h30(H: D95G) showing the most similarities to the binding pattern of h30-HlL4. PK properties were comparable to those of h30-HlL4, suggesting that the mutations did not have any impact on these parameters.

[0705] Example 24

[0706] Limited stability study and binding characteristics of h30-HlL4 mutants To confirm that the removal of potential manufacturability liabilities had improved the stability of h30-HlL4 at 40°C, a limited study was performed focusing on stability during incubation at 40°C and on target binding to aggregated TDP-43 by indirect ELISA. Materials and methods

[0707] Stability study: Under the conditions shown in Table 21, the tested antibodies were diluted to ~0.8 mg / ml in PBS and incubated at 40°C for 4 weeks

[0708] _ Table 21. Conditions for stability study

[0709] Temperature -80°C 40°C 40°C Time point TO T2W T4W h30-HlL4 X X X h30(H: D95G) X X X h30(L: G29Q) X X X

[0710]

[0711] h30(H: D95G-L: G29Q) X X X

[0712] Indirect ELISA: Antibodies were analyzed for binding to human TDP-43 aggregates using recombinant human TDP-43 protein. The procedure was essentially the same as described in Example 7 above, using 0.5 pg / ml of aggregated TDP-43 as coat.

[0713] Results

[0714] All three tested mutants showed an improved binding to target compared to h30-HlL4, after incubation at 40°C for 4 weeks (Figure 32). The measured EC50 values are given in Table 22.

[0715] Table 22. EC50 values by ELISA for binding of h30-HlL4 and mutants to aggregated TDP-43

[0716] EC50

[0717] Antibody, Conditions

[0718] (ng / ml))

[0719] h30-HlL4 3.9

[0720] h30-HlL4, 40°C, 4 w 193 h30(H: D95G) 4.4 h30(H: D95G), 40°C, 4w 44

[0721] h30(L: G29Q) 4.9 h30(L: G29Q), 40°C, 4w 61

[0722] h30(H: D95G-L: G29Q) 8.0

[0723]

[0724] h30(H: D95G-L: G29Q), 40°C, 4 w 38

[0725] Target binding was not restored to TO values, but improvements were observed for all mutants compared to h30-HlL4. The most pronounced improvement was observed for the isomerization single mutant h30(H: D95G) and the double mutant h30(H: D95G-L: G29Q). This strongly suggests that removal of the isomerization site drives the improvement in target binding, as no further improvement is seen for the double mutant.

[0726] Conclusion

[0727] The mutants improved binding to target after incubation at 40°C for 4 weeks compared to h30-HlL4, as analyzed by indirect ELISA. Ill

[0728] ITEMIZED LISTING OF EMBODIMENTS

[0729] 1. An antibody or antigen-binding fragment thereof, comprising an immunoglobulin heavy chain variable (VH) region and an immunoglobulin light chain variable (VL) region, said VH and VL regions forming a VH / VL pair comprising an antigen-binding surface, wherein said antigen-binding surface is composed of three complementarity-determining regions (CDRs) from said VH region and three CDRs from said VL region, and wherein said CDRs consist of the following amino acid sequences:

[0730] VH-CDR1: X1YX2X3X4 (SEQ ID NO:1), wherein

[0731] XI is selected from E, D and N,

[0732] X2 is selected from N and Y,

[0733] X3 is selected from M and I,

[0734] X4 is selected from S and H;

[0735] VH-CDR2: X5IX6X7X8X9X10X11TX12YX13X14X15X16X17X18 (SEQ ID NO:2), wherein

[0736] X5 is selected from V and A,

[0737] X6 is selected from N and S,

[0738] X7 is selected from P, T and N,

[0739] X8 is selected from Y, E and N, such as from Y and E,

[0740] X9 is selected from S and G,

[0741] X10 is selected from E, G and D,

[0742] XII is selected from N and S,

[0743] X12 is selected from N and H,

[0744] X13 is selected from N and P,

[0745] X14 is selected from P, Q and D,

[0746] X15 is selected from K and T,

[0747] X16 is selected from F and V,

[0748] X17 is selected from K and T,

[0749] X18 is selected from D and G;

[0750] VH-CDR3: X19X20GX21X22X23X24 (SEQ ID NO:3), wherein

[0751] X19 is A or missing, X20 is selected from D, E and G, such as from D and E,

[0752] X21 is selected from Y and S,

[0753] X22 is selected from F and L,

[0754] X23 is selected from D and A,

[0755] X24 is selected from F and Y;

[0756] VL-CDR1: X25SSQX26X27X28X29X30NX31X32X33X34YLX35 (SEQ ID NO:4), wherein

[0757] X25 is selected from R and K,

[0758] X26 is selected from I, Y and S,

[0759] X27 is selected from I and L,

[0760] X28 is selected from V and L,

[0761] X29 is selected from H and Y,

[0762] X30 is selected from S and T,

[0763] X31 is selected from G, N and Q, such as from G and N, X32 is Qor missing,

[0764] X33 is selected from A, D and K,

[0765] X34 is selected from N and T,

[0766] X35 is selected from E and A;

[0767] VL-CDR2: X36X37SX38RX39S (SEQ ID NO:5), wherein

[0768] X36 is selected from K and W,

[0769] X37 is selected from V and A,

[0770] X38 is selected from K, N and T,

[0771] X39 is selected from F and E;

[0772] VL-CDR3: X40QX41X42X43X44PX45T (SEQ ID NO:6), wherein

[0773] X40 is selected from Qand F,

[0774] X41 is selected from G and Y,

[0775] X42 is selected from Y and S,

[0776] X43 is selected from H and T,

[0777] X44 is selected from V and Y,

[0778] X45 is selected from L and P. 2. The antibody or antigen-binding fragment thereof according to item 1, wherein said VH-CDR1 consists of the following amino acid sequence

[0779] VH-CDR1: X1YNX3H (SEQ ID NO:7), wherein

[0780] XI is selected from E and D,

[0781] X3 is selected from M and I.

[0782] 3. The antibody or antigen-binding fragment thereof according to item 1, wherein said VH-CDR1 consists of the following amino acid sequence

[0783] VH-CDR1: X1YX2MX4 (SEQ ID NO:8), wherein

[0784] XI is selected from D and N,

[0785] X2 is selected from N and Y,

[0786] X4 is selected from S and H.

[0787] 4. The antibody or antigen-binding fragment thereof according to any one of items 1-3, wherein said VH-CDR2 consists of the following amino acid sequence VH-CDR2: VIX6X7YSX10NTNYNX14KFX17D (SEQID NO:9), wherein

[0788] X6 is selected from N and S,

[0789] X7 is selected from P and T,

[0790] X10 is selected from E and G,

[0791] X14 is selected from P and Q,

[0792] X17 is selected from K and T.

[0793] 5. The antibody or antigen-binding fragment thereof according to any one of items 1-3, wherein said VH-CDR2 consists of the following amino acid sequence VH-CDR2: X5IX6X7X8X9X10X11TX12YX13X14X15X16KX18 (SEQ ID NO:10), wherein

[0794] X5 is selected from V and A,

[0795] X6 is selected from N and S,

[0796] X7 is selected from T and N,

[0797] X8 is selected from Y, E and N, such as Y and E,

[0798] X9 is selected from S and G,

[0799] X10 is selected from G and D,

[0800] XII is selected from N and S, X12 is selected from N and H,

[0801] X13 is selected from N and P,

[0802] X14 is selected from Q and D,

[0803] X15 is selected from K and T,

[0804] X16 is selected from F and V,

[0805] X18 is selected from D and G.

[0806] 6. The antibody or antigen-binding fragment thereof according to any one of items 1-5, wherein said VH-CDR3 consists of the following amino acid sequence VH-CDR3: X19X20GYFDX24 (SEQ ID NO:11), wherein

[0807] X19 is missing,

[0808] X20 is selected from D and G,

[0809] X24 is selected from F and Y.

