AN ISOLATED TAU-BINDING ANTIBODY OR ITS BINDING FRAGMENT, A METHOD FOR ITS PRODUCTION, AN ISOLATED NUCLEIC ACID MOLECULE, AN ISOLATED CLONING OR EXPRESSION VECTOR, AND AN ISOLATED HOST CELL

Abstract

This refers to Tau-binding antibodies and their binding fragments. DNA molecules, host cell, and vector.

Description

DESCRIPTIVE MEMORY OF THE INVENTION PATENT EUROPE APPLICATION PRIORITY 15175519.6 ON: Tau-binding antibodies and their binding fragments REQUESTED BY: UCB Biopharma SPRL RESIDING IN: BELGIUM FOR THE TERM OF.....TWENTY.......YEARS The present invention relates, inter alia, to therapeutic and diagnostic antibodies binding to Tau and its binding fragments, methods for making said antibodies and their use for the treatment and / or diagnosis of tauopathies, such as Alzheimer's disease ; amyotrophic lateral sclerosis / dementia parkinsonism complex; argyrophilic granular disease (argyrophilia); chronic traumatic encephalopathy; corticobasal degeneration; diffuse neurofibrillary tangles with calcification; Down's Syndrome; familial dementia i British; danish familial dementia; frontotemporal dementia and parkinsonism associated with chromosome 17 caused by mutations in MAPT; Gerstmann Straussler-Scheinker disease; parkinsonism in Guadeloupe; myotonic dystrophy; neurodegeneration with brain accumulation of iron; Niemann-Pick disease, type C; non-Guámaniana motor neuron disease with neurofibrillary tangles; Pick's disease; post-encephalitic parkinsonism; cerebral amyloid angiopathy caused by prions; progressive subcortical gliosis; progressive supranuclear palsy; SLC9A6-related mental retardation; subacute sclerosing panencephalitis; tangie-only dementia; white matter tauopathy with globular glial inclusions (Clavaguera et al. Brain Pathology 23 (2013) 342349). The present invention also relates to methods for treating a human subject suffering from or suspected of being susceptible to the above-described taupathies, in particular, taupathies such as Alzheimer's disease and progressive supranuclear palsy. Alzheimer's disease (AD) and Progressive Supranuclear Palsy (PSP) are neurodegenerative diseases with high medical care needs, high cost for the health systems of societies, as well as a great burden for the affected families. The clinical signs of i AD include loss of memory, cognition, reasoning, judgment, and emotional stability, and eventually death. PSP includes severe and progressive problems with gait control and balance, falls, vertical eye movement disorders, cognitive problems, depression, apathy, and mild dementia. : Late symptoms include blurred vision, uncontrollable eye movement, slurred speech, difficulty swallowing, and death. For more than a decade, AD disease modification programs have targeted beta-amyloid peptide through various mechanisms. In contrast, much less progress has been made with a focus on intracellular Tau pathology, the second main hallmark of AD. Neurofibrillary tangles or inclusions containing aggregated hyperphosphorylated Tau define the hallmarks of AD pathology and a number of other tauopathies, including PSP. In these diseases there is a strong correlation between symptomatic progression and the level and distribution of intraneural Tau aggregates. In AD, Tau neuronal tangles appear first in the transentorhinal cortex, from where they spread to the hippocampus and neocortex. The tangles seen in AD neurons consist of aggregated insoluble hyperphosphorylated Tau. Direct toxic effects of pathological Tau species and / or loss of axonal transport due to uptake of functional Tau in the hyperphosphorylated and aggregated forms, which no longer have the ability to support axonal transport, have been proposed to constitute an i contribution to disease. In its non-pathological state, Tau is a highly soluble cytoplasmic microtubule-binding protein, which is produced in the human central nervous system (CNS) in 6 alternative major isoforms, comprised of amino acids ranging from 352 to 441. at length. These isoforms may have zero, one, or two N-terminal inserts (ON, due to splicing). IN, 2N), and three or four repeat sequences with C-terminals (3R or 4R). These C-terminal repeat sequences of amino acids 30-32, Rl, R2, R3, and R4, together constitute the Microtubule-Binding Tau Region (MTBR). Indeed, the main function of Tau is believed to be in the assembly and stabilization of axonal microtubules. Microtubules form tracks for axonal transport and cytoskeletal elements for cell growth (Clavaguera et al., Brain Pathology 23 (2013) 342-349). Three j Tau isoforms have been shown to contain three microtubule-binding regions (iMTBR): isoform 4, also referred to as 3R0N, NCBI Reference Sequence l(Wational Center for i Biotechnology Information, National Center for Biotechnology Information) NP_058525.1 (amino acid 352), isoform 7, also referred to as 3R1N, NCBI Sequence Reference NP 001190180.1 (amino acid 381) isoform 8, also referred to as 3R2N, NCBI Sequence Reference NP_0()1190181.1 (amino acid 410). While the other three Tau isoforms contain four MTBR: ! isoform 2, also referred to as 4R2N, NCBI Reference Sequence NP¡_005901.2 (amino acid 441), isoform 3, also designated as 4R0N, NCBI Reference Sequence NP;058518.1 (amino acid 383), and I isoform 5, also designated as 4R1N, NCBI Reference Sequence NCBI NP_001116539.1 (amino acid 412). Tau contains 85 possible phosphorylation sites on serine (S), threonine (T), and tyrosine (Y). Many of the phosphorylated residues on Tau are found in the proline-rich domain of Tau, flanking the microtubule-binding domain. All six Tau isoforms are present in the mature normal human brain, and at this step Tau phosphorylation is relatively reduced (Noble et al. Yo I al., 2013 Front Neurol. 2013; 4: 83). In the various taupathies, the Tau deposited in pathological lesions is invariably highly phosphorylated. Phosphoserine 202 and phosphothreonine 205 have been detected in aggregated Tau from brain samples and cerebrospinal fluid from PSP and AD patients (Buée et al., Brain Research Reviews 33 (2000) 95-130; Wray et al. J. Neurochem. 2008 Jun 1;105(6):2343-52,- Hanger et al., J. Biol Chem. 2007 Aug 10; 282(32):23645-54,- Maccioni et 2006 Feb; 27(2):237-44). al., Neurobiol Aging. Only symptomatic treatments are currently available for these diseases with little or no efficacy. There is no treatment currently available to slow or ideally stop the development of the disease. Therefore, there remains a need in the art for new compounds. I and compositions useful in the treatment of tauopathies. It is an object of the present invention, inter alia, to provide agents for treating or diagnosing tauopathies such as Alzheimer's disease (AD) or progressive supranuclear palsy (PSP). Furthermore] it is an object of the present invention to provide, inter alia, methods for treating or diagnosing tauopathies, such as Alzheimer's disease (AD) or progressive supranuclear palsy (PSP). These and other objects, as will become apparent from the following description, are achieved by the subject matter of the independent claims. Some of the specific aspects and embodiments thereof contemplated by the present description form the subject matter of the dependent claims. Still other aspects and embodiments thereof contemplated by the present description can be taken from the description below. In a first aspect, the present description provides an isolated antibody binding to Tau or its fragment i > linker, wherein said Tau-binding antibody or linker fragment thereof comprises the following: a light chain variable region comprising a CDR (Complementarity Determinin^ Region, complementarity determining region) 1 selected from SEQ ID No. (Sequence Identification Number): 1 or sequences at least % identical to she, a CDR2 selected from SEQ ID Yo No.: 2 or sequences at least 90% identical to it, and one CDR3 selected from SEQ ID No.: 3 or sequences at least 90% identical to it; and / or a heavy chain variable region comprising a CDR1 selected from SEQ ID No.: (4 or sequences at least 90% identical thereto, a CDR2 selected from i SEQ ID No.: 5 or at least 90% identical sequences thereto, and one CDR3 selected from SEQ ID No.: 6 or at least 90% identical sequences^ thereto. Yo In a second aspect, the present description provides I an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof comprises the following: a light chain variable region comprising SEQ ID No.: 7 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising the • _ I SEQ ID No.: 8 or sequences at least 80% identical to it. In a third aspect, the present disclosure provides an isolated !-binding antibody. Tau or its linking fragment, wherein said Tau-binding antibody or its linking fragment comprises the following: a light chain variable region comprising SEQ ID No.: 9 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 10 or at least 80 sequences % identical to her. In a fourth aspect, the present disclosure provides an isolated ί Tau binding antibody or its binding fragment, wherein said Tau binding antibody or its binding fragment comprises the following: a light chain variable region comprising SEQ ID No.: 13 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 16 or at least 80 sequences % identical to her. In a fifth aspect, the present disclosure provides an isolated Tau-binding antibody or -linking fragment thereof, wherein said Tau-binding antibody or -linking fragment is linked to a phosphorylated Tau fragment comprising amino acids from 197 to 206 of the SEQ ID NO: 55. As an embodiment of the first and fifth aspects, the present disclosure provides antibodies or binding fragments thereof, which may be chimeric, humanized or fully human antibodies or binding fragments thereof. As an embodiment of the second or third i aspect, the present disclosure provides antibodies or their binding fragments, which may be chimeric antibodies or their binding fragments^. As an embodiment of the fourth aspect, the present disclosure provides antibodies or binding fragments thereof, which may be humanized antibodies or binding fragments thereof. In a sixth aspect, the present disclosure provides an isolated Tau binding antibody or binding fragment thereof, wherein said Tau binding antibody or binding fragment thereof competes for Tau binding with a Tau binding antibody or binding fragment thereof. binding, according to any of the first through fourth aspects, as well as embodiments thereof. In a seventh aspect, the present disclosure provides an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof binds to substantially the same Tau epitope as a Tau-binding antibody. Tau or its linking fragment, according to any of the first through fifth aspects and embodiments thereof. As an embodiment of the sixth and seventh aspects, the present disclosure provides monoclonal antibodies or binding fragments thereof, which are humanized antibodies or binding fragments thereof. The antibodies and their binding fragments of the first through seventh aspects, as well as embodiments thereof, have the ability to bind to soluble forms of human Tau, paired helical filaments (PHF) of human Tau, or both. to soluble forms of human Tau such as human Tau paired helical filaments (PHF) comprising a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID NO: 55. In an eighth aspect, the present disclosure provides nucleic acid molecules comprising nucleic acid sequences, such as DNA sequences encoding the heavy and / or light chain of an antibody or binding fragment of the first to seventh aspects and forms of realization of them. In a ninth aspect, the present disclosure provides cloning or expression vectors comprising these aforementioned nucleic acid molecules. In a tenth aspect, the present disclosure provides host cells comprising these aforementioned nucleic acid molecules, cloning vectors or expression vectors. In an eleventh aspect, the present description provides methods for the production of antibodies and their binding fragments of the first to seventh aspects and embodiments thereof. ¡ A twelfth aspect of the present description relates to the use of antibodies and their binding fragments of the first to seventh aspects and the forms I of realizing them, to treat the taupatias such as, in particular, AD and PSP. Another aspect of the present disclosure relates to the use of the antibodies and their binding fragments of the first to seventh aspects as well as embodiments thereof for the diagnosis of taupathies such as, in particular, AD and PSP. Figure 1: Binding of AB1 having a rabbit VL sequence (VL_AB1) of SEQ ID No.: 7 and a rabbit VH sequence (VH_AB1) of SEQ ID No.: 8 for the biotinylated T197 peptide versus the binding to peptides • Τ174, Τ211, Τ230 and T396 biotinylated in the ELISA assay of Experiment 2.3. Figure 2: Diagram illustrating the cell aggregation assay of Experiment 3.1. Figure 3: Efficiency of binding antibodies to Tau having a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID No.: 20 (A), and a Tau-binding antibody having a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID NO: 21 (B) or a negative control IgG4 A33 antibody (C) in a cellular Tau aggregation assay using pathological human Tau fibrils recovered from human PSP patients (PSP -PHF8) as seeds. Figure 4: Western blot showing the binding properties of a Tau-binding ABl antibody having a light chain of SEQ ID No.: 9 and heavy chain of SEQ ID No.: 10, for those used contain Tau from AD or human PSP. Figure 5: A) illustrates the ABl donor VL (VL_AB1) of SEQ ID No.: 7 with CDRs 1 (SEQ ID No.: 1), 2 (SEQ ID No.: 2) and 3 (SEQ ID No.: 53) underlined. B) illustrates the human IGKVl39 acceptor region VL sequence of SEQ ID No.: 44 with acceptor CDRs 1, 2, and 3 underlined. C) illustrates the CDR-grafted sequence — gVL4 ABl of SEQ No.: 11 with CDRs 1 (SEQ ID No.: 1), 2 • _ (SEQ ID No.: 2) and 3 (SEQ ID No.: 53) underlined. D) illustrates the gVL9_ABl CDR-grafted sequence of SEQ No.: 12 with CDRs 1 (SEQ ID No.: 1), 2 (SEQ ID No.: 2) and 3 (SEQ ID No.: 54) underlined; CDR3 comprises an A91 mutation compared to VL_AB1. Figure 6: A) illustrates the AB1 VH donor (VH_AB1) i of SEQ ID No.: 8 with CDRs 1 (SEQ ID No.: 4), 2 (SEQ ID No.: 5) and 3 (SEQ ID No.: 48) underlined. b) illustrates the VH sequence of the human IGHV4-acceptor region 39 of SEQ ID No.: 45 with acceptor CDRs 1, 2, and 3 underlined. C) illustrates the gVH41_ABl CDR-grafted sequence of SEQ No.: 14 with CDRs 1 (SEQ ID No.: 4), 2 (SEQ ID No.: 5) and 3 (SEQ ID No.: 49) underlined. The donor residues are shown in bold cut: K71 and V78. Mutations in frame are highlighted (El). CDR3 comprises a N100Q substitution compared to VH_AB1. D) illustrates the coh CDR gVH49_ABl grafted sequence of SEQ No.: 15 with CDRs 1 (SEQ ID No.: 4), 2 (SEQ ID No.: 5) and 3 (SEQ ID No.: 50) underlined. Donor residues are shown in bold italics (K71 and V78). In-frame mutations are highlighted (El). CDR3 comprises an N100A substitution compared to VH_AB1. Figure 7: The efficacy of Tau-binding antibodies having a light chain of SEQ ID No.: 9 and i a heavy chain of SEQ ID NO:10, and that of a Tau AT8 binding antibody described in the literature as binding to an epitope comprising phosphorylated residues 202 and 205 of SEQ ID NO:55 or an antibody of negative control 101.4 in a cellular Tau aggregation assay using pathological human Tau fibrils recovered from human AD patients as seeds. Figure 8: Efficacy of Tau-binding antibodies having a light chain of SEQ ID NO: 17 and a I heavy chain of SEQ ID NO: ¡20 in a cellular Tau aggregation assay using human Tau pathological fibrils recovered from human AD patients or from human PSP patients or from human FTD patients (Fronto Temporal Dementia, dementia frontotemporal (Pick's disease)) as seeds. The present disclosure, as illustratively described below, may be suitably practiced in the absence of any element(s), limitation(s), or limitations, not specifically described herein. The present description will be detailed with respect to particular aspects and embodiments thereof and with reference to certain figures and examples, but the present invention is not limited thereby. Technical terms are used in their common sense unless otherwise indicated. If a specific • meaning is given to certain terms, definitions of the terms will be provided below in the context in which the terms are used. When the term comprising / n is used in the present description, as well as in the claims, it does not exclude other elements. For purposes of the present description, the term "consisting of" is considered to be a preferred embodiment of the term "compound(s)". If hereinafter a group is defined as comprising at least a certain number of embodiments, it should also be interpreted as describing a group preferably consisting only of these embodiments. For the purposes of the present description, the term "obtained" is considered to be a preferred embodiment of the term "obtainable". If hereinafter, for example, an antibody is defined to be obtainable from a specific source, it should also be interpreted as describing an antibody that is obtained from this source. When a definite or indefinite article is used to refer to a singular noun, for example un, una, el or la, it includes the plural of that article, unless otherwise specified. The terms "around" or "approximately" denote a range of precision that the person skilled in the art will understand to ensure the technical effect of the characteristic in question. The term generally indicates the deviation from the stated numerical value of ±10%, and preferably ±5%. Any reference to a Tau-binding antibody or its linking fragment as a preferred embodiment of the various aspects should be understood to contemplate monoclonal antibodies | binding to Tau or its linking fragments. For various aspects the present description mentions antibodies and their linking fragments comprising CDRs and variable regions of the respective light chain and / or heavy chain regions. Antibodies or their linking fragments comprising only a light chain variable region or heavy chain variable region may be useful, for example, for manufacturing methods or, for example, to detect variable regions that can, effectively, associate with another corresponding variable region. However, it should be understood that wherever reference is made to antibodies and their I linker fragments comprising CDRs and variable regions of the respective light chain and / or heavy chain regions, this always contemplates, as a preferred embodiment, antibodies and their fragments linker variable regions comprising the CDRs and respective regions of light chain and heavy chain. As used herein, the terms treatment, treating, and the like, refer to obtaining a desired pharmacological and / or physiological effect. The effect can be prophylactic, in terms of completely or partially preventing a disease or its symptom, and / or it can be therapeutic, in terms of a partial or complete cure for a disease. I and / or adverse effect attributable to the disease. Accordingly, a treatment covers any treatment of a disease in a mammal, particularly a human, and includes the following: (a) preventing the disease from manifesting in a subject that may be predisposed to the disease, but has not still been diagnosed with it; (b) inhibit the disease, that is, stop its development; and (c) alleviating the disease, ie, causing regression of the disease. A reference to a Tau-binding antibody or its binding fragment as a therapeutically active agent refers to the use of a Tau-binding antibody or its binding fragment in the treatment of a disease. A therapeutically effective amount refers to that amount of a Tau-binding antibody or its binding fragment that, when administered to a mammal or other subject to treat a disease, is sufficient to effect said treatment for the disease. The therapeutically effective amount will vary depending on the Tau-binding antibody or its binding fragment, the disease and its severity, as well as the age, weight, etc., of the subject to be treated. A reference to a Tau-binding antibody or its binding fragment as a diagnostically active agent refers to the use of a Tau-binding antibody or its binding fragment1 in the diagnosis of disease. A diagnostically effective amount refers to the amount of a Tau-binding antibody or its binding fragment that, when used in a diagnostic test on a biological sample, is sufficient to allow identification of a disease. or monitoring the amount of disease tissue as a means of monitoring the efficacy of therapeutic intervention. The present Application is based in part on the identification of an antibody designated AB1 that binds to human Tau. As is customary in the art, Tau residue numbering herein refers to Tau isoform 2 of SEQ ID No.: 55 (NCBI Reference Sequence: NP_005901.2). As will be established hereinafter ABl, was isolated from an immunized rabbit, and recognizes a phosphorylated Tau region within the range of amino acids between 197 and 206 of SEQ ID No.: 55. The examples establish that ABl is able to bind to human Tau Paired Helical Filaments (PHF) (see Example 2.4) and that ABl was able to detect Intraneuronal Neurofibrillary Tangle (NFT). , extraneuronal NFTs, neuritic plate-like structures, and neuropil threads in cryosections of human samples (see Example 3.2). It seems reasonable to consider that this behavior is, at least in part, mediated by the complementarity determining regions (CDRs) of the variable light chain region (VL) and the variable heavy chain region (VH). , Variable Heavy chain region) of ABl. In this context, the present description provides Tau-binding antibodies or their linking fragments comprising the CDRs or residues determining the specificity of the VL region of ABl (SEQ ID No.: 7) and / or the CDRs of the VH region of ABl (SEQ ID! No.: 8). Residues in antibody variable domains are conventionally numbered according to a system devised by Kabat et al. This system is reported in Kabat et al., 1987, in Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH, USA (hereinafter Kabat et al. (supra))|. This numbering system is used in the present specification, except where otherwise indicated. Kabat designations of residues do not always correspond directly to the linear numbering of amino acid residues. The actual linear amino acid sequence may contain fewer amino acids or additional amino acids than in the strict Kabat numbering that corresponds to a reduction or insertion, in a structural component, be it a framework region or a complementarity determining region (CDR), of the basic structure of the variable domain. The correct Kabat numbering of residues can be determined for a given antibody by aligning homology residues in the antibody sequence with a standard Kabat numbered sequence. However, according to Chothia (Chothia, C and Lesk, A.M. J. Mol. Biol., 196, 901-917 (1987)) the equivalent loop for the CDR-H1 extends from residue 26 to residue 32. • CDR1, CDR2, and CDR3 of VL of AB1 were thus identified to correspond to SEQ ID Nos.: 1, 2, and 53, respectively. The AB1 VH CDR1, CDR2, and CDR3 were thus identified to correspond to SEQ ID Nos.: 4, 5, and 48, respectively. The effect of amino acid substitutions, additions and / or deletions to the CDRs can be readily tested by a person skilled in the art, for example using the methods described in the examples. In the originally identified VH CDR3 (CDRH3), namely SEQ ID No.: 48, for example, a potential asparagine deamidation site was identified and modified by replacement of the asparagine residue with either glutamine , alanine, aspartic acid or serine.This led to sequences SEQ ID No.: 49, 50, 51 and 52 respectively for CDRH3.For brevity, the three sequences for CDRH3, namely SEQ ID Nos. :48, 49, 50, 51 and 52 were combined as SEQ ID No.: 6. Similarly, a potential glutamine deamidation site in CDR3 (CDRL3) was identified and modified with replacement of the contiguous glycine with alanine This led to the sequence SEQ ID NO: 54. For brevity, both sequences for CDRL3, namely SEQ ID NO: 53 and 54 were combined as SEQ ID NO: 3. It will be appreciated that the CDRs can be subjected to further modifications such as substitutions, additions and / or deletions without substantially changing, for example, binding properties compared to AB1. This can be achieved primarily, for example, by replacing amino acids in the CDRs with similar amino acids. The concept of similarity, as used herein, indicates that, at any particular position in the aligned sequences, the amino acid residue is of a similar type between the sequences. For example, leucine can be substituted for isoleucine or valine. Other amino acids that can be frequently substituted for one another include, but are not limited to, the following: phenylalanine, tyrosine, and tryptophan (amino acids having aromatic side chains); lysine, arginine, and histidine (amino acids having basic side chains); aspartate and glutamate (amino acids having acidic side chains); í asparagine and glutamine (amino acids having amide side chains); and cysteine ​​and methionine (amino acids having sulfur-containing side chains). In this context, the present description provides in one aspect an isolated Tau-binding antibody or its binding fragment •, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain variable region comprising a CDRl selected from SEQ ID No.: 1 or sequences at least 90% identical thereto, a CDR2 selected from SEQ ID No.: 2 or sequences at least 90% identical to her, and a CDR3 selected from SEQ ID No.: 3 or sequences at least 90% identical to her; and / or a heavy chain variable region comprising a CDR1 selected from SEQ ID No.: 4 or sequences at least 90% identical thereto, a CDR'2 selected from SEQ ID No.: 5 or sequences at least mdnos 90% identical to it, and / or a CDR3 selected from lia SEQ ID No.: 6 or sequences at least 90% identical to it. In