RU2795485C2 - Anti-il31 antibodies for veterinary use - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: recombinant production of an antibody or an isolated antibody fragment that binds to canine IL31, and can be used in veterinary medicine for the treatment of an IL31-induced condition.

EFFECT: obtaining an antibody or an isolated antibody fragment capable of binding to IL31 and reducing the signaling function of IL31 in cells.

53 cl, 13 dwg, 5 tbl, 12 ex

Description

[0001] This application claims priority over International Application PCT/US2017/023788, filed March 23, 2017, and US Provisional Application No. 62/463,543, filed February 24, 2017, each of which is incorporated herein in its entirety. as a link for any purpose.

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

[0002] The present invention relates to isolated anti-IL31 antibodies, e.g., binding to canine IL31, and methods of using the same, e.g., to treat IL31-induced conditions or to reduce IL31 signaling function in cells, e.g., in companion animals such as like dogs, cats and horses.

BACKGROUND OF THE INVENTION

[0003] Interleukin 31 (IL31) is a cytokine produced primarily by Th2 cells and is believed to be involved in the development of skin diseases such as pruritus and other forms of allergic diseases (eg, atopic dermatitis). IL31 functions through binding to its receptor and activating downstream signaling molecules, such as JAK1 activation, and is thought to be responsible for many of the clinical problems associated with dermatitis and other disorders.

[0004] Companion animals such as cats, dogs, and horses suffer from numerous skin diseases similar to human skin diseases, including atopic dermatitis. However, the sequences of IL31 in humans, cats, dogs and horses are different. Thus, there remains a need for methods and compounds that can be used exclusively to bind companion animal IL31, to treat IL31-induced conditions, and to reduce IL31 signaling.

SUMMARY OF THE INVENTION

[0005] In some embodiments, an isolated antibody is provided that binds to canine IL31. In some embodiments, the antibody binds to an epitope containing amino acids 34-50 of SEQ ID NO:22. In some embodiments, the antibody binds to an epitope containing the amino acid sequence of SEQ ID NO:23. In some embodiments, the antibody binds to an epitope containing the amino acid sequence PSDX ₁ X ₂ KI (SEQ ID NO:45), in which X is any amino acid residue. In some embodiments, X ₁ is a hydrophobic amino acid. In some embodiments, X ₁ is selected from A, V, I, and L. In some embodiments, X ₁ is selected from V and I. In some embodiments, X ₂ is a hydrophilic amino acid. In some embodiments, X ₂ is selected from A, R, K, Q, and N. In some embodiments, X ₂ is selected from R and Q. In some embodiments, X ₁ is V, and X ₂ is R. In some embodiments, X ₁ is I, and X ₂ is Q. In some embodiments, the antibody binds to an epitope containing the amino acid sequence of SEQ ID NO:88.

[0006] In some embodiments, the antibody binds to canine IL31 with a dissociation constant (Kd) less than 5×10 ^-6 M, less than 1×10 ^-6 M, less than 5×10 ^-7 M, less than 1×10 ^–7 M, less than 5× ^10–8 M, less than 1× ^10–8 M, ^less than 5×10–9 M, less than 1× ^10–9 M, less than 5× ^10–10 M, less less than 1× ^10–10 M, less than 5× ^10–11 M, less than 1× ^10–11 M, less than 5× ^10–12 M, or less than 1× ^10–12 M measured by bilayer interferometry.

[0007] In some embodiments, the antibody reduces IL31 signaling in the companion animal species, as measured by a decrease in STAT-3 phosphorylation. In some embodiments, the companion animal species is a canine, feline, or equine species.

[0008] In some embodiments, the antibody binds to feline IL31 or equine IL31 as determined by immunoblotting and/or bilayer interferometry analysis. In some embodiments, the antibody competes with the M14 monoclonal antibody for binding to canine IL31. In some embodiments, the antibody competes with the M14 monoclonal antibody for binding to feline IL31. In some embodiments, the antibody does not bind to human IL31 as determined by immunoblotting and/or bilayer interferometry analysis.

[0009] In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a canine, canine, feline, feline, equine, equine, or chimeric antibody. In some embodiments, the antibody is a chimeric antibody comprising a murine variable heavy chain framework or a murine variable light chain framework.

[0010] In some embodiments, the antibody comprises a heavy chain and a light chain, where:

A. the heavy chain contains a CDR-H1 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:1; a CDR-H2 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:2, 62, 89, or 87; and a CDR-H3 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:3, and

b. the light chain contains a CDR-L1 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:63; a CDR-L2 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:9; and a CDR-L3 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:10.

[0011] In some embodiments, the antibody comprises a heavy chain comprising (a) a CDR-H1 containing the amino acid sequence of SEQ ID NO:1, (b) a CDR-H2 containing the amino acid sequence of SEQ ID NO:2 or 89, and (c ) CDR-H3 containing the amino acid sequence of SEQ ID NO:3.

[0012] In some embodiments, the antibody comprises a heavy chain comprising (a) a CDR-H1 containing the amino acid sequence of SEQ ID NO:1, (b) a CDR-H2 containing the amino acid sequence of SEQ ID NO:62 or 87, and (c ) CDR-H3 containing the amino acid sequence of SEQ ID NO:3.

[0013] In some embodiments, the antibody comprises a light chain containing (a) a CDR-L1 containing the amino acid sequence of SEQ ID NO:8, (b) a CDR-L2 containing the amino acid sequence of SEQ ID NO:9, and (c) a CDR –L3 containing the amino acid sequence of SEQ ID NO:10.

[0014] In some embodiments, the antibody comprises a light chain comprising (a) a CDR-L1 containing the amino acid sequence of SEQ ID NO:63, (b) a CDR-L2 containing the amino acid sequence of SEQ ID NO:9, and (c) a CDR –L3 containing the amino acid sequence of SEQ ID NO:10.

[0015] In some embodiments, the antibody comprises one or more of: (a) heavy chain variable region framework region 1 (HC-FR1) sequence SEQ ID NO:4, 70, or 79, (b) HC-FR2 sequence SEQ ID NO :5, 71 or 80, (c) HC-FR3 sequences SEQ ID NO:6, 72, 73 or 81, (d) HC-FR4 sequences SEQ ID NO:7, 74, or 82, (e) framework sequences 1 light chain variable region (LC-FR1) SEQ ID NO:11, 75 or 83, (f) LC-FR2 sequence SEQ ID NO:12, 76 or 84, (g) LC-FR3 sequence SEQ ID NO:13, 77 or 85 or (h) LC-FR4 sequence SEQ ID NO:14, 78 or 86.

[0016] In some embodiments, the antibody comprises:

A. (i) a variable light chain sequence having at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:24; (ii) a variable heavy chain sequence having at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:25; or (iii) a variable light chain sequence as defined in (i) and a variable heavy chain sequence as defined in (ii); or

b. (i) a variable light chain sequence having at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:16; (ii) a heavy chain variable sequence having at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:15; or (iii) a variable light chain sequence as defined in (i) and a variable heavy chain sequence as defined in (ii); or

With. (i) a variable light chain sequence having at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:32; (ii) a heavy chain variable sequence having at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:33; or (iii) a variable light chain sequence as defined in (i) and a variable heavy chain sequence as defined in (ii).

[0017] In some embodiments, the antibody comprises the variable light chain sequence of SEQ ID NO:24; SEQ ID NO:16; or SEQ ID NO:32. In some embodiments, the antibody comprises the variable heavy chain sequence of SEQ ID NO:25; SEQ ID NO:15; or SEQ ID NO:33. In some embodiments, the antibody comprises: the variable light chain sequence of SEQ ID NO:24 and the variable heavy chain sequence of SEQ ID NO:25; the variable light chain sequence of SEQ ID NO:16 and the variable heavy chain sequence of SEQ ID NO:15; or a variable light chain sequence of SEQ ID NO:32 and a variable heavy chain sequence of SEQ ID NO:33.

[0018] In some embodiments, the antibody is a chimeric antibody containing a heavy chain constant region or a light chain constant region derived from a companion animal.

[0019] In some embodiments, the antibody comprises (a) a canine heavy chain constant region selected from IgG-A, IgG-B, IgG-C, and IgG-D constant regions; (b) a feline heavy chain constant region selected from IgG1, IgG2a and IgG2b constant regions; or (c) an equine heavy chain constant region selected from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 and IgG7 constant regions.

[0020] In some embodiments, the antibody comprises:

A. (i) the light chain amino acid sequence of SEQ ID NO:26; (ii) the amino acid sequence of the heavy chain of SEQ ID NO:27; or (iii) a light chain amino acid sequence as defined in (i) and a heavy chain amino acid sequence as defined in (ii); or

b. (i) the light chain amino acid sequence of SEQ ID NO:30; (ii) the amino acid sequence of the heavy chain of SEQ ID NO:31; or (iii) a light chain amino acid sequence as defined in (i) and a heavy chain amino acid sequence as defined in (ii); or

With. (i) the light chain amino acid sequence of SEQ ID NO:34; (ii) the amino acid sequence of the heavy chain of SEQ ID NO:35; or (iii) a light chain amino acid sequence as defined in (i) and a heavy chain amino acid sequence as defined in (ii).

[0021] In some embodiments, the antibody comprises the light chain amino acid sequence of SEQ ID NO:21. In some embodiments, the antibody comprises the heavy chain amino acid sequence of SEQ ID NO:17; SEQ ID NO:18; SEQ ID NO:19; or SEQ ID NO:20.

[0022] In some embodiments, the antibody is an antibody fragment selected from Fv, scFv, Fab, Fab', F(ab') ₂ and Fab'-SH.

[0023] In some embodiments, the antibody is bispecific, wherein the antibody binds to IL31 and one or more antigens selected from IL17, TNFα, CD20, CD19, CD25, IL4, IL13, IL23, IgE, CD11α, IL6R, α4-integrin , IL12, IL1β or BlyS.

[0024] In some embodiments, an isolated nucleic acid is provided that encodes an anti-IL31 antibody disclosed herein above. In some embodiments, a host cell is provided that contains a nucleic acid encoding an anti-IL-31 antibody disclosed herein above. In some embodiments, a method for producing an anti-IL31 antibody is provided, which comprises culturing a host cell that contains a nucleic acid encoding an anti-IL31 antibody as disclosed herein above and isolating the antibody.