[0810] 7. The antibody or antigen-binding fragment thereof according to any one of items 1-5, wherein said VH-CDR3 consists of the following amino acid sequence VH-CDR3: X19X20GX21X22X23Y (SEQ ID NO:12), wherein

[0811] X19 is A or missing,

[0812] X20 is selected from D, E and G, such as from D and E, X21 is selected from Y and S,

[0813] X22 is selected from F and L,

[0814] X23 is selected from D and A.

[0815] 8. The antibody or antigen-binding fragment thereof according to any one of items 1-7, wherein said VL-CDR1 consists of the following amino acid sequence VL-CDR1: RSSQX26IVHX30NX31X32X33TYLE (SEQ ID NO:13), wherein

[0816] X26 is selected from I and Y,

[0817] X30 is selected from S and T,

[0818] X31 is selected from G and Q,

[0819] X32 is missing,

[0820] X33 is selected from A and D.

[0821] 9. The antibody or antigen-binding fragment thereof according to any one of items 1-7, wherein said VL-CDR1 consists of the following amino acid sequence VL-CDR1: X25SSQX26X27X28X29SNX31X32X33X34YLX35 (SEQ ID NO:14), wherein

[0822] X25 is selected from R and K,

[0823] X26 is selected from Y and S,

[0824] X27 is selected from I and L,

[0825] X28 is selected from V and L,

[0826] X29 is selected from H and Y,

[0827] X31 is selected from G and N,

[0828] X32 is Qor missing,

[0829] X33 is selected from D and K,

[0830] X34 is selected from N and T,

[0831] X35 is selected from E and A.

[0832] 10. The antibody or antigen-binding fragment thereof according to any one of items 1-9, wherein said VL-CDR2 consists of the following amino acid sequence VL-CDR2: KVSX38RFS (SEQ ID NO:15), wherein

[0833] X38 is selected from K and N.

[0834] 11. The antibody or antigen-binding fragment thereof according to any one of items 1-9, wherein said VL-CDR2 consists of the following amino acid sequence VL-CDR2: X36X37SX38RX39S (SEQ ID NO:16), wherein

[0835] X36 is selected from K and W,

[0836] X37 is selected from V and A,

[0837] X38 is selected from N and T,

[0838] X39 is selected from F and E.

[0839] 12. The antibody or antigen-binding fragment thereof according to any one of items 1-11, wherein said VL-CDR3 consists of the following amino acid sequence VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0840] 13. The antibody or antigen-binding fragment thereof according to any one of items 1, 2, 4, 6, 8, 10 and 12, wherein said CDRs consist of the following amino acid sequences

[0841] VH-CDR1: X1YNX3H (SEQ ID NO:7), wherein XI is selected from E and D,

[0842] X3 is selected from M and I.

[0843] VH-CDR2: VIX6X7YSX10NTNYNX14KFX17D (SEQ ID NO:9), wherein

[0844] X6 is selected from N and S,

[0845] X7 is selected from P and T,

[0846] X10 is selected from E and G,

[0847] X14 is selected from P and Q,

[0848] X17 is selected from K and T;

[0849] VH-CDR3: X19X20GYFDX24 (SEQ ID NO:11), wherein

[0850] X19 is missing,

[0851] X20 is selected from D and G,

[0852] X24 is selected from F and Y.

[0853] VL-CDR1: RSSQX26IVHX30NX31X32X33TYLE (SEQ ID NO:13), wherein

[0854] X26 is selected from I and Y,

[0855] X30 is selected from S and T,

[0856] X31 is selected from G and Q,

[0857] X32 is missing,

[0858] X33 is selected from A and D.

[0859] VL-CDR2: KVSX38RFS (SEQ ID NO:15), wherein

[0860] X38 is selected from K and N;

[0861] VL-CDR3: FQGSHVPT (SEQ ID NO:17).

[0862] 14. The antibody or antigen-binding fragment thereof according to any one of items 1, 3, 5, 7, 9 and 11, wherein said CDRs consist of the following amino acid sequences

[0863] VH-CDR1: X1YX2MX4 (SEQ ID NO:8), wherein

[0864] XI is selected from D and N,

[0865] X2 is selected from N and Y,

[0866] X4 is selected from S and H; VH-CDR2: X5IX6X7X8X9X10X11TX12YX13X14X15X16KX18 (SEQ ID NO:10), wherein

[0867] X5 is selected from V and A,

[0868] X6 is selected from N and S,

[0869] X7 is selected from T and N,

[0870] X8 is selected from Y, E and N, such as Y and E,

[0871] X9 is selected from S and G,

[0872] X10 is selected from G and D,

[0873] Xll is selected from N and S,

[0874] X12 is selected from N and H,

[0875] X13 is selected from N and P,

[0876] X14 is selected from Q and D,

[0877] X15 is selected from K and T,

[0878] X16 is selected from F and V,

[0879] X18 is selected from D and G;

[0880] VH-CDR3: X19X20GX21X22X23Y (SEQ ID NO:12), wherein

[0881] X19 is A or missing,

[0882] X20 is selected from D and E,

[0883] X21 is selected from Y and S,

[0884] X22 is selected from F and L,

[0885] X23 is selected from D and A;

[0886] VL-CDR1: X25SSQX26X27X28X29SNX31X32X33X34YLX35 (SEQ ID NO:14), wherein

[0887] X25 is selected from R and K,

[0888] X26 is selected from Y and S,

[0889] X27 is selected from I and L,

[0890] X28 is selected from V and L,

[0891] X29 is selected from H and Y,

[0892] X31 is selected from G, N and Q, such as from G and N, X32 is Q or missing,

[0893] X33 is selected from D and K,

[0894] X34 is selected from N and T, X35 is selected from E and A;

[0895] VL-CDR2: X36X37SX38RX39S (SEQ ID NO:16), wherein

[0896] X36 is selected from K and W,

[0897] X37 is selected from V and A,

[0898] X38 is selected from N and T,

[0899] X39 is selected from F and E.

[0900] 15. The antibody or antigen-binding fragment thereof according to any one of items 1, wherein the amino acid sequence of said VH-CDR1 is selected from the group consisting of SEQ ID NO:18-20.

[0901] 16. The antibody or antigen-binding fragment thereof according to item 15, wherein the amino acid sequence of said VH-CDR1 is selected from the group consisting of SEQ ID NO:18 and SEQ ID NO:19.

[0902] 17. The antibody or antigen-binding fragment thereof according to item 15, wherein the amino acid sequence of said VH-CDR1 is selected from the group consisting of SEQ ID NO:19 and SEQ ID NO:20.

[0903] 18. The antibody or antigen-binding fragment thereof according to item 15 or 16, wherein the amino acid sequence of said VH-CDR1 is SEQ ID NO:18.

[0904] 19. The antibody or antigen-binding fragment thereof according to any one of items 15-17, wherein the amino acid sequence of said VH-CDR1 is SEQ ID NO:19.

[0905] 20. The antibody or antigen-binding fragment thereof according to item 15 or 17, wherein the amino acid sequence of said VH-CDR1 is SEQ ID NO:20.

[0906] 21. The antibody or antigen-binding fragment thereof according to any one of items 15-20, wherein the amino acid sequence of said VH-CDR2 is selected from the group consisting of SEQ ID NO:21-23 and SEQ ID NO:55, such as SEQ ID NO:21-22 and SEQ ID NO:55.

[0907] 22. The antibody or antigen-binding fragment thereof according to item 21, wherein the amino acid sequence of said VH-CDR2 is selected from the group consisting of SEQ ID NO:21 and SEQ ID NO:22.

[0908] 23. The antibody or antigen-binding fragment thereof according to item 21, wherein the amino acid sequence of said VH-CDR2 is selected from the group consisting of SEQ ID NO:22, SEQ ID NO:23 and SEQ ID NO:55, such as SEQ ID NO:22 and SEQ ID NO:55. 24. The antibody or antigen-binding fragment thereof according to item 21 or 22, wherein the amino acid sequence of said VH-CDR2 is SEQ ID NO:21.