a further aspect, the present description provides an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain comprising a CDRl selected from SEQ ID No.: 1 or sequences at least 90% identical thereto, a CDR2 selected from SEQ ID No.: 2 or sequences at least 90% identical thereto, and one CDR3 selected from SEQ ID No.: 3o sequences at least I % identical to it; and _ a heavy chain. In a further aspect, the present description provides an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain; and a heavy chain variable region comprising a CDRl selected from SEQ ID No.: 4 or sequences at least 90% identical thereto, a CDR2 selected from SEQ ID No.: 5 or sequences at least 90% identical to it, and / or a CDR3 selected from SEQ ID No.: 6 or sequences at least 90% identical to it. The term identity, as used herein, indicates that at any particular position in the aligned sequences, the amino acid residue is identical between the sequences. The degrees of identity can be easily calculated, for example, using the BLAST™ software available from NCBI (Altschul, S.F. et al., 1990, J. Mol. Biol. 215:403-410; Gish, W & States, D.J. 1993 Nature Genet 3:266-272 Madden T.L. et al., 1996 Meth Enzymol 266:131-141 Altschul S.F. et al., 1997 Nucleic Acids Res 25:3389-3402 Zhang J.k. Madden, T.L. 1997, Genome Res. 7:649-656). The identity of CDRL1, CDRL2, 'cDRL3, CDRH1, CDRH2, and CDRH3 for SEQ ID No.: 1, | 2, 3, 4, 5, and 6 _ respectively can be at least 90%, but can also be higher such as at least 95%, 96%, %, 98%, or 99% with an optional preference for identities greater. Positions of the different identity can be selected according to similarity considerations. ’ In this context, the present disclosure specifically considers Tau-binding antibodies or their linking fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 3, respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 6, respectively. The present description also considers binding antibodies to Tau or its linking fragments comprising a VL with jCDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 53 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 6 I respectively, binding antibodies to their Tau linker fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 54 respectively and a VH with CDRH1, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 6 respectively , Tau binding antibodies or their linking fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 3 respectively and a VH with CDRH1, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 48 respectively, binding antibodies to Tau or its linking fragments comprising a VL with CDRLl, • CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 3 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 4-9 respectively, Tau binding antibodies or their linking fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: lj. 2, 3 respectively and a VH with CDRHl, CDRH2, and CDRH3 left SEQ ID Nos: 4, 5, and 50 respectively, antibodies binding to Tau or its linking fragments comprising a VL with CDRLl, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 3 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 50 respectively, Tau-binding antibodies or their linking fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 3 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 51 respectively, and Tau binding antibodies or their linking fragments comprising a VL with CDRLI, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 3 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 52 respectively. Tau binding antibodies or their linking fragments comprising a VL with CDRLI, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 53 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 48 respectively, Tau-binding antibodies or their linking fragments comprising a VL with CDRLl, — CDRL2, and CDRL3 of SEQ ID Nos.: 1,,2, 53 respectively • and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 49 respectively , Tau-binding antibodies or their linking fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 53 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos. : 4, 5, and 50 respectively,, Tau-binding antibodies or their linking fragments comprising a VL with CDRLl, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 53 respectively and a VH with CDRHl, CDRH2 , and CDRH3 of SEQ ID Nos: 4, 5, and 51 respectively, and antibodies binding to Tau or its linking fragments comprising a VL with CDRLl, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 53 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 52 respectively; Tau binding antibodies or their linking fragments comprising a VL with CDRLI, CDRL2, and CDRL3 of SEQ ID Nos.: 1, |2, 54 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 48 respectively, Tau binding antibodies or their fragments linkers comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 54 respectively and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 49 respectively, Tau-binding antibodies or their linking fragments comprising a VL with CDRLl, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 54 respectively — and a VH with CDRHl, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, • and 50 respectively, Tau-binding antibodies or their linker fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 2, 54 respectively and a VH with CDRH1, CDRH2, and CDRH3 of SEQ ID Nos: 4, 5, and 51 respectively, and Tau binding antibodies or their linking fragments comprising a VL with CDRL1, CDRL2, and CDRL3 of SEQ ID Nos.: 1, 12, 54 respectively and a VH with CDRH1, CDRH2, and CDRH3 of SEQ ID Nos: 4 , 5, I and 52 respectively. Yo Tau-binding antibodies or their linking fragments, as contemplated in said first aspect, may comprise these CDRs incorporated into framework regions of different origin. Accordingly, the CDRs may be comprised within the original framework regions of AB1, namely the rabbit VL region of SEQ ID No.: 7 and the rabbit VH region of SEQ ID No.: 8. No However, CDRs can also i be incorporated into the framework regions of origin of I different species, such as a mouse or human framework region. Depending on the origin of the framework regions and constant regions, which can be combined with said framework regions, chimeric, murinized or humanized antibodies binding Tau or its linking fragments can be obtained. — Chimeric antibodies binding to Tau or its linking fragments will comprise CDRs within framework regions of non-human origin combined with constant regions of a different species, such as murine or human origin. Murinized antibodies binding to Tau or its binding fragments shall comprise the CDRs within framework regions of murine origin combined together with constant regions of human origin. Humanized Tau-binding antibodies or their linking fragments will comprise CDRs within conserved framework I regions of human origin combined together with constant regions of human origin. In this context, the present description provides, in another aspect, an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain variable region comprising SEQ ID No.: 7 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising the SEQ ID No.: 8 or sequences at least 80% identical to i she. The identity of VL and VH for SEQ ID respectively can be at least 80%, can be higher such as at least 80%, %, 90%, %, 96%, %, 98%, or 99% with an optional preference for larger identities. The different identity positions can be selected according to similarity considerations. It will be appreciated that, in terms of identity, there may be more flexibility for framework regions versus CDRs. In this context, the present description specifically considers Tau-binding antibodies or their linking fragments comprising a VL of SEQ ID No.: 7 and a VH of SEQ ID No.: é. Humanized Tau-binding antibodies or binding fragments thereof are particularly contemplated by the present disclosure. Yo For this purpose, CDRs can be grafted onto human framework regions. It will be appreciated that identification of such a CDR-grafted Tau-binding humanized antibody or its binding fragment can be achieved following art-established approaches. When the CDRs or specificity determining residues are grafted, any suitable acceptor human variable region framework region sequence can be used taking into account the key / type of the donor antibody from which the CDRs are derived: (see, for example, example, i Boss et al., US Patent No. 4,816,397; Boss et al., European Patent Document No. 0,120,694 Bl; Neuberger, M.S. et al., World Patent Document Number WO 86 / 01533; Neuberger, M.S. et al., European Patent Number 0,194,276 Bl; Winter, United States Patent Number 5,225,539; Winter, European Patent Document No. 0,239,400 Bl; Padlan, E.A. et al., European Patent Application No. 0,519,596 Al). Likewise, in a CDR-grafted antibody variable region of the present invention, the framework regions need not have exactly the same sequence as those of the acceptor antibody. CDRs can therefore be grafted with or without framework changes. Introducing changes to the framework based on a comparison between ! the framework regions of the donor variable regions and the acceptor framework regions may allow retention of, for example, the affinity of an antibody that may otherwise be reduced as a consequence of humanization. For example, unusual residues can be changed to more frequently occurring residues for that type or class of acceptor chain. Alternatively, selected residues in the acceptor framework regions can be changed so that they correspond to the residue found at the same position in the donor antibody (See Riechmann et al., 1998, Nature, 332, 323-324). Such changes should be kept to the minimum necessary to restore the affinity of the donor antibody. Residues for the switch can be selected using the protocol stipulated by Adair et al. (1991) (Humanised antibodies, World Patent Document No. WO91 / 09967). In a CDR-grafted antibody of the present invention, the acceptor heavy and light chains need not be derived from this antibody and may, if desired, comprise composite chains having framework regions that are derived from different chains. Examples of human acceptor framework regions that can be used in the present invention are KOL, NEWM, REI, EU, TUR, TEI, LAY, and POM (Kabat et al., supra). For example, KOL and NEWM can be used for the heavy chain, REI can be used for the light chain, and EU, LAY, and POM can be used for both the heavy and light chains. Alternatively, human germline sequences can be used; which are available from the following Internet sites: http: / / vbase.mrc-ce.cam.ac.uk / ). The present disclosure specifically contemplates using the human IGKVl-39 V region plus JK4 region of SEQ, ID No.: 44 (IMGT, http: / / www.imgt.org / ) as an acceptor framework region for CDRs. light chain and the human V region IGHV4-39 plus JH4 region of SEQ ID No.: 45 (IMGT, http: / / www.imgt.org / ) as an acceptor framework region for heavy chain CDRs. In SEQ ID No.: 45, positions 1, 73 and 80 can, for example, be considered for residue changes) in the framework regions. The glutamine residue at position 1 can be changed to glutamate. The valine residue at position 73 can be changed to lysine. The phenylalanine at position 80 can be changed to valine. Other positions in SEQ ID No. 45 for residue changes in the framework regions may be positions 39 and / or 75. For example, the isoleucine residue at position 39 of SEQ ID No: 45 may be changed to valine. The threonine residue at position 75 can be changed to serine. Positions in SEQ ID No.: 44 for residue changes in framework regions may be position 2 and / or 63. For example, the isoleucine residue in position 2 of SEQ ID No.: 44 can be changed to valine. The serine residue at position 63 of SEQ ID No.: 44 can be changed to lysine. In this context, the present disclosure provides, in another aspect, an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody Tau or its linking fragment comprises the following: a light chain variable region comprising the SEQ ID No.: 9 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 10 or sequences at least 80% identical thereto. The present description further provides, in another aspect, an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain variable region comprising the SEQ ID No.: 13 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 16 or sequences at least 80% identical thereto. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain variable region comprising SEQ ID No.: 13 or sequences at least 80% identical thereto, and / or J a heavy chain variable region comprising SEQ ID No.: 14, 15 or sequences thereof. least 80% identical to her. Furthermore, said isolated Tau-binding antibody or its-linking fragment may comprise the following: • a light chain variable region comprising SEQ ID No.: 11, 12 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 16 or sequences therefor. least 80% identical to her. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain variable region comprising SEQ ID No.: 11 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 14, 15 or sequences at least least 80% identical to it. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain variable region comprising SEQ ID No.: 12 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 14, 15 or sequences per ib least 80% identical to it. The identity of VL and VH for SEQ ID Nos.: 13 and 16 respectively can be at least 80%, but also — can be higher such as at least 80%, 85%, 90%, a VL of SEQ ID %, 96%, 97%, 98%, or 99% binding antibodies with an optional preference for higher identities. The different identity positions may be selected based on similarity considerations. It will be appreciated that, in terms of identity, there may be more flexibility for framework regions versus CDRs. In this context, the application specifically considers Tau-binding antibodies or their linking fragments comprising a VL of SEQ ID No.: 11 and a VH of SEQ ID No.: 14, Tau-binding antibodies or their fragments binders that comprise No.: 11 and a VH of SEQ ID No.: 15, to Tau or its linker fragments comprising a VL of SEQ ID No.: 12 and a VH of SEQ ID No.: 14, and binding antibodies to Tau or its linking fragments comprising a VL of SEQ ID No.: 12 and a VH of SEQ ID No.: 15. CDR-grafted humanized Tau-binding antibodies or linking fragments thereof may comprise constant regions of human origin. According to the amino acid sequence of the constant region of their heavy chains, antibodies or immunoglobulins j are divided into classes: IgA, IgD, IgE, IgG and IgM, and several of these can also be divided into subclasses (subtypes) , for example IgGl, IgG2, IgG3, and IgG4, IgAl, and IgA2. In particular, human IgG constant region domains, especially of the IgG1L and IgG3 isotypes, can be used when the antibody molecule is intended for therapeutic uses and antibody effector functions are required. Alternatively, the IgG2 and IgG4 isotypes can be used when the antibody molecule is intended for therapeutic purposes and effector functions of the antibody are not required. The present description specifically considers humanized antibodies of the IgG1 and IgG4 subtype. It will be appreciated that modifications of the sequences of these constant region domains may also be used. For example, one or more amino acids, such as 1 or 2 amino acid substitutions, additions and / or deletions, can also be made to the constant domains of the antibody, without significantly altering the ability of the antibody to bind Tau. IgG4 molecules in which the serine at position 241 has been changed to proline can also be used, as described in Angal et al., Molecular Immunology, 1993, 30(I), 105-108. Antibody effector functions include ADCC (Antibody-Dependent Cellular Cytotoxicity) and CDC (Complement-Dependent Cytotoxicity). ADCC refers to antibody-dependent cellular cytotoxicity. In order to determine whether an antibody is, in principle, capable of mediating ADDC, ADCC can be measured in vitro by, for example, the so-called 51 Cr, Eu, and 35 S release assays. A target cell containing the antigen of interest, ie Tau, can be labeled with these compounds. Following binding of therapeutic antibody, cells are washed and effector cells expressing Fc receptors, such as FcγRIII, are coincubated with antibody-labeled target cells, and lysis of target cells can be monitored by release of Fcγ. labels. Another methodology that is used is the so-called aCella TOX™ assay. The CDC refers i to complement-dependent cellular cytotoxicity. In order to determine whether an antibody is, in principle, capable of mediating CDC, CDC can be measured in vitro, as described, for example, in Delobel A et al., Methods Mol Biol. (2013); 988:115-43 or Current Protocols in Immunology, Supplement Chapter 13 (Print edition ISSN: 1934-3671). In this context, the present disclosure provides, in another aspect, an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof comprises! the next: a light chain comprising SEQ ID No.: 19 or sequences at least 70% identical thereto, and / or a heavy chain comprising SEQ ID No.: 22 or sequences at least 70% identical thereto. Said isolated Tau-binding antibody or its linking fragment may comprise the following: a light chain comprising SEQ ID No.: 19 or sequences at least 70% identical thereto, and / or a heavy chain comprising SEQ ID No.: 20, 21 or sequences at least 80% identical thereto. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain comprising SEQ ID No.: 17, 18 or sequences at least 70% identical to it, and / or a heavy chain comprising SEQ ID No.: 22 or sequences at least 80% identical to it . Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain comprising SEQ ID No.: 17, 18 or sequences at least 70% identical to it, and / or a heavy chain comprising SEQ ID No.: 20, 21 or sequences at least 80% identical to her. The light chain and heavy chain identity for SEQ ID Nos.: 19 and 22 respectively can be at least 70%, but can also be higher such as at least 70%, 75%, 80%, 85 %, 90%, 95%, 96%, 97%, 98%, or 99% with an optional preference for larger identities. The different identity positions can be selected according to similarity considerations. It will be appreciated that, in terms of identity, there may be greater flexibility for framework regions versus CDRs and even more flexibility for constant regions. Yo In this context, the application specifically considers Tau-binding antibodies or their linking fragments comprising a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID No.: 20, Tau-binding antibodies or their linking fragments comprising a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID No.: 21, Tau-binding antibodies or their linking fragments comprising a light chain of SEQ ID No. : 18 and a heavy chain of SEQ ID No.: 20, and Tau binding antibodies or their linking fragments comprising a light chain of SEQ ID No.: 18 and a heavy chain of SEQ ID No. : twenty-one. Furthermore, the present disclosure provides, in another aspect, an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof comprises the following: — a light chain comprising SEQ ID No.: 19 or sequences at least 70% identical to it, and / or a heavy chain comprising SEQ ID No.: 25 or sequences at least 70% identical to it. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain comprising SEQ ID No.: 19 or sequences at least 70% identical thereto, and / or a heavy chain comprising SEQ ID No.: 23 or SEQ ID No.: 24 or sequences at least least 70% identical to it. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain comprising SEQ ID No.: 19 or sequences at least 70% identical thereto, and / or a heavy chain comprising SEQ ID No.: 23 or SEQ ID No.: 24 or sequences at least least 80% identical to it. Said isolated Tau-binding antibody or binding fragment thereof may comprise the following: a light chain comprising SEQ ID No.: 17, 18 or sequences at least 70% identical thereto, and / or a heavy chain comprising SEQ ID No.: 23 or SEQ ID No.: 24 or sequences at least 80% identical to it. the request specifically In this context, I consider antibodies binding to Tau or its fragments linkers comprising a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID No. 23, Tau-binding antibodies or their linker fragments comprising a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID No.: 24, binding antibodies to Tau or its linking fragments comprising a light chain of SEQ ID No.: 18 and a heavy chain of SEQ ID No.: 23, and binding antibodies to Tau or its linking fragments comprising a light chain of SEQ ID No.: 18 and a heavy chain of SEQ ID No.: 24. The light chain and heavy chain identity for SEQ ID No.: 19 and SEQ ID Nos.: 23 or 24, respectively can be at least 70%, but can also be higher such as at least 70%. %, %, 90%, 95%, 96%, 97%, 98%, or 99% with an optional preference for larger identities. The different identity positions may be selected based on similarity considerations. It will be appreciated that, in terms of identity, there may be greater flexibility for framework regions versus CDRs and even more flexibility for constant regions. , Also, the present description provides a specific region or epitope of human Tau that is linked by a antibody or its linking fragment provided by the present description, in particular,! an antibody or its binding fragment comprising any one of CDR-Hl (SEQ ID No.: 4), CDR-H2 (SEQ ID No.: 5), CDR-H3 (SEQ ID No.: 6), CDR-L1 ( SEQ ID No.: 1), CDR-L2 (SEQ ID No.: 2) or CDR-L3 (SEQ ID No.: 3), for example, antibodies comprising the VL of SEQ ID No.: 7 and the SEQ ID VL No.: 8. The present description, furthermore, provides a specific region or epitope of human Tau, in particular, a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID NO.: 55, which is bound by an antibody or its linking fragment. , provided in the present description, in particular, an antibody or its linking fragment comprising the VL of SEQ ID No.: 7 and the VH of SEQ ID No.: 8. The Tau region within amino acids 197 to 206 of SEQ ID No.: 55 comprises four possible phosphorylation sites corresponding to serine residues at positions 198 (S198), 199 (S199), 202 (S202) and one threonine residue at position 205 (T205). The phrase "a phosphorylated Tau region within amino acids 197 through 206j of SEQ ID NO.: 55" refers to a Tau region within amino acids 197 through 206 of SEQ ID NO: 55 that comprises at least one phosphorylated residue selected from S198, S199, S202 and T205. As one skilled in the art will know, phosphorylated residues may also be referred to, for example, as Ser(PO3H2) or Thr(PO3H2)· Binding of a Tau-binding antibody to this specific region or epitope of Tau can be identified by any suitable epitope mapping method known in the art, in combination with any of the antibodies provided by the present disclosure. Examples of such methods include screening peptides of varying lengths derived from SEQ ID No.: 55 for binding to the Tau-binding antibodies or their binding fragments of the present disclosure with the smallest fragment that can specifically bind. to the antibody containing the sequence of the epitope recognized by the Tau-binding antibodies or their linking fragments. Given the existence of different isoforms of Tau in the central nervous system, it should be understood that any isoform can be used in the methods detailed herein. In a specific example, the longest isoform of Tau, i.e. isoform 2, as defined in SEQ ID No. 55 can be used. The Tau peptides of SEQ ID No. 55 can be produced from recombinantly, synthetically, or by proteolytic digestion of the Tau polypeptide. Antibody-binding peptides can be identified, for example, by Western blotting or mass spectrometry analysis. In another example, NMR (Nuclear Magnetic Resonance) spectroscopy or X-ray crystallography can be used to identify the epitope bound by a Tau-binding antibody or its binding fragment. Once identified, the epitope-binding fragment of an antibody of the present invention can be used, if necessary, as an immunogen to obtain additional antibodies that bind the same epitope. In addition, the antibody-binding epitope fragment of the present invention can be used to obtain epitope-binding proteins and, if necessary, inhibit at least Tau aggregation, such as polypeptide compounds or protein comprising more than 10 amino acids that are based on protein scaffolds, for example, lipocalin (anticalines), fibronectin (adnectins, trinectins), kunitz domains, C-type lectin, transferrin, gamma crystalline, cysteine ​​knots, repeats of ankyrin (DARPin) or protein A, (afibodies) as known in the art (Tomlinson, 2004; Mosavi et al., 2004; Gilí and Damle, 2006; Nilsson and Tolmachev, 2007; Binz et al., 2004). Additionally, the epitope-binding molecules further include organic molecules including peptides and cyclic peptides comprising not more than 10 amino acids, as well as peptidomimetics. Peptidomimetics are compounds that are based on amino acid sequences found at protein-protein interaction sites and are known in the art (Sillerud and Larson, 2005). j Yo In this context, the present disclosure provides, in another aspect, an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of the SEQ ID No.: 55. Such antibodies may be chimeric, murine, humanized, or fully human monoclonal antibodies or may be used to obtain chimeric, murine, humanized, or fully human monoclonal antibodies. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least one phosphorylated residue selected from S198, S199, S202 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least one phosphorylated residue selected from S198, and S199. In another aspect, the present disclosure provides an isolated Tau binding antibody or its linking fragment, wherein said L Tau binding antibody or its linking fragment binds to a phosphorylated Tau' region within amino acids 197 to 206 of the SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least one phosphorylated residue selected from S202 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ' ID No.: 55, wherein said phosphorylated Tau region comprises at least one phosphorylated residue comprising S198. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, where said phosphorylated Tau region comprises at least one phosphorylated residue comprising S199. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least one phosphorylated residue comprising S202. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least one phosphorylated residue comprising T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ; ID No.: 55, wherein said phosphorylated Tau region comprises at least two phosphorylated residues selected from S198, S199, S202 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of the Tau. SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least two phosphorylated residues comprising S198 and S199. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least two I phosphorylated residues comprising 3199 and S202. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least two phosphorylated residues comprising S202 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof binds to a phosphorylated Tau region within amino acids 197 to 206 of the SEQ ID No.: 55, wherein • said phosphorylated Tau region comprises at least two phosphorylated residues comprising S198 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least two phosphorylated residues comprising S198 and S202. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of the SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least two phosphorylated residues comprising S199 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three i phosphorylated residues selected from S198, S199, S202 and T205. In another aspect, the present disclosure provides an • isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of the SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three phosphorylated residues comprising S198 and S199. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three phosphorylated residues comprising S199 and S202. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three phosphorylated residues comprising S202 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof binds to a phosphorylated Tau region within amino acids 197 to 206 of the SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three phosphorylated residues comprising S198 and T205. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a j-phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three phosphorylated residues comprising S198 and S202. In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises at least three phosphorylated residues comprising S199 and T205. Yo In another aspect, the present disclosure provides an isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55, wherein said phosphorylated Tau region comprises the following four phosphorylated residues S198, S199, S202 and T205. Such antibodies may be chimeric, murine, humanized, or fully human monoclonal antibodies or they can be used to obtain human chimeric, murine, humanized, or fully monoclonal antibodies. In another aspect, the present disclosure provides an isolated neutralizing Tau binding antibody or binding fragment thereof, wherein said neutralizing Tau binding antibody or binding fragment thereof binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 35. Yo Such antibodies may be chimeric, murine, humanized, or fully human monoclonal antibodies or may be used to obtain chimeric, murine, humanized, or fully human monoclonal antibodies. In another aspect, the present disclosure provides an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof binds to substantially the same Tau epitope as a Tau-binding antibody or binding fragment thereof. binder described above. Epitope binding can be determined as described for epitope mapping using for example a Tau binding antibody or its linking fragment comprising a VL of SEQ ID No.: 7 and a VH of SEQ ID No. : 8 for reference. Such antibodies can be chimeric, humanized murine or fully human monoclonal antibodies or can be used to obtain chimeric antibodies, i murinized, humanized, or fully monoclonal human. In another aspect, the present disclosure provides an isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof competes for binding to Tau with a Tau-binding antibody described above. In this context, the present description specifically contemplates an isolated Tau-binding antibody or its binding fragment, wherein said Tau-binding antibody or its binding fragment compites by binding to Tau with a Tau-binding antibody or its binding fragment. comprising a VL of SEQ ID No.: 7 and a VH of SEQ ID No.: 8. Such antibodies may be chimeric, murine, humanized, or fully human monoclonal antibodies or may be used to obtain chimeric, murine, humanized, or fully human monoclonal antibodies. Competition for binding to Tau can be determined by a reduction in binding of the antibody or its binding fragment to Tau by at least about 50% or at least about 70% or at least about 80% or at least about 90% or at least about 95% or at least about 99% or about 100%, in the presence of the antibody to reference or its linker fragment which may comprise a VL of SEQ ID No.: 7 and a VH of SEQ ID No.: 8 or a VL of SEQ ID No.: 9 and a VH of SEQ ID No.: 10. Binding can be measured using surface plasmon resonance with the use of a BIAcore® instrument, various fluorescence detection technologies (e.g., fluorescence correlation spectroscopy, fluorescence cross-correlation, fluorescence lifetime measurements , etc.) or various types of radioimmunological assays or other assays used to follow antibody binding to a target molecule. The term Tau-binding antibody or its binding fragment means that the antibody or binding fragments thereof bind specifically to Tau via its variable regions, that is, they bind the Tau antigen with greater affinity than other antigens that are not Tau homologues. Tau. The Tau-binding antibody or its binding fragment binds Tau via its variable regions with at least twice, at least five times, at least 10, 20, 100, ΙΟ3, ΙΟ4, 105 or at least 106 times the affinity than other antigens that are not Tau homologues. It will be understood that Tau-binding antibodies and their linking fragments may, however, also interact with other proteins (eg, S. aureus protein A or other antibodies in the techniques). ELISA) through interactions with sequences outside the variable region of Tau-binding antibodies and their linking fragments. Such latter binding properties being mediated by sequences outside the variable regions of Tau-binding antibodies and their linking fragments and, in particular, by the constant regions of Tau-binding antibodies and their linking fragments, does not mean which are encompassed by the term Tau-binding antibody or its linking fragment. Screening assays for determining the binding specificity of an antibody are well known and routinely practiced in the art. Tau-binding antibodies or their binding fragments may have an equilibrium dissociation constant (KD) for the binding affinity of the antibody (or its binding fragment) to its antigen in the nanomolar range. Therefore, KD may be below about 1*10-6, for example below about 5*10-7' such as about 2*10~7 or less, and can be measured using, for example, surface plasmon resonance and the BIAcore device, as described in the examples. As mentioned above, the present disclosure provides antibodies binding to Tau or its linking fragments. A full-length antibody — includes a constant domain and a variable region. The • constant region may not need to be present to its full extent in an antigen-binding fragment of an antibody. However, it should be understood that wherever the application contemplates the use of antibodies that mediate ADCC and / or CDC, a linking fragment must comprise a constant region of enough extension so that it is still able to mediate the ADCC and / or the CDC. As mentioned above, the present description also relates to human Tau-binding antibodies or their binding fragments, which can be generated as an alternative to humanization. For example, transgenic animals (eg mice) are known I in the art because they are capable, after immunization, of producing a full repertoire of human antibodies in the absence of endogenous murine antibody production. For example, it has been described that homozygous deletion of the antibody Heavy-chain Joining Region (JH) gene in chimeric and germline mutant mice results in complete inhibition of antibody production. endogenous. Transfer of the human germline immunoglobulin gene array into such germline mutant mice will result in the production of human antibodies with specificity against a particular antigen, following immunization of the transgenic animal carrying the immunoglobulin genes. germline cells with said antigen. Technologies for producing such transgenic animals, as well as technologies for isolating and producing human antibodies from such transgenic animals, are known in the art (Lonberg, 2005; Green, 1999; Kellermann and Green, 2002; Nicholson et al., 1999). ). Alternatively, in the transgenic animal, eg, the mouse, only the immunoglobulin genes encoding the mouse antibody variable regions are replaced with the corresponding human variable immunoglobulin gene sequences. The mouse germline immunoglobulin genes, which encode the constant regions of the antibody, remain unchanged. In this way, the effector functions of the antibody in the transgenic mouse immune system, and consequently the development of B cells, remain essentially unchanged, which may lead to an enhanced antibody response, upon antigen challenge in vivo. . Once the genes encoding a particular antibody of interest have been isolated from such transgenic animals, the genes encoding the constant regions can be replaced with human genes from the constant region, in order to obtain a fully human antibody. Other methods for obtaining antibody fragments from human antibodies in vitro are based on display technologies, such as phage display or ribosome display technology, where recombinant DNA libraries are used, which are either generated, by at least in part, artificially or from donor repertoires of immunoglobulin variable domain (V) genes. Phage and ribosome display technologies for the generation of human antibodies are well known in the art (Winter et al., 1994; Hoogenboom, 2002; Kretzschmar and von Ruden, 2002; Groves and Osbourn, 2005; Dufner et al. , 2006). Human antibodies can also be generated from isolated human B cells that are immunized ex vivo with an antigen of interest and subsequently fused to generate hybridomas that can then be screened for optimal human antibody (Grasso et al., 2004; Li et'al., 2006). The term Tau-binding neutralizing antibody, as used herein, refers to an antibody that binds to Tau and inhibits at least one of its biological activities. In a particular embodiment, a Tau-binding neutralizing antibody, as used herein, refers to — an antibody that binds to Tau and inhibits • Tau aggregation in an in vitro assay, such as , for example, in an in vitro assay, as described in experiment 3.1 below. The term "antibody" as used herein generally refers to intact (whole, full-length) antibodies, ie, comprising two heavy chain and two light chain elements. The antibody may further comprise additional binding domains, for example according to the DVD-Ig molecule, as described in world patent document number WO 2007 / 024715 or so-called (FabFv)2Fc , which is described in world patent document number WO2011 / 030107. Thus, antibody as used herein includes full-length bi-, tri-, or tetra-valent antibodies. Antibody-binding fragments include single-chain (ie, full-length heavy chain and light chain) antibodies; Fab, modified Fab, Fab', modified Fab', F(ab')2, Fv, Fab-Fv, Fab-dsFv, FabscFv, Fab-scFc, disulfide-stabilized Fab-scFv, single domain antibodies (eg VH or VL or VHH) , scFv, scFv-scFc, dsscFv, dsscFv-scFc, bi, tri or tetravalent antibodies, Bis-scFv, diabodies, tribodies, — triabodies, tetrabodies, domain antibodies (dAb), such as sdAbs, VHH, and VNAR fragments, and epitope-binding fragments of any of the aforementioned (see, eg, Holliger and Hudson, 2005, Nature Biotech, 23(9):1126-1136; Adair and Lawson, 2005, Drug Design Reviews, Online 2(3), 209-217). Methods for creating and making these antibody fragments are well known in the art (see, eg, Verma et al., 1998, Journal of Immunological Methods, 216, 165-181). Fab-Fv format was first described in world patent document number W02009 / 040562 and disulfide stabilized versions of them, Fabi dsFv was first described in patent document I worldwide with the number W02010 / 035012. A disulfide stabilized form of Fab-scFv was described in World Patent No. WO2013 / 068571. Antibody formats comprising scFc formats were first described in world patent document number WO2008 / 012543. Other antibody fragments for use in the present invention include Fab and Fab' fragments which are described in World Patent Application Nos. W02005 / 003169, W02005 / 003170 and W02005 / 003171. Multivalent antibodies may comprise I multiple specificities for example bispecific or may be monospecific (see, for example, world patent documents under numbers WO92 / 22583 and W005 / 113605). An example of the latter is a Tri-Fab (or TFM), as described in World Patent Document No. WO92 / 22583. In one embodiment, a Fab fragment is provided. In one embodiment, a Fab' fragment is provided. A typical Fab' molecule comprises a pair of heavy and light chains in which the heavy chain comprises a VH variable region, a CH1 constant domain, and a natural or modified hinge region and the light chain comprises a VL variable region and a CL constant domain. . Yo In one embodiment a dimer of a Fab' according to the present disclosure is provided to create an F(ab')2 eg the dimerization may be through the hinge. In one embodiment, the antibody or its binding fragment comprises a binding domain. A binding domain will generally comprise 6 CDRs, three from a heavy chain and three from a light chain. In one embodiment, the CDRs are in a framework region and together form a variable region. Accordingly, in one embodiment, an antibody or binding fragment comprises an antigen-specific binding domain comprising a light chain variable region and a heavy chain variable region. It will be appreciated that the affinity of the binding antibodies to Tau or their linking fragments provided by the present disclosure may be altered using suitable methods known in the art. The present description therefore also relates to variants of the antibody molecules of the present invention, which have an improved affinity for Tau. Such variants can be obtained by a number of affinity maturation protocols including CDR mutation (Yang et al., J. Mol. Biol., 254, 392-403, 1995), strand swapping ( Marks et al., Bio / Technology, 10, 779-783, 1992), use of strains [ E. coli mutators (Low et al.^ J. Mol. Biol., 250, 359368, 1996), DNA recombination (Patten et al., Curr. opinion Biotechnol., 8, 724-733, 1997), phage display (Thompson et al., J. Mol. Biol., 256, 77-88, 1996) and PCR (Polymerase Chain Reaction, polymerase chain reaction) (Crameri et al., Nature, 391, 288-291, 1998). Vaughan et al. (supra) discusses these affinity maturation methods. Tau-binding antibodies and their linking fragments may thus also encompass any of the above specifically mentioned amino acid sequences of the heavy or light chains with one or more conservative substitutions (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 conservative substitutions). Positions in an amino acid sequence that are candidates for conservative substitutions can be determined, and I select synthetic and naturally occurring amino acids that make conservative substitutions for any particular amino acids. Consideration for the selection of conservative substitutions includes the context in which any particular amino acid substitution is made, the hydrophobicity or polarity of the side chain, the general size of the side chain, and the pK value of the side chains with character. acidic or basic under physiological conditions. For example, lysine, arginine, and histidine are often suitably substituted one with the and another. As is known in the art, this is because all three amino acids have basic side chains, whereas the pK value for the lysine and arginine side chains are much closer to each other (about 10 and 12) than for the lysine and arginine side chains. histidine (about 6) . Similarly, glycine, alanine, valine, leucine, and isoleucine are often suitably substituted one with the another, with the proviso that glycine is often not adequately substituted by the other members of the group. Other amino acid groups frequently suitably substituted for one another include, but are not limited to, the group consisting of glutamic and aspartic acids; the group consisting of phenylalanine, tyrosine, and tryptophan; and the group consisting of serine, threonine, and optionally tyrosine. Tau-binding antibodies and their linking fragments as mentioned in the context of the present invention may encompass derivatives of the exemplary antibodies, fragments and sequences described herein. The term "derivatives" includes Tau-binding antibodies and their linking fragments, which have been chemically modified. Examples of chemical modification include covalent attachment of one or more polymers, such as water-soluble polymers, N-bonded or 0-bonded carbohydrates, sugars, phosphates, and / or other molecules, such as detectable labels such as, for example, fluorophores. n Tau-binding antibody or its for use in the present invention be conjugated to one or more molecules effectors. It will be appreciated that the effector molecule may comprise a single effector molecule or two or more such molecules so linked, in order to form a single moiety which may be linked to the antibodies of the present invention. Where it is desired to obtain an antibody fragment linked to an effector molecule, this can be prepared by standard chemical or recombinant DNA procedures in which the antibody fragment is linked, either directly or via a coupling agent, to the target molecule. effector molecule. Techniques for conjugating such effector molecules to antibodies are well known in the art (see, Hellstrom et al., Controlled Drug Delivery, 2nd Ed., Robinson et al., eds., 1987, pp. '623-53; Thorpe et al., 1982, Immunol.Rev., 62:119-58 and Dubowchik et al., 1999, Pharmacology and Therapeutics, 83, 67-123). These techniques for conjugating the effector molecules may include site-specific or non-site-specific conjugation or random conjugation. Particular chemical procedures include, for example, those described in World Patent Nos. WO 93 / 06231, WO 92 / 22583, WO 89 / 00195, WO 89 / 01476 and WO 03 / 031581. Alternatively, when the effector molecule is a protein or polypeptide, binding may be achieved using recombinant DNA methods, for example, as described in World Patent Number WO 86 / 01533 and EP0392745. Alternatively, a particular binding site for the effector molecule may be engineered into the antigen-binding antibody or antibody fragment of the present invention, for example, as described in World Patent No. WO 2008 / 038024. In addition, a coupling agent may be used to link the effector molecule to the antigen-binding antibody or antibody fragment of the present invention, for example, as described in World Patent No. WO 2005 / 113605. It will be understood by the person skilled in the art that the aforementioned possibilities can be used individually or in combination. Yo The term effector molecule, as used herein, includes, for example, drugs, toxins, biologically effective proteins, eg, enzymes, other antibody or antibody fragments, polymers that are synthetic or produced from naturally occurring nucleic acids and fragments thereof for example DNA, RNA and fragments thereof, radionuclides, particularly radioiodide, radioisotopes, chelated metals, nanoparticles and reporter groups such fluorescent compounds or compounds that can be detected by NMR or ESR spectroscopy. The effector molecule, as used herein, it also includes therapeutic agents such as chemotherapeutic agents, therapeutic polypeptides, nanoparticles, liposomes, or therapeutic nucleic acids. Other effector molecules may include chelated radionuclides such as luIn and 90Y, Lu177, Bismuth213, Californium252, Iridium192, and Tungsten188 / Rhenium188; or drugs such as, but not limited to, alkyl phosphocholines, topoisomerase I inhibitors, taxoids, and suramin. Other effector molecules include proteins, peptides, and enzymes. Enzymes of interest include, but are not limited to, proteolytic enzymes, hydrolases, lyases, isomerases, transferases. Proteins, polypeptides, and peptides of interest include, but are not limited to, immunoglobulins, toxins such as abrin, ricin A, Pseudomonas exotoxin, or diphtheria toxin, a protein such as insulin, tumor necrosis factor, otinterferon, β-interferon, nerve growth factor, platelet-derived growth factor or tissue plasminogen activator, a thrombotic agent or an anti-angiogenic agent, eg, angiostatin or endostatin, or, a biological response modifier, such as a lymphokine, interleukin-1 ( IL-1), interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte-colony stimulating factor (G-CSF), nerve growth factor (NGF) ΊΟ or other growth factors and immunoglobulins, or other protein or polypeptide compounds comprising more than 10 amino acids that are based on protein scaffolds, for example, of lipocalin (anticalines), fibronectin (adnectins, trinectins), Kunitz domains, C-type lectin, transferrin, gamma-crystallin, cysteine ​​knots, ankyrin repeats (DARPins), Fyn SH3 domains (phinomers), or protein A (afibodies), as known in the art (Tomlinson, 2004; Mosavi et al. , 2004; Gilí and Damle, 2006; Nilsson and Tolmachev, 2007; Binz et al., 2004; Silacci et al. 2014). Other effector molecules include peptides and proteins that enhance or facilitate penetration of the blood-brain barrier. For example, world patent documents numbered W02010 / 043047, W02010 / 063122, W02010 / 063123 or W02011 / 041897 describe i peptides or polypeptides that can act as a vector that has the ability to transport a therapeutic molecule across the blood-brain barrier and a method of conjugating them to a therapeutic molecule. Peptides and proteins of interest in the context of blood-brain barrier penetration include, but are not limited to, peptides and proteins that bind to a blood-brain barrier receptor, such as transferrin receptor, glucose receptor, insulin receptor, insulin-like growth factor, low-density lipoprotein-related protein receptor 8, low-density lipoprotein-related protein receptor 1, and heparin-binding epidermal growth factor-like growth factor. Alternatively, the effector molecule is an antibody fragment such as a domain antibody, camelid antibody, or shark-derived antibody (vNAR) that specifically binds to the aforementioned blood-brain barrier receptors. Other effector molecules may include detectable substances useful, for example, in diagnosis. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive nuclides, positron-emitting metals, such as can be used in positron emission tomography or photon emission computed tomography i single, and non-radioactive paramagnetic metal ions. See generally US Patent No. 4,741,900 for metal ions that can be conjugated to antibodies for diagnostic use. Suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase • acetylcholinesterase; suitable prosthetic groups include streptavidin, avidin, and biotin; suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, fluorescein dichlorotriazinylamine, dansyl chloride, and phycoerythrin; suitable luminescent materials include luminol; suitable bioluminescent materials include luciferase, luciferin, and aequorin; and suitable radioactive nuclides include 124I, 125I, 131I, luIn, 99Tc, 89Zr, 90Y, 64Cu, 68Ga, and 18F. A particular class of effector molecules suitable as diagnostically useful detectable substances include electron-deficient tetrazines and trans-cyclooctene (TCO), as described in Wyffels et al. 2014, Nuclear Medicine and biology 41 (2014):513-523, wherein a Tau-binding antibody of the present invention linked to tetrazine can be administered and allowed to achieve maximal uptake and sufficient clearance from non-target sites. , followed by subsequent administration of TCO or an optimized TCO analogue, labeled with a suitable radioactive nuclide, such that the TCO will covalently bind tetrazine in the Tau-binding antibody of the present invention, and allow its detection, by For example, using positron emission tomography or single photon emission computed tomography. In one embodiment, a Tau, Fab', or scFv-binding Fab is provided attached to a radioactive nuclide or tetrazine. Links to a radioactive nuclide or to tetrazine can be made by coupling, through any available amino acid side chain or terminal amino acid functional group located on the antibody fragment, for example, any amino, imino, thiol, hydroxyl group. or free carboxyl. Such amino acids may occur naturally in the antibody fragment or may be engineered into the fragment using recombinant DNA methods (see, for example, US Patent Nos. 5,219,996 and 5,667. 