[0025] In some embodiments, a pharmaceutical composition is provided that comprises an anti-IL31 antibody disclosed herein and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition is provided that comprises an anti-IL-31 antibody disclosed herein and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises L-histidine, sodium chloride, and polysorbate 80.

[0026] In some embodiments, the implementation of the pharmaceutical composition has a pH in the range from 5.0 to 6.2. In some embodiments, the implementation of the pharmaceutical composition has a pH in the range from 5.0 to 6.0 or from 5.3 to 5.7 or equal to 5.5.

[0027] In some embodiments, the pharmaceutical composition contains L-histidine at a concentration of 5 mM to 100 mM, 10 mM to 50 mM, 20 mM to 30 mM, 10 mM to 30 mM, or 20 mM.

[0028] In some embodiments, the pharmaceutical composition contains sodium chloride at a concentration of 80 to 200 mM, 100 to 175 mM, 120 to 150 mM, or 140 mM.

[0029] In some embodiments, the pharmaceutical composition comprises polysorbate 80 at a concentration of 0.005 mg/mL to 0.5 mg/mL, 0.01 mg/mL to 0.1 mg/mL, or 0.05 mg/mL.

[0030] In some embodiments, the pharmaceutically acceptable carrier contains at least one sugar. In some embodiments, the pharmaceutical composition contains at least one sugar at a concentration of 0.5% to 20%, 1% to 10%, 1% to 5%, or 1% to 3%. In some embodiments, the pharmaceutically acceptable carrier comprises sucrose, trehalose, D-mannitol, maltose, and/or sorbitol.

[0031] In some embodiments, the pharmaceutically acceptable carrier contains an antibacterial agent. In some embodiments, the implementation of the pharmaceutical composition contains cresol or methylparaben. In some embodiments, the pharmaceutical composition contains 0.2% m-cresol and/or 0.9% methyl paraben.

The use of antibodies and pharmaceutical compositions

[0032] In some embodiments, methods for treating a companion animal species having an IL31-induced condition are provided, comprising administering to the companion animal species a therapeutically effective amount of an anti-Il31 antibody disclosed herein or a pharmaceutical composition comprising an antibody disclosed herein. description. In some embodiments, the companion animal species is a canine, feline, or equine species. In some embodiments, the IL31-induced condition is an pruritic or allergic condition. In some embodiments, the IL31-induced condition is selected from atopic dermatitis, pruritus, asthma, psoriasis, scleroderma, and eczema.

[0033] In some embodiments, the anti-IL31 antibody or pharmaceutical composition is administered parenterally. In some embodiments, the anti-IL31 antibody or pharmaceutical composition is administered intramuscularly, intraperitoneally, intracerebrospinally, subcutaneously, intraarterially, intrasynovially, intrathecally, or by inhalation.

[0034] In some embodiments, the method comprises administering an anti-IL31 antibody or pharmaceutical composition in combination with a Jak inhibitor, a PI3K inhibitor, an AKT inhibitor, or a MAPK inhibitor. In some embodiments, the method comprises administering an anti-IL31 antibody or pharmaceutical composition in combination with one or more antibodies selected from anti-IL17 antibody, anti-TNFα antibody, anti-CD20 antibody, anti-CD19 antibody, anti-CD25 antibody, anti-IL4 antibodies, anti-IL13 antibodies, anti-IL23 antibodies, anti-IgE antibodies, anti-CD11α antibodies, anti-IL6R antibodies, anti-α4-integrin antibodies, anti-IL12 antibodies, anti-IL1β antibodies and anti-BlyS antibodies .

[0035] In some embodiments, methods are provided for reducing IL31 signaling in a cell, comprising exposing the cell to an anti-IL31 antibody, a pharmaceutical composition disclosed herein, under conditions that allow the antibody to bind to extracellular IL31, thereby reducing binding to IL31 receptor and/or reducing the signaling function of IL31 in the cell. In some embodiments, the cell is exposed to the antibody or pharmaceutical composition ex vivo . In some embodiments, the cell is exposed to the antibody or pharmaceutical composition in vivo . In some embodiments, the cage is a dog cage, a cat cage, or a horse cage.

[0036] In some embodiments, a method is provided for detecting IL31 in a sample obtained from a companion animal species, comprising contacting the sample with an anti-IL31 antibody or pharmaceutical composition disclosed herein under conditions to allow binding of the antibody to IL31, and detection in the sample of the formation of a complex of antibodies with IL31. In some embodiments, the sample is a biological sample obtained from a dog, cat, or horse.

BRIEF DESCRIPTION OF THE DRAWINGS

[0037] FIG. 1A shows the sequence alignment of the variable light chains of the M14, M18, M19 and M87 mouse monoclonal antibody clones. In FIG. 1B shows sequence alignment of the variable heavy chains of M14, M18, M19 and M87 mouse monoclonal antibody clones.

[0038] FIG. 2A and FIG. 2B shows plots of canine IL31 binding to chimeric M14 antibody at various concentrations.

[0039] FIG. 3A and FIG. 3B shows plots of canine IL31 binding to canine M14 antibody at various concentrations.

[0040] In FIG. 4 shows immunoblots demonstrating suppressed canine IL31 signaling at various concentrations of caninized M14 antibody.

[0041] FIG. 5A and 5B show immunoblots for GST-canine IL31 deletions probed with M14 antibody and anti-GST antibody, respectively.

[0042] FIG. 6A and 6B show immunoblots for GST-canine IL31 deletions probed with M14 antibody and anti-GST antibody, respectively.

[0043] FIG. 7A and 7B show immunoblots of feline and equine IL-31 proteins fused to human Fc probed with M14 antibody and anti-FC antibody, respectively.

[0044] FIG. 8 shows the results of immunoblot analysis for detailed epitope mapping and alanine scan of mature canine IL31 epitope with anti-canine IL31 antibody (upper panel) and anti-GST antibody (lower panel).

[0045] FIG. 9 shows the results of immunoblot analysis of detailed epitope mapping and alanine scan of mature canine IL31 epitope with anti-canine IL31 antibody (upper panel) and anti-GST antibody (lower panel).

[0046] FIG. 10 shows the results of a detailed epitope mapping immunoblot analysis and alanine scan of a mature canine IL31 epitope with an anti-canine IL31 antibody (upper panel) and an anti-GST antibody (lower panel).

[0047] FIG. 11 shows the results of a detailed epitope mapping immunoblot analysis and alanine scan of a mature canine IL31 epitope with an anti-canine IL31 antibody (upper panel) and an anti-GST antibody (lower panel).

[0048] FIG. 12 shows the results of detailed epitope mapping immunoblot analysis and alanine scan of mature canine IL31 epitope with anti-canine IL31 antibody (upper panel) and anti-GST antibody (lower panel).

[0049] FIG. 13 shows an immunoblot demonstrating cross-reactivity of the M14 antibody to canine IL31 with walrus IL31.

DESCRIPTION OF SOME SEQUENCES

[0050] Table 1 lists some of the sequences mentioned in the present description.