[0909] 25. The binding molecule according to any one of items 21-23, wherein the amino acid sequence of said VH-CDR2 is SEQ ID NO:22.

[0910] 26. The antibody or antigen-binding fragment thereof according to item 21 or 23, wherein the amino acid sequence of said VH-CDR2 is SEQ ID NO:23 or SEQ ID NO:55, such as wherein the amino acid sequence of said VH-CDR2 is SEQ ID NO:55.

[0911] 27. The antibody or antigen-binding fragment thereof according to any one of items 15-26, wherein the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:24-26 and 60, such as the group consisting of SEQ ID NO:24-26.

[0912] 28. The antibody or antigen-binding fragment thereof according to item 27, wherein the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:24, 25 and 60, such as the group consisting of SEQ ID NO:24 and SEQ ID NO:25 or the group consisting of SEQ ID NO:25 and SEQ ID NO:60.

[0913] 29. The antibody or antigen-binding fragment thereof according to item 27, wherein the amino acid sequence of said VH-CDR3 is selected from the group consisting of SEQ ID NO:25 and SEQ ID NO:26.

[0914] 30. The antibody or antigen-binding fragment thereof according to item 27 or 28, wherein the amino acid sequence of said VH-CDR3 is SEQ ID NO:24.

[0915] 31. The antibody or antigen-binding fragment thereof according to any one of items 27-29, wherein the amino acid sequence of said VH-CDR3 is SEQ ID NO:25.

[0916] 32. The antibody or antigen-binding fragment thereof according to item 27 or 29, wherein the amino acid sequence of said VH-CDR3 is SEQ ID NO:26.

[0917] 33. The antibody or antigen-binding fragment thereof according to item 27 or 28, wherein the amino acid sequence of said VH-CDR3 is SEQ ID NO:60.

[0918] 34. The antibody or antigen-binding fragment thereof according to any one of items 15-33, wherein the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:27-29 and 62, such as the group consisting of SEQ ID NO:27-29.

[0919] 35. The antibody or antigen-binding fragment thereof according to item 34, wherein the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:27, 28 and 62, such as the group consisting of SEQ ID NO:27 and SEQ ID NO:28 or the group consisting of SEQ ID NO:28 and SEQ ID NO:62.

[0920] 36. The antibody or antigen-binding fragment thereof according to item 34, wherein the amino acid sequence of said VL-CDR1 is selected from the group consisting of SEQ ID NO:28 and SEQ ID NO:29.

[0921] 37. The antibody or antigen-binding fragment thereof according to item 34 or 35, wherein the amino acid sequence of said VL-CDR1 is SEQ ID NO:27.

[0922] 38. The antibody or antigen-binding fragment thereof according to any one of items 34-36, wherein the amino acid sequence of said VL-CDR1 is SEQ ID NO:28.

[0923] 39. The antibody or antigen-binding fragment thereof according to item 34 or 36, wherein the amino acid sequence of said VL-CDR1 is SEQ ID NO:29.

[0924] 41. The antibody or antigen-binding fragment thereof according to item 34 or 35, wherein the amino acid sequence of said VL-CDR1 is SEQ ID NO:62.

[0925] 42. The antibody or antigen-binding fragment thereof according to any one of items 15-41, wherein the amino acid sequence of said VL-CDR2 is selected from the group consisting of SEQ ID NO:30-32.

[0926] 43. The antibody or antigen-binding fragment thereof according to item 42, wherein the amino acid sequence of said VL-CDR2 is selected from the group consisting of SEQ ID NO:30 and SEQ ID NO:31.

[0927] 44. The antibody or antigen-binding fragment thereof according to item 42, wherein the amino acid sequence of said VL-CDR2 is selected from the group consisting of SEQ ID NO:31 and SEQ ID NO:32.

[0928] 45. The antibody or antigen-binding fragment thereof according to item 42 or 43, wherein the amino acid sequence of said VL-CDR2 is SEQ ID NO:30.

[0929] 46. The antibody or antigen-binding fragment thereof according to any one of items 42-44, wherein the amino acid sequence of said VL-CDR2 is SEQ ID NO:31.

[0930] 47. The antibody or antigen-binding fragment thereof according to item 42 or 44, wherein the amino acid sequence of said VL-CDR2 is SEQ ID NO:32.

[0931] 48. The antibody or antigen-binding fragment thereof according to any one of items 15-47, wherein the amino acid sequence of said VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33.

[0932] 49. The antibody or antigen-binding fragment thereof according to item 48, wherein the amino acid sequence of said VL-CDR3 is SEQ ID NO:17. 50. The antibody or antigen-binding fragment thereof according to item 48, wherein the amino acid sequence of said VL-CDR3 is SEQ ID NO:33.

[0933] 51. The antibody or antigen-binding fragment thereof according to any one of items 15-50, wherein the amino acid sequences of the 6 CDRs are the following: the VH-CDR1 is selected from the group consisting of SEQ ID NO:18-20;

[0934] the VH-CDR2 is selected from the group consisting of SEQ ID NO:21-23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:21-22 and SEQ ID NO:55; the VH-CDR3 is selected from the group consisting of SEQ ID NO:24-26 and 60, such as the group consisting of SEQ ID NO:24-26 or the group consisting of SEQ ID NO:25 and SEQ ID NO:60;

[0935] the VL-CDR1 is selected from the group consisting of SEQ ID NO:27-29 and 62, such as the group consisting of SEQ ID NO:27-29 or the group consisting of SEQ ID NO:28 and SEQ ID NO:62;

[0936] the VL-CDR2 is selected from the group consisting of SEQ ID NO:30-32; and the VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33.

[0937] 52. The antibody or antigen-binding fragment thereof according to item 51, wherein the amino acid sequences of the 6 CDRs are the following:

[0938] the VH-CDR1 is selected from the group consisting of SEQ ID NO:18 and SEQ ID NO:19,

[0939] the VH-CDR2 is selected from the group consisting of SEQ ID NO:21 and SEQ ID NO:22,

[0940] the VH-CDR3 is selected from the group consisting of SEQ ID NO:24 and SEQ ID NO:25,

[0941] the VL-CDR1 is selected from the group consisting of SEQ ID NO:27 and SEQ ID NO:28,

[0942] the VL-CDR2 is selected from the group consisting of SEQ ID NO:30 and SEQ ID NO:31, and

[0943] the VL-CDR3 is SEQ ID NO:17.

[0944] 53. The antibody or antigen-binding fragment thereof according to item 51, wherein the amino acid sequences of the 6 CDRs are the following:

[0945] the VH-CDR1 is selected from the group consisting of SEQ ID NO:19 and SEQ ID NO:20; the VH-CDR2 is selected from the group consisting of SEQ ID NO:22, SEQ ID NO:23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:22 and SEQ ID NO:55; the VH-CDR3 is selected from the group consisting of SEQ ID NO:25 and SEQ ID NO:26;

[0946] the VL-CDR1 is selected from the group consisting of SEQ ID NO:28 and SEQ ID NO:29;

[0947] the VL-CDR2 is selected from the group consisting of SEQ ID NO:31 and SEQ ID NO:32; and

[0948] the VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33.

[0949] 54. The antibody or antigen-binding fragment thereof according to item 51 or 52, wherein the amino acid sequences of the 6 CDRs are the following:

[0950] VH-CDR1: EYNIH (SEQ ID NO:18), VH-CDR2: VINPYSENTNYNPKFTD (SEQ ID NO:21), VH-CDR3: DGYFDF (SEQ ID NO:24), VL-CDR1: RSSQIIVHTNGATYLE (SEQ ID NO:27), VL-CDR2: KVSKRFS (SEQ ID NO:30), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0951] 55. The antibody or antigen-binding fragment thereof according to any one of items 51-53, wherein the amino acid sequences of the 6 CDRs are the following: VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: DGYFDY (SEQ ID NO:25), VL-CDR1: RSSQYIVHSNGDTYLE (SEQ ID NO:28), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0952] 56. The antibody or antigen-binding fragment thereof according to item 51, wherein the amino acid sequences of the 6 CDRs are the following:

[0953] VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: GGYFDY (SEQ ID NO:60), VL-CDR1: RSSQYIVHSNGDTYLE (SEQ ID NO:28), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0954] 57. The antibody or antigen-binding fragment thereof according to item 51, wherein the amino acid sequences of the 6 CDRs are the following:

[0955] VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: DGYFDY (SEQ ID NO:25), VL-CDR1: RSSQYIVHSNQDTYLE (SEQ ID NO:62), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17).