425; World Patent Nos. WO98 / 25971 and W02008 / 038024). In one example, the Tau-binding antibody or its linking fragment of the present invention is a modified Fab fragment, wherein the modification is the addition of one or more amino acids to the C-terminus of the heavy chain to allow coupling of an effector molecule. Suitably, the additional amino acids form a modified hinge region containing one or more cysteine ​​residues, to which the effector molecule can bind. In one embodiment, if the radionuclide is a metal ion, such as 111In, 99Tc, 89Zr, 90Y, 64Cu, or 68Ga, it may be bound via a macrocycle chelator, for example, as described in Turner et al. (Br. J. Cancer, 1994, 70:35-41; Comparative biodistribution of indium-lll-labelled macrocycle chimeric B72.3 antibody conjugates in tumour-bearing mice) whereby the latter is, in turn, covalently bound to the amino acid side chain or amino acid terminal functional group(s) of the aforementioned antibody or antibody fragment. In a further embodiment, the latter radionuclide-bound macrocycle chelate may be the effector molecule described in World Patent No. WO05 / 113605, which is part of a cross-linking agent linking two or more anti-antibodies. -Tau or fragments of them. In another example, the effector molecule may increase the useful half-life of the antibody in vivo, and / or reduce the immunogenicity of the antibody, and / or enhance the delivery of an antibody across an epithelial barrier to the immune system. Examples of suitable effector molecules of this type include polymers, albumin, and albumin-binding proteins or albumin-binding compounds such as those described in World Patent Number WO05 / 117984. When said effector molecule is a polymer it may, in general, be a synthetic polymer or one that occurs naturally, for example, an optionally substituted straight or branched chain polyalkylene, polyalkenylene or polyoxyalkylene polymer or a branched or branched polysaccharide. unbranched, eg, a homo- or hetero-polysaccharide. Specific optional substituents that may be present on the aforementioned synthetic polymers include one or more hydroxy, methyl, or methoxy groups. Specific examples of synthetic polymers include poly(ethylene glycol), optionally substituted straight or branched chain poly(propylene glycol) poly(vinyl alcohol) or derivatives thereof, especially poly(ethylene glycol^) such as optionally substituted methoxypoly(ethylene glycol) or derivatives of they. Specific naturally occurring polymers include lactose, amylose, dextran, glycogen, or derivatives thereof. In one embodiment, the effector molecule is albumin or a fragment thereof, such as human serum albumin or a fragment thereof. The size of the polymer can be varied as desired, but will generally be on average molecular weight. in the range from 500 Da to 50,000 Da, for example, from 5,000 to 40,000 Da, such as from 20,000 to 40,000 Da. The size of the polymer may be, in particular, selected on the basis of the product's intended use, for example, the ability to localize to certain tissues, such as the brain, or to extend the half-life in the circulation (for a review see Chapman, 2002, Advanced Drug Delivery Reviews, 54, 531545). Thus, for example, when the product is designed to leave the circulation and penetrate the tissue. Suitable polymers include a polyalkylene polymer, such as a polyethylene glycol or, especially, a methoxypoly(ethylene glycol) or a derivative thereof, and especially with a molecular weight in the range of from about 15,000 Da to about 40,000 Da. . In one example, the present invention is poly(ethylene glycol) (PEG). antibodies for use in binding to fractions of In a particular example, the antibody is a Tau-binding antibody or its linking fragment, and the PEG molecules may be linked through any available amino acid side chain or terminal amino acid functional group located on the antibody fragment, for example, any group — amino, imino, free thiol, hydroxyl or carboxyl. Such amino acids may occur naturally in the antibody fragment or may be engineered into the fragment using recombinant DNA methods (see, for example, US Patent Nos. 5,219,996 and 5,667. 425; World Patent Nos. WO98 / 25971 and W02008 / 038024). In one example, the Tau-binding antibody or its linking fragment of the present invention is a modified Fab fragment wherein the modification is the addition of one or more amino acids to the C-terminus of the heavy chain to allow coupling. of an effector molecule. Suitably, the additional amino acids form a modified hinge region containing one or more cysteine ​​residues to which the effector molecule can bind. Multiple sites can be used to link two or more PEG molecules. Suitably, the PEG molecules are covalently linked through a thiol group of at least one cysteine ​​residue located on the antibody fragment. Each polymer molecule bound to the modified antibody fragment can be covalently bound to the sulfur atom of a cysteine ​​residue located in the fragment. The covalent bond will generally be a disulfide bond or, in particular, a sulfur-78-carbon bond. When a thiol group is used as the point of attachment, activated effector molecules, for example, thiol-selective derivatives such as maleimides and cysteine ​​derivatives, can be suitably used. An activated polymer can be used as the starting material in the preparation of the polymer-modified antibody fragments, as described above. The activated polymer can be any polymer containing a reactive thiol group, such as an ester or α-halocarboxylic acid, eg, iodoacetamide, an imide, eg, maleimide, a vinyl sulfone, or a disulfide. Such starting materials may be obtained commercially (eg, through Nektar, formerly Shearwater Polymers Inc., Huntsville, AL, USA) or may be prepared from commercially available starting materials using conventional chemical procedures. Particular PEG molecules include 20K methoxy-PEG-amine (available from Nektar, formerly Shearwater; Rapp Polimere; and SunBio) and M-PEG-SPA (available from Nektar, formerly Shearwater). In another aspect, the present disclosure provides nucleic acid molecules comprising nucleic acid sequences encoding Tau-binding antibodies and their linking fragments, for nucleic acid molecules comprising nucleic acid sequences encoding their heavy variable chains and / or or light and for nucleic acid molecules comprising nucleic acid sequences encoding the CDR1, CDR2 and / or CDR3 of their variable heavy and / or light chains. By way of example, the VL of AB1 (SEQ ID No.: 7) can be encoded by SEQ ID No.: 26). The ABl VH (SEQ ID No.: 8) can be encoded by SEQ ID No.: 27). The humanized VL of SEQ ID No.: 12 may be encoded by SEQ ID No.: 30. The humanized VL of SEQ ID No.: 13 may be encoded by SEQ ID No.: 31. The humanized VH of SEQ ID No.: 14 may be encoded by SEQ ID No.: 32 and the humanized VH of SEQ ID No.: 15 may be encoded by SEQ ID No.: 33. The humanized light chain of SEQ ID No.: 17 may be encoded by SEQ ID No.: 30. The humanized light chain of SEQ ID No.: 18 may be encoded by SEQ ID No.: 31. The chain The humanized heavy chain of SEQ ID No.: 20 may be encoded by SEQ ID No.: 36 and the humanized heavy chain of SEQ ID No.: 21 may be encoded by SEQ ID No.: 37. The humanized heavy chain of SEQ ID No.: 23 may be encoded by SEQ ID No.: 38 and the humanized heavy chain of SEQ ID No.: 24 may be encoded by SEQ ID No.: 39. Tau-binding antibodies and their linking fragments may be encoded by a single nucleic acid (eg, a single nucleic acid comprising the nucleotide sequences encoding the heavy and light chain polypeptides of the antibody) or by two or more acids. separate nucleic acids, each of which encodes a different part of the antibody or antibody fragment. In this regard, the present disclosure provides one or more nucleic acids encoding any of the aforementioned antibodies or binding fragments. Nucleic acid molecules can be DNA, cDNA, RNA, and the like. For example, the DNA sequences encoding the antibody in whole or in part for the heavy and light chains can be synthesized, as desired, from the determined DNA sequences or on the basis of the corresponding amino acid sequences. DNA encoding acceptor framework region sequences is widely available to those skilled in the art and can be readily synthesized based on their known amino acid sequences. Conventional molecular biology techniques can be used to prepare the DNA sequences encoding the antibody molecule of the present invention. The desired DNA sequences can be synthesized in whole or in part using oligonucleotide synthesis techniques. As appropriate, site-directed mutagenesis and polymerase chain reaction (PCR) techniques may be used. Preferably, the coding nucleic acid sequences are operably linked to the expression control sequences allowing expression in prokaryotic or eukaryotic cells. Expression of said polynucleotide comprises transcription of the polynucleotide into a translatable mRNA. Elements of expression in mammalian cells are eukaryotic, preferably, ensuring regulators well known to those skilled in the art. They generally comprise regulatory sequences that ensure transcription initiation and, optionally, poly-A signals that ensure transcription termination as well as transcript stabilization. Additional regulatory elements may include transcriptional enhancers, as well as translational enhancers and / or heterologous or naturally associated promoter regions. In a further aspect, the present disclosure thus provides cloning or expression vectors comprising said nucleic acid sequences encoding Tau-binding antibodies, as well as linking fragments thereof. A vector is any molecule or composition that has the ability to transport a nucleic acid sequence into a suitable host cell where, for example, synthesis of the encoded polypeptide may take place. Generally, and preferably, a vector is a nucleic acid that has been genetically engineered using recombinant DNA techniques that are known in the art to incorporate a desired nucleic acid sequence (eg, a nucleic acid). nucleic of the present invention). Expression vectors typically contain one or more of the following components (if they are not already provided by the nucleic acid molecules): a promoter, one or more enhancer sequences, an origin of replication, a termination sequence transcription, a complete intron sequence containing a donor and acceptor splice site, a leader sequence for secretion, a ribosome binding site, a polyadenylation sequence, a polylinker region for inserting the nucleic acid encoding the polypeptide a be expressed, and a selectable marker element. Vectors are generally selected to be functional in the host cell in which the vector is to be used (the vector is compatible with the host cell's machinery such that amplification of the gene can occur and / or gene expression). The present disclosure, in a further aspect, thus provides host cells comprising cloning or expression vectors, as described above, and / or nucleic acid sequences encoding Tau-binding antibodies and fragments thereof. binders, as previously described. The host cell may be any type of cell that has the ability to be transformed with the nucleic acid or vector, in order to thereby produce a binding antibody to Tau or its encoded linker fragment. The host cell comprising the nucleic acid or vector can be used to produce the Tau-binding antibody or its linking fragment or a portion thereof (eg, a heavy chain sequence, or a light chain sequence encoded by Tau). nucleic or vector). After introducing the nucleic acid or vector into the cell, the cell is cultured under conditions suitable for expression of the encoded sequence. The antibody, antigen-binding fragment, or portion of the antibody can then be isolated from the cell. • Host cells can be prokaryotic host cells (such as E. coli) or eukaryotic host cells (such as a yeast cell, an insect cell, or a vertebrate cell). The host cell, when cultured under the right conditions, expresses an antibody or its binding fragment which can subsequently be harvested from the culture medium (if the host cell secretes it into the medium) or directly from the host cell that produces it (if is not secreted). Selection of a suitable host cell will depend on a number of factors, such as desired expression levels, polypeptide modifications that are desirable or necessary for their activity, such as glycosylation or phosphorylation, and ease of folding into an active molecule from the point of activation. biological view. Selection of the host cell will depend in part on whether the antibody or its binding fragment is to be post-transcriptionally modified (eg, glycosated and / or phosphorylated). If this is the case, yeast, insect or mammalian host cells are preferred. Suitable mammalian host cells include CHO, myeloma or hybridoma cells. Suitable types of Chinese Hamster Ovary cells (CHO cells) for use in the present invention may include — CHO and CHO-K1 cells including dhfr-CHO cells, such as CHO-DG44 cells and CHODXB11 cells and which can be used with a selectable DHFR marker or CHOKI-SV cells which can be used with a selectable glutamine synthetase marker. Many are available through the American Type Culture Collection (ATCC), Manassas, Va. Examples include mammalian cells, such as Chinese hamster ovary (CHO) cells (ATCC No. CCL61), human embryonic kidney (HEK) 293 or 293T cells (ATCC No. CRL1573), 3T3 cells (ATCC No. CCL92 ) or PER.C6 cells. Other cell types for use in expressing the antibodies include lymphocytic cell lines, eg NSO myeloma cells and SP2 cells, COS cells. Another aspect of the present description provides a process for the production of a binding antibody to Tau or its linking fragment comprising culturing a host cell containing, for example, a vector under conditions suitable to lead to the expression of a Tau antibody. binding to Tau or its binding fragment, eg, from DNA encoding the Tau-binding antibody or its binding fragment, and isolating the antibody molecule. The Tau-binding antibody or its linking fragment may comprise only one heavy or light chain polypeptide, in which case only one heavy chain or light chain polypeptide encoding sequence needs to be used to transfect host cells. For the production of products comprising both heavy and light chains, the cell line can be transfected with two vectors, a first vector encoding a light chain polypeptide and a second vector encoding a heavy chain polypeptide. Alternatively, a single vector may be used, the vector including the light chain and heavy chain polypeptide-encoding sequences. Tau binding antibody or its binding fragment antibodies and fragments according to the present disclosure are expressed at good levels from host cells. In this way, the properties of the antibodies and / or fragments are favorable for commercial processing. Thus, a process is provided for culturing a host cell and expressing the Tau-binding antibody or its binding fragment, isolating the latter, and optionally purifying it to provide an isolated Tau-binding antibody or its binding fragment. In one of the embodiments, the process further comprises the step of conjugating an effector molecule with an isolated antibody or fragment, for example, conjugating with a particular PEG polymer, as described herein. document. The Tau-binding antibody or its binding fragment can be formulated into compositions, especially pharmaceutical or diagnostic compositions. Pharmaceutical compositions comprise a therapeutically or prophylactically effective amount of a Tau-binding antibody or binding fragment thereof in admixture with a suitable carrier, eg, a pharmaceutically acceptable agent. Diagnostic compositions comprise a diagnostically effective amount of a Tau-binding antibody or binding fragment thereof in admixture with a suitable carrier, eg, a diagnostically acceptable agent. Pharmaceutically acceptable agents for use in the present pharmaceutical compositions include carriers, excipients, diluents, antioxidants, preservatives, coloring, flavoring and diluting agents, emulsifying agents, suspending agents, solvents, fillers, bulking agents, buffer solutions, release, tonicity agents, co-solvents, wetting agents, complexing agents, regulating agents, antimicrobials and surface active agents. The composition may be in liquid form or in freeze-dried or lyophilized form and may include one or more lyoprotectants, excipients, surface-active agents, high molecular weight structural additives and / or bulking agents (see, for example, the US Pat. US Patent Nos. 6,685,940, 6,566,329, and 6,372,716). The compositions may be suitable for parenteral administration. Exemplary compositions are suitable for injection or infusion into an animal by any route available to the skilled worker, such as intraarticular, subcutaneous, intravenous, intramuscular, intraperitoneal, intracerebral (intraparenchymal), intracerebroventricular, intramuscular, infraocular, intraarterial, or intralesional routes. A parenteral formulation will generally be a sterile, pyrogen-free, isotonic aqueous solution, optionally containing pharmaceutically acceptable preservatives. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, include saline and buffer medium. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, Ringer's lactate, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer's dextrose, and the like. Preservatives and other additives may also be present, such as, for example, antimicrobial agents, antioxidants, chelating agents, inert gases, and the like. See generally Remington's Pharmaceutical Science, 16th Ed., Mack Eds., 1980, which is incorporated herein by reference. The pharmaceutical compositions described herein may be formulated for sustained or controlled release in a manner that provides for local concentration of the product (eg, bolus, depot effect) and / or increased half-life or stability in a particular local environment. . The compositions may include the formulation of the antibodies, linker fragments, nucleic acids, or vectors of the present invention with particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, etc., as well as agents, such as a biodegradable matrix, particles microcapsules, microcapsules, microspheres, microspheres and bioerodible release particles, implantable injectables that provide controlled or sustained release of the active agent that can then be released as a depot injection. Alternatively or additionally, the compositions may be administered locally through implantation into the affected area of ​​a membrane, sponge, or other suitable material onto which an antibody, binding fragment, nucleic acid, or vector herein has been absorbed or encapsulated. invention. When an implantation device is used, the device may be implanted into any suitable tissue or organ, and delivery of an antibody, linker fragment, nucleic acid, or vector of the present invention may be [SIC] directly through the device via bolus or through continuous administration or through a catheter by continuous infusion. A pharmaceutical composition comprising a Tau-binding antibody or binding fragment thereof may be formulated for inhalation, such as, for example, as a dry powder. Inhalation solutions may also be formulated in a suitable propellant for aerosol delivery. In yet another formulation, the solutions can be nebulized. One aspect of the present disclosure relates to the use of Tau-binding antibodies and their binding fragments as a therapeutically active agent in the treatment of disease. • Another aspect of the present disclosure relates to the use of Tau-binding antibodies and their binding fragments in the treatment of tauopathies. Tauopathies that have been described as containing Tau inclusions (Clavaguera et al., Brain Pathology 23 (2013) 342-349) include Alzheimer's disease (AD); amyotrophic lateral sclerosis / parkinsonism-dementia complex; argyrophilic granular disease; chronic traumatic encephalopathy; corticobasal degeneration; diffuse neurofibrillary tangles with calcification; Down's Syndrome; British familial dementia; danish familial dementia; frontotemporal dementia and parkinsonism associated with chromosome 17 caused by mutations in MAPT; Gerstmann-Straussler-Scheinker disease; parkinsonism in Guadeloupe; myotonic dystrophy; neurodegeneration with brain accumulation of iron; Niemann-Pick disease, type C; non-Guamanian motor neuron disease with neurofibrillary tangles; Pick's disease; Postencephalitic parkinsonism; cerebral amyloid angiopathy caused by prions; progressive subcortical gliosis; progressive supranuclear palsy (PSP); SLC9A6-related mental retardation; subacute sclerosing panencephalitis; dementia with only neurofibrillary degeneration; and white matter tauopathy with globular glial inclusions. Another aspect of the present description relates thus with the use of Tau-binding antibodies and their linking fragments in the treatment of Alzheimer's disease and / or progressive supranuclear palsy. Correspondingly, the present disclosure also relates to methods for treating tauopathies, in particular Alzheimer's disease and / or progressive supranuclear palsy, by administering a therapeutically active amount of a Tau-binding antibody or its linking fragment, to a subject that needs it. The present disclosure also relates to the use of a Tau-binding antibody or its binding fragment in the manufacture of a medicament for the treatment of tauopathies, in particular Alzheimer's disease and / or progressive supranuclear palsy. In another aspect of the present disclosure the Tau-binding antibody or its binding fragment can be used either alone or in combination with other agents in a therapy. For example, the Tau-binding antibody or its binding fragment can be co-administered with at least one additional therapeutic agent. In certain aspects, an additional therapeutic agent is a therapeutic agent effective for treating the same or a different disorder as the one being treated with the Tau-binding antibody or its linking fragment. The agents Exemplary additional therapeutics include, but are not limited to, the following: cholinesterase inhibitors (such as donepezil, galantamine, rovastigmine, and tacrine), NMDA receptor antagonists (such as memantine), amyloid beta peptide aggregation inhibitors, antioxidants, modulators gamma-secretase inhibitors, NGF gene therapy or nerve growth factor (NGF) mimics, PPARγ agonists, HMS-CoA reductase inhibitors (statins), ampakines, calcium channel blockers, GABA receptor antagonists, glycogen inhibitors synthase kinase, intravenous immunoglobulin, muscarinic receptor agonists, nicotinic receptor modulators, active or passive amyloid beta peptide immunization, phosphodiesterase inhibitors, serotonin receptor antagonists, and additional anti-amyloid beta peptide antibodies or anti-Tau antibodies . Exemplary additional neurological drugs may be selected from a growth hormone or neurotrophic factor; examples include, but are not limited to, Brain-Derived Neurotrophic Factor (BDNF), Nerve Growth Factor (NGF), Neurotrophin-4 / 5, Fibroblast Growth Factor ( FGF, Fibroblast Growth Factor)-2 and other FGF, neurotrophin (NT)-3, erythropoietin (EPO), hepatocyte growth factor (HGF, Hepatocyte Growth Factor), Epidermal Growth Factor (EGF), Transforming Growth Factor (TGF)-al ha, TGFbeta, Vascular Endothelial Growth Factor (VEGF), Interleukin-1 Receptor Antagonist ( IL-lra), Ciliary Neurotrophic Factor (CNTF), Glial Derived Neurotrophic Factor (GDNF), Neurturin, Platelet-Derived Growth Factor (PDGF), Heregulin, neuregulin, artemin, persephin, interleukins, Glial cell line-derived neurotrophic factor (GFR), Granulocyte-Colony Stimulating Factor (CSF), granulocyte-macrophage-CSF, netrins, cardiotrophin-1, hedgehogs, leukemia inhibitory factor (LIF), midkine, pleiotrophin, bone morphogenetic protein (BMP), netrin as, saposins, semaphorins, and stem cell factor (SCF). In certain embodiments, at least one additional therapeutic agent is selected for its ability to mitigate one or more side effects of the neurological drug. Such combination therapies as noted above encompass combination administration (wherein two or more therapeutic agents are included in the same formulation or in separate formulations), and the separate administration, in that case, the administration of the Tau-binding antibody or its binding fragment, may occur before, simultaneously with, and / or following the administration of the additional therapeutic agent and / or adjuvant. Tau-binding antibodies or their binding fragments may also be used in combination with other interventional therapies such as, but not limited to, radiation therapy, behavioral therapy, or other therapies known in the art and appropriate for the neurological disorder being treated. or prevented. Another aspect of the present disclosure relates to the use of Tau binding antibodies and their binding fragments as an active diagnostic agent. An aspect of the present disclosure also relates to the use of Tau binding antibodies and their binding fragments in the diagnosis of tauopathies, in particular Alzheimer's disease and / or progressive supranuclear palsy. Said diagnostic tests can preferably be carried out on the biological samples. A biological sample encompasses a variety of sample types obtained from an individual and may be used in a diagnostic assay or for monitoring. The definition • encompasses cerebrospinal fluid, blood, as well as other liquid samples of biological origin, solid tissue samples, such as a biopsy specimen or tissue cultures, or. cells derived from them and their progeny. The definition also includes samples that have been manipulated in any way after they are obtained, such as by reagent treatment, solubilization, or enrichment for certain components, such as polynucleotides. The term "biological sample" encompasses a clinical sample and also includes cells in culture, cell supernatants, cell samples, serum, plasma, biological fluid, and tissue samples. The term biological sample includes urine, saliva, cerebrospinal fluid, blood fractions such as plasma and serum, and the like. Diagnostic tests can preferably be carried out on biological samples that are not in contact with the human or animal body. Such diagnostic tests are also called in vitro tests. In vitro diagnostic tests can rely on a method for in vitro detection of Tau in a biological sample that has been obtained from an individual that comprises the steps that consist of the following: i) contacting the biological sample with a binding antibody a — Tau or its linking moiety, as described herein; and ii) detecting the binding of the Tau-binding antibody or its Tau-binding fragment, as described herein. By comparing the detected level of Tau to a suitable control, one can then diagnose the presence—or possible presence—of a tauopathy such as Alzheimer's disease and / or progressive supranuclear palsy. Said detection method can then be used to determine whether a subject suffers from, or is at risk of developing, a tauopathy, including determining the pitch (severity) of a tauopathy. Therefore, the present description provides an in vitro diagnostic method of a taupatia such as Alzheimer's disease and / or progressive supranuclear palsy in a subject, comprising the steps that include i) evaluating the level or state of Tau in a biological sample obtained from the subject using a Tau-binding antibody or its binding fragment, as described herein; and ii) comparing the level or status of Tau to a reference, standard or normal control value, which indicates the level or status of Tau in normal control subjects. A significant difference between the level and / or status of the Tau polypeptide in the biological sample and the normal control value indicates that the individual suffers from a tauopathy such as Alzheimer's and / or progressive supranuclear disease. With respect to these various aspects and embodiments that have been described herein, the present description contemplates, inter alia: document forms paralysis, alia, lo 1. An isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof comprises the following: a light chain variable region comprising a CDR1 selected from SEQ ID No.: 1 or sequences at least 90% identical thereto, a CDR2 selected from SEQ ID No.: 2 or sequences at least 90% identical thereto , and a CDR3 selected from SEQ ID No.: 3 or sequences at least 90% identical thereto; and / or a heavy chain variable region comprising a CDRl selected from SEQ ID No.: 4 or sequences at least 90% identical thereto, a CDR2 selected from SEQ ID No.: 5 or sequences at least 90 % identical to it, and / or a CDR3 selected from SEQ ID No.: 6 or sequences at least [SIC] 0% identical to it. 2. A Tau-binding antibody or its binding fragment according to embodiment 1, wherein said Tau-binding antibody or its binding fragment comprises the following: a CDR1 chain variable region selected from SEQ selected from SEQ ID No.: 2, light comprising an ID No.: 1, a CDR2 and a heavy selected CDR3 comprising an ID No.: 4, a CDR2 and / or or a CDR3 selected to Tau or its fragment of embodiment 1 or 2, in a Tau or its fragment of embodiment 1 or 2, in a Tau or its fragment of embodiment 1 or 2, in a Tau or its fragment of embodiment 1 or 2 , in Tau or its embodiment fragment 1 or 2, in Tau or its embodiment fragment 1 or 2, in SEQ ID No.: 3; and a chain variable region CDRl selected from SEQ selected from SEQ ID No.: 5, from SEQ ID No.: 6. 