Table 1: Description of some sequences SEQID NO: Subsequence Description 1 GDSITSGYW Amino acid sequence of the CDR-H1 variable heavy chain of mouse antibody clones M14, M18 and M19 2 YISYSGITDYNPSLKS Amino acid sequence of the CDR-H2 variable heavy chain of mouse antibody clones M14 and M19 62 YISYSGITYYNPSLKS Amino acid sequence of the CDR-H2 variable heavy chain of mouse antibody clone M18 89 YISYSGITDY Alternative amino acid sequence of the CDR-H2 variable heavy chain of mouse antibody clones M14 and M19 87 YISYSGITYY Alternative amino acid sequence of the CDR-H2 variable heavy chain of mouse antibody clone M18 3 ARYGNYGYAMDY Amino acid sequence of the CDR-H3 variable heavy chain of mouse antibody clones M14, M18 and M19 4 EVQLQESGPSLVKPSQTLSLTCSVT Amino acid sequence of the HC-FR1 framework region of the heavy chain variable region of mouse antibody clone M14 5 NWIRKFPGNNKLEYMG Amino acid sequence of the HC-FR2 framework region of the heavy chain variable region of mouse antibody clone M14 6 RISITRDTSKNQYYLQLNSVTTEDTATYYC Amino acid sequence of the HC-FR3 framework region of the heavy chain variable region of mouse antibody clone M14 7 WGQGTSVTVSS Amino acid sequence of the HC-FR4 framework region of the heavy chain variable region of mouse antibody clone M14 8 RASESVDTYGNSFMH Amino acid sequence of the CDR-L1 variable light chain of mouse antibody clones M14 and M19 63 RASESVDTYGNSFIH Amino acid sequence of the CDR-L1 variable light chain of M18 mouse antibody clones 9 RASNLES Amino acid sequence of the CDR-L2 variable light chain of mouse antibody clones M14, M18 and M19 10 QQSYEDPWT Amino acid sequence of the CDR-L3 variable light chain of mouse antibody clones M14, M18 and M19 eleven DIVLTQSPASLAVSLGQRATISC Amino acid sequence of the LC-FR1 framework region of the light chain variable region of mouse antibody clone M14 12 WYQQKSGQSPKLLIY Amino acid sequence of the LC-FR2 framework region of the light chain variable region of mouse antibody clone M14 13 GIPARFGGSGSRTDFFTLTIDPVEADDVATYYC Amino acid sequence of the LC-FR3 framework region of the light chain variable region of mouse antibody clone M14 14 FGGGTKLEIK Amino acid sequence of the LC-FR4 framework region of the light chain variable region of mouse antibody clone M14 15 EVQLVESGPSLVKPGGSLRLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRITISRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTLVTVSS Caninized amino acid sequence of the variable heavy chain of mouse antibody clone M14 16 DIVMTQSPASLSVSLGQRATISCRASESVDTYGNSFMHWYQQKPGQSPKLLIYRASNLESGIPARFGGSGSGTDFTLTIDPVQADDVATYYCQQSYEDPWTFGGGTKLEIK Caninized amino acid sequence of the variable light chain of mouse antibody clone M14 70 EVQLVESGPSLVKPGGSLRLTCSVT Caninized HC–FR1 M14 71 NWIRKFPGNNKLEYMG Caninized HC–FR2 M14 72 RITISRDTSKNQYYLQLNSVTTEDTATYYC Caninized HC–FR3 M14 73 NPSLKSRITISRDTSKNQYYLQLNSVTTEDTATYYC Alternative Canned HC–FR3 M14 74 WGQGTLVTVSS Caninized HC–FR4 M14 75 DIVMTQSPASLSVSLGQRATISC Caninized LC–FR1 M14 76 WYQQKPGQSPKLLIY Caninized LC–FR2 M14 77 GIPARFGGSGSGTDFTLTIDPVQADDVATYYC Caninized LC–FR3 M14 78 FGGGTKLEIK Caninized LC–FR4 M14 17 EVQLVESGPSLVKPGGSLRLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRITISRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTLVTVSSASTTAPSVFPLAPSCGSTSGSTVALACLVSGYFPEPVTVSWNSGSLTSGVHTFPSVLQSSGLHSLSSMVTVPSSRWPSETFTCNVV HPASNTKVDKPVFNECRCTDTPCPVPEPLGGPSVLIFPPKPKDILRITRTPEVTCVVLDLGREDPEVQISWFVDGKEVHTAKTQSREQQFNGTYRVVSVLPIEHQDWLTGKEFKCRVNHIDLPSPIERTISKARGRAHKPSVYVLPPSPKELSSSDTVSITCLIKDFYPPDIDVEWQSNGQQEPERKMTHRTPPQ LDEDGSYFLYSKLSVDKSRWQQGDPFTCAVMHETLQNHYTDLSLSHSPGK Caninized Heavy Chain Sequence of Mouse Antibody Clone M14 and Canine IgG-A 18 EVQLVESGPSLVKPGGSLRLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRITISRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTLVTVSSASTTAPSVFPLAPSCGSTSGSTVALACLVSGYFPEPVTVSWNSGSLTSGVHTFPSVLQSSGLYSLSSMVPSSRWPSETFTCNVAH PASKTKVDKPVPKRENGRVPRPPDCPKCPAPEMLGGPSVFIFPPKPKDTLLIARTPEVTCVVVDLDPEDPEVQISWFVDGKQMQTAKTQPREEQFNGTYRVVSVLPIGHQDWLKGKQFTCKVNNKALPSPIERTISKARGQAHQPSVYVLPPSREELSKNTVSLTCLIKDFFPPDIDVEWQSNGQQEPESKY RTTPPQLDEDGSYFLYSKLSVDKSRWQRGDTFICAVMHEALHNHYTQESLSHSPGK Caninized Heavy Chain Sequence of Mouse Antibody Clone M14 and Canine IgG-B 19 EVQLVESGPSLVKPGGSLRLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRITISRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTLVTVSSASTTAPSVFPLAPSCGSQSGSTVALACLVSGYIPEPVTVSWNSVSLTSGVHTFPSVLQSSGLYSLSSMVPSSRWPSETFTCNVA HPATNTKVDKPVAKECEKCNCNNCPCPGCGLLGGPSVFIFPPKPKDILVTARTPTVTCVVVDLDPENPEVQISWFVDSKQVQTANTQPREEQSNGTYRVVSVLPIGHQDWLSGKQFKCKVNNKALPSPIEEIISKTPGQAHQPNVYVLPPSRDEMSKNTVTLTCLVKDFFPPEIDVEWQSNGQQEPESKYRM TPPQLDEDGSYFLYSKLSVDKSRWQRGDTFICAVMHEALHNHYTQISLSHSPGK Caninized Heavy Chain Sequence of Mouse Antibody Clone M14 and Canine IgG–C 20 EVQLVESGPSLVKPGGSLRLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRITISRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTLVTVSSASTTAPSVFPLAPSCGSTSGSTVALACLVSGYFPEPVTVSWNSGSLTSGVHTFPSVLQSSGLYSLSSTVPSSRWPSETFTCNVV HPASNTKVDKPVPKESTCKCISPPCPVPESLGGPSVFIFPPKPKDILRITRTPEITCVVLDLGREDPEVQISWFVDGKEVHTAKTQPREQQFNSTYRVVSVLPIEHQDWLTGKEFKCRVNHIGLPSPIERTISKARGQAHQPSVYVLPPSPKELSSSDTVTLTCLIKDFFPPEIDVEWQSNGQPEPESKYHTTAPQ LDEDGSYFLYSKLSVDKSRWQQGDTFTCAVMHEALQNHYTDLSLSHSPGK Caninized Heavy Chain Sequence of Mouse Antibody Clone M14 and Canine IgG–D 21 DIVMTQSPASLSVSLGQRATISCRASESVDTYGNSFMHWYQQKPGQSPKLLIYRASNLESGIPARFGGSGSGTDFTLTIDPVQADDVATYYCQQSYEDPWTFGGGTKLEIKRNDAQPAVYLFQPSPDQLHTGSASVVCLLNSFYPKDINVKWKVDGVIQDTGIQESVTEQDKDSTYSLSSTLTM SSTEYLSHELYSCEITHKSLPSTLIKSFQRSECQRVD Caninized Light Chain Sequence of Mouse Antibody Clone M14 and Canine Light Chain Constant Region 22 MLSHTGPSRFALFLLCSMETLLSSHMAPTHQLPPSDVRKIILELQPLSRGLLEDYQKKETGVPESNRTLLLCLTSDSQPPRLNSSAILPYFRAIRPLSDKNIIDKIIEQLDKLKFQHEPETEISVPADTFECKSFILTILQQFSACLESVFKSLNSGPQ Amino acid sequence of canine IL31 23 PSDVRKIILELQPLSRG Canine IL31 epitope 24 DIVLTQSPASLAVSLGQRATISCRASESVDTYGNSFMHWYQQKSGQSPKLLIYRASNLESGIPARFGGSGSRTDFTLTIDPVEADDVATYYCQQSYEDPWTFGGGTKLEIK Amino acid sequence of the variable light chain of mouse antibody clone M14 25 EVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRISITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSS The amino acid sequence of the variable heavy chain of the mouse antibody clone M14 26 DIVLTQSPASLAVSLGQRATISCRASESVDTYGNSFMHWYQQKSGQSPKLLIYRASNLESGIPARFGGSGSRTDFTLTIDPVEADDVATYYCQQSYEDPWTFGGGTKLEIKRNDAQPAVYLFQPSPDQLHTGSASVVCLLNSFYPKDINVKWKVDGVIQDTGIQESVTEQDKDSTYSLSSTLTMSS TEYLSHELYSCEITHKSLPSTLIKSFQRSECQRVD Murine antibody clone M14 variable light chain chimeric sequence and canine light chain constant region 27 EVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRISITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSSASTTAPSVFPLAPSCGSTSGSTVALACLVSGYFPEPVTVSWNSGSLTSGVHTFPSVLQSSGLYSLSSMVPSSRWPSETFTCN VAHPASKTKVDKPVPKRENGRVPRPPDCPKCPAPEMLGGPSVFIFPPKPKDTLLIARTPEVTCVVVDLDPEDPEVQISWFVDGKQMQTAKTQPREEQFNGTYRVVSVLPIGHQDWLKGKQFTCKVNNKALPSPIERTISKARGQAHQPSVYVLPPSREELSKNTVSLTCLIKDFFPPDIDVEWQSNGQQEP ESKYRTTPPQLDEDGSYFLYSKLSVDKSRWQRGDTFICAVMHEALHNHYTQESLSHSPGK The chimeric sequence of the variable heavy chain of the mouse antibody clone M14 and canine IgG-B 28 MLSHAGPARFALFLLCCMETLLPSHMAPAHRLQ PSDVRKI ILELRPMSKGLLQDYLKKEIGLPESNHSSLPCLSSDSQLPHINGSAILPYFRAIRPLSDKNTIDKIIEQLDKLKFQREPEAKVSMPADNFERKNFILAVLQQFSACLEHVLQSLNSGPQ Amino acid sequence of feline IL31
NCBI No: XP_011286140.1
[felis catus] 29 MVSHIGSTRFALFLLCCLGTLMFSHTGPIYQLQPKEIQAIIVELQNLSKKLLDDYLNKEKGVQKFDSDLPSCFTSDSQAPGNINSSAILPYFKAISPSLNNDKSLYIIEQLDKLNFQNAPETEVSMPTDNFERKRFILTILRWFSNCLELAMKTLTTAEQALPPLDPSTPHAGAVALTHHQQDRTALDRAVFPFVWAAPRGGEV GDGGH Amino acid sequence of equine IL31 thirty DIVLTQSPASLAVSLGQRATISCRASESVDTYGNSFMHWYQQKSGQSPKLLIYRASNLESGIPARFGGSGSRTDFTLTIDPVEADDVATYYCQQSYEDPWTFGGGTKLEIKRSDAQPSVFLFQPSLDELHTGSASIVCILNDFYPKEVNVKWKVDGVVQNKGIQESTTEQNSKDSTYSLSSTLTMSSTEY QSHEKFSCEVTHKSLASTLVKSFNRSECQRE Murine antibody clone M14 variable light chain chimeric sequence and feline light chain constant region 31 EVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRISITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSSASTTAPSVFPLAPSCGTTSGATVALACLVLGYFPEPVTVSWNSGALTSGVHTFPAVLQASGLYSLSSMVTVPSSRWLSDTFT CNVAHPPSNTKVDKTVRKTDHPPGPKPCDCPKCPPPEMLGGPSIFIFPPKPKDTLSISRTPEVTCLVVDLGPDDSDVQITWFVDNTQVYTAKTSPREEQFNSTYRVVSVLPILHQDWLKGKEFKCKVNSKSLPSPIERTISKAKGQPHEPQVYVLPPAQEELSRNKVSVTCLIKSFHPPDIAVEWEITGQPEP ENNYRTTPPQLDSDGTYFVYSKLSVDRSHWQRGNTYTCSVSHEALHHTQKSLTQSPGK Murine antibody clone M14 chimeric heavy chain variable sequence and feline heavy chain constant region 32 EIQMTQSPSSLSASPGDRVTISCRASESVDTYGNSFMHWYQQKPGQSPKLLIYRASNLESGVPSRFSGSGSGTDFTLTISSLEPEDAATYYCQQSYEDPWTFGGGTKLEIK Felinized variable light chain sequence of mouse antibody clone M14 33 DVQLVESGGDLVKPGGSLRLTCSVTGDSITSGYWNWVRQAPGKGLQWVAYISYSGITDYADSVKGRFTISRDNAKNTLYLQLNNLKAEDTATYYCARYGNYGYAMDYWGQGTLVTVSS Felinized variable heavy chain sequence of mouse antibody clone M14 79 DVQLVESGGDLVKPGGSLRLTCSVT Felined HC–FR1 M14 80 NWVRQAPGKGLQWVA Felined HC-FR2 M14 81 ADSVKGRFTISRDNAKNTLYLQLNNLKAEDTATYYCA Felined HC–FR3 M14 82 WGQGTLVTVSS Felined HC–FR4 M14 83 EIQMTQSPSSLSASPGDRVTISC Felined LC–FR1 M14 84 WYQQKPGQSPKLLIY Felined LC–FR2 M14 85 GVPSRFSGSGSGTDFTLTISSLEPEDAATYYC Felined LC–FR3 M14 86 