[0956] 58. The antibody or antigen-binding fragment thereof according to item 51 or 53, wherein the amino acid sequences of the 6 CDRs are the following:

[0957] VH-CDR1: NYYMS (SEQ ID NO:20), VH-CDR2: AINNEGDSTHYPDTVKG (SEQ ID NO:55), VH-CDR3: AEGSLAY (SEQ ID NO:26), VL-CDR1: KSSQSLLYSNNQKNYLA (SEQ ID NO:29), VL-CDR2: WASTRES (SEQ ID NO:32), VL-CDR3: QQYYTYPLT (SEQ ID NO:33).

[0958] 59. The antibody or antigen-binding fragment thereof according to any one of the preceding items, which is selected from the group consisting of recombinant human antibodies and fragments thereof; humanized antibodies and fragments thereof; and antibodies and fragments thereof that have been mutated to reduce antigenicity thereof in humans.

[0959] 60. The antibody or antigen-binding fragment thereof according to item 59, which is a recombinant human antibody or fragment thereof.

[0960] 61. The antibody or antigen-binding fragment thereof according to item 59, which is a humanized antibody or fragment thereof.

[0961] 62. The antibody or antigen-binding fragment thereof according to item 59, wherein said antibody or fragment thereof has been mutated to reduce antigenicity thereof in humans.

[0962] 63. The antibody or antigen-binding fragment thereof according to any one of items 59-62, which is of IgG class.

[0963] 64. The antibody or antigen-binding fragment thereof according to item 63, wherein said IgG class is selected from the group consisting of IgG1 and IgG4. 65. The antibody or antigen-binding fragment thereof according to item 64, wherein said IgG class is IgG1.

[0964] 66. The antibody or antigen-binding fragment thereof according to item 64, wherein said IgG class is IgG4.

[0965] 67. The antibody or antigen-binding fragment thereof according to any one of items 1-66, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of

[0966] (i) SEQ ID NO:40-44 and 61, such as consisting of SEQ ID NO:40-44; and

[0967] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0968] 68. The antibody or antigen-binding fragment thereof according to item 67, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of

[0969] (i) SEQ ID NO:40, 41 and 61, such as consisting of SEQ ID NO:40 and SEQ ID NO:41; and

[0970] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0971] 69. The antibody or antigen-binding fragment thereof according to item 67, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of

[0972] (i) SEQ ID NO:42-44; and

[0973] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0974] 70. The antibody or antigen-binding fragment thereof according to item 67 or 69, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of

[0975] (i) SEQ ID NO:43 and SEQ ID NO:44; and

[0976] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

[0977] 71. The antibody or antigen-binding fragment thereof according to item 67 or 68, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:40; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:40, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:40.

[0978] 72. The antibody or antigen-binding fragment thereof according to any one of items 67, 68 and 71, wherein said VH region comprises or consists of SEQ ID NO:40.

[0979] 73. The antibody or antigen-binding fragment thereof according to item 67 or 68, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:41; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:41, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:41.

[0980] 74. The antibody or antigen-binding fragment thereof according to any one of items 67, 68 and 73, wherein said VH region comprises or consists of SEQ ID NO:41. 75. The antibody or antigen-binding fragment thereof according to item 67 or 68, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:61; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:61, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:61.

[0981] 76. The antibody or antigen-binding fragment thereof according to any one of items 67, 68 and 75, wherein said VH region comprises or consists of SEQ ID NO:61.

[0982] 77. The antibody or antigen-binding fragment thereof according to item 67 or 69, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:42; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:42, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:42.

[0983] 78. The antibody or antigen-binding fragment thereof according to any one of items 67, 69 and 77, wherein said VH region comprises or consists of SEQ ID NO:42.

[0984] 79. The antibody or antigen-binding fragment thereof according to any one of items 67, 69 and 70, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:43; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:43, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:43.

[0985] 80. The antibody or antigen-binding fragment thereof according to any one of items 67, 69, 70 and 79, wherein said VH region comprises or consists of SEQ ID NO:43.

[0986] 81. The antibody or antigen-binding fragment thereof according to any one of items 67, 69 and 70, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:44; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:44, provided that the sequences of the VH-CDRs are 100% identical to those of SEQ ID NO:44.

[0987] 82. The antibody or antigen-binding fragment thereof according to any one of items 67, 69, 70 and 81, wherein said VH region comprises or consists of SEQ ID NO:44.

[0988] 83. The antibody or antigen-binding fragment thereof according to any one of items 1-82, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of

[0989] (i) SEQ ID NO:45-48 and 63, such as consisting of SEQ ID NO:45-48; and

[0990] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i).

[0991] 84. The antibody or antigen-binding fragment thereof according to item 83, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of

[0992] (i) SEQ ID NO:45, 46 and 63, such as consisting of SEQ ID NO:45 and SEQ ID NO:46; and

[0993] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i).

[0994] 85. The antibody or antigen-binding fragment thereof according to item 83, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of (i) SEQ ID NO:47 and SEQ ID NO:48; and

[0995] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of items 1-58, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i).

[0996] 86. The antibody or antigen-binding fragment thereof according to item 83 or 84, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:45; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:45, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:45.

[0997] 87. The antibody or antigen-binding fragment thereof according to any one of items 83, 84 and 86, wherein said VL region comprises or consists of SEQ ID NO:45.

[0998] 88. The antibody or antigen-binding fragment thereof according to item 83 or 84, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:46; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:46, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:46.

[0999] 89. The antibody or antigen-binding fragment thereof according to any one of items 83, 84 and 88, wherein said VL region comprises or consists of SEQ ID NO:46.

[1000] 90. The antibody or antigen-binding fragment thereof according to item 83 or 84, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:63; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:63, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:63.

[1001] 91. The antibody or antigen-binding fragment thereof according to any one of items 83, 84 and 90, wherein said VL region comprises or consists of SEQ ID NO:63.

[1002] 92. The antibody or antigen-binding fragment thereof according to item 83 or 85, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:47; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:47, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:47.

[1003] 93. The antibody or antigen-binding fragment thereof according to any one of items 83, 85 and 92, wherein said VL region comprises or consists of SEQ ID NO:47.

[1004] 94. The antibody or antigen-binding fragment thereof according to item 83 or 85, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:48; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:48, provided that the sequences of the VL-CDRs are 100% identical to those of SEQ ID NO:48.

[1005] 95. The antibody or antigen-binding fragment thereof according to any one of items 83, 85 and 94, wherein said VL region comprises or consists of SEQ ID NO:48.

[1006] 96. The antibody or antigen-binding fragment thereof according to any one of items 67-95, wherein said VH region is as defined in any one of items 67-82 and said VL region is as defined in any one of items 83-95.

[1007] 97. The antibody or antigen-binding fragment thereof according to item 96, wherein said VH region is as defined in item 68 and said VL region is as defined in item 84. 98. The antibody or antigen-binding fragment thereof according to item 96 or 97, wherein said VH region is as defined in any one of items 71-76 and said VL region is as defined in any one of items 86-91.

[1008] 99. The antibody or antigen-binding fragment thereof according to any one of items 96-98, wherein said VH region is as defined in any one of items 71, 73 and 75 and said VL region is as defined in any one of items 86, 88 and 90.

[1009] 100. The antibody or antigen-binding fragment thereof according to any one of items 96-99, wherein said VH region is as defined in any one of items 72, 74 and 76 and said VL region is as defined in any one of items 87, 89 and 91.