3. A linker-binding antibody according to the form where Xi of SEQ ID No.: 3 is A. 4. A linker binding antibody according to the form where Χχ of SEQ ID No.: 3 is G. 5. A linker binding antibody according to the form where X2 of SEQ ID No.: 6 is A. 6. A linker binding antibody according to the form where X2 of SEQ ID No.: 6 is Q. 7. A linker binding antibody according to the form where X2 of SEQ ID No.: 6 is N. 8. A bond-binding antibody according to the form 100 • where X2 of SEQ ID No.: 6 is D. 9. A Tau-binding antibody or its linking fragment according to embodiment 1 or 2, wherein X2 of SEQ ID No.: 6 is S. 10. A Tau-binding antibody or its binding fragment according to any of embodiments 1, 2, 3, 4, 5, 6, 7, 8 or 9, wherein said Tau-binding antibody or its binding fragment binder is a monoclonal antibody. 11. A Tau-binding antibody or its binding fragment according to embodiment 10, in wherein said Tau-binding antibody or its linking fragment is a chimeric, humanized or fully human antibody. 12. An antibody binding to Tau or its linking fragment according to embodiment 11, wherein said antibody binding to Tau or its linking fragment is a humanized antibody of the IgGl or IgG4 subtype. 13. An antibody binding to Tau or its linking fragment according to any of embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12, wherein said antibody of Tau O binding its linking fragment binds to a phosphorylated Tau region within — amino acids 197 to 206 of SEQ ID No.: 55 101 ♦ 14. A Tau binding antibody or its linking fragment according to any one of embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13, in wherein said Tau-binding antibody or linking fragment binds to soluble forms of human Tau, paired helical filaments (PHF) of human Tau, or both soluble forms of human Tau and paired helical filaments (PHF) of human Tau. 15. An isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof comprises the following: a light chain variable region comprising SEQ ID No.: 7 or sequences at least 80% identical thereto, and / or a heavy chain variable region comprising SEQ ID No.: 8 or at least 80 sequences % identical to her. 16. A Tau-binding antibody or its binding fragment according to embodiment 15, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain variable region comprising SEQ ID No.: 7, and a heavy chain variable region comprising — SEQ ID No.: 8. 102 17. A binding antibody to Tau or its binding fragment according to embodiment 15 or 16, wherein Xx of : SEQ ID No. : 7 is A. 18. A binding antibody to Tau or its fragment binder according to i embodiment 15 or 16, where Χχ of ; SEQ ID No.: 7 is G. 19. A Tau binding antibody or its linking fragment according to embodiment 15 or 16, wherein X2 of SEQ ID No.: 8 is A. 20 A Tau-binding antibody or its linking fragment according to embodiment 15 or 16, wherein X 2 of : SEQ ID No. : 8 is Q. .21. A Tau-binding antibody or its linking fragment according to embodiment 15 or 16, wherein X2 of SEQ ID No. : 8 is N. 22. A Tau-binding antibody or its linking fragment according with embodiment 15 or 16, wherein X2 of i SEQ ID No.: 8 is D. 23. A binding antibody to Tau or its linking fragment according to embodiment 15 or 16, wherein X2 de : SEQ ID No. : [SIC] s. 24. A Tau-binding antibody or its binding fragment according to any of embodiments 15, 16, 17, 18, 19, 20, 21, 22 or 23, wherein said Tau-binding antibody or its binding fragment binding is 103 a monoclonal antibody. Tau binding antibody or its binding fragment 25. A binding antibody according to embodiment 24, wherein said Tau binding antibody or its binding fragment is a chimeric antibody • 26. A Tau binding antibody or its binding fragment according to any of embodiments 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25, wherein said Tau-binding antibody or its linking fragment binds to a region of Tau phosphorylated within amino acids 197 to 206 of SEQ ID No.: 55. 27. An antibody binding to Tau or its linking fragment according to any of embodiments 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or 26, wherein said Tau-binding antibody or binding fragment binds to soluble forms of human Tau, paired helical filaments (PHF) of human Tau, or to both soluble forms of human Tau as the paired helical filaments (PHF) of human Tau. 28. An isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment comprises the following: a light chain variable region comprising SEQ ID No.: 9 or sequences of at least 80% identical to it, and / or a heavy chain variable region comprising SEQ ID No.: 10 or sequences at least 80% identical to it. 29. An isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment comprises the following: a light chain variable region comprising SEQ ID No.: 13 or sequences of at least 80% identical to it, and / or a heavy chain variable region comprising SEQ ID No.: 16 or sequences at least 80% identical to it. 30. A Tau-binding antibody or its binding fragment according to embodiment 29, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain variable region comprising SEQ ID No. : 13, and a heavy chain variable region comprising SEQ ID No. : 16. 31. A Tau-binding antibody or its linking fragment according to embodiment 29 or 30, wherein Xi of SEQ ID No.: 13 is A. 32. A Tau-binding antibody or its linking fragment according to embodiment 29 or 30, 105 wherein Xi of SEQ ID NO: 13 is G-junction. 33. An antibody to Tau or its linking fragment according to embodiment 29 or 30 , where X 2 of SEQ ID No. : 16 is A. 34 . An antibody binding to Tau or its linking fragment according to Embodiment 29 or 30, wherein X2 de ; SEQ ID No.: 16 is Q. 35. A Tau-binding antibody or its linking fragment according to embodiment 29 or 30, wherein X2 of SEQ ID No.: 16 is N. 36. An antibody binding to Tau 0 its linking fragment according to embodiment 29 0 30, wherein X2 of i SEQ ID No.: 16 is D. 37. An antibody binding to Tau 0 its linking fragment according with embodiment 29 or 30, wherein X2 of SEQ ID No.: 16 is S. 38. A Tau-binding antibody or its linking fragment according to embodiment 29, wherein the variable region of heavy chain comprises SEQ ID No.: 14 or 15. 39. A Tau-binding antibody or its linking fragment according to embodiment 29, wherein the light chain variable region comprises SEQ ID No.: 11 or 12. 40. A Tau binding antibody or its 106 linker fragment according to any of embodiments 29, 30, 31, 32, 33, 34, 35, 36, 37, 38 or 39, wherein di This Tau-binding antibody or its binding fragment is a monoclonal antibody. 41. A Tau-binding antibody or its binding fragment according to embodiment 40, wherein said Tau-binding antibody or its binding fragment is a humanized antibody. 42. A Tau-binding antibody or its binding fragment according to embodiment 41, wherein said Tau-binding antibody or its binding fragment is of the IgG1 or IgG4 subtype. 43. A Tau binding antibody or its linking fragment according to any of embodiments 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 or 42, in wherein said γ i Tau binding antibody or its linking fragment binds to a phosphorylated Tau region within amino acids 197 to 206 of SEQ ID No.: 55.44.A Tau-binding antibody or its binding fragment according to any of embodiments 29, 30, 31, 32, 33, 34, 35, , 36, 37, : 38, 39, 40, 41, 42 or 43 , wherein said Tau-binding antibody or linker fragment binds to soluble forms of human Tau, paired helical filaments (PHF) of Tau — human or both soluble forms of human Tau and 107 paired helical filaments (PHF) of human Tau. 45. An isolated Tau-binding antibody or its linking fragment, wherein said Tau-binding antibody or its linking fragment comprises the following: a light chain comprising SEQ ID No.: 19 or at least 70% identical sequences to it, and / or a heavy chain comprising SEQ ID No.: 22 or sequences at least 70% identical to it. 46. ​​A Tau-binding antibody or its binding fragment according to embodiment 45, wherein said Tau-binding antibody or its binding fragment comprises the following: a light chain comprising the . SEQ ID No.: 19, and a heavy chain comprising i SEQ ID No.: 22. 47. A binding antibody to Tau or its linking fragment according to embodiment 45 or 46, wherein Xi of SEQ ID No.: 19 is A. 48. A binding antibody to Tau or its linking fragment according to embodiment 45 or 46, wherein Xl of SEQ ID No.: 19 is G. 49. A binding antibody to Tau or its linking fragment according to embodiment 45 or 46, wherein X2 of SEQ ID No.: 22 is A. 50. A Tau-binding antibody or its linking fragment according to embodiment 45 or 46, 108 wherein X2 of SEQ ID No.: 22 is Q. 51. A Tau 0 binding antibody its linking fragment according to embodiment 45 0 46, wherein X2 of SEQ ID No.: 22 is N. 52. A Tau 0 binding antibody its linking fragment according to embodiment embodiment 45 or 46, wherein X2 of SEQ ID No. : 22 is D. 53. An antibody binding to Tau or its linking fragment according to embodiment 45 or 46, wherein X2 of : l a SEQ ID No. : 22 is S. 54 . A Tau-binding antibody or its linking fragment according to embodiment 45, wherein the heavy chain variable region comprises SEQ ID No.: 14 or 15. 55. A Tau-binding antibody or its linking fragment according to embodiment 45, wherein the light chain variable region comprises SEQ ID No.: 11 or 12 56. A Tau-binding antibody or its linking fragment according to any of embodiments 45, 46, 47, 48, 49, 50, 51, 52, 53, 54 or 55, wherein said Tau-binding antibody or binding fragment thereof is a humanized monoclonal antibody. 57. A Tau-binding antibody or its binding fragment — according to embodiment 56, at 109 • wherein said Tau-binding antibody or its binding fragment is of the IgGl or IgG4 subtype. 58. A Tau binding antibody or its linking fragment according to any one of embodiments 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56 or 57, wherein said Tau-binding antibody or its linking fragment binds to a phosphorylated Tau region comprising within amino acids 197 to 206 of SEQ ID No.: 55. 59. A Tau-binding antibody or its linking fragment according to any of embodiments 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57 or 58, wherein said Tau-binding antibody or linker fragment binds to forms soluble forms of human Tau, paired helical filaments (PHF) of human Tau or both soluble forms of human Tau and paired helical filaments (PHF) of human Tau. 60. An isolated Tau-binding antibody or linking fragment thereof, wherein said Tau-binding antibody or linking fragment binds to a phosphorylated Tau region comprising within amino acids 197 to 206 of SEQ ID No.: 55 61. A Tau-binding antibody or its linking fragment according to embodiment 60, wherein - said Tau-binding antibody or its 110 * linking fragment is a monoclonal antibody. 62. A Tau-binding antibody or its binding fragment according to embodiment 60 or 61, wherein said Tau-binding antibody or its binding fragment is a chimeric, humanized, or fully human antibody. 63.A Tau-binding antibody or its binding fragment according to embodiment 62, wherein said Tau-binding antibody or its binding fragment is a humanized monoclonal antibody or its binding fragment of subtype IgGl or IgG4. 64. A Tau binding antibody or binding fragment thereof according to any one of embodiments 60, 61, 62 or 63, wherein said Tau binding antibody or binding fragment binds to soluble forms of human Tau, paired helical filaments (PHF) of human Tau or to both the soluble forms of human Tau and the paired helical filaments (PHF) of human Tau. 65. An isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof competes for binding to Tau with a Tau-binding antibody or binding fragment thereof according to any of the forms of realization 1, 2, — 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 111 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63 or 64. 66. An antibody binding to Tau or its linking fragment according to the shape of embodiment 65, wherein said Tau binding antibody or binding fragment thereof competes for Tau binding with a Tau binding antibody or binding fragment comprising the following: a light chain variable region comprising SEQ ID No.: 11 or 12, and a heavy chain variable region comprising SEQ ID No. 67. : 14 or 15. An isolated Tau-binding antibody or binding fragment thereof, wherein said Tau-binding antibody or binding fragment thereof binds to substantially the same Tau epitope as a Tau-binding antibody or binding fragment thereof, of according to any of embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66 or [SIC] 67. An antibody binding to Tau or its binding fragment 112 e according to embodiment 67, wherein said Tau-binding antibody or its binding fragment binds to substantially the same Tau epitope as a Tau-binding antibody or binding fragment a binding antibody a Tau or linker fragment comprising the following: a light chain variable region comprising SEQ ID No.: 11 or 12, and a heavy chain variable region comprising SEQ ID No.: 14 or 15. 69. An isolated Tau-binding antibody or its linking fragment according to any one of embodiments 65, 66, 67 or 68, wherein said Tau-binding antibody or its binding fragment is a monoclonal antibody. Tau-binding antibody or its linking fragment 70. A linker according to embodiment 69, wherein said Tau-binding antibody or its linking fragment is a chimeric, humanized, or fully human antibody. 71. A Tau-binding antibody or its binding fragment according to embodiment 70, wherein said Tau-binding antibody or its binding fragment is a humanized antibody of the IgGl or —IgG4 subtype. 113 72. A Tau-binding antibody or its binding fragment according to any of embodiments 65, 66, 67, 68, 69, 70 or 71, wherein said Tau-binding antibody or its binding fragment binds to a phosphorylated Tau region [SIC] amino acids 197 to 206 of SEQ ID No.: 55. 73. A Tau-binding antibody or its linking fragment according to any of embodiments 65, 66, 67, 68 , 69, 70, 71 or 72, wherein said Tau-binding antibody or binding fragment binds to soluble forms of human Tau, paired helical filaments (PHF) of human Tau, or to both soluble forms of human Tau and paired helical filaments (PHF) of human Tau. 74.An isolated Tau-binding antibody or its binding fragment according to any one of embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72 or 73 wherein said Tau-binding antibody or binding fragment thereof is a Fab, Fab', an F(ab')2, an Fd, and an Fv, a scFv , a Fab-Fv, Fab-scFv, Fab-dsFv, Fab-scFc, scFv-scFc, dsscFv, dsscFv-scFc, a diabody, a triabody, a 114 tetrabody, a linear antibody, or an antibody containing VHH (variable domain heavy chain). 75. An isolated nucleic acid molecule encoding the heavy and / or light chain of a Tau-binding antibody or its linking fragment according to any of embodiments 1, 2, 3, 4, 5, 6, 7 , 8, 9 , 10 r 11 , 12 1 3, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 , 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57 , 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, or 74. 76. A cloning vector or expression comprising a or more nucleic acid sequences according to embodiment 75. 77. A host cell comprising one or more nucleic acid sequences according to embodiment 75 or one or more cloning or expression vectors according to embodiment 75. embodiment 76. 78. A host cell according to embodiment 77 that is not a human embryonic stem cell. 79. A method for the production of a Tau-binding antibody or its binding fragment according to any of embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 21, 115 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, . 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73 or 74 comprising, at least, the steps consisting of the following: a) cultivating a cell according to embodiment 77 or 78, and b) isolating said Tau-binding antibody or binding fragment thereof. 80. An isolated Tau-binding antibody or its linking fragment according to any of embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, , 65, 66, 67, 68, 69, 70, 71, 72, 73 or 74 for use as a therapeutically active agent. 81. An isolated Tau-binding antibody or its linking fragment according to any of embodiments 1, 2, 3, 4, 5, i 6, 7, 8, 9, 10, 11, 12, 13, 14 , 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 , 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 , 65, 66, 67, 68, 69, 70, 71, 72, 73 or 74 for use in treating a taupatia. 116 • 82. An isolated Tau-binding antibody or its binding fragment for use according to embodiment 81, wherein said tauopathy is Alzheimer's disease. 83. An isolated Tau-binding antibody or its binding fragment for use according to embodiment 81, wherein said tauopathy is progressive supranuclear palsy. 84. A method for the treatment of a tauopathy comprising the step of administering a binding antibody to Tau or its binding fragment according to any of the forms of: embodiment 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, or 74 to a subject in need. 85. A method according to embodiment 84, wherein said tauopathy is Alzheimer's disease. 86. A method according to embodiment 85, wherein said taupaty is progressive supranuclear palsy. 87.An isolated Tau-binding antibody or its binding fragment according to any of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 117, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73 or 74 for use as a diagnostic agent. 88. An isolated Tau-binding antibody or its linking fragment according to any of embodiments 1, 2, 3, 4, 5, '6, 7, 8, 9, 10, 11, 12, 13, 14 , 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 , 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 , 65, 66, 67, 68, 69, 70, 71, 72, 73 or 74 for use in diagnosing a tauopathy. 89. An isolated Tau-binding antibody or binding fragment thereof for use according to embodiment 88, wherein said tauopathy is Alzheimer's disease. 90. An isolated Tau-binding antibody or its binding fragment for use according to embodiment 88, wherein said tauopathy is progressive supranuclear palsy. In the following, the present invention is described with respect to some examples, which, however, are not intended to be limiting. Experiments 118 Experiment 1 - Generation of Tau-binding Antibodies 1.1 Design and Production of Tau Peptides Peptides and immunogens were provided by Peptide Protein Research Ltd., Bishop's Waltham, R.Ü., and were synthesized by in-phase Fmoc peptide chemistry. solid according to the method of Atherton and Sheppard. (Ref: Atherton, E.; Sheppard, R.C. (1989). Solid Phase peptide synthesis: a practical approach. Oxford, England: IRL Press). Peptides containing phosphoserine (pSer), phosphothreonine (pThr), 3-nitrotyrosine (nTyr), and Ν-ε-acetyl lysine (aLys) were synthesized using the Fmoc-protected precursors FmocSer(PO(OBzl)OH)-OH, Fmoc -Thr(PO(OBzl)OH)-OH, Fmoc-Tyr(3NO2)-OH and Fmoc-Lys(Ac)-OH. A peptide was designed and used to produce Tau-binding antibodies that can recognize all Tau isoforms that are post-translationally modified with phosphoserine, phosphothreonine, and / or nitrosotyrosine; it represents residues 197 to 206 as they align to Tau isoform 2 (SEQ ID NO: 55, üniprot code: P10636-8, NCBI ref: NP_005901.2): N-acetyl-nTyr pSer pSer Pro Cys* pSer Pro Gly pThr Pro-amide (peptide designated T197, and defined in SEQ ID NO: 119-56). The N and C termini of peptide were capped with acetyl and amide groups respectively and cys* means that the thiol group, with cysteine ​​side chain, is the conjugation point for binding to carrier protein or biotin for preparation of immunogens. or assay reagent respectively, as described below. Assay reagent for monitoring antiserum titers was prepared by reacting equal masses of maleimido-PEG-biotin and peptide. Three different immunogens were prepared by reacting the peptide with the following carrier proteins that were substituted with maleimide groups on the ε-amino lysine side chains: Hemocyanin extracted from Keyhole Limpet Haemocyanin (KLH), bovine serum albumin ( BSA, Bove Serum Albumin) and ovalbumin (OVA, OVAlbulmin). 1.2 Immunization Two female New Zealand White rabbits (>2 kg) were immunized subcutaneously with 500 pg of total peptide mixture (T197 and T211) emulsified in an equal volume of Complete Freund's Adjuvant (CFA) by mixing vigorously. with a syringe. Peptides were designed conjugated to KLH, OVA and BSA and alternately immunized. The rabbits received 2. 120 booster injections at 21-day intervals using Incomplete Freund's Adjuvant (IFA) with bleeds taken, from the ear, 14 days after immunization. Termination occurred 14 days after the final boost with single cell suspensions of spleen, peripheral blood mononuclear cells, and bone marrow prepared and frozen in DMSO / 10% FCS at -80°C. 1.3 B cell culture B cell cultures were prepared using a method similar to that described by Zubler et al. (1985). Briefly, Peripheral Blood Mononuclear Cell (PBMC)-derived B cells from immunized rabbits were cultured at a density of approximately 3000 cells per well in 200-barcoded 96-well tissue culture plates. μΐ / well of RPMI medium (Roswell Park Memorial Institute Medium, Roswell Park Memorial Institute medium) 1640 (Gibco BRL) supplemented with FCS (Fetal Calf Serum) 10% (PAA laboratories ltd), 2% HEPES ( 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) (Sigma Aldrich), 1% L-Glutamine (Gibco BRL), 1% penicillin / streptomycin solution (Gibco BRL), 0 .1% β-mercaptoethanol (Gibco BRL), 3% 121 culture supernatant of activated splenocytes and mutant EL4 murine thymoma cells irradiated with gamma rays (5xl04 / well) for seven days at 37°C in a 5% CO2 atmosphere. In total, approximately 1.2 x 107 B cells were sampled. 1.4 Primary detection The presence of T197 peptide-specific antibodies in B-cell culture supernatants was determined using a homogeneous fluorescence-based binding assay using Superavidin™ beads (Bangs Laboratories) coated with biotinylated T197 peptide as a source of target antigen. Screening included transfer of 10 ul of supernatant from barcoded 96-well tissue culture plates to barcoded black-walled 384-well assay plates containing immobilized T197 on beads (10 ul / cavity) using a Matrix Platemate liquid handler. Binding was revealed with a goat anti-rabbit IgG Fcy-specific Cy-5 conjugate (Jackson). Plates were read on an Applied Biosystems 8200 Cell Detection System. 1.5 Secondary detection Following primary screening, positive supernatants were consolidated into standard 96-well barcode plates using a scanning robot. 