FGGGTKLEIK Felined LC–FR4 M14 34 EIQMTQSPSSLSASPGDRVTISCRASESVDTYGNSFMHWYQKPGQSPKLLIYRASNLESGVPSRFSGSGSGTDFTLTISSLEPEDAATYYCQQSYEDPWTFGGGTKLEIKRSDAQPSVFLFQPSLDELHTGSASIVCILNDFYPKEVNVKWKVDGVVQNKGIQESTTEQNSKDSTYSLSSTLTMSSTEYQSH EKFSCEVTHKSLASTLVKSFNRSECQRE Felinized light chain sequence of mouse antibody clone M14 35 DVQLVESGGDLVKPGGSLRLTCSVTGDSITSGYWNWVRQAPGKGLQWVAYISYSGITDYADSVKGRFTISRDNAKNTLYLQLNNLKAEDTATYYCARYGNYGYAMDYWGQGTLVTVSSASTTAPSVFPLAPSCGTTSGATVALACLVLGYFPEPVTVSWNSGALTSGVHTFPAVLQASGLYSLSSMVTVPSSRWLSDT FTCNVAHPPSNTKVDKTVRKTDHPPGPKPCDCPKCPPPEMLGGPSIFIFPPKPKDTLSISRTPEVTCLVVDLGPDDSDVQITWFVDNTQVYTAKTSPREEQFNSTYRVVSVLPILHQDWLKGKEFKCKVNSKSLPSPIERTISKAKGQPHEPQVYVLPPAQEELSRNKVSVTCLIKSFHPPDIAVEWEITGQPE PENNYRTTPPQLDSDGTYFVYSKLSVDRSHWQRGNTYTCSVSHEALHSHHTQKSLTQSPGK Felinized heavy chain sequence of mouse antibody clone M14 36 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISCRASESVDTYGNSFMHWYQQKSGQSPKLLIYRASNLESGIPARFGGSGSRTDFTLTIDPVEADDVATYYCQQSYEDPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSW TDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC Light chain amino acid sequence of mouse antibody clone M14 37 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSPGQRATISCRASESVDTYGNSFIHWYQQKPGQSPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVETDDVATYYCQQSYEDPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQ DSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC Light chain amino acid sequence of mouse antibody clone M18 38 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISCRASESVDTYGNSFMHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDIATYYCQQSYEDPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTD QDSKDSTYSMSSTLLTTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC Light chain amino acid sequence of mouse antibody clone M19 39 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISYRASKSVSTSGYSYMHWNQQKPGQPPRLLIYLVSNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHIRELTRSFGGGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQ DSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC Light chain amino acid sequence of mouse antibody clone M87 64 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSPGQRATISCRASESVDTYGNSFIHWYQQKPGQSPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVETDDVATYYCQQSYEDPWTFGGGTKLEIK Amino acid sequence of the variable light chain of the mouse antibody clone M18 65 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISCRASESVDTYGNSFMHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDIATYYCQQSYEDPWTFGGGTKLEIK Amino acid sequence of the variable light chain of mouse antibody clone M19 66 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLGQRATISYRASKSVSTSGYSYMHWNQQKPGQPPRLLIYLVSNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHIRELTRSFGGGTKLEIK Amino acid sequence of the variable light chain of mouse antibody clone M87 40 MAVLGLLLCLVTFPSCVLSEVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRISITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSSAKTTPPSVYPLAPGS Amino acid sequence of the heavy chain variable and hinge regions of mouse antibody clone M14 41 MAVLGLLFCLVTFPSCVLSEVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRISITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSSAKTTPPSVYPLAPGS Amino acid sequence of the heavy chain variable and hinge regions of mouse antibody clone M18 42 MAVLGLLFCLVTFPSCVLSEVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNELEYMGYISYSGITYYNPSLKSRFSITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSSAKTTPPSVYPLAPGS Amino acid sequence of the heavy chain variable and hinge regions of mouse antibody clone M19 43 MAVLGLLFCLVTFPSCVLSEVKLVESGGGLVQPGGSLRLSCATSGFTFTDYYMNWVRQPPGKALEWLGFIRNKANGYTTEYSASVKGRFTISRDNSQSILYLQMNTLRAEDSATYYCARDYYGSCFDYWGQGTTLTVSSAKTTPPSVYPLAPGS Amino acid sequence of the heavy chain variable and hinge regions of mouse antibody clone M87 67 MAVLGLLFCLVTFPSCVLSEVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNKLEYMGYISYSGITDYNPSLKSRISITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSS The amino acid sequence of the variable heavy chain of the mouse antibody clone M18 68 MAVLGLLFCLVTFPSCVLSEVQLQESGPSLVKPSQTLSLTCSVTGDSITSGYWNWIRKFPGNELEYMGYISYSGITYYNPSLKSRFSITRDTSKNQYYLQLNSVTTEDTATYYCARYGNYGYAMDYWGQGTSVTVSS The amino acid sequence of the variable heavy chain of the mouse antibody clone M19 69 MAVLGLLFCLVTFPSCVLSEVKLVESGGGLVQPGGSLRLSCATSGFTFTDYYMNWVRQPPGKALEWLGFIRNKANGYTTEYSASVKGRFTISRDNSQSILYLQMNTLRAEDSATYYCARDYYGSCFDYWGQGTTLTVSS Amino acid sequence of the variable heavy chain of mouse antibody clone M87 44 SSHMAPTHQLP PSDVRKI ILELQPLSRGLLEDYQKKETGVPESNRTLLLCLTSDSQPPRLNSSAILPYFRAIRPLSDKNIIDKIIEQLDKLKFQHEPETEISVPADTFECKSFILTILQQFSACLESVFKSLNSGPQ Amino acid sequence of mature canine IL31 45 PSDX ₁ X ₂ KI, where X ₁ and X ₂ represent any amino acid; or where X ₁ is a hydrophobic amino acid, or where X ₁ is selected from A, V, I and L, or where X ₁ is selected from V and I; and where X ₂ represents a hydrophilic amino acid, or where X ₂ is selected from A, R, K, Q and N; or where X ₂ is selected from R and Q. Epitope IL31 88 PSDVRKI Alternative canine IL31 epitope 46 MASHSGPSTSVLFLFCCLGGWLASHTLPVRLLRPSDDVQKIVEELQSLSKMLLKDVEEEKGVLVSQNYTLPCLSPDAQPPNNIHSPAIRAYLKTIRQLDNKSVIDEIIEHLDKLIFQDAPETNISVPTDTHECKRFILTISQQFSECMDLALKSLTSGAQQATT Amino acid sequence of human IL31 precursor
NCBI No: NP_001014358.1 47 MLSHAGPARFALFLLCFMGTSLTSQTAPIHQLH PSDVRKI ILELQPLSKGLLEDYLKKEMGVPESNHFLLPCLTSDSQPPRINSSAILPYFRAIRPLSDKNTINKIIEQLDKLKFQHEPETEVSVPADTFESKSFILAILQQFSACLDHVFKSLNPGPQQVMQGHLIEPIPSGTADV Predicted amino acid sequence of walrus IL31
NCBI No: XP_004395998.1
[odobenus rosmarus divergens] 48 MLSHAGPARFALFLLCFMGTSLTSQTAPIHQLH PSDVRKI ILELQPLSKGLLEDYLKKEMGVPESNHFLLPCLTSDSQPPRINSSAILPYFRAIRPLSDKNTINKIIEQLDKLKFQHEPETEVSVPADTFESKSFILAILQQFSACLDHVFKSLNPGPQQVMQGHLIEPIPSGTADV GSGSHHHHHH Walrus_IL31_His6 49 MASHSGPATSVLFLLCCLGGWLTSHTLPVHFLQ PSDIQKI VEELQSLSKMLLKDVKEDKGVLVSQNYTLPCLTPDAQPPNIIHSPAIRAYLKTIRELDNKSVIDEIIEHLDKLIFQDAPETNISVPTDTHECKRFILTISQQFSECMDLALKSLTSGAQQATT Predicted amino acid sequence of olive baboon IL31
NCBI No: XP_003907358.1
[papio anubis] 50 MASHSGPATSVLFLLCCLGGWLTSHTLPVHFLQ PSDIQKI VEELQSLSKMLLKDVKEDKGVLVSQNYTLPCLTPDAQPPNIIHSPAIRAYLKTIRELDNKSVIDEIIEHLDKLIFQDAPETNISVPTDTHECKRFILTISQQFSECMDLALKSLTSGAQQATT GSGSHHHHHH Olive baboon_IL31_His6 51 MLSRAVPAGFALFLLCYMETLLTSHTAPTHRLP PSDVRKI ILELQPLSKGLLEDYLKKETGLPESNHSLVPCLTSDSESEAPHINSSAILPYFRAIRPLSDKNVIDKIIEQLDKLKFQHEPETEVSVPADTFEGKSFILTILQQFSACLERVFKSLNPGAQ Predicted amino acid sequence of polar bear IL31
NCBI No: XP_008687166.1
[ursus maritimus] 52 mlshagparfalfllcfmetsltsqtvpihqlq psdvrki ilelqplskglledylkkemgvpesnhfllpcltsdsqpprinssailpyfrairplsdkniinkiieqldklkfqhepetevsvpadtfesksfiltilqqfsaclghvlkslnpgpqqvmqghlakpipsgtadmyetlrhhh Predicted amino acid sequence of Weddell seal IL31
NCBI No: XP_006746595.1
[leptonychotes weddellii] 53 mlshagparfalfllccmetllpshmapthrlq psdvrki ilelrpmskgllqdylkkemglpesnhsslpclssdsqlphingsailpyfrairplsdkntidkiieqldklkfqrepeaevsmpadnferknfilavlqqfsaclehvlqslnsgpq Predicted amino acid sequence of Amur tiger IL31
NCBI No: XP_007079636.1
[panthera tigris altaica] 54 mlshagparfalfllccmetllpshmapthrlq psdvrki ilelrpmskgllqdylkkemglpesnhsslpclssdsqlphingsailpyfrairplsdkntidkiieqldklkfqrepeaevsmpadnferknfilavlqqfsaclehvlqslnsgsq Predicted amino acid sequence of cheetah IL31
NCBI No: XP_014919275.1
[acinonyx jubatus] 55 mashsgpatsvlfllcclggwltshtlpvhflq psdiqki veelqslskmllkdvkedkgvlvsqnytlpcltpdaqppniihspairaylktirqldnksvideiiehldklifqdapetnisvptdtheckrfiltisqqfsecmdlalksltsgaqqatt Predicted amino acid sequence of cynomolgus monkey IL31
GenBank No: EHH66805.1
[macaca fascicularis] 56 gpatsvlfllcclggwltshtlpvhflq psdiqki veelqslskmllkdvkedkgvlvsqnytlpcltpdaqppniihspairaylktirqldnksvideiiehldklifqdapetnisvptdtheckrfiltisqqfsecmdlalksltsgaqqatt Predicted amino acid sequence of IL31 rhesus poppy (partial)
GenBank No: EHH21279.1
[macaca mulatta] 57 mashsgpatsvlfllcclggwltshtlpvhflq psdiqki vkelqslskmllkdvkedkgvlvsqnytlpcltpdaqppniihspairaylktirqldnrsvideiiehldklifqdapetnisvptdtheckrfiltisqqfsecmdlalksltsgaqqatt Predicted amino acid sequence of mandrill IL31
NCBI No: XP_011819882.1
[mandrillus leucophaeus] 58 mashsgpatsvlfllcclggwltshtlpvhflq psdiqki veelqslskmllkdvkedkgvlvsqnyklpcltpdaqppniihspairaylktirqldnksvideiiehldklifqdapetnisvptdtheckrfiltisqqfsecmdlalksltsgaqqatt Predicted amino acid sequence of green macaque IL31
NCBI No: XP_008003211.1
[chlorocebus sabaeus] 59 mashsgpatsvlfllcclggwltshtlpvhflq psdiqki veelqslskmllkdvkedkgvlvsqnytlpcltpdaqppniihspairaylktirqldnrsvideiiehldklifqdapetnisvptdtheckrfiltisqqfsecmdlalksltsgaqqatt Predicted amino acid sequence of smoky mangabey IL31
NCBI No: XP_011926625.1
[cercocebus atys] 60 mashsgpttsvlfllcclggwltshtlpvhflr psdiqki veelqslskmllkdveedkgvlvsqnytlpcltpdaqppniihspairaylktirqldnksvideiiehldklifqdapetnisvptdtheckrfiltisqqfsecmdlalksltsgaqqatt Predicted amino acid sequence of golden snub-nosed monkey IL31
NCBI No: XP_010366647.1
[rhinopithecus roxellana] 61 MIFHTGTTKPTLVLLCCIGTWLATCSLSFGAPISKEDLRTTIDLLKQESQDLYNNYSIKQASGMSADESIQLPCFSLDREALTNISVIIAHLEKVKVLSENTVDTSWVIRWLTNISCFNPLNLNISVPGNTDESYDCKVFVLTVLKQFSNCMAELQAKDNTTC Amino acid sequence of the mouse IL31 precursor
NCBI No.: NP_083870
[musmusculus]