[1010] 101. The antibody or antigen-binding fragment thereof according to any one of items 96-99, wherein said VH region is as defined in item 71 and said VL region is as defined in item 86.

[1011] 102. The antibody or antigen-binding fragment thereof according to any one of items 96-101, wherein said VH region is as defined in item 72 and said VL region is as defined in item 87.

[1012] 103. The antibody or antigen-binding fragment thereof according to any one of items 96-99, wherein said VH region is as defined in item 75 and said VL region is as defined in item 86.

[1013] 104. The antibody or antigen-binding fragment thereof according to any one of items 96-101, wherein said VH region is as defined in item 76 and said VL region is as defined in item 87.

[1014] 105. The antibody or antigen-binding fragment thereof according to any one of items 96-99, wherein said VH region is as defined in item 71 and said VL region is as defined in item 90.

[1015] 106. The antibody or antigen-binding fragment thereof according to any one of items 96-101, wherein said VH region is as defined in item 72 and said VL region is as defined in item 91.

[1016] 107. The antibody or antigen-binding fragment thereof according to any one of items 96-99, wherein said VH region is as defined in item 75 and said VL region is as defined in item 90.

[1017] 108. The antibody or antigen-binding fragment thereof according to any one of items 96-101, wherein said VH region is as defined in item 76 and said VL region is as defined in item 91. 109. The antibody or antigen-binding fragment thereof according to any one of items 96-99, wherein said VH region is as defined in item 73 and said VL region is as defined in item 88.

[1018] 110. The antibody or antigen-binding fragment thereof according to any one of items 96-99 and 109, wherein said VH region is as defined in item 74 and said VL region is as defined in item 89.

[1019] 111. The antibody or antigen-binding fragment thereof according to item 96, wherein said VH region is as defined in item 69 or 70 and said VL region is as defined in item 85.

[1020] 112. The antibody or antigen-binding fragment thereof according to item 111, wherein said VH region is as defined in any one of items 77-82 and said VL region is as defined in any one of items 92-95.

[1021] 113. The antibody or antigen-binding fragment thereof according to item 111 or 112, wherein said VH region is as defined in any one of items 77-80 and said VL region is as defined in any one of items 92 and 93.

[1022] 114. The antibody or antigen-binding fragment thereof according to item 111 or 112, wherein said VH region is as defined in any one of items 77-82 and said VL region is as defined in any one of items 94 and 95.

[1023] 115. The antibody or antigen-binding fragment thereof according to item 111 or 112, wherein said VH region is as defined in any one of items 77, 79 and 81 and said VL region is as defined in any one of items 92 and 94.

[1024] 116. The antibody or antigen-binding fragment thereof according to any one of items 111, 112 and 115, wherein said VH region is as defined in any one of items 78, 80 and 82 and said VL region is as defined in any one of items 93 and 95.

[1025] 117. The antibody or antigen-binding fragment thereof according to any one of items 111-113 and 115, wherein said VH region is as defined in any one of items 77, 79 and 81 and said VL region is as defined in item 92.

[1026] 118. The antibody or antigen-binding fragment thereof according to any one of items 111, 112, 114 and 115, wherein said VH region is as defined in any one of items 77, 79 and 81 and said VL region is as defined in item 94.

[1027] 119. The antibody or antigen-binding fragment thereof according to any one of items 111-113 and 115-117, wherein said VH region is as defined in any one of items 78, 80 and 82 and said VL region is as defined in item 93. 120. The antibody or antigen-binding fragment thereof according to any one of items 111, 112, 114-116 and 118, wherein said VH region is as defined in any one of items 78, 80 and 82 and said VL region is as defined in item 95.

[1028] 121. The antibody or antigen-binding fragment thereof according to any one of items 111-113, 115 and 117, wherein said VH region is as defined in item 77 or 78 and said VL region is as defined in item 92 or 93.

[1029] 122. The antibody or antigen-binding fragment thereof according to any one of items 111-113, 115, 117 and 121, wherein said VH region is as defined in item 77 and said VL region is as defined in item 92.

[1030] 123. The antibody or antigen-binding fragment thereof according to item 111-113, 115, 117 and 121, wherein said VH region is as defined in item 78 and said VL region is as defined in item 93.

[1031] 124. The antibody or antigen-binding fragment thereof according to item 111-113, 115, 117 and 121, wherein said VH region is as defined in item 70 and said VL region is as defined in item 85.

[1032] 125. The antibody or antigen-binding fragment thereof according to item 111 or 124, wherein said VH region is as defined in any one of items 79-82 and said VL region is as defined in any one of items 92-95.

[1033] 126. The antibody or antigen-binding fragment thereof according to any one of items 111, 124 and 125, wherein said VH region is as defined in any one of items 79-82 and said VL region is as defined in item 92 or 93.

[1034] 127. The antibody or antigen-binding fragment thereof according to any one of items 111 and 124-126, wherein said VH region is as defined in item 79 or 81 and said VL region is as defined in item 92.

[1035] 128. The antibody or antigen-binding fragment thereof according to any one of items 111 and 124-127, wherein said VH region is as defined in item 80 or 82 and said VL region is as defined in item 93.

[1036] 129. The antibody or antigen-binding fragment thereof according to any one of items 111, 124 and 125, wherein said VH region is as defined in any one of items 79-82 and said VL region is as defined in item 94 or 95.

[1037] 130. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125 and 129, wherein said VH region is as defined in item 79 or 81 and said VL region is as defined in item 94. 131. The antibody or antigen-binding fragment thereof according to any one of items 111, 122, 125, 129 and 130, wherein said VH region is as defined in item 80 or 81 and said VL region is as defined in item 95.

[1038] 132. The antibody or antigen-binding fragment thereof according to any one of items 111, 124 and 125, wherein said VH region is as defined in item 79 or 80 and said VL region is as defined in any one of items 92-95.

[1039] 133. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125 and 132, wherein said VH region is as defined in item 79 and said VL region is as defined in item 92 or 94.

[1040] 134. The antibody or antigen-binding fragment thereof according to any one of items 111, 124-127, 132 and 133, wherein said VH region is as defined in item 79 and said VL region is as defined in item 92.

[1041] 135. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125, 129, 130, 132 and 133, wherein said VH region is as defined in item 79 and said VL region is as defined in item 94.

[1042] 136. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125, 132 and 133, wherein said VH region is as defined in item 80 and said VL region is as defined in item 93 or 95.

[1043] 137. The antibody or antigen-binding fragment thereof according to any one of items 111, 124-128, 132-134 and 136, wherein said VH region is as defined in item 80 and said VL region is as defined in item 93.

[1044] 138. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125, 129-133, 135 and 136, wherein said VH region is as defined in item 80 and said VL region is as defined in item 95.

[1045] 139. The antibody or antigen-binding fragment thereof according to any one of items 111, 124 and 125, wherein said VH region is as defined in item 81 or 82 and said VL region is as defined in any one of items 92-95.

[1046] 140. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125 and 139, wherein said VH region is as defined in item 81 and said VL region is as defined in item 92 or 94.

[1047] 141. The antibody or antigen-binding fragment thereof according to any one of items 111, 124-127, 139 and 140, wherein said VH region is as defined in item 81 and said VL region is as defined in item 92. 142. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125, 129, 130, 139 and 140, wherein said VH region is as defined in item 81 and said VL region is as defined in item 94.

[1048] 143. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125, 139 and 140, wherein said VH region is as defined in item 82 and said VL region is as defined in item 93 or 95.

[1049] 144. The antibody or antigen-binding fragment thereof according to any one of items 111, 124-128, 139-141 and 143, wherein said VH region is as defined in item 82 and said VL region is as defined in item 93.

[1050] 145. The antibody or antigen-binding fragment thereof according to any one of items 111, 124, 125, 129-131, 139, 140, 142 and 143, wherein said VH region is as defined in item 82 and said VL region is as defined in item 95.

[1051] 146. The antibody or antigen-binding fragment thereof according to any one of the preceding items further comprising a heavy chain constant (CH) region.