122 Notice Onyx sample rearrangement and B cells were frozen in the cell culture plates at [SIC] 800C. The plates were then subjected to an ELISA assay on peptide T197 and also on streptavidin alone. This was done in order to determine the peptide specificity for each well, and to exclude false positive wells showing non-specific binding to Superavidin beads. The ELISA assay included capture of biotinylated T197 on streptavidin-coated Maxisorp 384-well plates (ThermoScientific / Nunc) in carbonate coating buffer (dH2O + 0.16% Na2CO3 + 0.3% NaHCO3). Plates were blocked with 1% w / v PEG / PBS and then incubated with 10 ul / well of B cell culture supernatant (diluted 1:1 with blocking buffer). HRP-conjugated goat anti-rabbit IgG fe secondary antibody (Stratech Scientific Ltd / Jackson ImmunoResearch) was added to the plates, followed by visualization of binding with a TMB substrate (3,3',5,5'-Tetramethylbenzidine , from EMD Millipore; 10 μΙ / well). Optical density was measured at 630 nM using a BioTek Synergy 2 microplate reader. Primary binding assay identified 880 hits and after ELISA detection, 406 of them were shown to bind specifically to T197. B-cell supernatants that demonstrate specificity 123 to T197 were selected for further analysis by Biacore to identify those showing the best affinity. 1.6 Variable region recovery To allow recovery of antibody variable region genes from a selection of wells of interest, a deconvolution step had to be performed to allow identification of antigen-specific B cells in a given well containing a population of B cells. heterogeneous B cells. This was achieved using the Fluorescent Spotlight method (Clargo et al., 2014). Briefly, immunoglobulin-secreting B cells from a positive well were mixed with streptavidin beads (New England Biolabs) coated with biotinylated peptide T197 and a final 1:1200 dilution of a specific FITC (Fluorescein IsoThioCyanate) conjugate. to the goat anti-rabbit Fcy fragment (Jackson). After static incubation at 37°C for 1 hour, antigen-specific B cells could be identified due to the presence of a fluorescent halo surrounding the B cell. A number of these individual B cell clones, identified using an Olympus microscope , was then collected with an Eppendorf micromanipulator and deposited in a PCR tube. 124 Antibody variable region genes were recovered from single cells by reverse transcription (RT)-PCR using heavy and light chain variable region specific primers. Two rounds of PCR were performed on a Aviso Onyx liquid handler robot, with restriction sites incorporating 2nd PCR nested at the 3' and 5' ends, which allowed cloning of the variable region into a mammalian expression vector with Rabbit IgG (VH) or rabbit kappa (VL). Anti-T197 antibody genes from 31 different wells were successfully cloned into expression vectors. The heavy and light chain constructs were co-transfected into HEK-293 cells using Fectin 293 (Invitrogen) and expressed recombinant antibody in a 125 ml Erlenmeyer flask in a 30 ml volume. After 5-7 days of expression, supernatants were collected and purified using affinity chromatography. Experiment 2 - Additional detection of identified antibodies 2.1 Tau production in E. coli Genes encoding the different Tau isoforms were synthetically generated and codon optimized for expression in E. coli. Conventional molecular biology techniques were used to sub-clone into a 125 modified pET32 vector engineered to produce Tau with a 6His-TEV tag at the N-terminus. E. coli BL 21 (DE3) cells were transformed with the aforementioned vector, and the protein was expressed by conventional techniques. E. coli cells were then recovered by centrifugation, lysed and Tau protein was captured from the soluble fraction by affinity chromatography using NiNTA (Qiagen). The 6His tag was removed using TEV protease followed by a second NiNTA chromatography step. The purified Tau was buffer exchanged into appropriate buffer solutions depending on the application. Endotoxin was removed from samples generated for immunizations using Proteus NoEndo™ columns (Vivaproducts). Generation of Isotopically Labeled Tau for Nuclear Magnetic Resonance (NMR) Studies: Protein expression was carried out as described above, except that minimal media was used for incorporation of 15N, 13C and 2H into the protein. E. coli cell pellets were lysed and the Tau protein was purified using a NiNTA affinity chromatography step (Qiagen), the 6His tag was removed with TEV protease, and then the Tau protein was purified by gel filtration using a Superdex 200 unit (GE-Healthcare). 126 2.2 Tau production in HEK293 A gene encoding Tau isoform 2 was synthetically generated using the wild-type DNA sequence. Standard molecular biology techniques were used to sub-clone it into a pMV-IOHisTEV expression vector (containing a CMV (CytoMegaloVirus, cytomegalovirus) promoter) engineered to produce Tau with an lOHis-TEV tag at the N-terminus. The resulting vector was transfected using the Expi293™ Expression System (Invitrogen) following the manufacturer's protocols. This system uses human Expi293F cells derived from the HEK293 cell line. Tau protein was accumulated in the culture medium from which it had been recovered using immobilized metal ion affinity chromatography on Ni Sepharose Excel column (GE Healthcare). The lOHis tag was then removed using TEV protease before reapplying it to the Ni Sepharose column and the flow-through cleaved Tau collected. The purified Tau was exchanged from the buffer solution into the appropriate buffer solutions depending on the application. 2.3 Preparation of Tau PHF fibrils from human brain samples 127 Paired helical filament (PHF) Tau protein was purified from the brain samples of donors with Alzheimer's disease or progressive supranuclear palsy, according to the protocol published by Ksiezak-Reding and Wall (Neurobiology of Aging 15, 11-19 , 1994). Fractions 8 (equivalent to sucrose gradient centrifugation before crude PHF-Tau in this reference) and 11 (equivalent to fraction A2, with soluble PHF in SDS as described in this reference) which have been previously described as enriched in PHF-Tau were recovered and used for the BIAcore assay and the cell assay of Experiment 3 2.4 Detection by ELISA The purified antibody was then subjected to further detection by ELISA and by Biacore to confirm the activity of the recombinant antibody and to select the highest affinity and most specific antibody. Again, the ELISA assay included capture of biotinylated T197 in Maxisorp 384-well plates (ThermoScientific / Nunc) coated with streptavidin in carbonate coating buffer (dH2O + 0.16% Na2CO3 + 0.3% NaHCO3). Separate plates were also coated with different Tau peptides with mapping of alternative regions of the Tau molecule to verify binding specificity to the T197 sequence only. 128 Plates were blocked with 1% w / v PEG / PBS and then incubated with various dilutions of purified transient supernatant. Secondary HRP-conjugated goat anti-rabbit IgG fe antibody (Stratech Scientific Ltd / Jackson ImmunoResearch) was added to the plates, followed by visualization of binding with TMB substrate (3,3',5,5'Tetramethylbenzidine, from EMD Millipore ; 10 μΙ / well). Optical density was measured at 630 nM using the BioTek Synergy 2 microplate reader. Data for the selected antibody, ABl with rabbit VL of SEQ ID No. 7 and rabbit VH of SEQ ID No. 8, are shown in Figure 1. As can be seen, ABl shows highly selective binding only to T197 and not to the four peptides that correspond to other regions of the molecule. Tau: T174 N-acetyl-C*K pTPPAPKpTPP amide (SEQ ID No. 62) ; T211 N-acetyl- R pT P pS L P pT P C* amide (SEQ ID No. 63) ; T230 N-acetyl-R pT P P K pS P pS SC* amide (SEQ ID No. 57); Y T396 N-acetyl-C*pS P V V pS G D pT pS amide (SEQ ID No. 64) ; position of biotin conjugation. 2.5 Detection using BIAcore 129 Selected rabbit anti-Tau IgG antibody clones were transiently expressed, purified and analyzed using the SPR Biacore T200 platform. First, antibodies were captured on a CM5 sensor chip using immobilized goat anti-rabbit Fe F(ab') 2 gamma reagent. Flow cells 2, 3 and 4 showed immobilization levels between 5600 and 6100 RU, while flow cell 1 blocked (dextran only) served as a control. Purified IgGs were diluted to 0.5 gg / ml in HBS-EP+ buffer from GE Healthcare and captured on the chip at a flow rate of 10 μΐ / min. Each capture step was followed by 180 s injections of analyte, with a peptide / streptavidin complex, control buffers, and the dephosphorylated peptide / streptavidin complex. Peptides were dephosphorylated using 10 μΐ of a 100 μΜ peptide solution in HBS-EP+ containing 3 mM EDTA and 1% DMSO, diluted 1:10 in HBS-N buffer (GE Healthcare). containing 0.3 mM EDTA and saturating EDTA with 2 μΐ of 0.5M MgC12. 2 μΐ of a 1:10 dilution (in HBS-N) of Calf Intestine Alkaline Phosphatase (NEB, catalog number M0290S) was added and incubated at 37°C for one hour, then stored at 4°C. The phosphatase reaction was inhibited by adding 1 μΐ of 0.5M EDTA to the 130 mix. The solution was used for the preparation of the dephosphorylated peptide / streptavidin complex. For the rabbit antibody with a VL region of SEQ ID No.: 7 and a VH of SEQ ID No.: 8, the T230 peptide from the Tau region 230-238 with the amino acid sequence of SEQ ID No. No.: 57 as a negative control. The data was fitted with a bivalent analyte model. Table 1 shows the measured affinity values ​​for this antibody when bound to phosphorylated and dephosphorylated T197 and T230 peptide / streptavidin complexes. Tau Peptide Binding (RU) Ka (1 / Ms) Kd (1 / s) KD (nM) T197 61 9.2E+05 2.6E-03 3 T197 deph. 1 No significant binding T230 0 No significant binding T230 desfos. 0 No significant union Table 1 Selected monoclonal Fab fragments (mFab) were prepared from murine mABl antibody with light chain of SEQ ID No.: 58 and heavy chain of SEQ ID No.: 59 using the Pierce Ficin Cleavage Kit (Catalog Number 44980). , Thermo Scientific) according to the manufacturer's protocol. Absorption at 280 nm was used to determine the concentration of the Fab stock solutions for Biacore analysis. A 131 insoluble Tau protein preparation from patients with Alzheimer's disease (AD-PHF, fraction 11), HEK-derived Tau isoform 2 monomers (amino acids 1-441), and isoform 2 monomers expressed in E. coli were amine-immobilized on the CM5 chip, and the binding of anti-Tau mFabs was measured with the Biacore T200 instrument. GE Healthcare HBS-EP buffer was used for immobilizations apart from AD-PHF for which 10 mM acetic acid (pH 3.0) was used. HBS-EP+ buffer was supplemented with 300 mM NaCl and 1.25% CM-Dextran (Sigma) and used as the assay buffer. While using (Fe)1 flow cell as a reference, the following RU values ​​were obtained for Fc2-4: 44 RU with 5 ug / mL Tau E. coli, 56 RU with 5 ug / m Tau HEK , and 500 RU with a 1:20 diluted solution of the AD-PHF material. Two cycles of 60s of 10 mM Glycine (pH 1.7) were used for regeneration. Flow rates of 10 ul / min were used for immobilization and regeneration, while a flow rate of 30 ul / min was used for analyte binding. For AD-PHF, multiple manual injections were applied up to 500 RU, including EDC / NHS and EtoA covers. Five start-up cycles and 12 control cycles per mFab or buffer sample were applied, using 90 ul analyte injections for either 180s or 300s. 132 for dissociation. Eleven 1:3 dilutions of 600 nM solution plus buffer were used for each mFab. Sample Rmax ka (1 / Ms) kd KD (RU) (1 / s) (M)* 101.4 E. coli iso-2 No significant binding (control HEK iso-2 No significant binding isotype) AD-PHF No significant binding mABl E. coli iso-2 No significant binding HEK iso-2 No significant binding AD-PHF 5 7.96E+04 4.70E-02 5.90E-07 Table 2. Binding of mFab from mABl and control antibody 101.4 to monomeric Tau isoform 2 expressed in E. coli, mammalian HEK293 cells, and Tau PHF fibrils isolated from Alzheimer's disease patients. 2.6 Epitope mapping using BIAcore Recombinant Tau isoform 2 monomer (amino acids 1 to 441), expressed and purified from HEK cells, and insoluble Tau protein in the form of a paired helical filament preparation isolated from brains of patients with Alzheimer's disease (AD-PHF), were immobilized with amine on the CM5 chip with the use of a Biacore 3000 instrument and HBS-EP (GE Healthcare) as a passing buffer. The former was amine-coupled to flow cells 2 and 4 respectively, after surface activation with carboxymethyldextran. 133 «I of these and reference flow cells, by injecting 70 pl of a fresh mixture of 50 mM N-hydroxysuccimide and 200 mM l-ethyl-3-(3-dimethylaminopropyl)carbodiimide, with a flow rate of 10 µΐ / min. Tau monomer immobilization was achieved by injecting 160 pl at 50 pg / ml in 10 mM acetate buffered at pH 5.0, while AD-PHF was immobilized by injecting twenty 160 pl aliquots at 2 pg / ml. in 10 mM acetic acid (pH 3.0). Control and test flow cell surfaces were quenched with a 50 µl pulse of 1 M ethanolamine.HCI, pH 8.5. Epitope mapping was carried out using preincubation solutions of a humanized Tau-binding antibody having a light chain of SEQ ID No.: 17 and a heavy chain of SEQ ID No.: 20 (L17H20) and peptides test or buffer controls, prepared in passing buffer at 200 nM and 5000 nM respectively. These were tested individually in a series of sensorgram cycles with a constant flow rate of 10 μl / min by injecting 50 μl, over control, monomer tau and AD-PHF flow cells. Units of antibody binding response were recorded on spot reports taken 15 seconds after completion of each injection as the difference between test values ​​and reference flow cells. The 134 ChIP was regenerated at the end of each cycle by two 20 µl injections of 1.5 M guanidine in phosphate buffered saline. Table 3. Keys: nY is nitrotyrosine, pS is phosphoserine; pT is phosphothreonine; n.s. not significant to a confidence level of 95%. Peptide Tau Sequence Percent Inhibition ID 197 198 199 200 201 202 203 204 205 206 HEK PHF iso-2 T197 nY pS pS P G pS P G pT P 54 100 T197B Y S S P n.s. n.s. T197C Y pS T197E nY pS T197F nY pS T197G nY pS T197H nY pS T197I pS P G pS P S P G pS P pS P G S P pS P G pS P pS P G G pS P G pT P 49 G pT P 52 G pT P 30 G Τ P n.s n. s G pT P 55 101 102 n.s. n.s. 101 135 Antibody reactivity to a given test peptide was evident by the percentage level of inhibition of antibody binding to either immobilized Tau monomer or AD-PHF relative to the average value of antibody binding calculated for control cycles. Test peptides were prepared and analyzed on the basis of peptide-containing residues 196 to 206 of Tau protein (SEQ ID No.: 65), and the role of post-translational modification with phosphoserine, phosphothreonine and / or nitrotyrosine, as defined in Table 3. Based on this analysis, it is concluded that the minimal epitope is the phosphorylated Tau region defined by amino acids 201 to 206 of the SEQ ID No.: 55, (corresponding to the predominant Gly pSer Pro Gly pThr Pro), and that said epitope has an absolute requirement for the presence of phosphothreonine at position 205 of SEQ ID No.: 55, and of phosphoserine at position 202 of SEQ ID No.: 55. Experiment 3 - Further characterization of the antibodies identified 3.1 Cellular assay Preparation of Crude Soluble and Insoluble Fractions of Tau Transgenic Mice to Induce Tau Aggregation — Mice were used for these experiments. 136 transgenics expressing human Tau P301S (Alien et to the. , 2002 J. Neurosci. 22 (21) :9340-51, and P301L (Lewis al., 2000 Nat Genet. (4):402-5.,- Gótz J, et al., 2001 Biol Chem. 276(1):529-34) . The crude soluble and insoluble fractions prepared from the brain of P301S and P301L Tau transgenic mice by differential centrifugation. Briefly, brain tissues from P301S (spinal cord and brainstem) and P301L (midbrain and brainstem) Tau transgenic mice were homogenized in cold TBS (Fisher Scientific) using the Pellet Pestle Motor handheld homogenizer (Kontes) at 1 .5 mL microfuge tubes on ice. The homogenates (H) were then centrifuged at 4000 g for 10 min, at 'C, to remove tissue debris. The resulting supernatants (SO) were centrifuged at 20,000 g for 20 min, at 4 °C, to obtain supernatants corresponding to the crude soluble fraction (SI). The remaining pellets (Pl) were resuspended in 1 ml of 1% sarcosyl solution prepared in TBS, incubated for 1 h at room temperature and then centrifuged at 100,000 g for 1 h at 4 °C. The supernatants (S2) were discarded. The pellets (P2) were washed with 5 ml of cold TBS, and then resuspended in TBS to obtain the crude insoluble fraction (P2'). 137 Preparation of HEK-293-F cells expressing human Tau with P301S mutation HEK-293-F cells (Life Technologies) were transfected with the pDNA3.1(+) cDNA vector expressing human Tau isoform 2 with a P301S mutation, using 293 Fectin (Life Technologies) according to the manufacturer's instructions. Aliquots of transfected cells were stored in liquid nitrogen. Induction of Tau aggregation Figure 2 illustrates the different steps of the cell aggregation assay used to characterize the activity of therapeutic Tau antibodies. On day 1, HEK-293-F cells expressing P301S-mutated human Tau isoform 2 (P301S-tau) were thawed at 37 °C and diluted in 293 expression medium (Life Technologies) containing 10% fetal bovine serum and 1% Penicillin-Streptomycin (FFBS). Cells were counted using an automated cell counter (Vi-CELL XR, Beckman Coulter), and then plated onto poly-D-lysine precoated 96-well plates (Greiner Bio-One) at a density of 25,000. living cells per cavity. Cells were kept at 37°C in 5% CO2. On the same day, sonicated insoluble human Tau obtained from Alzheimer's disease patients (AD-PHF, fraction 8) 0 was incubated with 138 progressive supranuclear palsy (PSP-PHF, fraction 8) or with frontotemporal dementia (FTD-PHF) or the brain fractions of P301S or P301L transgenic mouse brains, (used as seeds to induce Tau aggregation), with or without antibodies anti-Tau in FFBS medium at 4 °C with gentle shaking overnight. AD-PHF, fraction 8 at 80 ng / μΙ and 60 ng / μΐ was used for the AD and PSP samples, respectively; the soluble brain fraction from P301S and P301L transgenic mice was used at 0.1 pg / pl to 1.2 pg / pl, respectively. On day 2, the seeds or seed / antibody mixtures were applied to the cells for 24 h. On day 3, the culture medium was replaced with fresh FFBS medium containing the antibody, and the cells were maintained in culture for another 24 h. On day 4, Tau aggregation was measured using a Tau aggregation assay kit (Cisbio) on the basis of Homogenous Time-Resolved Fluorescence (HTRF) energy transfer, according to the instructions. manufacturer. Fluorescence was measured with SpectraMax Paradigm (Molecular Devices). Aggregation was reported as percent aggregation relative to control (-) corresponding to the maximal aggregation response induced by fibrils or exogenous fractions in the absence of antibody. 139 The effect of ABI and other prior art Tau-binding antibodies on induced Tau aggregation was tested. The prior art antibodies were IPN002, World Patent Number WO2014 / 028777A2, PT3, World Patent Number WO2013 / 096380A2, and mAb2.10.3, World Patent Number WO2010 / 142423A2. The results of this trial are summarized in Table 3 and Figure 3. Table 4 summarizes the potency (IC50) and maximum efficacy (Imax at 300 nM) of ABl having a murine VL of SEQ ID No.: 9 and a murine VH of SEQ ID No.: 10 (VL9VH10), of a Tau-binding antibody having the light chain of SEQ ID No.: 17 and the heavy chain of SEQ ID No.:23 (L17H23), a Tau-binding antibody having the light chain of SEQ ID No.: 17 and the heavy chain of SEQ ID No.:24 (L17H24), a Tau-binding antibody having the light chain of SEQ ID No.: 17 and the heavy chain of SEQ ID No. :20 (L17H20), a Tau-binding antibody having the light chain of SEQ ID No.: 17 and the heavy chain of SEQ ID No.:21 (L17H21), and competing antibodies against a range of Tau seed from various brain extracts. While Figure 3 shows the efficacy of a Tau-binding antibody that 140 • _ has the light chain of SEQ ID No.: 17 and the heavy chain of SEQ ID No.: 20 (L14H20), and from a Tau-binding antibody that has the light chain of SEQ ID No. :17 and the heavy chain of SEQ ID No.:21 (L17H21) in a cell aggregation assay using pathological human Tau fibrils from human PSP patients. Table 4 Experiment 3.1 τιΑΒ Tg Mice (P301S) IC50 / Imax Tg Mice (P301L) IC50 Human PSP Samples Human FTD VL9VH10 IC50 Samples: 8 nM IC50:9 nM IC50: 10 nM IC50: ND IC50: InM lmax: 62 % Imax : 81% Imax * 7 9% Imax 54% Imax: 81% L17H23 Not Tested Not Tested Not Tested IC50: ND Not Tested IgGl Imax: 49% L17H24 Not Tested Not Tested Not Tested IC50: ND Not Tested IgGl Imax: 4 7% L17H20 Not tested Not tested Not tested IC50: 42 nM IC50: InM IgG4 Imax 65 or Imax: 79% L17H21 Not tested Not tested Not tested IC50'' 66 nM Not tested IgG4 Imax: 47% IPN002 ic50:nd IC50 :122 nM IC50:ND ICso:2O7 nM IC50: ND Imax 22 % Imax: 73 % Imax: 19 % Imax: 64 % Imax: 50 % PT3 IC50 : 3 5 0 nM IC5o:26 nM 5o ic:32 nM IC50:4 7 nM IC50: InM Imax 56% Imax* 69% Imax: 69% τ R S 5xmax 0 Imax: 80% Mab2.10. 2IC50:ND ic50:nd IC50:ND IC50:ND IC50: ND max 25 ΐ Imax: 29% Imax: 16% Imax: 28%<*) Imax: 30% 141 AT8 Not tested Not tested IC50: ND IC50: ND IC50: ND (MN1020) -^max 19% max 25% -^-max 36 HJ8.5 IC50: ND Not tested IC50: ND IC50: 73 nM IC50: ND Imax: 4 3% -í-max 46% -^-max · 79% -J-max · 67% ND: Not determined. (*) maximum efficiency at 100 nM Other experiments were carried out in the cell assay to determine the activity of ABl having a murine VL of SEQ ID No.: 9 and a murine VH of SEQ ID No.: 10 ( VL9VH10) with the use of Tau seed of P301L (n=2), AD (n=3) and PSP (n=3) to obtain final ID50 values ​​of 15nM, 27nM and 70nM, respectively, and values ​​of Imax of 79%, 68% and 57%, respectively. 3.2 Histological analysis ABI having a rabbit VL of SEQ ID NO: 7 and a rabbit VH of SEQ ID NO.:8, the humanized ABI having a VL of SEQ ID NO.: 8, were tested. 11 and VH of SEQ ID No.: 14 (VL11VH14) or a VL of SEQ ID No.: 11 and VH of SEQ ID No.: 15 (VL11VH15) and the antibodies IPN002, PT3 and Mab2.10.3 of prior art and the optimal concentration was determined using cryosections of human hippocampus from an Alzheimer's disease donor, which had previously been shown to contain pathological Tau structures, using AT8 immunostaining (as described in Braak & Braak, 1995, Neurobiol Aging; 16( 3):271-8). ABI and all 142 prior art antibodies exhibited concentration-dependent specific immunoreactivity apart from 101.4 (negative control antibody). From these data, a single optimal concentration of antibody was selected and used for detection in a panel of six human brain samples. Three samples from donors with Alzheimer's disease or very elderly donors exhibited high levels of Tau pathology (positive Tau pathology detected using AT8 immunostaining), and three from donors without Tau pathology (negative Tau pathology detected using AT8 immunostaining). AB1, VL11VH14, VL11VH15, PT3 and Mab2.10.3 showed a similar pattern of immunostaining in the AT8 positive samples. Specific immunostaining of neurofibrillary tangles (intraneuronal NFT), cytoplasmic Tau, neuritic plaque-like structures, and neuropil strands were observed within the hippocampus and temporal cortex of Tau-positive pathology specimens. However, much less immunostaining was detected in AT8 negative tissues. This result suggests that these antibodies preferentially recognize pathological Tau compared to non-pathological Tau. A similar signal was provided by IPN002 in both positive and negative Tau pathology samples. 143 > _ 3.3 Western Transfer Western blots were carried out using a chemiluminescent readout: Used AD preparations, human PSPs were loaded onto 10% polyacrylamide gels (20 gg protein per lane). Proteins were separated by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and electroblotted onto a PVDF (Polyvinylidene Fluoride) membrane. Membranes were blocked in 4% BSA (bovine serum albumin (in TBST: 50 mM Tris, 150 mM NaCl, 0.05% Tween 20, pH adjusted with HCl to pH 7.6). Membranes were incubated overnight at 4°C with primary antibody or non-immune IgG control antibody, rinsed in TBST, incubated with secondary antibody for 1 hour (mouse antibiotin), rinsed in TBST (Tris~Buffered Saline Tris) were incubated with tertiary antibody for 1 hour (anti-mouse IgG-peroxidase), rinsed in TBST, and developed using ECL (enhanced chemiluminescence). The mABl having a murine VL of SEQ ID No: 9 and a murine VH of SEQ ID No.: 10 binds to pathological Tau from human AD samples, but weakly to PSP. See figure 4. — Experiment 4 - Humanization of antibodies 144 • identified AB1 with VL of SEQ ID No.: 7 and VH of SEQ ID No.: 8 was humanized by grafting the CDRs of the rabbit antibody V region into the framework regions of the rabbit antibody V region. the human germ line. In order to recover antibody activity, a number of framework region residues from the rabbit V regions were also retained in the humanized sequence. These residues were selected using the protocol indicated by Adair et al. (1991) (Humanised antibodies, World Patent Document No. WO91 / 09967). The alignments of the rabbit antibody V region (donor) sequences with the human germline V region (acceptor) sequences are shown in Figures 5 and 6, along with the designed humanized sequences. CDRs grafted from donor to acceptor sequence are as defined by Kabat (Kabat et al., 1987), with the exception of CDR-H1 where the combined Chothia / Kabat definition is used (See Adair et al., 1991 Humanized antibodies, world patent document number WO91 / 09967). The human IGKV1-39 V region plus the JK4 V region (IMGT, http: / / www.imgt.org / ) was selected as the acceptor for the AB1 antibody light chain CDRs. The — light chain framework residues in the gL4 grafts and 145 • gL9 are all from the human germline gene. CDRL3 was mutated in the gL9 graft to modify a possible deamidation site. The human IGHV4-39 V region plus the JH4 V region (IMGT, http: / / www.imgt.org / ) was selected as the acceptor for the ABl antibody heavy chain CDRs. In common with many rabbit antibodies, the AB1 antibody VH gene is shorter than the selected human acceptor. When aligned with the human acceptor sequence, ABl antibody VH region framework 1 lacks the N-terminal residue, which is retained in the humanized antibody (Figure 6>). ABl VH region framework 3 Rabbit also lacks two residues (75 and 76) in the loop between beta sheet strands D and E: in grafts gH41 and gH49 the space is filled by the corresponding residues (Lysine 75, K75; Asparagine 76, N76) of the selected human acceptor sequence (Figure 6).The heavy chain framework region residues in the gH41 and gH4 9 grafts are all from the human germline gene, with the exception of one or more residues from the group comprising the residues 71 and 78 (Kabat numbering), where the donor residues Lysine (K71) and Valine (V78) are retained, respectively Retention of residues K71 and V78 was essential for the full potency of the humanized antibody Glutamine residue in position 1 of the 146 • human framework region was replaced with glutamic acid (El) to obtain expression and purification of a homogeneous product: conversion of Glutamine to pyroGlutamate at the N-terminus of antibodies and antibody fragments is widely reported. CDRH3 was mutated in the gH41 and gH49 grafts to modify a possible deamidation site. Genes encoding a number of variant heavy and light chain V region sequences for each antibody were designed and constructed using an automated synthesis approach using DNA2.0 Inc. Additional variant heavy and light chain V regions were created by modifying the VH and VK genes by oligonucleotide-directed mutagenesis, including, in some cases, mutations within the CDRs to modify potential deamidation sites. For transient expression, the humanized light chain V region genes were cloned into the UCB human light chain expression vector pMhCK, which contains the DNA encoding the human Kappa chain constant region (Km3 allotype). The humanized heavy chain V region genes were cloned into the UCB human gamma-4 heavy chain expression vector pMhy4P FL, which contains the DNA encoding the human gamma-4 heavy chain constant region—with the stabilizing hinge. the S241P mutation 147 • (Angal et al., Mol Immunol. 