DESCRIPTION OF SOME EMBODIMENTS

[0051] Antibodies are provided that bind to canine IL31, feline IL31, or equine IL31. Also provided are heavy chains and light chains of antibodies that are capable of forming antibodies that bind to IL31. In addition, antibodies, heavy chains and light chains containing one or more specific complementarity determining regions (CDRs) are provided. Polynucleotides encoding antibodies to canine IL31 are provided. Methods for making or purifying anti-canine IL31 antibodies are also provided. Methods of treatment with anti-canine IL31 antibodies are provided. Such methods include, without limitation, methods for treating IL31-induced conditions in a companion animal species. Methods for detecting IL31 in a sample obtained from a companion animal species are provided.

[0052] To facilitate understanding in the present description, the following are definitions of terms used in the present description.

[0053] As used herein, numerical terms such as Kd are calculated from scientific measurements and are therefore subject to a corresponding measurement error. In some cases, a numeric term may include numeric values rounded to the nearest significant number.

[0054] As used herein, the singular means "at least one" or "one or more" unless otherwise indicated. Used in the present description, the term "or" means "and/or" unless otherwise indicated. In the context of a multiple-dependency claim, the use of "or" when referring to another claim refers only to those claims.

Anti-IL31 antibodies

[0055] Novel antibodies directed against IL31 are provided, such as antibodies that bind to canine IL31, feline IL31, and/or equine IL31. Anti-IL31 antibodies provided herein include, without limitation, monoclonal antibodies, mouse antibodies, chimeric antibodies, caninized antibodies, felinized antibodies, and equinized antibodies. In some embodiments, the anti-IL31 antibody is an isolated mouse monoclonal antibody such as M14, M18, M19 and M87.

[0056] Monoclonal antibodies M14, M18, M19 and M87 were isolated as follows. Briefly, mice were immunized with canine IL31 and mouse monoclonal antibody clones were generated using standard hybridoma technology. For screening hybridoma clones producing antibodies that bind to IL31, used enzyme-linked immunosorbent assay (ELISA). Based on the binding affinity and cell functional analysis described herein, hybridoma clones producing monoclonal antibodies M14, M18, M19 and M87 were selected for further study. The variable heavy chain (VH) and variable light chain (VL) of each of the four clones were sequenced and analyzed by sequence alignment (FIG. 1).

[0057] The amino acid sequences of the M14 monoclonal antibody are also provided herein. For example, for the monoclonal antibody M14, variable heavy chain CDRs (SEQ ID NOs:1-3; SEQ ID NO:89 is an alternative definition of CDR-H2), light chain variable region CDRs (SEQ ID NOs:8-10), framework sequences heavy chain variable region regions (SEQ ID NOS:4-7); and light chain variable region framework sequences (SEQ ID NOS:11-14). The amino acid sequences of the variable light chain, light chain, variable heavy chain, and heavy chain variable and hinge regions of the M14 monoclonal antibody are shown (SEQ ID NOS: 24, 36, 25, and 40, respectively).

[0058] In addition, the CDR amino acid sequences, framework sequences, variable light chain, variable heavy chain sequences of the M18, M19 and M87 monoclonal antibodies are provided (see, for example, FIG. 1). Variable Light Chain CDR (SEQ ID NO:1-3; SEQ ID NO:89 is an alternative definition of CDR-H2), Variable Light Chain CDR (SEQ ID NO:8-10), Variable Light Chain (SEQ ID NO:65) provided , light chain (SEQ ID NO:38), heavy chain variable and hinge regions (SEQ ID NO:42), and heavy chain variable (SEQ ID NO:68) of the M19 monoclonal antibody. Variable heavy chain CDRs (SEQ ID NOs: 1, 62 and 3; SEQ ID NO: 87 is an alternative definition of CDR-H2), variable light chain CDRs (SEQ ID NOs: 63, 9 and 10), variable light chain (SEQ ID NO:64), light chain (SEQ ID NO:37), heavy chain variable and hinge regions (SEQ ID NO:41) and variable heavy chain (SEQ ID NO:67) of the M18 monoclonal antibody. The variable light chain (SEQ ID NO:66), light chain (SEQ ID NO:39), variable and heavy chain hinge regions (SEQ ID NO:43), and variable heavy chain (SEQ ID NO:69) of the M87 monoclonal antibody are provided.

[0059] This application also provides chimeric, caninized, felinized and equinized antibodies derived from monoclonal antibodies M14, M18, M19 and M87. In some embodiments, the amino acid sequences of the caninized M14 monoclonal antibody are provided, such as SEQ ID NOS: 15-21 and 70-78. In some embodiments, the amino acid sequences of felinized antibodies derived from the M14 monoclonal antibody are provided, such as SEQ ID NOS:32-35 and 79-86. In some embodiments, the amino acid sequences of chimeric antibodies derived from the M14 monoclonal antibody are provided, such as SEQ ID NOS: 26, 27, 30, and 31.

[0060] The term "antibody" is used herein in its broadest sense and encompasses various antibody structures, including, without limitation, monoclonal antibodies, polyclonal antibodies, polyspecific antibodies (e.g., bispecific (such as bispecific T cell recruiters) and trispecific antibodies ) and antibody fragments (such as Fab, F(ab') ₂ , ScFv, mini-body, dia-body, tri-body and tetra-body), provided they exhibit the required antigen-binding activity. Canine, feline, and equine species have different varieties (classes) of antibodies that are common to many mammals.