[1052] 147. The antibody or antigen-binding fragment thereof according to item 146, wherein said CH region comprises or consists of an amino acid sequence selected from the group consisting of

[1053] (i) SEQ ID NO:49 and SEQ ID NO:50; and

[1054] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i).

[1055] 148. The antibody or antigen-binding fragment thereof according to item 146 or 147, wherein said CH region comprises or consists of SEQ ID NO:49 or SEQ ID NO:50.

[1056] 149. The antibody or antigen-binding fragment thereof according to item 146 or 147, wherein said CH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:49; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:49.

[1057] 150. The antibody or antigen-binding fragment thereof according to any one of items 146-149, wherein said CH region comprises or consists of SEQ ID NO:49. 151. The antibody or antigen-binding fragment thereof according to item 146 or 147, wherein said CH region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:50; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:50.

[1058] 152. The antibody or antigen-binding fragment thereof according to any one of items 146-147 and 151, wherein said CH region comprises or consists of SEQ ID NO:50.

[1059] 153. The antibody or antigen-binding fragment thereof according to any one of the preceding items further comprising a light chain constant (CL) region.

[1060] 154. The antibody or antigen-binding fragment thereof according to item 153, wherein said CL region comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NO:51; and amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to SEQ ID NO:51.

[1061] 155. The antibody or antigen-binding fragment thereof according to item 153 or 154, wherein said CL region comprises or consists of SEQ ID NO:51.

[1062] 156. The antibody or antigen-binding fragment thereof according to any one of items 146-155, wherein the antibody or antigen-binding fragment thereof comprises a CH region as defined in any one of items 146-152 and a CL region as defined in any one of items 153-155.

[1063] 157. The antibody or antigen-binding fragment thereof according to item 156, wherein the antibody or antigen-binding fragment thereof comprises a CH region as defined in any one of items 147-152 and a CL region as defined in any one of items 154-155, respectively.

[1064] 158. The antibody or antigen-binding fragment thereof according to item 156 or 157, wherein the CH region is as defined in item 149 and the CL region is as defined in item 154.

[1065] 159. The antibody or antigen-binding fragment thereof according to item 158, wherein the CH region comprises or consists of SEQ ID NO:49 and CL region comprises or consists of SEQ ID NO:51. 160. The antibody or antigen-binding fragment thereof according to item 156 or 157, wherein the CH region is as defined in item 151 and the CL region is as defined in item 154.

[1066] 161. The antibody or antigen-binding fragment thereof according to item 160, wherein CH region comprises or consists of SEQ ID NO:50 and CL region comprises or consists of SEQ ID NO:51.

[1067] 162. The antibody or antigen-binding fragment thereof according to any one of the preceding items, wherein said antigen-binding surface provides the antibody or antigen-binding fragment thereof with the capacity to bind selectively to an epitope of TDP-43.

[1068] 163. The antibody or antigen-binding fragment thereof according to item 162, wherein said epitope comprises or consists of SEQ ID NO:52.

[1069] 164. The antibody or antigen-binding fragment thereof according to item 162 or 163, wherein said TDP-43 comprises or consists of an amino acid sequence selected from the group consisting of

[1070] (i) SEQ ID NO:53; and

[1071] (ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99% identity to SEQ ID NO:53, provided that said amino acid sequence comprises SEQ ID NO:52.

[1072] 165. The antibody or antigen-binding fragment thereof according to any one of items 162-164, wherein said TDP-43 comprises or consists of SEQ ID NO:53.

[1073] 166. The antibody or antigen-binding fragment thereof according to any one of items 162-165, wherein said TDP-43 is pathological TDP-43.

[1074] 167. The antibody or antigen-binding fragment thereof according to any one of the preceding items that has a higher binding affinity for pathological TDP-43 than for physiological TDP-43 and / or is capable of selective binding of pathological TDP-43 over physiological TDP-43.

[1075] 168. The antibody or antigen-binding fragment thereof according to item 166 or 167, wherein said pathological TDP-43 is a TDP-43 aggregate, such as a non-reversible TDP-43 aggregate. 169. The antibody or antigen-binding fragment thereof according to item 168, wherein the molecular weight of said TDP-43 aggregate is higher than the molecular weight of a TDP-43 monomer, wherein said molecular weight of said TDP-43 aggregate is at least about 2x higher, such as at least about 3x higher, such as at least about 4x higher, such as at least about 5x higher, such as at least about 6x higher, such as at least about 7x higher, such as at least about 8x higher, such as at least about 9x higher, such as at least about 10x higher, such as at least about 20x higher, such as at least about 30x higher, such as at least about 40x higher, such as at least about 50x higher, such as at least about 100x higher when compared to the molecular weight of said TPD-43 monomer, optionally as determined by SDS-PAGE and Western blot, wherein said TPD-43 monomer has a molecular weight of about 49 kDa.

[1076] 170. The antibody or antigen-binding fragment thereof according to item 168 or 169, wherein the size of said TDP-43 aggregate is greater than the size of a TDP-43 monomer, wherein said size of said TDP-43 aggregate is at least about 2x greater, such as at least about 3x greater, such as at least about 4x greater, such as at least about 5x greater, such as at least about 6x greater, such as at least about 7x greater, such as at least about 8x greater, such as at least about 9x greater, such as at least about 10x greater when compared to the size of said TPD-43 monomer, optionally as determined by HPLC and / or transmission electron microscopy (TEM).

[1077] 171. The antibody or antigen-binding fragment thereof according to any one of items 167-170 that has at least 2x higher binding affinity for pathological TDP-43 than for physiological TDP-43, such as at least about 10x higher, such as at least about 20x higher, such as at least about 30x higher, such as at least about 40x higher, such as at least about 50x higher, such as at least about 60x higher, such as at least about 70x higher, such as at least about 80x higher, such as at least about 90x higher, such as at least about 100x higher, such as at least about 200x higher, such as at least about 300x higher, such as at least about 400x higher, such as at least about 500x higher, such as at least about 600x higher, such as at least about 700x higher, such as at least about 800x higher, such as at least about 900x higher, such as at least about lOOOx higher, such as at least about 2000x higher, such as at least about 3000x higher, such as at least about 4000x higher, such as at least about 5000x higher, such as at least about 6000x higher, such as at least about 7000x higher, such as at least about 8000x higher, such as at least about 8500x higher binding affinity.

[1078] 172. The antibody or antigen-binding fragment thereof according to any one of items 166-171 that has essentially no binding affinity for physiological TDP-43, for example TDP-43 monomers, non-disease associated TDP-43 oligomers and / or nondisease associated reversible TDP-43 aggregates.

[1079] 173. The antibody or antigen-binding fragment thereof according to any one of items 167-172, wherein said binding affinity is determined by surface plasmon resonance.

[1080] 174. The antibody or antigen-binding fragment thereof according to any one of items 166-173 that has a binding affinity for pathological TDP-43 that corresponds to a KDvalue of at most about 1 nM, such as at most about 900 pM, such as at most about 800 pM, such as at most about 700 pM, such as at most about 600 pM, such as at most about 500 pM, such as at most about 400 pM, such as at most about 300 pM, such as at most about 200 pM, such as at most about 190 pM, such as at most about 180 pM, such as at most about 170 pM, such as at most about 160 pM, such as at most about 150 pM, such as at most about 140 pM, such as at most about 130 pM, such as at most about 120 pM, such as at most about 110 pM, such as at most about 100, such as at most about 90 pM, such as at most about 80 pM, such as at most about 70 pM, such as at most about 63 pM;

[1081] or such as a KDvalue of from about 1 pM to about 200 pM, such as from about 10 pM to about 190 pM, such as from about 20 pM to about 180 pM, such as from about 30 pM to about 170 pM, such as from about 40 pM to about 160 pM, such as from about 50 pM to about 150 pM, such as from about 60 pM to about 140 pM, such as from about 63 pM to about 140 pM.

[1082] 175. The antibody or antigen-binding fragment thereof according to any one of the preceding items that has a lower half maximal effective concentration (EC50) for binding pathological TDP-43 than for binding physiological TDP-43.