1993, 30(1):105-8). Alternatively, the humanized VH genes were cloned into the UCB gamma-1 heavy chain expression vector pMhylFL, which contains the DNA encoding the human gamma1 constant region (Glml7, 1 allotype). In order to assess the monovalent binding kinetics of humanized antibodies, humanized VH genes were also cloned into the human UCB Fab-HIS expression vector pMhFablOHIS, which contains DNA encoding the hinge domain of human CHl gamma-1 with a C-terminal tag of ten histidine residues: The histidine tag facilitates purification of expressed Fabs by affinity chromatography. Cotransfection of the resulting heavy and light chain vectors into HEK293 cells in suspension was achieved using 293 Fectin (12347-019 Invitrogen), and expression of humanized recombinant antibodies was obtained in IgG4P or Fab-HIS formats. humans. Humanized antibody chain variants, as well as their combinations, were expressed and evaluated according to their potency relative to the parent antibody, their biophysical properties, and their suitability for further processing. For stable expression of humanized recombinant antibodies in mammalian cells, the gene for 148 the humanized light chain V region was ligated to a DNA sequence encoding the human C-Kappa constant region (Km3 allotype), to create a contiguous light chain gene. The humanized heavy chain genes were ligated to DNA encoding either the human gamma-4P heavy chain constant region or the human gamma-1 heavy chain constant region (Glml7, allotype 1), to create the contiguous heavy chain genes. . The heavy and light chain genes were cloned into a mammalian expression vector. 149 LIST OF SEQUENCES <110> UCB BIOPHARMA sprl <120> Tau-binding antibo drer <230> PFO31-EP-EPA <170> Patentln version 3 5 <210> 1 <211> 11 <2I2> PRT <213> y, 4 4X0 . .. 2 1 5** - ill. L X i 1 Ló d i .2' tr. quenc <22 0 <22 0 CDR-L1 <400> I Gln Al a Ser Gln Ser Val S c, r c; i 3 <210 - <211> 7 < *' 1 > PRT <21 3> Artificial Se quenc 6 <220> <223> CDR-L2 < 4 0 0 > 2 Ala Al ¿t Χ. <8υ Ala S Λ . >* 1 5 <21 0 > 25 < 211 > 1 P <212> PM <210 Artificial Se quenc e <22 0> <22 3> CDR-L3 <220 MISC FEATURE <222> (3) . . (3) <22 3> Xaa can be A or i. G* 150 Gln Gln Xaa Tyr Thr Aro Thr Aso lie Aso Asn Thr 15 1 <210 4 < 211 > 1 v <212> PRT < 213 > Artifi <220> <223> CDR-H1 lequence <4 00> 4 Gly He Asp Leu Ser Thr Trp Arg Met Asn 1 S 1L <210> 5 <211> 16 <212> PR <213> rni Á X ί X i 1C i 3 i. S β Cj U β H C *5 [le lie Gly Thr Gly Gly Arg Thr Tyr Tyr Ala Asn Trp Ala Lys i 5 10 15 Gl < 210 > 6 <213> Artificial Sequencí <220 > <223> CDR-H3 <22i> KISC FEATVRE <222 > (6)..(6) <2^s> xad can be A, Q, N, E' or S <400> 6 Leu Gly Ala Asn Asn Xaa Gly Tyr Pro Leu Asp Le 15 10 151 210> 2 i ¿ < 3> PRT Rabfcit <22 0 > <221> < 2 5* <22 3> <4 C ¿ > Asp Val 1 MISC FEAT URE ) e A T iir x* ox G Gln Thr •srr¡ Ala iZ' e x 10 Val Glu Ala Ala Val 15 Gly (91) · X cí ¿i C 7 val . <91 an b Met Gly Thr Val Thr lie Lys Cys Gln Ala Ser Gln e X Val 3$ r· Ser Tyr 20 25 30 Leu Ala 1 Y- Gln Gln Lys •a x y Gln Pm Pro Ly e Leu Leu lie .5 u 4 ·' c. *3 -U Tyr Ala A.X <3 Ser Tyr Leu Ala C ¿a v- Gly Val Pro Ser Arg Ph e Lys Gly 50 55 60 Ser Gly v 21V Thr 2lu Pile Thr Leu Thr lie Ser a <> v Leu Glu cys c X 7 B / X) 80 Ala Asp .Ale Ala Thr Tyr Tyr cys Gln Gln Xa a Tyr Thr Arg Thr Asp 85 90 95 lie Asp Asn Thr Phe Gly Gly G1 y τ n r* Lys val Val Val 1.4 X U. i ? <.· 10 5 lio <210 > 8 <2il> 117 <212> PRT <213> Rabbi t <22 2 > <22r > ΜI .4 < FEAS URE < ( i ··- 0) . . (x oó) <22 3> Xaa c •an b e A, Q, N, 2 s or r s <400> 152 Gln Si su Glu Glu Ser Gly Gly Arg Leu Val Thr ero Gly Thr Pro 5 10 15 Leu Thr Leu Thr Cys Thr Val Ser Gly lie Aso Leu Tnr Trp Arg 3 0 Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp lie Gly 3 5 4 0 4 5 lie He Gly Th.i ;ly Gly Arg Thr Tyr Tyr Ala Asn Trp Ala Lys Gly 55 60 Arg Phe Thr lie 65 jr Lys Thr Ser Thr Thr Val Asp Leu Lys Val Th Ί0 7 5 3 0 Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyi 85 ’ 90 Phe Cys Ala Arg Leu Gly $5 .a Asn Asn Xaa Gly Tyr P. 100 :o Leu Asp Leu Trp Gly ero Gly Thr Leu 10' 5 110 val Thr Val ser Ser 115 < 210 > 9 <211> 110 <212> PRT <213> Artificial Dry? <223> rnurinised <4 00'** 9 Asp Val Val Met Thr Gln Se< a Ser r e u Ala Va. r S e r rsu G1 * 10 i 5 Gln Arg Ala Thr Ile Ser Cys; Gln Wing Ser Gln 20 25 Val Ser Ser Tyr 30 ,eu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pr; 4 ' Lys Leu Leu He 45 153 Tvr Ala Ala Ser Tyr Leu Ala 5 O 5 5 ily Val Pro Ser Arg ene Lys Gly 60 Gly Ser Gly Thr Glu ?h< 70 Fhr Leu Asn lie His Pro Val Glu Glu 7 5 SO Glu Asp Ala Ala Thr ryr Tyr Cys Gln Gln Gly Tyr ihr Arg Thr Asp I :le Asp Asn Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110 <210> 1.0 <211> 113 <212> PRT <213: Artificial Security <22ú> <223> murinised <400> ΙΟ In Val Gln Leu Lys Glu Giy :o Gly Leu Val Wing 10 gln Ser Leu Ser lie Thr Cys Thr Val Ser Gly lie Asp Leu Ser Thr Trp 20 25 30 :g Mer. Asn Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 11« ?ly lie lie Gly Thr Gly Gly Arg Thr Tyr Tyr Ala Asn Trp Ala Lys 50 55 60 Gly Arg Phe Yes 65 .le Ser Lys Asp Se: 70 Thr Gln Val Phe Leu Lys Met 75 80 Asn Ser Leu Gln. rhr Glu Asp rhr Ala Thr Tyr Phe 8 5 P 0 Cys Ala Arg Leu r. c Pro Leu Asp Leu 105 Gly Wing Gly 110 Gly Ala Asn Asn Asn Gly Tyi 100 154 Thr Val Thr Val Ser Ser <210> 11 <2il> 11.0 <212> PRT <213> Artificial sauencs <22 ú> < 2 2 3 > h urna ñire d <400> 11 Asp He Gln Met Thr Gln 1 5 go Pro be Leu Ser Ala er Val Gly Asp Arg Val Thr lie Thr Cys Gln Ala Ser Gln Ser Val .Ser Ser Ty: Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu lie 35* 40 45 Tyr Ala Ala Ser Tyr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Gly Ser Gly Th he Thr Lí / 0 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Thr Arg Thr Asp 85 90 S 5 lie Asp Asn rhr Phe Gly Gly Gly Thr Lys Val Glu He Lys 10 0 105 110 <210> 12 <21i> 110 <212> ' Pggy <213 > Artificial Sequenci < 2 2 3 .> h urna n i z e d <400> 12 Asp lie Gln Met Thr Gln Yes 1 5 Pro Being Leu ser Ara Ser Val Gly 15 155 Asp Arg Val Thr lie Thr Cys Gln Ala Ser Gln Ser Val Se: 25 30 Leu Ala Trp Tyr Gln Gin Lys Pro Gly Lys Ala Pro Lys Le' 35 4 0 45 Tyr Ala Ala Ser Tyr Leu Ala Ser Gly Val Pro Ser Are; PL 5 0 55 6 0 r Gly Ser Gly Thr Asp ene Thr Leu Thr lie Ser Ser Le to be Tyr leu lie Being Gly σι φ Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Tyr Thr Ar 5 5 90 Thr Asp 95 and Gly Thr Ly 105 Va 1 lu lie Lys <210> 13 <211> 110 <212> PRT <213> Ar t i fi c i a1 Sequence <22 n> < 2 2 3 > Il U1TUH π i Z “ cl <220> <221> MISC_FEATURE <222> (91)..(91) <22 3> Xaa can Pe A or G Asp lie Gln Met Thr Gln S 1 5 u Ser Ala S Val Gly 15 Asp Arg Val Th Thr Cys Gln Ala Ser Gln S 25 Val Ser 0 Tyr Leu Ara Trp n Gln Pro Gly Lys Wing Lys I Ili so 156 Tyr Ala Ala Se: 50 'yr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 55 60 Asp Phe Thr Leu Thr 11« 7 0 7 5 Ser Leu Gln P: Glu Asp » Phe Ala Thr Tyr Tyr 3 t¡> lie Asp^ > Asn Thr Phe Gly Gly 1 0 2 <210 14 <211> 12 0 <210 PRT <213> Artificial Sequence <220> <223 > humanized < 4 0 0 > 14 Slu Leu Gln Leu Gln Glu :io Gly Pro Gly Leu val Lys 10 Glu ihr Leu Ser Leu Thr Cys Thr Val Ser Gly lie Asp Leu Ser Thr Trp 2 o 2 5 3 0 Arg Met Asn 3 5 :le Arg Gln Prc Pro Gly Lys Gly Leu Glu Trp lie 4 V 4 3 Gly lie lie Gly 50 Gly Gly Arg Th 55 Tyr Tyr Wing As 60 . rp Ala Lys Gly Arg Val rhr lie Ser Lys Asp Thr Ser Lys Asn Gln Val Ser 65 70 75 Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 6 9 9 0 9 5 Leu Gi.y Ala Asn Asn Gj Leu A' Leu Trp Gly 110 10: 157 Gly Thr Leu Val Thr Val Ser Ser 115 120 < 2 r * ? > la <21i> 12ú <212> PRT <213> Artificial Sequence < 2 2* O > <223> humane <400 Glu Leu e χ η G1 υ s e r o i y e r o G x y Leu Val x? y s P r c S ¿ 5 10 1: ;hr Leu Ser Leu Thr Cys Thr Val Ser Gly lie Aso Leu Ser Ti 20 25 * 30 Arg Met Asn Tro lie Arg Gln Pro Pro Glv Lví Gly Leu Glu Ti 4 5 Gly lie lie Gly Tin 50 Gly Gly Arg Thr Tyr Tyr Ala Asn xrp to: ου Gly Arg Val Thr 65 Ser Lys Asp xhr Ser Lys Asn Gln Val Yes 70 75 Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cl Arg Leu Gly Ala Asn Asn Ala Gly Tyr 100 105 Pro Leu Asp Leu Trp rr Glu ir Trp :p lie L 3 Lys sr Leu 8 0 / s Ala y Ly Gln ;ly Thr Leu Val 115 'br Val Se: <210> 16 < 211 > 12 0 <212> PRT <213> Artificial Dry* 120 human x thirsty 158 <220> <221 > < 2 2 2 > <22 5> <40 > Glu Leu 1 MISO (10 3) Xaa c G1 n FEATURE . . (103) άπ me A, y, R, D oí s Leu Gln Glu ser Gly P r o Gly 10 Leu Val Lys Pro 15 Glu Thr Leu Ser Leu Thr Cys Thr Val C*p. TU y He Asp Leu Ser- Thr Trp 2 0 2 5 30 Arg Met Asn τ rp ríe Ar y Gln Pro Gly Lys Gly Leu Glu T'-n lie 3 s á V 4.5 Gly lie lie Gly Thr Gly Gly Arg Thr Tyr Tyr Ala Asn Trp Ara Lys 50 €0 Gly Arg V a 1 Thr lie Gcx Lys Asp Tnr S e r Lys Asn G1 n Val Ser Leu & I ,< i o 7 5 í? 0 Lys Leu Ser Ser Val. Thr A i. a Ara Asp Thr Ala Val Tyr Tyr C y s Ala 2 5 9 0 95 Are? Leu Gly Ala Asn Asn Xaa Gly Tyr r—.-, Leu Asp Leu Trp Gly -5 10 ΰ 105 110 Gly Thr Leu Val Thr val Ser Ser i 15 120 <210> 17 <211> 217 <212> PRT <21 3 > Artificial Sequence <22 3> humar izad <4 00> 17 Asp lie Gln Met Thr Gln Ser Pro <ζ »Λ I Ser Leu Ser Ala ser Va 1 Gly ini bin ser rro yes 5 it 159 Aso Aro val Thr lie Thr Cys Gln Ala ¡r Gln Ser Val Se: 0 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu . y x Ai¿t h~¿t. -Yes ;.'yX ΙίβΛΙ Ai.3 SE1 5 5 y v ¿i. Yo Pne Hoop Yes Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Se: 65 70 75 Ser Leu Gln Glu Aunty Thr Tyr Tyr Cys Gln Gly Tyr Thr Arg Th: C¿5 lie Ase Asn Thr Phe Gly' Gly Gly Thr Lys Vs 100 105 5lu lie Lys Arg 1 or Val Ala Ala ero Ser Val Pne lie en 115 120 »r Asp Glu Gln 125 ory Thr Ara Val Val Cys Leu Leu Asn Asn Phe Tyr 135 140 Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 145 '150. 1" Ace: Glu Ser Val Thr Glu Gln Ase1 7 ú lys ace Thr 17 5 be leu ISO leu Thr Leu Ser Lys Ala Asp Tyr Glu Lyí 185 190 Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Se: tyr lie gly asp Thr Leu Pro gly 160 Tyr His Val lvs se: Asn Arg Gly Glu Cy: 21.5 <210> 18 <211> 217 160 <> <2i2> PRT <21.3> Artificial Sequen.cs <223 > h urna n i 2ed <4 00> 18 Asü lie Gln Ket Th val Gly 15 Asp Arg Val Thr lie Thr Cys Gln Ala Ser Gln Ser Val Ser Ser Tyr 20 2 5 3 0 Leu Ala Trp ’iyr Gln Gln Lys Prc Gly Lys Ala Pro Lys Leu Leu ríe 35 40 45 yr Ala Ala 50 íly Val Pro Ser Arg Phe Ser Gly eú Be Gly 65 Glv Thr As Ί»' :*he Thr Leu Thr 11« 75 Ser Leu Gln Pr ílu Asp Phe Ala hr Tyr Tyr Cys Gln Gln Ala Tyr Thr Arg Thr Ae>p =, c¡ c, ς He Asp Asn Thr Phe Gly Gly Gly Thr Lys Val Glulie Lys Arg Ths 100 105 110 Val Wing Wing Pro Ss Val Phe lie Phe Pro Pro 1 ñ Asp. Glu Gln Leu 1 - ς Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 1 3 0 13 5 14 0 3rd Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly 145 150 155 160 Asn Ser Gln Glu Gln Asp 170 Tnr Tyr 175 105 Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His 180 185 ’ 190 161 Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 1.59 <211> 217 <212> PRT <213> Art <220> <223> humanized <2 2 0> <22 i > MISC_PEATURE <' 2 2 2 > (91)..(91) <22Xaa can be A <400> IS Asp lie Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val 1 5 10 15 Asp Arg Val Thr lie Thr Cys Gln Ala Ser Gln Ser Val Leu Ala Trp Tyr Gln Gln Lys Pro GIv Lys Ala Pro Lys Leu Leu 35 40 45 Tyr Ala Al¿ *yr Leu Ala Ser Gly Val 5 5 o s&r Arg me sej 6 f! Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 11« 65 70* 75 Ser Leu Gln Jlu Aso Phe Ala Thr Tyr Tyr Cys Gln Gln Xaa Tyr Thr Arg Thr y V D ll£ Asp Asn Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg 10 5 110 gly TyG lie gly Pro 0 A.s.p. Thr 162 Val Ala Ala Prc Ser Val Phe lie Phe Prc 115 120 :o Ser Asp Glu Gln 12 5 Lys Ser Gly Thr Ala 130 Val Val Cys Leu Leu Asn Asn Phe Tyr 135 140 Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Sei 145 15 0 .15 5 Asn Ser Gln Glu ex Val Thr Glu Gln Asr 6 5 llí :er Lys Asp Ser Th 11!. ser reu Ser Ser rhr ;u Thr Leu Ser Lys Ala Asp Tyr Glu Lys 185 190 L and s Val Tyr Ara c and s 18 5 Glu Val Tm His Gln Gly Leu Ser Ser Pro **> Γ'. c. Leu Fxo gly 160 Tyr His Val hr Lys Yes 210 re Asn Ai Gly Glu Cy; 215 <210 20 <211> 447 <212> PRT <213> Artificial Jeauern <__3> number <400> 20 Glu Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys P ero Se: 15 .su Tnr cys Thr Val ser dy lie Asp Leu Se. ‘0 25 30 Thr Arg Met Asn Trp lie Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp .i 5 4 0 45 rly lie Ilí -r ni. Thr Tyr Tyr Ala Asn Trp Ala 6C glu Trp lie Lys 163 Gly Arg Val Thr lie Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 7 5 Lys Leu Ser Ser Val The Wing Wing Asp Thr Wing Val Tyr Tyr Cys 90 95 Arg Leu Gly Ala Asn Asn Gln Gly Tyr 100 105 so Leu Asp leu Trp Gly 110 Gly Thr Leu Val Thr Val Ser Ser Ala 115 120 ¡r Thr Lys Gly I 5 •ro Leu Ala ero Cys Ser Arg se Λ 1 ? C V x y hr Ser Glu Ser Thr Ala 14 v Leu Gly Cvs 145 Leu Val Lys Asp Tyr Phe Pro Glu ero Val Thr Val i o O 15 5 Tro Asn Ser Glv Ala Leu 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu 160 185 Val Val Thr Val Ser Ser Ser Leu Gly Thr Lvs Thr Tyr Thr Cys Asn Val Aso His ’ro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly 210 215 220 Pro Cys Pro 225 ;o Cys Pro Wing Prc Glu Phe Leu Gly Gly 230 235 rhe Leu Pne ''4: Lys Pro Lys Asp Thr Leu Metlie Ser Arg - c f, — c c: Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 sr Gln Glu Asp Pro 270 r p T y r Val Asp G1y Va1 lu Val His Asn Al« 0 SLeu a? To gln Val To Be 160 goes 1 P 1' C» Lys Val 2 4 0 Thr glu Lys 164 Lys 290 Pr o Arg Glu Glu Gln 2 95 phe Asn Ser Thr Tyr 3 0 0 Arg Val Val. ser Val 3 0 5 Leu Thr Val Leu His Gln 3 1 ·.' Asp Trp Leu As n Gly Lys Glu Tyr Lys 320 Cys Lys Val Ser Asn Lys Gly Leu 3 3 or 3rd lie Glu Lys Tnx lie 335 Ser Lys Ala Lys 3 4 0 Gly Gln. Pro Arg 345 Gln Val Tyr Thr Leu Pro 350 Pro Ser Gln 35 5 Glu Glu Lys 3 6 0 Asn Gln Val Ser Leu 365 Thr Cys Leu Val Lys 3 7 0 Gly Phe Tyr Pro Ser 37 5 Asp lie Ala Val Glu 33 0 Trp Glu Ser Asn Gly 305 Gln Pro Glu A r n Asn Tyr 3 S 0 Lys Thr Thr P r o 3 9 5 Pro Val Leu Asp Ser 4 00 Asp Gly Se X Phe Phe 4 0 5 Leu Tyr Ser Arg Leu / I ,*( Φ H v Va i A sp Lys Ser Arg 415 τρ: ΐ -ρ Gln GlU yiy 41 3 Asn Val Phe Ser Cys Ser 425 Val Met His Glu Ala Leu 4 30 His Asn His Tyr Thr Gln Lys Leu Ser Leu Ser Leu Gly Lys 4.35 44 0 44 5 <21Ο> 21 <211> 4 4 Z r RT Art i fic i a1 Sequence s_ J. <21 ?> n > <22 3 hunianired < 4 0 0 > 1 Glu Le u. Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 10 1 o 165 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly He Asp Leu Ser Thr Trp 2 O 2’ 5 3 O Arg Met. Asn Trp He Arg Gln Pro Prc Gly Ly: Gly Leu Glu Trp 11« 45 Gly Thr 'yr Tyr Ala Asn Trp Ala Ly Glv Arg Val Th :le Ser Lys Asp» Thr Ser Lys Asn Gln Val Ser Leu 70 * 75 80 Lys Leu Se Val Thr Ala Ala A.sp Thr Ala Val Tyr Tyr Cys; Wing 3 5 95; 95 Arg Leu Gly Ala Asn Asn Ala Gly Tyx 10 0 1 «15 ro Leu Asp Leu Trp Gly Gln liü Gly Thr Leu Var Tnr Car Ser Ser Wing S: r Thr Lys Gly Pro Ser Val ?h.e Pro Leu Ala 130 <io Cys ser Arg se: 135 Ser Glu Ser Thr Ala Ala 140 Leu Gly Cys Leu Val Lys Asp Tyr Phí 145 150 Pro Glu Pro Val Thr Val S 15 5 1 . mop; asp. ly Ala Leu Thr Ser Gly Val His Thr 165 170 Ala Val 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser S 18 0 18 5 Val Val Thr Val i 9 0 sr Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Ly: 195 200 * 205 :o ser Asn Thr Lys Val Asp Lys; Arg val Glu Ser Lys Tyr Gly fro .0 215 220 *> 166 Pro Cys Pro íys Pro Ala Pro Glu Phe Leu Gly Gly Pro :so ¡35 Val 240 Phe reu me fr< 'ro i.vs Lys Asp Thr Leu Met Il< ;r Arg Thr c c Gilí Var Thr Cys Val Vdi Val B.S'p Va.i Ser Gln Glu. aso; 6 0 265 2 7 or Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Ly; 275 280 285 hr Lys ring Arg Glu Glu Gln Phe Asn 290 295 Ser Thr'iyr Arg Val Val Se: 300 Val Leu Thr Val Leu His Gln Asp 3 0 5 310 p Leu Asn Gly Lys Glu Tyr Lys 315 320 Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Se: 325 330 He Glu Lys Thr He Lys. Ala Lys 340 o Gln Val Tvr Thr Leu Pro ;o Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 eu Thr Cys Leu pj yes Σ A S O 375 Val Glu Trp Glu Ser Asn 33 0 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 ,su Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg d 1 0 á I S Trp Gln Glu Gly Asn Val Phe Ser Cys S« 4 2 0 4 2 3 Val Met his Glu Ala Leu ¿30 His Asn His Tvr of him go Gln Lys Ser Leu r r i? Leu Gly Ly; 167 <210> AΣ uX Γ XC ίctΪ. S £ U βΩ Ü * <22 0> <223> humanized <22 0> <222> (10ϊ)..(103) <22 3> Xaa can be A, Q, N, D or: <400> 22 Glu Leu Gln Leu Gln Giu Ser Gly 1 5 or Gly Leu Val Lys Pro Ser Glu 10 15 Leu Ser Leu τη or hr Val Ser Gly He Asp Leu Ser Thr Trp 2 or 3 0 Arg Met Asn 35 -P He Arg Gln Prc 4 0 ;o Gly Lys Gly Leu Glu Trp Ilí 4 'le He Gly Thr Gly Gly Arg Thr Tyr Tyr Ala Asn Trp Ala Ly; Gly Arg Val Tr 65 ir laughs be Lys Asp Tnr iex Lys Asn Gln Val Ser L? 75 8C Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 6 5 S 0 9 5 Arg Leu Gly Ala Asn or 0 Asn Xaa Gly Tyr Pro Leu Asp Leu Trp Gly Gln 10 5 110 Gly Thr Leu Val Thr Val ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Leu Ala Pro Cys Ser Arg 1 . / Glu Ser Thr Ala Alt I 4 or 168 Leu Gly Cys 145 su Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Asn Ser Gly Ala Leu Thr Ser Gly Val Hís Thr Phe Pro Ala i oo 4 7 ·} i 7 9 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 * 185 190 •Y Ti ir Lys Thr ’hr Cys A.sn Val Ase His Be 160 val hoop Lys ;o Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly 210 215 220 Pro Hoop Wing Glu Phe Leu Gly Gly Pro Yes Val 2 40 Phe Leu Phe er< Pro. Lyí 245 ,ys Asp 'Thr Leu Met lie Ser Arg Thr ;lu Val Thr Cys Val Val Val Ase Val :er Gln Glu Asp Pro 270 glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 27 5 2 8 0 2 85 Lys Thr Lys Pro Arg Glu Glu Gln Phe As rhr Tyr Arg Val Val 3 0 0 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 ’ 315 Lys 320 ::ys Lys Val Ser Asn Lys Gly Leu Pro, Ser Ser lie Glu Lys Thr 325 330 335 I le Ser Lys Aia Lys Gxy Gln ?o Arg Glu aro Gln Val Tyr Tnr Leu 3 4 5 35 0 Pro Gln Glu Glu Met 355 ir Lys Asn Gln Val Ser Leu Thr Cys .8 60 3 c 9 Leu 169 Val Lys Gly Phe Tyr P 370 rc- Ser Asp lie Ala Val Glu Trp Glu Ser Asn 37 5 3 8ú Gly Gln u Asn Asn Tyr 9 0 : Lys Thr Thr Prc 355 Pro Val Leu Asp 00 A s or G .X. And be Phe Phe Leu Tyr Ser Arg Leu Thr Val 405 410 Lys Ser Arg 415 Trp» Gln Glu Gly sn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 4 2 5 4 3 0 His Asn His Ty: 4 35 'hr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys <210> 23 <211> 450 <212> PRT <213> Artificial Sequenee <22 3> humanxzed <400> 23 Glu Leu Gln Leu Gln Glu Ser Gly Pró Gly Leu Val Lys Pro Ser Glr i 0 15 Thr Leu Ser Leu Thr Cvs Thr Val Ser Gis Asp Leu Ser Thr Trp 30 Arg Met Asn Trp lie Arg Gln Pro Pro Gly Lys 3 5 4 0 Gly Leu Glu Trp i; **} 5ly lie lie Gly Thr Gly Gly Arg Thr Tyr Tyr Ala. Asn Trp Ala Lys 50 55 ¢0 Gly Arg Val Th 65 :.le Ser Lys Asp Thr Ser Lys Asn Gln Val 70 ' 7 5 Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Z 5 0 Z¡ 9 5 170 Arg Leu Gly Ala Asn Asn Gln Gly Tyr Pro Leu Asp Leu Trp Gly Sin 110 Ly Thr Leu Val Thr Val 115 Ala Ser shr Lys Gly Prc Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser TI 130 135 Ser Gly Gly Thr Wing Wing 2 4 0 Leu Gly Cys Leu Val Lys 145 150 sp Tyr Phe Pro Gli Val Thr Val Ser i O :o Asn Ser Gly Ala Leu Thr Ser .y Val His Thr phe Pro Ala Val 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leí. Ser Val Val Thr Val Pro 1 Qñ 195 GlnTnrTyr Cys Asr Val Asn His Ly; :er Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 010 215 22 0 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met xle 24 5 2 30 2 5 5 ser Aro Lu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Tyr Val Aso Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn. Ser Thr Tyr ?f i i ύ 171 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys no 315 320 ys Lys Vaj 3 - 5 Asn Lys Ala Leu Pi .5 3v Wing Pro lie Glu 3 3 u Lys Thr lie Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln. Ty Val: 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 3 2) 5 3 6 0' 3 6 5 Fhr Gys Leu Val Lys Gly Phe Tyr 370 * ' 375 !sr Asp lie Ala Vaj 3 0 o lu Tro Glu 3S 5 Asn Gly Gln Pro Glu Asn As 350 Fvr Lys rhr ihr 3 55 Val 4 00 Leu Aso Ser Asp Gly Ser Phe Phe Leu Tyr £405 410 3rd Lys Leu Thr Val Asp 415 Lys Ser Arg Trp G 420 Ln Gln Gly Asn Val 425 ?he Ser Cys Ser Val Met His 4 30 u Ala Leu His Asn 4 3 5 -S Tyr Thr Gln Ly: 4 4 0 Leu Ser Leu Ser 445 Gly Lys 4 50 < 210 > 2 4 <211> 450 < 2 12 > PP.T <21 3> Artificial Sequence <22 0> <22 3> humanized < 4 u ύ> 2 4 Glu Leu Gln Leu Gln Glu Ser Glv P: o ΐ ~ 1y Leu Val L y: 10 ’ro Ser Glu 15 172 Thr Leu Arg Met Asn Trp lie Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp He 3 5 40 4 5 Gly He He Gly Thr Gly Gly Arg Thr xyr Tyr Ala Asn Trp Ala Lys Gly Arg Val Thr He Ser Lys Asp Thr Ser Lys Asn Gln Val Ser Leu / tí .ys Leu ser Ser val. Thr Are: Axa. Asir inr Ala. Val Tyr Tyr Cys-Ala Ara Leu Gly Ala Asn Asn Ala Gly Tyr 100 105 Leu Asp. Leu Trp Gly Glr 110 Gly Thr Leu Val Thr Val Ser 11.5 Wing Ser Thr L: Gly Pro 12 5 Val Pro Leu Axa ero Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 α> μ' * u Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 5 150 155 Val Thr Val Asn Ser Glv Ala Leu Thr Ser Glv Val His Thr Phe Pro Ala Val 175 Leu Gln Ser Ser ,i P 0 .y Leu Tyr Ser Leu S< Val Val Thr Val Pro li»0 Gln Thr Tyr He Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lvs Thr His Ti Cys Pro Pro Cys 2 3 0 ?ro Ala i'ro Glu Leu Leu Gly Gly 235 240 173 Pro Ser Val Phe Leu Phe 2 4 5 Pro Pro Lys Pro Lys Asp Thr Leu Met II; 5 0 255 Ser Arq Thr Pro Glu Val Thr Cys Val Val Val Aso» Val Ser His Glu / ». Asp Pro Glu Var Ly? 275 ?he Asn Trp Tyr Val Asp Gly Val Glu Val H; 280'’85 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Va Val Leu Thr Val Leu Hi 310 Glr» asp Trp Leu Asn Gly Ly: 315 32( Giu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ara Pro Iré Glu 325 330 335 Lys Thr lie Ser L\ A 4 u Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tvr ihr Leu Pro Pro •5· q; c Ser Arg Aso Glu Leu Thr Lvs Asn Gln Val Ser Leu hr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp He Ala Val Glu Trp» 5 Glu Ser Asn Gly Gln Pro Glu Asn 385 390 Asn Tyr Lys Thr Ti Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 ~ 415 Lys Ser Hoop Trp Glu Gln Gly Asn Val phe S¿ Val Met Hi) Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 440 174 Lys 450 <21C <211 4 - --- A < ? . > 4 5 0 :> PRT }> Sequenc: !> 'í> huiaanized L> MISC FEATURE :> (103?..(103) )> Xaa can be A, Q, N, D oj <4 0< Glu Thr A i. g Gly Gly Lys Ar g Gly Phe ·> Leu Gln Leu Gln Glu Ser Gly 5 Gly Leu Val Lys 1C Leu Ser Leu Thr Cys Thr Val Ser Gly lie Asp Leu Ser Thr Trp _ í? _3u Met. Asn Trp lie Arg Gln Pro 3 5 4 0 or Gly Lys Gly Leu Glu Trp lie 4 5 lie lie Gl Gly Gly Arg Thr Tyr Tyr Ala Asn Trp Ala Lyí 5 5 6 0 Arg Val Thr lie Ser Ly= 7fi Asp Thr Ser Lys Asn Gln Val Ser Leu Leu Se: Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 8 5 í>0 δ 5 <eu Gly Ala Asn Asn Xaa Gly Tyr Prc Leu Asp Leu Trp Gly Gln 1.0 0 10 5 110 hr Leu Val Thr Val Ser Ser 115 120 Ser Thr Lys Gly P: Ser Val o Leu Ala Pro Ser Ser Lys Ser Tlu 13 0 üly Gly Tlix AIh ai..