[0061] The term "antibody" includes, without limitation, fragments that are capable of binding to an antigen, such as Fv, single chain Fv (scFv), Fab, Fab', di-scFv, sdAb (single domain antibody) and (Fab') ₂ (including the chemically bonded F(ab') ₂ ). Cleavage of antibodies with papain produces two identical antigen-binding fragments, called "Fab" fragments, each of which has one antigen-binding site, and the remaining "Fc" fragment, whose name reflects its ability to easily crystallize. Treatment with pepsin produces an F(ab') ₂ fragment that has two antigen binding sites and is still capable of antigen crosslinking. The term antibody also includes, without limitation, chimeric antibodies, humanized antibodies, and antibodies of various species such as mouse, human, cynomolgus monkey, dog, cat, horse, and the like. In addition, for all antibody constructs provided herein, variants are also contemplated having sequences from other organisms. Thus, if a mouse variant of an antibody is disclosed, one skilled in the art will understand how to convert an antibody based on a mouse sequence into an antibody with a cat, dog, horse or the like sequence. Antibody fragments also include any orientation of single chain scFv, tandem di-scFv, diabody, tandem tri-sdcFv, minibody, and the like. Antibody fragments also include nanobodies (sdAb, an antibody having a single monomeric domain, such as a pair of heavy chain variable domains, without a light chain). In some embodiments, an antibody fragment may be referred to as being species-specific (eg, mouse scFv or canine scFv). In this case, the sequence of at least a portion of the non-CDR regions is meant, not the origin of the construct. In some embodiments, the antibodies are labeled or conjugated to a second fragment.

[0062] The terms "label" and "detectable label" mean a fragment attached to an antibody or its analyte that allows detection of a reaction (eg, binding) between members of a specific binding pair. A labeled member of a specific binding pair is referred to as "detectable labeled". Thus, the term "labeled binding protein" refers to a protein with an embedded label that allows identification of the binding protein. In some embodiments, the label is a detectable marker that can produce a visually or instrumentally detectable signal, such as incorporating a radiolabeled amino acid or attaching biotinylated fragments to a polypeptide that can be detected using labeled avidin (e.g., streptavidin containing fluorescently labeled or having enzymatic activity that can be detected by optical or colorimetric methods). Examples of labels for polypeptides include, without limitation, the following: radioisotopes or radionuclides (e.g. ³ H, ¹⁴ C, ³⁵ S, ⁹⁰ Y, ⁹⁹ Tc, ¹¹¹ In, ¹²⁵ I, ¹³¹ I, ¹⁷⁷ Lu, ¹⁶⁶ Ho, or ¹⁵³ Sm) ; chromogens, fluorescent labels (eg, FITC, rhodamine, lanthanide phosphors), enzymatic labels (eg, horseradish peroxidase, luciferase, alkaline phosphatase); chemiluminescent markers; biotinylated groups; predetermined polypeptide epitopes recognized by the secondary reporter (eg, leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags); and magnetic agents such as gadolinium chelates. Typical examples of labels commonly used for immunoassays include moieties that generate light, such as acridinium compounds, and moieties that generate fluorescence, such as fluorescein. In this regard, the fragment itself may have an undetectable label, but may become detectable upon reaction with another fragment.

[0063] The term "monoclonal antibody" refers to an antibody of a substantially homogeneous population of antibodies, i. the individual antibodies that make up a population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific and directed against a single antigenic site. In addition, unlike polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. Thus, a monoclonal antibody sample can bind to the same epitope on an antigen. The modifier "monoclonal" indicates the nature of the antibody derived from a substantially homogeneous population of antibodies and should not be construed as requiring the production of the antibody by any particular method. For example, monoclonal antibodies can be made by the hybridoma method first described by Kohler and Milstein, 1975, Nature 256:495, or can be made by recombinant DNA methods such as those described in US Pat. No. 4,816,567. Monoclonal antibodies can also be isolated from phage libraries prepared by the methods described in, for example, McCafferty et al., 1990, Nature 348: 552-554.

[0064] In some embodiments, the monoclonal antibody is an isolated mouse antibody selected from clones M14, M18, M19, and M87.

[0065] "Amino acid sequence" means the sequence of amino acid residues in a peptide or protein. The terms "polypeptide" and "protein" are used interchangeably to refer to a polymer composed of amino acid residues and are not limited to a minimum length. Such polymers of amino acid residues may contain natural or non-natural amino acid residues and include, without limitation, peptides, oligopeptides, dimers, trimers and multimers of amino acid residues. This definition covers both full-length proteins and their fragments. The terms also include modifications to the polypeptide after expression, such as glycosylation, sialylation, acetylation, phosphorylation, and the like. In addition, for the purposes of the present invention, the term "polypeptide" refers to a protein that includes modifications such as deletions, additions and substitutions (usually conservative in nature) in the native sequence, provided that the protein retains the desired activity. These modifications may be intentional, such as those resulting from site-directed mutagenesis, or may be accidental, such as resulting from mutations in hosts that produce proteins, or errors due to PCR amplification.

[0066] As used herein, the term "IL31" refers to any native IL31 that results from the expression and processing of IL31 in a cell. The term includes IL31 from any vertebrate source, including mammals such as primates (eg, humans and cynomolgus monkeys), rodents (eg, mice and rats), and companion animals (eg, dogs, cats, and horses), unless otherwise noted. . The term also includes naturally occurring variants of IL31, such as splice variants or allelic variants.

[0067] In some embodiments, canine IL31 comprises the amino acid sequence of SEQ ID NO:22 or SEQ ID NO:44. In some embodiments, feline IL31 comprises the amino acid sequence of SEQ ID NO:28. In some embodiments, equine IL31 comprises the amino acid sequence of SEQ ID NO:29. In some embodiments, human IL31 contains the amino acid sequence of SEQ ID NO:46. In some embodiments, walrus IL31 comprises the amino acid sequence of SEQ ID. NO: 47. In some embodiments, mouse IL31 comprises the amino acid sequence of SEQ ID NO:61. In other embodiments, IL31 comprises the amino acid sequence of SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, or SEQ ID NO:60.

[0068] The term "IL31-binding domain" of an antibody means a binding domain formed by the light chain and heavy chain of an anti-IL31 antibody that binds to IL31.

[0069] In some embodiments, the IL31-binding domain binds to canine IL31 with a higher affinity than it binds to human IL31. In some embodiments, the IL31-binding domain binds to IL31 of other companion animals, such as feline IL31 or equine IL31. In some embodiments, the IL31 binding domain does not bind to human IL31.

[0070] As used herein, the term "epitope" refers to the site on a target molecule (e.g., an antigen, such as a protein, nucleic acid, carbohydrate, or lipid) to which an antigen-binding molecule (e.g., an antibody, antibody fragment, or scaffold protein) binds. containing antibody binding sites). Epitopes often include a reactive surface grouping molecules such as amino acids, polypeptides, or carbohydrate side chains, and have specific three-dimensional structural characteristics as well as specific charge characteristics. Epitopes can be formed from both adjacent and adjacent non-adjacent residues (eg, amino acids, nucleotides, sugars, lipid fragment) of the target molecule. Epitopes formed from contiguous residues (eg, amino acids, nucleotides, sugars, lipid moiety) are usually retained by denaturing solvents, while folded epitopes are usually "lost" upon treatment with denaturing solvents. An epitope may include, without limitation, at least 3, at least 5, or 8-10 residues (eg, amino acids or nucleotides). In some examples, an epitope is less than 20 residues (eg, amino acids or nucleotides) long, less than 15 residues, or less than 12 residues long. Two antibodies can bind to the same epitope on an antigen if they exhibit competitive binding to the antigen. In some embodiments, an epitope can be identified by a certain minimum distance to a residue in a CDR on an antigen-binding molecule. In some embodiments, an epitope can be identified by the above distance and further limited to those residues involved in a bond (eg, hydrogen bond) between an antibody residue and an antigen residue. An epitope can also be identified using various scans, for example an alanine or arginine scan can indicate one or more residues with which an antigen-binding molecule can interact. Unless explicitly stated, the set of residues as an epitope does not exclude that other residues may also be part of the epitope for a particular antibody. Rather, the presence of such a set signifies a minimal repertoire (or set of species) of epitopes. Thus, in some embodiments, the set of residues identified as an epitope denotes the minimum significant epitope for an antigen, rather than a list of exclusion residues for an epitope on an antigen.

[0071] In some embodiments, the implementation of the epitope contains the amino acid sequence PSDX ₁ X ₂ KI (SEQ ID NO:45), where X is any amino acid residue. In some embodiments, X ₁ is a hydrophobic amino acid. In some embodiments, X ₁ is selected from A, V, I, and L. In some embodiments, X ₁ is selected from V and I. In some embodiments, X ₂ is a hydrophilic amino acid. In some embodiments, X ₂ is selected from A, R, K, Q, and N. In some embodiments, X ₂ is selected from R and Q. In some embodiments, X ₁ is V, and X ₂ is R. In some embodiments, implementation X ₁ represents I, and X ₂ represents Q. In some embodiments, the implementation of the epitope contains the amino acid sequence of SEQ ID NO:88. In some embodiments, the implementation of the epitope contains the amino acid sequence of SEQ ID NO:23. In some embodiments, the epitope is within amino acids 34-50 of SEQ ID NO:22. In some embodiments, the epitope comprises amino acids 34-50 of SEQ ID NO:22.

[0072] The term "CDR" means a complementarity determining region defined by at least one identification method known to one of skill in the art. In some embodiments, CDRs may be defined according to any of Chothia numbering schemes, Kabat numbering schemes, a combination of Kabat and Chothia, an AbM definition, a contact definition, or a combination of Kabat, Chothia, AbM, or contact definitions. The various CDRs in an antibody may be designated by their respective chain number and type, including, but not limited to, CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3. As used herein, the term "CDR" also encompasses "hypervariable region" or HVR, including hypervariable loops.

[0073] In some embodiments, the anti-IL31 antibody comprises a heavy chain containing (a) a CDR-H1 containing the amino acid sequence of SEQ ID NO:1; (b) a CDR-H2 containing the amino acid sequence of SEQ ID NO:2, SEQ ID NO:62, SEQ ID NO:89, or SEQ ID NO:87; or (c) a CDR-H3 containing the amino acid sequence of SEQ ID NO:3. In some embodiments, the anti-IL31 antibody comprises a light chain containing (a) a CDR-L1 containing the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:63; (b) CDR-L2 containing the amino acid sequence of SEQ ID NO:9; or (c) a CDR-L3 containing the amino acid sequence of SEQ ID NO:10.