[1083] 176. The antibody or antigen-binding fragment thereof according to item 175 that has at least about 1.5x lower half maximal effective concentration (EC50) for binding pathological TDP-43 than for binding physiological TDP-43, such as at least about 1.6x lower, such as at least about 1.7x lower, such as at least about 1.8x lower, such as at least about 1.9x lower, such as at least about 2x lower, such as at least about 3x lower, such as at least about 4x lower, such as at least about 5x lower, such as at least about 6x lower, such as at least about 7x lower, such as at least about 8x lower, such as at least about 9x lower, such as at least about 10x lower, such as at least about 11x lower half maximal effective concentration (EC50).

[1084] 177. The antibody or antigen-binding fragment thereof according to item 175 or 176, wherein said half maximal effective concentration (EC50) is determined by ELISA, such as indirect ELISA.

[1085] 178. The antibody or antigen-binding fragment thereof according to any one of items 166-177 that has a half maximal effective concentration (EC50) for binding pathological TDP-43 that corresponds to an EC50value of at most about 20 ng / ml, such as at most about 19 ng / ml, such as at most about 18 ng / ml, such as at most about 17 ng / ml, such as at most about 16 ng / ml, such as at most about 15 ng / ml, such as at most about 14 ng / ml, such as at most about 13 ng / ml, such as at most about 12 ng / ml, such as at most about 11 ng / ml, such as at most about 10 ng / ml, such as at most about 9 ng / ml, such as at most about 8 ng / ml, such as at most about 7.5 ng / ml, such as at most about 7 ng / ml, such as at most about 6 ng / ml, such as at most about 5 ng / ml, such as at most about 4 ng / ml, such as at most about 3 ng / ml, optionally as determined by ELISA, such as indirect ELISA;

[1086] such as an EC₅₀ value of from 1 ng / ml to about 20 ng / ml, such as from about 1 ng / ml to about 15 ng / ml, such as from about 1 ng / ml to about 14 ng / ml, such as from about 1 ng / ml to about 13 ng / ml, such as from about 1 ng / ml to about 12 ng / ml, such as from about 1 ng / ml to about 11 ng / ml, such from about 1 ng / ml to about 12 ng / ml, such as from about 1 ng / ml to about 10 ng / ml, such as from about 1 ng / ml to about 9 ng / ml, such as from about 1 ng / ml to about 8 ng / ml, such as from about 2 ng / ml to about 8 ng / ml, such as from about 2.5 ng / ml to about 8 ng / ml, such as from about 3 ng / ml to about 7.5 ng / ml, optionally as determined by ELISA, such as indirect ELISA.

[1087] 179. The antibody or antigen-binding fragment thereof according to any one of items 166-178 that has a half maximal effective concentration (EC50) for binding physiological TDP-43 that corresponds to an EC50 value of at least about 4 ng / ml, such as at least about 5 ng / ml, such as at least about 6 ng / ml, such as at least about 7 ng / ml, such as at least about 8 ng / ml, such as at least about 9 ng / ml, such as at least about 10 ng / ml, such as at least about 20 ng / ml, such as at least about 25 ng / ml, such as at least about 30 ng / ml, such as at least about 35 ng / ml, such as at least about 40 ng / ml, such as at least about 45 ng / ml, such as at least about 50 ng / ml, such as at least about 55 ng / ml, such as at least about 57 ng / ml, such as at least about 60 ng / ml, optionally as determined by ELISA, such as indirect ELISA.

[1088] 180. The antibody or antigen-binding fragment thereof according to any one of items 166-179 that has a lower half maximal inhibitory concentration (IC50) for binding pathological TDP-43 than for binding physiological TDP-43.

[1089] 181. The antibody or antigen-binding fragment thereof according to item 180 that has at least about 50x lower half maximal inhibitory concentration (IC50) for binding pathological TDP-43 than for binding physiological TDP-43, such at least about 100x lower, such as at least about 150x lower, such as at least about 200x lower, such as at least about 250x lower, such as at least about 300x lower, such as at least about 310x lower, such at least about 314x lower, such as at least about 315x, such at least about 350x lower, such at least about 400x lower, such at least about 450x lower, such at least about 500x lower, such at least about 550x lower, such at least about 580x lower, such at least about 583x lower, such at least about 585x lower, such at least about 600x lower, such at least about 630x lower, such at least about 635x lower, such at least about 636x lower, such at least about 640x lower, such at least about 650x lower half maximal inhibitory concentration (IC50).

[1090] 182. The antibody or antigen-binding fragment thereof according to item 180 or 181, wherein said half maximal inhibitory concentration (IC50) is determined by ELISA, such as by inhibition ELISA.

[1091] 183. The antibody or antigen-binding fragment thereof according to any one of items 166-182 that has a half maximal inhibitory concentration (IC50) for binding pathological TDP-43 that corresponds to an IC50value of at most about 10 ng / ml, such as at most about 9 ng / ml, such as at most about 8 ng / ml, such as at most about 7 ng / ml, such as at most about 6 ng / ml, such as at most about 5 ng / ml, such as at most about 4 ng / ml, such as at most about 3 ng / ml, such as at most about 2 ng / ml, such as at most about 1.9 ng / ml, such as at most about 1.8 ng / ml, such as at most about 1.7 ng / ml, such as at most about 1.6 ng / ml, such as at most about 1.5 ng / ml, such as at most about 1.4 ng / ml, such as at most about 1.3 ng / ml, such as at most about 1.2 ng / ml, such as at most about 1.1 ng / ml, such as at most about 1 ng / ml, such as at most about 0.9 ng / ml, optionally as d...

Claims

CLAIMS1. An antibody or antigen-binding fragment thereof, comprising an immunoglobulin heavy chain variable (VH) region and an immunoglobulin light chain variable (VL) region, said VH and VL regions forming a VH / VL pair comprising an antigen-binding surface, wherein said antigen-binding surface is composed of three complementarity-determining regions (CDRs) from said VH region and three CDRs from said VL region, and wherein said CDRs consist of the following amino acid sequences:VH-CDR1: X1YX2X3X4 (SEQ ID NO:1), whereinXI is selected from E, D and N,X2 is selected from N and Y,X3 is selected from M and I,X4 is selected from S and H;VH-CDR2: X5IX6X7X8X9X10X11TX12YX13X14X15X16X17X18 (SEQ ID NO:2), whereinX5 is selected from V and A,X6 is selected from N and S,X7 is selected from P, T and N,X8 is selected from Y, E and N, such as from Y and E,X9 is selected from S and G,X10 is selected from E, G and D,XII is selected from N and S,X12 is selected from N and H,X13 is selected from N and P,X14 is selected from P, Q and D,X15 is selected from K and T,X16 is selected from F and V,X17 is selected from K and T,X18 is selected from D and G;VH-CDR3: X19X20GX21X22X23X24 (SEQ ID NO:3), whereinX19 is A or missing,X20 is selected from D, E and G, such as from D and E,X21 is selected from Y and S,X22 is selected from F and L,X23 is selected from D and A,X24 is selected from F and Y;VL-CDR1: X25SSQX26X27X28X29X30NX31X32X33X34YLX35 (SEQ ID NO:4), whereinX25 is selected from R and K,X26 is selected from I, Y and S,X27 is selected from I and L,X28 is selected from V and L,X29 is selected from H and Y,X30 is selected from S and T,X31 is selected from G, N and Q, such as from G and N, X32 is Qor missing,X33 is selected from A, D and K,X34 is selected from N and T,X35 is selected from E and A;VL-CDR2: X36X37SX38RX39S (SEQ ID NO:5), whereinX36 is selected from K and W,X37 is selected from V and A,X38 is selected from K, N and T,X39 is selected from F and E;VL-CDR3: X40QX41X42X43X44PX45T (SEQ ID NO:6), whereinX40 is selected from Q and F,X41 is selected from G and Y,X42 is selected from Y and S,X43 is selected from H and T,X44 is selected from V and Y,X45 is selected from L and P.