¿i 14 0 175 φ '7' ' u Gly Cys Leu val Lys Asp Tyr Phe Pro Glu 5 150 155 Val Thr Val Ser 160 Tro Asi Ly Ala Leu Thr Ser Gly Val 165 170 Pro Ala Val 17 5 ;u Gln Ser Ser Gly Leu Tyr Ser Leu Ser 180 185 Ser Val Val Thr Val Pro 150 Ser Ser Leu Gly Thr Gln Thr Tyr lie Cys Asn Val Asn. His Lyj 195 2.0 c 2ür Ser Asn 210 Lys val Asp Lys: Lys Car olu 215 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 'Leu Leu Gly Gly Ί 5 ** 4 > > Pro Ser Val Phe Le ?ro Lys Pro Lys Asp Tfsr Leu Met 11« 2 5 0 2 5 5 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 255 Asn Ala Lys Thr Lys Arg Glu Glu Gln Tyr Asn Ser 2 95 3 0 0 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 32C u Tyr Lys cys Lys Var Ssr Asn Lys Ara Leu la Pro lie Glu 3 5 .ys Thr lie Ser Lys Ala Lys Gly Gln P: 340 345 Arg Glu Pro Gln Val Ty: 176 Tnr Leu Pro Pro Ser Arg Asp Glu Leu Thr Ly; 3 5 5 3 6 ύ Asn. Gln Val Ser Leí 3 6 5 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp 3 7 0 37 5 3 3 0 Asn llv Fin. Fxc Flu Asn Asn Tyx Fys Th 3 9 C* 3 9 5 Pro Val 4 ύ 0 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 405 410 Lys Leu Thr Val Asp 415 Lys Ser Arg rrp Gln Gln Gly Asn Val Ph 410 425 Cys Ser Val Ket 4.3 ¡) Glu Ala Leu His Asn His Tyr Thr Gln Lys 435 440 Leu Ser Leu Ser 445 Gly Lys 3 5 0 <210 > 36 <2il> 330 <212: DNA <213> Rabbit < ¿¿ o v > gatgttgrga 6 0 ; ceagac tccagcctcc gt. yyacocao ctgtgggagg falls ¿tgt cace atcaagtgcc aggccagtca. gagcgttagt agttacttag cctggtatca gcagaaacca 0 gggcagcctc ccaagctcct gatetatget gcatcctatc tggcatccgg ggtcccatca c g g 1.1 c a a a g g c a g t g g a t c t g g g a c a g a g 11 c a c t. c t c a c c a t. c a g c g a c (; t g g a g t ¡g t. 24 0 gceyatgctg ccacttatta ctgtcaacag ggttatacta. ggactgatat tgataa.ta.ct 3 0 0 ttcggcggag ggaccaaggt ggtggtcgaa 3 3 0 177 < 4 0 0 > 2 Ί Q 3. Q t C £ί C Γ. G C? 60 3.! í g 3. g 5 c c g g g g m t coco tv? atcacgcctg ggaea eecct g a. c a c t c a. C C T OF C 3 C 3 θ 'T C T I 2 0 CTGGAATCGA CCTCAGTACT TGY¿IQ3 31 G3 ACTGGGTCCG CCAGGCTCCA GGGGGGGC 18 or' ~ TGGAATGGAT CGGAATCATT GGTACTGGTG G 13 G 3 3 C 3T3 CTACGCGAAC i acctcgacca cggtggatct gaaggtc ac c agtccgacaa í . > . ’ C C Of 3 Q CJ 3 C 3 C ggccacctat ttctgtgcca gattgggtgc t33t33t331 ct o 1131 c c ts 11 351 t g vi 3 c o t g t g gggcccgggg accctggtca c c g v c t c g a g z <21ύ> 28 <211> 330 <212> DNA * χ 3 s Axtxfxcxax sequence <r 2 2 'j > <223> isuri niveo < 4 0 0 > 2 3 gatgtcgtga 60 tgactcacitc acccgcaagc ct.ggctgtgt cgctgggaca gcgggctact atctcotcfcc •t -< J.. — a a g cgt oc c 3 gtctgtctcc tcgtacttgg cctggtatca gcagaaggac ggaeagceet catctacgcc g c c t c a t a c c ttgcgtccgg agtgccttcg agat tcaaag 2 4 0 y aa gc ggaa g cggcactgag tr.tacacr.ga a c a t c c a c c c g g t g g a a g a a gaggacgcag O !? eeacgtacta ctgtcaacaa ggíít.aoaccc gcaccgatat tgacaatacc ttcggtggcg cr g a c 13 a g c t g g a a a x c a a g <21O> 29 <;ιι> -3 34 178 <212> 2ΝΑ <213> Artificial Sequence <223> murinised <4ύό> 29 caagtgcagc tgaaagaatc aggaccggga ctggtggcac caagccagtc 0 acctgtactg tgtccggaat cgacctctcc acctggcgca tgaattgggt 120 cccggaaagg gcctcgaatg gattggcatc atcggtactg ggggcagaac 130 aattgggcga aggggcgctt ctcaatctcg aaagactcga cccaagtgtt 240 aacagccttc agaccgagga taccctctacc tacttttgcg cqaggctggg '5 ; c c tg t c ga t c ccggcagcct cttgaagatg 3gCC33C33C aacggttacc ccíctcgarct. qtqqqqagcc ggaacgac ”'354 isactovctc gagc 20> 30 )11> 33 0 ;i2> DNA I13> Artificial Sequence <22 3> humanized <400> 30 gatatccaaa tgacccagtc cccatcctcc ctgtccgctt ctgtcggtga. 60 attacttgtc aagccagcca gtcggtgtcc tcatacctcg cctggtatca ccgcgtgacc gcagaagccg q g ¿í a ¿i g g c g c c c a a a c 01t1 g 3 rc t a c g c c g c c t c g t a c c t g g c a r. c* c g g 190 agattcagcg ggtcgggctc cqgaactgac ttcaccctga ccatcagcag 240 gaggactttg ccacttacta ctgccagcag ggatacaccc ggaecgatat 30 0 c c t g c* a g c c t ttcggcgggg gaactaaggt cgaaatcaag 3 3 q 179 <210> 31 <21i> 3 30 < 212 > DHA <213> Artificial Sequence. <22 0> <2 23> humanized <400> 31 gatatc caaa tgacccagtc cccatcctcc attacttgtc aagccagcca gtcggtgtcc 120 ggaaaggcgc 100 ccaaactxtt gatctacgcc agattcagcg 40 gaggactttg 300 ttcggcgggg 30 ggtcgggctc ccacttacta gaactaaggt. cggaactgac ctgccagcag cgaaatcaag ctgtccgctt teataceteg gcctcgtacc ttcaceetga geetacaccc ctgtcggtga cctggtatca tggcatccgg ceateagcag ggaccgatat ccgcgtgacc gcagaagccg cgtgccgtca cctgcagcct tgacaacacc <210> 32 <211> 360 <212s DNA <213> Artificial sequence <220> <22 3 > humanization <400> 32 gagctccaac tgcaagaatc cggccctggt ctggtcaagc cctccgaaac acctgtactg tgtccgggat cgacttgagc acctggagga tgaactggat 12 0 ccgggaaagg gactggagtg gatcggcatt atcggtactg ggggacggac 180 aattgggcaa agggcagagt gacgatttca aaggaccacc 0 aaactgtcct ccgtgactgc ggctgacacc gccgtgtact attgcgcccg 300 aacaaceagg gctacccgct ggatctttgg ggacagggaa ccctcgtgac 3 60 cctgagcctc tcgccagcca ctactacgcc ggtgtccctg gctgggagcc tgtctcgagt 180 <» <21Ο> 33 <211> 360 <212> DNA <213> Artificial Sequence humanised < 4 0 0 > 33 gagctccaac. tgcaagaatc cggccctggt 60 c t. g g t o a a g c ccc o c ga a a c cctgagcctc acctgtactg tgtecgggat cgacttgagc acctggagga tgaactggat tccrccagcca ccgggaaagg gactggagtg gatcggcatt a t cggtactg ggggacggac tí O aattgggcaa agggcagagt gacgatttca aaggaacccacct cgaagaaccacct 240 c t actac gc c ggtgtccctg aaactgtcct ccgtgactgc ggctgacacc gccgtgtact attgcgcccg 0 aacaaegccg gct acccgc. t g ga t ct ttgg gga ca ggg a a ccc t cgtgac 3 6 0 gctgggagcc tgtctcgagt <210> 34 <2.11> 651 <212 > DNA <213> Artif; .al Seque nc« <223> humanizad <400> 34 gatatccaaa tgacccagtc cccatcctcc ctgtccgctt ctgtcggtga at tacttgtc aagccagcca gtcggtgtcc teatacct cg cctggtatca 0 ggaaaggcgc ccaaactttt gatctacgcc gcctcgtacc tggcatccgg ccgcgtgacc gcagaagccg c g t. ge c g t c a. agatteageg ggtcgggctc cggaactgac ttcaccctga ccatcagcag cctgcagcct gaggactttg ccacttacta ctgccagcag gga.taca.ccc cigaccgatai 300 181 ttcggcgggg c, a a c t a a g g t. 31 <' C Q a a ei X £ ¿i ¿i. £í c cr g a c <? g X g g ccgícxccctc c g X. g t x c a x c ttcceaccct ccgacgagca 420 geXta>cc g g c a c c g c c X ccgXegtgXg c c X g c t cí a a c aacttctacc cccgegaggc <1 C f C ei a g £ξ t- 0- C ¿i O tggaaggtgg a e a a c g c c c x gcagxccggc aactcccagg aatccgtcac 540 cgagcagg&c T. x c . a a c?crac a gcacctactc cctgtcctcc accctgaccc tgtcca aggc 600) cgactacgag aagcacaagg tgtacgcctg cgaagxgacc caccagggcc tgtccagccc cgtgaccaag tccttcaacc ggggcgagtg c <210> 35 <211> 651 <212> DNA <213> Artificial Sequ ? 3 5 gatatccaaa tgacccagtc 6 Oí cccatcctcc c X g x c c g c X t ctgtcggtga ccgcgxgacc attacttgt c a age c a gc c a 120? gtcggtgtcc x c a x a c ex c g cctggtatca gcagaagecg g g a a a g g c g c c c a a a c 1.111. 13 2g &t -? ί ή c ce c gcctccftacc tggcatccgg cgtgccgtca agatteageg gg1.cgggetc 240 c gg¿t a c xc? & c x x c a c c c X g a ccatcagcag cctgcagcct. gaggactttg ccacttacta 3 0' 0 ctgccacrcéicf gcctacaccc g ga c c ga t at X g a e a a c a c c ttcggcgggg gaactaaggt 3 fc I heard C g he. ei ei X, C & ¿i M c g x a c g g X a g cggc c ccate xgtc x x. ca x c 11 c c c g c c a t e t. g a t g a g c a 42 0 g x. X. g <3, ¿i a x c x ggaactgcct ctgttgtgtg cctgcxgaat aaettetate ccagagaggc 4 0 0 c β η β. g X3 c a g x g g a a g g x g g st ¿segcc c t c £' a a x c g g g X aactcccagg agagtgtcac íi.'S agagcaggac a.g c a a g g a c a. gcacctacac c c X e a g c a c* c 182 acccxgacgc 6 ? v tgagcaaagc agactacgag aaac ac aaa g tctacgcctcj caica c t? ci c c 651 tgagctcgcc c g t c a e a a a g agetteaa ca agggagagtg <210> 36 <211> 1341 < 12 > uNA <2.2 3> Art. i .ficial Seqi <22 0> <22.3> humanized < 4 0 0 > 36 gagctccaac 60 tgcaagaatc ccrggccgacaggt acctgtactg 120 tgtccgggat c tí a e 11 g a. g c ae etgga gga t g a a e t g g a t ccgggaaagg 180 gactggagtg gatcggcatt atcggtaetg ggggacggac aattgggcaa 240 agggcagagt gacgatttca aaggacacct cgaagaae c a ¿1 3 3 Ct t C C t ccgtgactgc g g c1 g a e ¿t c c g c e g t g t a c t attgcgcccg β 3 C ct 3 € C ct O C» 3 6 6 gctacccgct ggatctttgg gq a c a g g g a a ccctcgtgac gcctccacca 420 agggcccctc cgtgttccct ctggcccctt gctcccggtc tctaecgeeg 4 3 2 ctctgggctg c c 11( g t c a a g cía c t a e 11 c c ccgagcccgt tgg-aaet etg t 4 0 gcgccctgac ctccggcgtg cacaccttcc ctgccgtgc.t gocetgtact 6 00 ccctgtcctc cgtcgtgacc gtgccctcct ccagcctggg 13 e 3 e e t g t a 66 2 acgtggacca caagccctcc a a e a c c a a g g tggaeaa geg a a g t a c g a c c ~t 2 0 ctccctgccc cccctgccct gcccctgaat ttctgggcgg ttcctgttcc c c c c a a a. or c c c a a g g a c a c c c t ga t g a. t c t cccggacccc cgaagtcacc t c ctgagcctc tcgccagcca c tacta cgc c ggtgtccctg gctgggagcc tgtctcgagt cacctccgag gacagtgtcc gcagtcctcc c a c C- a a g a c c ggtggaatct acct tcccítg c ga ag t ga c c 183 30 tgcgtggtgg 6 4 0 tggaegtqte ccaggaagat cccgaggtcc agttcaattq gtacgtcfg&e ggcgtgcíaací 9 0 0 tgcacastgc casga c c aag c c c a g a g a g g aa.cagt teas C t C C 3. C C t el C cgggtggtgt 9 60 c c g t g c t cí a c cgtgctgcac caggactggc t gaacggcaa agagtacaag tgcaaggtgt 102 0 c c a a c a a g g g cctgccctcc ageapegaaa agaceatete caacígccaag ggccagcccc 1080 ge gag c c cca ggtgtacacc ctgcccccta gccaggaaga gatgaccaag aaccaggtgt 1140 ccc t. gazette t c t g g t c aa g ggcttctacc c c t c c ga c a t tgecgtggaa tgggagtcca 12 00 a c g g c c a g c* c c. cí p a ci ~· a a s ta esagseca ccccccctgt gctggacagc gacggctcct 1260 tcttcctgta ctctcggctg acegtggaca agtcccggtg gcaggaaggc a a c g t c 11 c T. 1320 cetgctccgt gatgeaegag gccctgcaca accactacac egg agg agt ccat gt4cc ct4cc <21 ύ> <211> <212> <213> 37 1341 DNA Arti fie i al Sequence <220> <22 3 > humanized <. 4 u 0 '> 37 era ge te é ?aae tge^ísgaate eggecetgqt etggteaage c o t c c g a a ¿i c acetgtacto tcftccgggat cgacttgctGO acctggagga 120 t g a a c c ef g a t ccgggaaagg gactggagtg gatcggcatt ateggtactg 16 0 ggggacggac aattgg gcaa agggcagagt gaegatttca aaggacacct c cí a a g a a c c a 4 OR cctgagcetc tcgccagcca ctactacgcc ggtgtccctg 184 aaactgtcc t 300 ccgtgactgc ggctgacacc gecgtgtact attgcgcccg gc.tgggagcc άάίάάί' gC C G 360 gctacccgct ggatctttgg ggacagggaa ccctcgtgac tgtctcgagt gectecaeca 4 2 0 agggecaete cgtgttccct c. t g g c c c c tt q c t c c c g g t c c a c c t c c g a g tctaccgccg 430 ctctgggctg cctggtcaag gactacttcc ccgagcccgt gac ag t g tc c tggaactctg kj C’· G C G C X 9 3 O ctccggcgtg cacaccttcc ctgccgtgct g cagt.ce te c g y e c i y t a e t 600 C C C X 9 1?. C C X C c g t c g t g a c c g c g e c c t c c t ccagcctggg cac caayace tSCSCCtQtH 660 BC Ci L.CÍCÍ 3 C C'cl caa gccct ce a a c a c c a ac g tggacaagcg ggtggaatct a a g t a c o g c e 7 2 C ct.cc ctgccc e e c c t g c c c t gcccctgaat 1i c t g g g c g g accttccgtg ttcctgttcc 700 ccccaaagcc caaggacace ctgatgatct cccggacccc eyaagtgacc tgcg tggtgg 8 4 0 t 99 3 c 9x ei t. e ccaggaagat cccgaggtcc agttcaattg gtaegtggac ggcgtggaag 500 tgcacaatgc caagaccaag cccagagagg a a c a g 11 c a a ctccacctac cgggtggtgt 960 c c σχ g c t. ga c cgtcictgcac caggactggc X Cf 3 3 c 99 £33 ag a g t a c a a g tgcaaggtcft 1020 c c 3 3 c 3.3 g g g cctgccctcc a g c a t c g a a a 3 G 3 C C 3 X C t C c a a g g c c a a g ggccagcccc 10 3 0' g o g a 9 £ e c c a ggtgtacacc ctgcccccta gccagga&ga gatgaecaag a a c c a g g t g t 114 0 cect -gacctcj t c t g g t ca a g ggcttctacc cctccgacat tgccgtggaa t gggagt c ca 1200 a c 9 g c. c a: 3 c c c gagaacaac tacaagacca ccccccctgt g e t g g a c a g c* gacggctcct 1260 x c t x c c x g x a e t, ~ t e g g,-1 g accgtggaca agtcccggtg gcaggaaggc a a e g t c 11 c t 1320 cctgctccgt. g a. t ge ac g ag g c c c t e c a c a 3 € C 3 C X el. C 3 G c e a g a a g t c c 185 ct gtccctga gcctgggcaa g <2I0> 38 <2T1> 1350 <212> DNA <213> Artificial Sequence ^-22.3^ Π UFuci Π ¿ Z£*G <4ú0> .3 3 gagctccaac 60 tgeaagaatc cggccctggt ctggtcaagc cctoegaaac cctgagcctc acctgtactg 120 tgtccgggat cgacttgagc acctggagga tgaactggat tcgccagcca c c g g g a a a g y gactggagtg g ate g gca 11 atcggtactg ggggacggac ctactacgcc aattgggcaa 2 4 0 agggeagagt gacgatttca aaggacacct cgaagaacca ggtgtccctg aaactgtcct Q O f, ccgtgactgc ggc tga core gccgtgtact attgcgcccg gctgggagcc a a c a a c c a g g 3 6 ? gctacccgct. ggatctttgg ggacagggaa ccctcgtgac tgtctcgagt gcctccacca 420 agggcccctc cgtgttcccg ctcgctccat catcgaagte taccagcgga ggcactgcgg 4 8 0 ctctcggttg c c t c g t q a y gactacttcc c g g a g o c g g t g a c c y c g tg y ta 4 . y a g c c c tg a c eagcggggtg c a c a c c 111 c cggccgtctt gcagtcaagc ggcctttact 600 ccctgtcatc agtggtgact gtcccgtcca gctcattggg aacccaaacc tacatctgca 660 atgtgaatca c aasc c t age aacaceaagg 11 g a c a a gt a a a agt a agtacgagccc! a c a a g a c t c a cacttgtccg ccgtgcccgg cacccgaact gctgggaggt cccagcgtct 7 0 u ttctgttccc tccaaagccg aaagacacgc tgatgatctc ccgcaccccg gaggtcactt gcgtggtcgt ggacgtgtca catigaggatc cagaggtgaa «gttcsattgg fiáñ 186 tac'gtggatg gcgtcgaagr ccecáñtqcc aaaaetaagc ccagagaaga acagtaca at t c g a c e t a c c g c g t c g t g t c c g t g c t c a c g g c g 11 g c a t. c a g g a 11 o g c t 9 € G gaatacaagt gcaaagtgt.c caacaaggícg ctgecggcac cgatcgagaa 10 2 O aaagcgaagg gacagcctag ggaaccteaa gtctacacgc tgccaccatc 8 O ctgactaaga atcaagrctc actgacttgt ctggtgaagg ggttttaccc 1140 gccgtggagt gggaatccaa cggccagcca gagaacaact acaagactac 12 u O ctcgactcgg atggatcgtt cttcctttac tcgaagctca ccgtggataa 1260 c a g c a g« g a a a c g t. ~j tt c f. C C T G C Z C G G T G A TOCA T GA A G E C C T C C A T A A A I32Ü ”CA A A A A A GT C GE T G T C C C T G T C G C C GGA A A A A G 1350 GAACGGGAA G ACGGTGAA RAGCGACATT CCCTCGTG 3 9 gagctccaac tg lagaatc cggccetggt ctggtcaagc cctccgaaac cctgagcctc acctgtactg tgtccgggat cgacttgagc acctggagga tgaactggat tegecageca ccgggaaagg gactggagtg gatcggcatt atcggtactg ggggacggac 180 aattgggcaa agggcagagt gacgatrtca aaggacacct egaagaacca 4 OR aaactgtcct ccgtgactgc ggctgacacc gccgtgtact attgcgcccg 2> V V ctactacgcc ggtgtccctg gctgggagcc aacaacgccg gctacccgct ggatctttgg ggacagggaa ccctcgtgac tgtctcgagt 187 360 g c c t ccs c c a a. g g g c c c c X c c g t g t1. c c c g ctcgctccat c 3X c g 3 3 g X c taccagcgga ggcactgcgg 40 0 ​​c X c X ego X Xg cctegtgaací gactacttcc cqy3qoeggx gaccgtgtcg tggaacagcg 54 0 g 3. g c c e X g & c cageggggtg ca cacct11 c <. Choose v or x v,. xx. g c ag tc aag c ggcctttact 6 0 0 cectgxeaxc agtggtgact gtcccgtcca gexeatxggg aacccaaacc tacatctgca 6 6 0 3 X Cí X Q 3 3 X C ¿i caaacctagc a a c a cea aqg t X g 3C3 3 03 3 agtcgagccc aagt Íct"Oct'caga 720-Oct'caga I heard ccgtgcccgg cacccgaact gctgggaggt c c c ag c g t c t 7S0 x. X ct x, or X x c* e e. tccaaagecg ssa^scsc^c X Q 3 X g 3 X C X C cc geaccccg gaggtcactt >· ~ -t ·_' gcgXggXcgt ggacgtgtca catgaggacc c 3 g 3 g g x g o 3 gttcaatt g g tacgtggatg 90 0 gcgtcqaagx ccacaatgcc aaaactaagc ccagag&aga acagtacaat tegacctace S6ü gcgXcqXciXe cgtgctcacg gtgttgcatc aggatxggct. «aaccíggaag g a a t a c a a g t 102 0 gcaaagXcrte caacaaggcg c f g c c g g c a c c g 3 X c g 3 g 3 3 a a c t a t c t c c a a a g c g a a g g 1 0 8 0 g- ¿i e 3 o c e x a g ggaacctcaa g t c t a c a c g c Xgceaccaxc acggcfatgaa ctgactaaga 114 0 atcaagtctc actgacttgt ctggtgaagg ggtxxx&cec tagcgacatt gccgtggagt 12 0 0 o g g aa t c c a a c ggccageca gagaaca a c t 3 C 3 3 Q 3 C X- 3 C ccctccagtg c t c g a c t c ο ό atggatcgtt cttcctttac tcgaagctca c c g x g a x 3 3 gtcccggtgg cagcagggaa 1320 acgtgttctc ctgctcggtg atgcatgaag ccctccataa ccactatacc caaaagtcgc gtccgggactagc 188 <210> 40 <211> 132 <212» PRT <213> Rabbit Ci> MI3C_FEATCRE <222> (i) . . (22 ) <223> signal sequence <4 00> 40 Met. Asp- Met Arg Ala Pro Th: 1 3 Gln Leu Leu Gly Leu Leu Leu Leu 10 15 Leu Pro Gly Ala Arg Cys Asp Val Val Met Thr Gln Thx Pro Ala Serval Glu Ala Ala Val Gly Gly Thr Val Th 35 40 le Lys Cys Gln Ala Se: 4 5 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 55 60 ro Pro Lys Leu Leu lie Tyr Ala Ala 5 70 er Tyr Leu Ara 7 5 (er Gly Val ;c> Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Th 05 lie Ser Asp Leu Glu Cys Ala Asp Ala Ala Thr Ty: 100 105 Fyr Cys Gln Gln Gly Tyr Thr Arg 115 Asp Asn Thr - 0 Phe Gly Gly Gly Thr Ly' 125 Va r Va I Va i G1 u ! '3. ñ 0 > 41 1> 1 3 6 2 > PRT 3> Rao 189 < 2 2 (J > <221> MTSC FEATURE <222> (1)-.(10) < 2 2 3 > Signa! sequence <4 00> 41 Met Gl? .3 i.AX i'xv Ltrll Αχ g Trp Leu Leu Leu Val Ala Val Leu Lys Gly i O 15 Val Gln Cys Gln Ser Leu Glu Glu Ser Gly xi Leu Val Thi 30 Gly Thr Pro 35 Leu Thr Leu Thr cyj 4Q Val Gly lie Asp Leu Ser 45 Thr Trp Arg Met A« 5 0 ;c Val Arg Gln Ala. 55 :x Gly Lys Gly Leu Glu 6 0 Trp lie Gly lie I) ex Gly Thr Gly Gly Arg Th: 70 7 5 Tyr Tyr Ala Asn Trp 80 iia rys Giy Arg me Thr Lys Thr Ser Thr Thr Val Asp Leu SO 05 Lys Val Thr Ser 100 or Thr Thr Gj Asp Thr Wing Thr 7y.: Pray Cys Ala s or Gly Ala Asn Asn As I : S Gry Tyi 12 € Leu Asp Leu Trp Gly Pro 12 5 Gly Thr Leu Val Thr Val 130 135 <210> 42 <211> 3S¿ <2i2> DNí < ' i < R¿í Jod i χ rnxsc___reature (1)..(66) Signal sequence 190 < 4 0 0 > a fe qgac a tefa 60 y y y c c c c c a c tcagctgctg gggctcctgc tgctctggct cccaggtgcc a ya fe y 0 y a x y 120 ttgtgatgac c c a y a c fe c c a gcctccgtgg aggcagctgt gggaggcaca gtcaccatca 180 agtgccaggc c a y fe. c ay a and c •y fe. faith to and faith to and faith. fe a c fe · fe a y c* c fe- y y fe a fe c a y c a y aaaccagggc agcctcccaa gctcctgatc tatgctgcat cctatctggc atccggggtc ccatcacggt 30 0 ú Ccíd a y y c a y tggatctggg a c a y ay fe, fe: c a. c faith. c fe c a c c a l cagcgacctg gaytytyccy 3 fe 0 atgctgccac fe t a o Ό a. c fe y fe caacagggtt atactaggac tgatattgat a a fe. a c faith faith faith c y 396 y c y o a y ex ex a c caaggtggtg yfecyaa <21O> 43 <211> 40$ <212> DNA <213> Rafobit <221> mise feature <222> (1)..(57) <22 3> Signal sequence < 4 0 0 > 4 3 to faith. y y a y a c faith. y 60 y y c v y c y c fe. y gcttctcctg yfecycfeyfegc ucaaaggtg— ccagtgtcag fecgcfeayaay a a fe c c c? o y o o tcgcctggtc aogccfegyga cacccctgac actcacctgc a c a y fe c fe c fe- y 180 yaafecyaccfe cagtacttgg aya a feyaac fe g g g t c c g c c a ggctccaggg a&gyggetgy 24 0 aatgyafecgg aatcattggt a efe y y feg g fea gaacatacta cgcgaactgg y c a a a.a y y c c gattcacca- ctccaaaacc fecgaccacgy tggatctgaa ggtcaccagt c c y a c a* a c c y 4 ú, ί) a y y a e a c y y c C á C C131t1C tgfegeeagafe tgggtgctaa taataatggt 191 tatcctttgg c 406 ¡cttgtgggg cceggggacc ctggtcaccg tetegagt. <210 4 4 <211> 107 <212> PRT <213> He tao sapiens < 4 0 0 > 4 4 Asplie Gln. Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 c 10 15 Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gln Ser lie Ser Ser Tyr '> β r c óf-, •O V Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu lie 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 ü z> 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Gln Pro 65 7 0* 75 80 Glu A.sp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Leu 65 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glulie Lys 10 '0 105 <210> 45 <211> 114 <212> PRT <213> Homo sapiens <40 ύ> 45 Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro ser Glu - 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser lie Ser Ser Ser 25 3 0 192 Ser Tyr Tyr Trp 3s Ly Trp lie Arg Gln Pro Pr Gly Lys 45 Glv Leu Glu Trp lie Gly 50 ¡•yr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Sei fcu Leu Lys A r es Va 1 r h r lis o ñ Val Asp TI TLys Asn Gln Phe Ser Leu Lys Leu .Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Tyi ?he Asp Tyr rrp Gly Gln Gly Thr Leu Val Thr Val 115 110 < 210 > 4 6 <211> 321 <212> DNA nGiiiO sapiens <400> 4€ gacatccaga t.gacccagr.c tccatcctcc ctgtctgcat ctgtaggaga cagagtcace eo atcacttgcc gggcaagtca gagea.tt.agc: agctatttaa attggtatca gcagaaacca 12 0 gggaaagccc ctaagctcct gatetatget gcatecagtt tgcaaagtgg ggtcccatca aggttcagtg gcagtggatc tgggacagat ttcactetca ceatcagcag tctgcaacct 0 ga.agat.tttg caacttacta ctgtcaacag agttacagta cccctctcac tttcggcgga .3 0 0 gggaccaagg tggagatcaa a 321 <210> 47 <211> 342 <212> DNA <213> Homo sacien; 193 cagctgcagc tgcaggagttc gggcccaggu ctggtgaagc cttcggagac acctgcactg tctctggtgg ctccatcagc agtagtagtt actactgggg 120 cagcccccag ggaaggggct ggaggtggatt gggagtatct attatagtgg ISO ......... tacaacccgt ccctcaagag tcgagtcacc atatccgtag acacgtccaa ctggatccgc gagcacctac gaaccagttc tccctgaagc tgagctctgt gaccgccgca gacacggctg tgtattactg 300 tttgactact gggaccaagg aaccctggtc accgtctcct ca 342 <211> 12 <213> Rafobit <400> 43 Leu Gly Ala Asn Asn Asn Gly Tyr Pro Leu Aso Leu 1 5 10 tgcgagatac <210 <211 > <212> <213> 4 S 12 PRT Artificial : Sequence <22 0> <22 3> humanized <400 49 Leu Gly Ala A s n a. yes i Gln Gly Tyr Pro Leu 1 5 10 <210> 5 0 <211> 12 <21 3> Artificial SO <4 0ü> 194 su Glv Wing Asn Asn Wing Glv Tn Lo u .ηsx' Le u 1 i : > o 1 <211> 12 <212> PRT <213> Artificial .Sequence <__s> humancreate < 4 '3 V: 1 Leu Gly Ala Asn Asn Asn Gly Tyr 1 5 Pro Leu Asp Leu 10 <21.0> <^12> Ppm <213> Artificial Sequete* <22 0> <2 2 3> human i 2 ed <400> 52 Leu Gly Ala Asn Asn. Be Gly Tyj 1 * 5 Pro Leu Asp Leu 10 11> rabbi i Gln Gln Gly Tyr Thr Arg Thr Asp 1 r Tle A s o A .s n a h r 10 < 2 r> > 9 4 <2il> 12 <212> PRT <213> Artificial Sequence 195 Gln Gln Ala Tyr Thr Arg Th: 1 5 isp lie Asp Asn Thr 1C! <210 > 55 < 2 1.1 > 4 4 2 <213> Home sapiens <400> 55 Met. Ala Glu Pro Arg· Gln Glu Phe Glu Val Met Glu Asp His Ala Gl; 1 5 10 15 Fyr Gly Leu Gly Asp Arg Lys Asp Gln Gly Gly Tyr Thr Met Rj Gln Asp Gln Glu Gly Aí 35 Thr Asp Ala Gly Leu Lys Glu Yes 40 45 Pro Leu Gln .hr biu Asp Gly o o Glu Glu iii and Ser c-iu τη fi 0 Asp Ala Lys Ser Thr Pro Thr Ala Glu Asp Val Thr Ala Pro Leu Val 65 7 0 75 50 Asd Glu g1y Ai :o Gly i»ys Gln Ara Ala Ala Gln Pro Hrs Tnr Glu i 00 0 5 &¿u Gry Tnr. Tnr Ala Glu Glu Ala Gly laughs Gly Assisi Tnr Pr 100 105 * i ib Leu Glu Asp Glu Ala Ala Gly His Val Thr Gln Ala Arg Met Vai 115 120 125 Ser Lys Ser Lys Asp Gly Thr Gly Ser Asp Asp Lys Lys Ala Lys Gly 135 Wing Asp Gly Lys Ti 145 rys lie Wing Thr Pro Arg Gly Wing Wing Pro P 150 155 1 ΟιK Gly Gln Lys íly Gln Ala Asn Ala Thr Ara He Pro Ala Lys Thr Pro 165 170 175 196th Wing pro Lys i S O -Y Lys Ser Gly Asp Arg Ser Gly Tyr Ser Ser Pro Gly Se 195 200 Pro Gly Thr Pro Gly Ser 210 215 Thr Arg Glu Pro Lys 22 0 Lys Val Ala Val Val Ara Thr Pro Pro Lys Ser 225 230 235 Yes?·?· Ala r»ys '¿i,eu Gln Thr Ala Pro Val .ero Me- ero Asr Leu Lys Asn Val 255 Lys Ser Lys He Gly Ser Thr Glu Asn Leu Lys H: 2 6 and 2 6 5 Gln Pro Gly G i ’JlJ ?iy Lys Val Gin lie lie Asn Lys Lys Leu Asp Leu Ser Asn Val Gil 275 250 205 Jer Lys Cys Gly 2 9 0 »r Lys Asp Asn 295 .ys His Val h D ’ro Gly Gly Gly Ser Val Gln He Val Tyr Lys 305 310 ’ro Val Asp Leu 3 i 5 Lys Val Thr Ser 320 Lys Cys Gly Ser Leu Gly Asn 1.1« í .3 o Lys Pro Gly Gly Gly Gl: 3 5 Vai Glu Val uys Ser Glu Lys Leu Asp me Lys Asp» Arg Var Gln Ser Lys lie Gly 3s 3 5 5 r Leu Asp Asn He Th: 60 lis Val Pro Gly Gly Gly Asn 365 Lys Lys lie Glu Thr His Lys Leu Thr 370 375 Arg Glu Asn Ala Lys Al 3 o ñ Lys Thr Asp His Gly Ala Glu He Val Tyr Lys Ser Pro Val Val Ser Ser Pro Arg His Leu Ser Asn Val Ser Se 405 385 Gly Asp Thr I1e Asp Met S&r Wing Ser 4 35 <2iQ> 56 <211> 10 197 Val Asp Ser Pro Gln Leu Ala Thr Leu Al 4 2 0 4 25 Leu Ala Lys Gln Gly Leu 4 4 0 <212> PRT <213> Artificial Sequence <22 0> < 2 2 3 > ei o d i f í. e d <221> MISC_F£ATURE <222> (1)..(1} <223> ACETYLATION and NYTROSYLATION < 2 2 0 > <221> MISC_FEATURE <222> (1) . . (1) <223> NYTROSYLATION <22 0 > <221> MISC FEATURE <221> MISC_FEATURE <222> (3)..(3) <223> PHOSPHORYLATTON < 2 2 0 > <221> MISC_FEATURE <222> (5)..(0) <22 3> Position of Biotin Conjugation <22 0> <221> MISC_FEATURE <222> (6)..(6) <223> PKOSPHORYLATION Asp Glu val 4 30 198 <220 MISe_FEATURE <222> (9)..(9) <22 3> PHGSPHORYLATIGN <220> <221> MISC FEATURE <222 > (18) . . (18) <22 3 > AMIDATION < 4 0 0 > 5 6 Tyr Se r Ser Pro Cys X z> Gly Thr Prc 10 <210> 57 < 2Ί l > 10 <212 > 12 ρπ- \ -i. <21 3> Artificial <22 0> <22 3> modified <2 0> <22 1?> MISC FEATURE <22 0 (1)..(1) < 2 2 3> ACETYLAT10N <22 0> <22 1 > MISC FEATURE < 2.2 2' R (2} . . (2 ) or ** 3> PHOSPKORYLAT <22 0> < 2 2: 1> MISC_FEATURE < 2 2 2 > ( or) . . ( G ) <223> PHOSPHORYLATION <22 0> <221> MISC_FEATURE <222> (8) . . (8) <223> PHOSPHORYLATION <220> <221> MISC FEATURE <222> (18)..(18) <22 0> <22i> MISO FEATURE <222> (10)7.(10) <223> AMIDATION I ίΰθ > 57 199 Ser Set Cys 1C <210 > 5 8 <211> 217 <212> PRT <2Ι3> Mouse < 4 0Ο 5 8 Asp Val Val Met Th: 1 5 Gíb Ser Prc, Aia Ser Leu Ara Val Ser L Gln Arg Ala Thr He Ser Cys Gln Ala Ser Gln Veri Ser Ser Ty: 3 0 Leu Ara Tr * .vs Pro GiV Gln 4 (> Tyr Ala Ala Ser Tyr Leu Ala Ser Gly Val Pro Ser Arg Phe Lys Gly 50 55 60 Thr Glu Phe Thr Leu Asn He Hi 10 7 5 Pro Val Glu Gl' 8 0 Glu Asp Ala Ala Thr Tyr Tyr cys Gln Gln Gly Tyr Thr Ara Thr Asr He Asp Asn ?he Gly Gly Gly Thr Lys Leu 105 lu He Lys Arg Thr 11C Asp Wing Wing Pro Th 115 Ok Yes Glu Gln Leu 125 Thr Ser Gly Glv Al, Val Val Cys Phe Leu Asn Asn Phe Tyr Pro 135 140 Lys 145 A s ρ ríe A sn Val L v s T r d L v s I. eru Ara Gln ¡Π.Ζ: i 16i 200 Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser 165 170 Lys Asp Ser Thr Ty.i 175 Ser Ket Asn Ser Tyr 195 Val Lys *2* x— 1 <210> 59 <211> 4 42 <212> PRT <213> Mouse < < 0 0 > 59 0 rhr Leu Thr Leu Thr Lys Asp lí 5 Cys Glu Ala Thr 200 Asn Arg Asn Glu Cys 'yr Glu Arg Hi; Lys Th; rnr .3« 205 lie Gln Val Gln 1 Val Ala Prc Ser Gln Ser Leu Ser lie Thr Cys Thr Val Ser Gly lie Asp Leu Ser Thr Trp Ara Ket Asn Tro Val Ara 33 Gly Ly; i.y Leu Gl' 4 5 ctr lie Gly lie lie Gly Thr Gly Gly Arg Thr Tyr Tyr Wing Asn Trp Wing Lys 50 55 60 Gly Arg Phe Ser lie Ser Lys Asp ser Thr Gin Val Phe Leu Lys Met Ser Leu Gln Thr Gl 8 5 Asp Thr Wing Thr Ty: Pne Cys Ala Arg Lí u 5 Giy Ala Asn Asn Asn Gry Tyr íu 1 0 ñ Leu Asp Leu 105 Gly Ara Gly Thr Val Thr Val Se. 115 Wing Lys Thr Thr 120 R Σ O 3 3 V 3.. 125 Tyr Pro 201 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 13 0 135 14 0 'ys Leu Val Lys Gly Tyr .45 150 Glu Pro Val Thr Val Thr Trp Asn 155 16 0 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 165 170 Wing Val Leu Gln 175 Ser Asp Leu Tyr Thr Leu Ser Ser Ser 180 185 Val rhr Val -ro ser Se: 190 ;p Prc Ser Glu 195 Val Thr Cys Asn Val Ala His Pro Ala S« se 3 Thr Lys Val Asp Lys Lys lie Val Pro Arg Asp 210 215 .ί ζ- y O X X* V i- Ύ S P X O ys He Cys Thr Val .Ό Glu Vaj >0 ser se: Go] 2 3= -he he Pro Pro & l'l Lys Pro Lys Asp Val Leu Thr 15 Leu Thr 2 5 0 Lys Val Thr Cys 2 55 Val Val Val Asp He ser Lys Asp» Asp» Pro Glu Val Gln Phe Ser Trp 260 265 270 Phe Val Asp Asp Val Glu Val His Thr Ala Gln 275 280 sin nos ;o Arg Glu Glu Gln Ph 290 Asn Ser Thr Phe Arg Ser Val Ser Glu Leu ero lie Met 2 9 5 3 0 0 Leu Asn Gly l ys Glu Lys Cys Arg Val Asn Se: 315 325 Ala Ala Phe Pro Ala ero He Glu Lys Thr He ser Lys Thr Lys Glv 3 2 or 3 3 0 3 3 5 Arg Pro Lys Ala Pro Gln Val T”, :o Pro Lys Glu Gln 0 202 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met He Thr Asp Ph 355 3€v 365 ;o Glu Asp He 37 0 Thr Val Glu Trp Gln. Trp Asn Gly Gln Pro ala <slu 375 380 Asn Tyr Lys Asn Thr Gln Pro lie Met Asp Thr 385 390 3S5 Gly Ser Tyr che 400 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly As 05 Τ Κ V P ri Thr Cys Ser V i 2 0 ..eu His Glu Gly Leu His Asn His His Th: 425 430 ;lu Lys Ser Leu Ser His Ser Pro Gly Lyí 4 35 4 4 0 _ i i t ? ¿ L212> DNA í213> Mous <400> 60 gatgtcgtga tgactcagtc acccgcaagc ctggctgtgt cgctgggaca gcgggctact atctcgtgcc aac< 120 g t c t g t. c t c o t cg t a c 11 gg o c t. gg t a t c a g c a g a a g o c a ggacagccgc cgaaactcct catctacgcc gcctcatacc ttgcgtccgg aqtgccttcg

Claims

1. An isolated Tau-binding antibody or its binding fragment, characterized in that said Tau-binding antibody or its binding fragment comprises the following: a light-chain variable region comprising SEQ ID No.: 11, and a heavy-chain variable region comprising SEQ ID No.:

14. 13 Claims follow