[0074] In some embodiments, the anti-IL31 antibody comprises a heavy chain comprising (a) a CDR-H1 containing the amino acid sequence of SEQ ID NO:1, (b) a CDR-H2 containing the amino acid sequence of SEQ ID NO:2 or 89, and (c) CDR-H3 containing the amino acid sequence of SEQ ID NO:3; and a light chain containing (a) a CDR-L1 containing the amino acid sequence of SEQ ID NO:8 or 63, (b) a CDR-L2 containing the amino acid sequence of SEQ ID NO:9, and (c) a CDR-L3 containing the amino acid sequence SEQ ID NO:10.

[0075] In some embodiments, the anti-IL31 antibody comprises a heavy chain comprising (a) a CDR-H1 containing the amino acid sequence of SEQ ID NO:1, (b) a CDR-H2 containing the amino acid sequence of SEQ ID NO:62 or 87, and (c) a CDR-H3 containing the amino acid sequence of SEQ ID NO:3; and a light chain containing (a) a CDR-L1 containing the amino acid sequence of SEQ ID NO:8 or 63, (b) a CDR-L2 containing the amino acid sequence of SEQ ID NO:9, and (c) a CDR-L3 containing the amino acid sequence SEQ ID NO:10.

[0076] As used herein, the term "variable region" refers to a region containing at least three CDRs. In some embodiments, the variable region includes three CDRs and at least one framework region ("FR"). The terms "heavy chain variable region" or "heavy chain variable region" are used interchangeably to refer to a region containing at least three heavy chain CDRs. The terms “light chain variable region” or “light chain variable region” are used interchangeably to refer to a region containing at least three light chain CDRs. In some embodiments, the variable heavy chain or variable light chain comprises at least one framework region. In some embodiments, the antibody comprises at least one heavy chain backbone selected from HC-FR1, HC-FR2, HC-FR3, and HC-FR4. In some embodiments, the antibody comprises at least one light chain framework selected from LC-FR1, LC-FR2, LC-FR3, and LC-FR4. The framework regions can be located between light chain CDRs or between heavy chain CDRs. For example, an antibody may contain a heavy chain variable having the following structure: (HC-FR1)-(CDR-H1)-(HC-FR2)-(CDR-H2)-(HC-FR3)-(CDR-H3)-( HC–FR4). The antibody may contain a variable heavy chain having the following structure: (CDR-H1)-(HC-FR2)-(CDR-H2)-(HC-FR3)-(CDR-H3). The antibody may also contain a variable light chain having the following structure: (LC-FR1)-(CDR-L1)-(LC-FR2)-(CDR-L2)-(LC-FR3)-(CDR-L3)-(LC -FR4). The antibody may also contain a variable light chain having the following structure: (CDR-L1)-(LC-FR2)-(CDR-L2)-(LC-FR3)-(CDR-L3).

[0077] In some embodiments, the anti-IL31 antibody comprises one or more of: (a) heavy chain variable region framework 1 (HC-FR1) sequence of SEQ ID NO:4, (b) HC-FR2 sequence of SEQ ID NO: 5, (c) HC-FR3 sequences SEQ ID NO:6, (d) HC-FR4 sequences SEQ ID NO:7, (e) light chain variable region framework 1 (LC-FR1) sequences SEQ ID NO:11, (f) LC-FR2 sequence SEQ ID NO:12, (g) LC-FR3 sequence SEQ ID NO:13, or (h) LC-FR4 sequence SEQ ID NO:14. In some embodiments, the anti-IL31 antibody comprises the variable light chain sequence of (a) SEQ ID NO:16, (b) SEQ ID NO:24, or (c) SEQ ID NO:32. In some embodiments, the anti-IL31 antibody comprises the variable heavy chain sequence of (a) SEQ ID NO:15; (b) SEQ ID NO:25; or (c) SEQ ID NO:33. In some embodiments, the anti-IL31 antibody comprises (a) the variable light chain sequence of SEQ ID NO:16 and the variable heavy chain sequence of SEQ ID NO:15; (b) the variable light chain sequence of SEQ ID NO:24 and the variable heavy chain sequence of SEQ ID NO:25; or (c) the variable light chain sequence of SEQ ID NO:32 and the variable heavy chain sequence of SEQ ID NO:33.

[0078] As used herein, the term "constant region" refers to a region containing at least three constant domains. The terms "heavy chain constant region" or "heavy chain constant region" are used interchangeably to refer to a region containing at least three heavy chain constant domains, CH1, CH2 and CH3. Non-limiting examples of heavy chain constant regions include γ, δ, α, ε, and μ. Each heavy chain constant region corresponds to an antibody isotype. For example, an antibody containing constant region γ is an IgG antibody, an antibody containing constant region δ is an IgD antibody, an antibody containing constant region α is an IgA antibody, an antibody containing constant region μ is an IgM antibody, and the antibody containing the ε constant region is an IgE antibody. Certain isotypes can be further subdivided into subclasses. For example, IgG antibodies include, without limitation, IgG1 (containing the γ1 constant region), IgG2 (containing the γ2 constant region), IgG3 (containing the γ3 constant region), and IgG4 (containing the γ4 constant region); IgA antibodies include, without limitation, IgA1 antibodies (containing the α1 constant region) and IgA2 antibodies (containing the α2 constant region); and IgM antibodies include, without limitation, IgM1 and IgM2. The terms “light chain constant region” or “light chain constant region” are used interchangeably to refer to the region containing the light chain constant domain, CL. Non-limiting examples of light chain constant regions include λ and κ. The scope of the term "constant region" includes deletions and substitutions within domains that do not change function, unless otherwise indicated. Dogs, cats, and horses have antibody classes such as IgG, IgA, IgD, IgE, and IgM. The class of canine IgG antibodies includes IgG-A, IgG-B, IgG-C and IgG-D. The class of feline IgG antibodies includes IgG1a, IgG1b, and IgG2. The class of equine IgG antibodies includes IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 and IgG7.

[0079] The term "chimeric antibody" or "chimeric" refers to an antibody in which part of the heavy chain or light chain is from a particular source or species, while at least part of the remaining heavy chain or light chain is from another source or kind. In some embodiments, a chimeric antibody refers to an antibody containing at least one variable region from a first species (such as mouse, rat, cynomolgus monkey, and the like.) and at least one constant region from a second species (such as human , dog, cat, horse, etc.). In some embodiments, the chimeric antibody comprises at least one murine variable region and at least one canine constant region. In some embodiments, the chimeric antibody comprises at least one murine variable region and at least one feline constant region. In some embodiments, all variable regions of the chimeric antibody are of the first species and all constant regions of the chimeric antibody are of the second species. In some embodiments, the chimeric antibody comprises a heavy chain constant region or a light chain constant region of a companion animal. In some embodiments, the chimeric antibody comprises murine variable heavy and light chains and constant heavy and light chains of a companion animal. For example, a chimeric antibody may comprise a murine heavy and light chain variable and a canine heavy and light chain constant; the chimeric antibody may comprise a murine heavy and light chain variable and a feline heavy and light chain constant; or the chimeric antibody may comprise a mouse heavy and light chain variable and an equine heavy and light chain constant.

[0080] In some embodiments, the anti-IL31 antibody comprises a chimeric antibody comprising:

A. (i) the light chain amino acid sequence of SEQ ID NO:26; (ii) the amino acid sequence of the heavy chain of SEQ ID NO:27; or (iii) a light chain amino acid sequence as defined in (i) and a heavy chain sequence as defined in (ii); or

b. (i) the light chain amino acid sequence of SEQ ID NO:30; (ii) the amino acid sequence of the heavy chain of SEQ ID NO:31; or (iii) a light chain amino acid sequence as defined in (i) and a heavy chain sequence as defined in (ii).

[0081] "Canine chimera" or "canine chimeric antibody" refers to a chimeric antibody having at least a heavy chain portion or a light chain portion derived from a dog. "Feline chimera" or "feline chimeric antibody" refers to a chimeric antibody having at least a heavy chain portion or a light chain portion derived from a cat. An "equine chimera" or "equine chimeric antibody" refers to a chimeric antibody having at least a heavy chain portion or a light chain portion derived from a horse. In some embodiments, the canine chimeric antibody comprises a mouse heavy and light chain variable and a canine heavy and light chain constant. In some embodiments, the feline chimeric antibody comprises a murine heavy and light chain variable and a feline heavy and light chain constant. In some embodiments, the equine chimeric antibody comprises a murine heavy and light chain variable and an equine heavy and light chain constant. In some embodiments, the antibody is a chimeric antibody comprising a murine variable heavy chain framework or a murine variable light chain framework.

[0082] As used herein, "canine antibody" encompasses antibodies produced in dogs; antibodies produced in non-canine animals that contain canine immunoglobulin genes or contain canine immunoglobulin peptides; or antibodies selected by in vitro methods such as phage display, where the antibody repertoire is based on a canine immunoglobulin sequence. The term "canine antibody" refers to the genus of sequences that are canine sequences. Thus, the term refers not to the process by which an antibody was created, but to the genus of the corresponding sequences.

[0083] In some embodiments, the anti-IL31 antibody comprises a canine heavy chain constant region selected from IgG-A, IgG-B, IgG-C, and IgG-D constant regions. In some embodiments, the anti-IL31 antibody is a canine IgG-A, IgG-B, IgG-C, or IgG-D antibody. In some embodiments, the anti-IL31 antibody is (a) a canine IgG-A antibody comprising the heavy chain amino acid sequence of SEQ ID NO:17; (b) a canine IgG-B antibody comprising the heavy chain amino acid sequence of SEQ ID NO:18; (c) a canine IgG-C antibody comprising the heavy chain amino acid sequence of SEQ ID NO:19; or (d) a canine IgG-D antibody comprising the heavy chain amino acid sequence of SEQ ID NO:20.