2. The antibody or antigen-binding fragment thereof according claim 1, wherein the amino acid sequences of the 6 CDRs are the following:the VH-CDR1 is selected from the group consisting of SEQ ID NQ:18-20;the VH-CDR2 is selected from the group consisting of SEQ ID NO:21-23 and SEQ ID NO:55, such as the group consisting of SEQ ID NO:21-22 and SEQ ID NO:55; the VH-CDR3 is selected from the group consisting of SEQ ID NO:24-26 and 60, such as the group consisting of SEQ ID NO:24-26 or the group consisting of SEQ ID NO:25 and SEQ ID NO:60;the VL-CDR1 is selected from the group consisting of SEQ ID NO:27-29 and 62, such as the group consisting of SEQ ID NO:27-29 or the group consisting of SEQ ID NO:28 and SEQ ID NO:62;the VL-CDR2 is selected from the group consisting of SEQ ID NO:30-32; and the VL-CDR3 is selected from the group consisting of SEQ ID NO:17 and SEQ ID NO:33.

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the amino acid sequences of the 6 CDRs are the following:VH-CDR1: EYNIH (SEQ ID NO:18), VH-CDR2: VINPYSENTNYNPKFTD (SEQ ID NO:21), VH-CDR3: DGYFDF (SEQ ID NO:24), VL-CDR1: RSSQIIVHTNGATYLE (SEQ ID NO:27), VL-CDR2: KVSKRFS (SEQ ID NO:30), VL-CDR3: FQGSHVPPT (SEQ ID NO:17); or wherein the amino acid sequences of the 6 CDRs are the following:VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: DGYFDY (SEQ ID NO:25), VL-CDR1: RSSQYIVHSNGDTYLE (SEQ ID NO:28), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17); or wherein the amino acid sequences of the 6 CDRs are the following: VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22),VH-CDR3: GGYFDY (SEQ ID NO:60), VL-CDR1: RSSQYIVHSNGDTYLE (SEQ ID NO:28), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17); or wherein the amino acid sequences of the 6 CDRs are the following:VH-CDR1: DYNMH (SEQ ID NO:19), VH-CDR2: VISTYSGNTNYNQKFKD (SEQ ID NO:22), VH-CDR3: DGYFDY (SEQ ID NO:25), VL-CDR1: RSSQYIVHSNQDTYLE (SEQ ID NO:62), VL-CDR2: KVSNRFS (SEQ ID NO:31), VL-CDR3: FQGSHVPPT (SEQ ID NO:17); or wherein the amino acid sequences of the 6 CDRs are the following:VH-CDR1: NYYMS (SEQ ID NO:20), VH-CDR2: AINNEGDSTHYPDTVKG (SEQ ID NO:55), VH-CDR3: AEGSLAY (SEQ ID NO:26), VL-CDR1: KSSQSLLYSNNQKNYLA (SEQ ID NO:29), VL-CDR2: WASTRES (SEQ ID NO:32), VL-CDR3: QQYYTYPLT (SEQ ID NO:33).

4. The antibody or antigen-binding fragment thereof according to any one of claims 1-3, wherein said VH region comprises or consists of an amino acid sequence selected from the group consisting of(i) SEQ ID NO:40-44 and 61, such as consisting of SEQ ID NO:40-44; and(ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VH-CDRs are as defined in any one of claims 1-3, such as wherein the sequences of the VH-CDRs are 100% identical to those of the sequence defined in (i).

5. The antibody or antigen-binding fragment thereof according to any one of claims 1-4, wherein said VL region comprises or consists of an amino acid sequence selected from the group consisting of(i) SEQ ID NO:45-48 and 63, such as consisting of SEQ ID NO:45-48; and(ii) amino acid sequences having at least 70%, such as at least 80%, such as at least 90%, such as at least 92%, such as at least 94%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, such as at least 99%, identity to a sequence defined in (i), provided that the sequences of the VL-CDRs are as defined in any one of claims 1-3, such as wherein the sequences of the VL-CDRs are 100% identical to those of the sequence defined in (i).

6. The antibody or antigen-binding fragment thereof according to claims 4 and 5, wherein said VH region is as defined in claim 4 and said VL region is as defined in claim 5.

7. The antibody or antigen-binding fragment thereof according to any one of claims 4-6, wherein said VH region comprises the amino acid sequence SEQ ID NO:40 and said VL region comprises the amino acid sequence SEQ ID NO:45; or wherein said VH region comprises the amino acid sequence SEQ ID NO:40 and said VL region comprises the amino acid sequence SEQ ID NO:63; or wherein said VH region comprises the amino acid sequence SEQ ID NO:61 and said VL region comprises the amino acid sequence SEQ ID NO:45; or wherein said VH region comprises the amino acid sequence SEQ ID NO:61 and said VL region comprises the amino acid sequence SEQ ID NO:63;or wherein said VH region comprises the amino acid sequence SEQ ID NO:41 and said VL region comprises the amino acid sequence SEQ ID NO:46; or wherein said VH region comprises the amino acid sequence SEQ ID NO:42 and said VL region comprises the amino acid sequence SEQ ID NO:47; or wherein said VH region comprises the amino acid sequence SEQ ID NO:43 and said VL region comprises the amino acid sequence SEQ ID NO:48; or wherein said VH region comprises the amino acid sequence SEQ ID NO:43 and said VL region comprises the amino acid sequence SEQ ID NO:47; or wherein said VH region comprises the amino acid sequence SEQ ID NO:44 and said VL region comprises the amino acid sequence SEQ ID NO:48; or wherein said VH region comprises the amino acid sequence SEQ ID NO:44 and said VL region comprises the amino acid sequence SEQ ID NO:47.

8. The antibody or antigen-binding fragment thereof according to any one of claims 1-7, wherein said antigen-binding surface provides the antibody or antigenbinding fragment thereof with the capacity to bind selectively to an epitope of TDP-43, wherein said epitope comprises or consists of SEQ ID NO:52.

9. The antibody or antigen-binding fragment thereof according to any one of claims 1-8, wherein the antibody or antigen-binding fragment thereof has a higher binding affinity for pathological TDP-43 than for physiological TDP-43 and / or is capable of selective binding of pathological TDP-43 over physiological TDP-43.

10. The antibody or antigen-binding fragment thereof according to claim 9, wherein said pathological TDP-43 is a TDP-43 aggregate, such as a non-reversible TDP-43 aggregate.

11. A pharmaceutical composition comprising the antibody or antigenbinding fragment thereof as defined in any one of claims 1-10 and a pharmaceutically acceptable carrier and / or excipient.

12. An antibody or antigen-binding fragment thereof as defined in any one of claims 1-10 or the pharmaceutical composition as defined in claim 11 for use as a medicament.

13. A antibody or antigen-binding fragment thereof as defined in any one of claims 1-10 or the pharmaceutical composition as defined in claim 11 for use in therapeutic treatment or for use in prophylactic treatment with respect to a neurodegenerative disorder associated with a TDP-43 proteopathy, such as a disorder associated with non-reversible aggregation of TDP-43.

14. The antibody or antigen-binding fragment thereof or the pharmaceutical composition for use according to claim 13, wherein said disorder is selected from the group consisting of amyotrophic lateral sclerosis (ALS); Alzheimer's disease (AD), optionally wherein said AD is selected from the group consisting of familial AD andsporadic AD; limbic-predominant age-related TDP-43 encephalopathy (LATE); and frontotemporal dementia (FTD).

15. A method of detecting pathological TDP-43, such as non-reversible TDP-43 aggregates, said method comprising the steps ofproviding a sample suspected to contain pathological TDP-43, such as non-reversible TDP-43 aggregates,contacting said sample with the antibody or antigen-binding fragment thereof as defined in any one of claims 1-10 or the pharmaceutical composition as defined in claim 11, anddetecting the binding of said antibody or antigen-binding fragment thereof to indicate the presence of pathological TDP-43, such as non-reversible TDP-43 aggregates, in said sample.

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