[0084] The term "feline antibody" as used herein encompasses antibodies produced in a cat; antibodies produced in non-feline animals that contain feline immunoglobulin genes or contain feline immunoglobulin peptides; or antibodies selected by in vitro methods such as phage display, where the antibody repertoire is based on the feline immunoglobulin sequence. The term "feline antibody" refers to a genus of sequences that are feline sequences. Thus, the term refers not to the process by which an antibody was created, but to the genus of the corresponding sequences.

[0085] In some embodiments, the anti-IL31 antibody comprises a feline heavy chain constant region selected from IgG1, IgG2a, and IgG2b constant regions. In some embodiments, the anti-IL31 antibody is a feline IgG1, IgG2a, or IgG2b antibody.

[0086] As used herein, "equine antibody" includes antibodies derived from horses; antibodies produced in non-equine animals that contain equine immunoglobulin genes or contain equine immunoglobulin peptides; or antibodies selected by in vitro methods such as phage display, where the antibody repertoire is based on the equine immunoglobulin sequence. The term "equine antibody" refers to a genus of sequences that are equine sequences. Thus, the term refers not to the process by which an antibody was created, but to the genus of the corresponding sequences.

[0087] In some embodiments, the anti-IL31 antibody comprises an equine heavy chain constant region selected from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, and IgG7 constant regions. In some embodiments, the anti-IL31 antibody is an equine IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, or IgG7 antibody.

[0088] "Caninized antibody" means an antibody in which at least one amino acid in a portion of a non-canine variable region has been replaced with a corresponding amino acid from a canine variable region. In some embodiments, the caninized antibody comprises at least one canine constant region (eg, γ constant region, α constant region, δ constant region, ε constant region, μ constant region, and the like) or a fragment thereof. In some embodiments, the caninized antibody is an antibody fragment such as a Fab, scFv, (Fab') ₂ and the like. The term "caninized" also refers to forms of non-canine (e.g., murine) antibodies that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab') ₂ or other antibody antigen-binding sequences ) that contain the minimum sequence of non-canine immunoglobulin. Canine antibodies can include canine immunoglobulins (recipient antibody) in which residues from the CDR of the recipient are replaced by residues from a CDR of a non-canine species (donor antibody) such as a mouse, rat or rabbit having the desired specificity, affinity and potency. In some cases, canine immunoglobulin Fv framework region (FR) residues are replaced by corresponding non-canine residues. In addition, the caninized antibody may contain residues that are not found in either the recipient antibody or the imported CDR or framework sequences, but which are included for further purification and optimization of antibody performance.

[0089] In some embodiments, at least one amino acid residue in the mouse variable heavy chain or mouse variable light chain portion is replaced with a corresponding amino acid from the canine variable region. In some embodiments, the modified chain is fused to a canine constant heavy chain or canine constant light chain. In some embodiments, the anti-IL31 antibody is a caninized antibody comprising (a) the heavy chain sequence of SEQ ID NO:15, (b) the heavy chain sequence of SEQ ID NO:17, (c) the heavy chain sequence of SEQ ID NO:18, (d) the heavy chain sequence of SEQ ID NO:19, (e) the heavy chain sequence of SEQ ID NO:20, (f) the light chain sequence of SEQ ID NO:16, or (g) the light chain sequence of SEQ ID NO:21.

[0090] The term "felinized antibody" means an antibody in which at least one amino acid in a portion of the non-feline variable region is replaced with a corresponding amino acid from the feline variable region. In some embodiments, the felineized antibody comprises at least one feline constant region (eg, γ constant region, α constant region, δ constant region, ε constant region, μ constant region, and the like) or a fragment thereof. In some embodiments, the felinized antibody is an antibody fragment such as a Fab, scFv, (Fab') ₂ and the like. The term "felinized" also refers to non-feline (e.g., murine) forms of antibodies that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab') ₂ or other antibody antigen-binding sequences ) that contain minimal non-feline immunoglobulin sequence. Felinized antibodies may include feline immunoglobulins (recipient antibody) in which residues from the CDR of the recipient are replaced by residues from a CDR of a non-feline species (donor antibody) such as a mouse, rat or rabbit having the desired specificity, affinity and potency. In some cases, feline immunoglobulin Fv framework region (FR) residues are replaced by corresponding non-feline residues. In addition, the felinized antibody may contain residues that are not found in either the recipient antibody or the imported CDR or framework sequences, but which are included for further purification and optimization of antibody performance.

[0091] In some embodiments, at least one amino acid residue in the mouse variable heavy chain or mouse variable light chain portion is replaced with a corresponding amino acid from the feline variable region. In some embodiments, the modified chain is fused to a feline constant heavy chain or a canine constant light chain. In some embodiments, the anti-IL31 antibody is a felinized antibody comprising (a) the light chain sequence of SEQ ID NO:32, (b) the light chain sequence of SEQ ID NO:34, (c) the heavy chain sequence of SEQ ID NO:33, or (d) the heavy chain sequence of SEQ ID NO:35.

[0092] "Equinized antibody" means an antibody in which at least one amino acid in a portion of the non-equine variable region is replaced by a corresponding equine amino acid from the variable region. In some embodiments, the equinized antibody comprises at least one equine constant region (eg, γ constant region, α constant region, δ constant region, ε constant region, μ constant region, and the like) or a fragment thereof. In some embodiments, the implementation of the equinized antibody is an antibody fragment, such as Fab, scFv, (Fab') ₂ and the like. The term "equinized" also refers to forms of non-equine (eg, mouse) antibodies that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab') ₂ or other antibody antigen-binding sequences ) that contain the minimum sequence of non-equine immunoglobulin. Equinized antibodies may include equine immunoglobulins (recipient antibody) in which residues from the CDR of the recipient are replaced by residues from a CDR of a non-equine species (donor antibody) such as a mouse, rat or rabbit having the desired specificity, affinity and potency. In some cases, equine immunoglobulin Fv framework region (FR) residues are replaced by corresponding non-equine residues. In addition, the equine antibody may contain residues that are not found in either the recipient antibody or the imported CDR sequences or framework regions, but which are included for further purification and optimization of antibody performance.

[0093] In some embodiments, at least one amino acid residue in the mouse variable heavy chain or mouse variable light chain portion is replaced with a corresponding amino acid from the equine variable region. In some embodiments, the modified chain is fused to an equine constant heavy chain or canine constant light chain.

[0094] The term "IgX Fc" means that the Fc region is derived from a specific antibody isotype (eg, IgG, IgA, IgD, IgE, IgM, and the like.), where "X" denotes the isotype of the antibody. Thus, "IgG Fc" denotes the Fc region of the γ chain, "IgA Fc" denotes the Fc region of the α chain, "IgD Fc" denotes the Fc region of the δ chain, "IgE Fc" denotes the Fc region of the ε chain , "IgM Fc" refers to the Fc region of the μ chain and the like. In some embodiments, the IgG Fc region includes CH1, hinge region, CH2, CH3, and CL1. "IgX-N-Fc" means that the Fc region is derived from a specific subclass of an antibody isotype (such as canine IgG subclass A, B, C, or D; feline IgG subclass 1, 2a, or 2b; or IgG1, IgG2, IgG3, IgG4 subclass , IgG5, IgG6 or IgG7 and the like equine IgG), where "N" denotes a subclass. In some embodiments, the IgX Fc or IgX-N-Fc regions are from a companion animal such as a dog, cat, or horse. In some embodiments, IgG Fc regions are isolated from canine γ heavy chains such as IgG-A, IgG-B, IgG-C, or IgG-D. In some cases, IgG Fc regions are isolated from feline γ heavy chains such as IgG1, IgG2a, or IgG2b. Antibodies containing the Fc-region of IgG-A, IgG-B, IgG-C or IgG-D can provide higher levels of expression in recombinant production systems.

[0095] The term "affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (eg, an antibody) and its binding partner (eg, an antigen). The affinity of a molecule X for its partner Y can generally be represented by a dissociation constant (K _D ). Affinity can be measured by conventional methods known in the art such as, for example, immunoblot assay, KD ELISA, KinEx A, Bilayer Interferometry (BLI) or surface plasmon resonance devices.

[0096] The terms "K _D ", "K _d ", "Kd" or "Kd value" are used interchangeably to refer to the equilibrium dissociation constant of an antibody-antigen interaction. In some embodiments, the K _d of the antibody is measured by bilayer interferometry using a biosensor such as the Octet® system (Pall ForteBio LLC, Fremont, CA) according to the manufacturer's instructions. Briefly, the biotinylated antigen is immobilized on the sensor tip and antibody association is monitored for ninety seconds and dissociation is monitored for 600 seconds. The buffer for dilutions and binding steps is 20 mM phosphate, 150 mM NaCl, pH 7.2. The background curve corresponding to the buffer only is subtracted to correct for any deviation. The data are fit to a 2:1 binding model using ForteBio data analysis software to determine the association rate constant (k _on ), dissociation rate constant (k _off ) and K _d . The equilibrium dissociation constant (K _d ) is calculated as the ratio k _off /k _on . The term "k _on " refers to the rate constant for association of an antibody with an antigen, and the term "k _off " refers to the rate constant for the dissociation of an antibody from an antibody/antigen complex.

[0097] The term "binds" to an antigen or epitope is a term that is well known in the art, and methods for determining such binding are also well known in the art. A molecule is said to exhibit "binding" if it reacts, associates, or has an affinity for a particular cell or substance, and the reaction, association, or affinity can be detected by one or more methods known in the art, such as, for example, immunoblot assay, ELISA KD, KinEx A, Bilayer Interferometry (BLI), Surface Plasmon Resonance or others.

[0098] "Surface plasmon resonance" means an optical phenomenon that allows the analysis of biospecific interactions in real time by detecting changes in protein concentration in the biosensor matrix, for example, using the BIAcore ^TM system (BIAcore International AB, GE Healthcare, Uppsala, Sweden and Piscataway, NJ). For additional description, see Jonsson et al. (1993) Ann. Biol. Clin. 51:19-26.

[0099] The term "bilayer interferometry" refers to an optical analytical technique that analyzes the interference pattern of light reflected from the immobilized protein layer on the biosensor tip and the internal reference layer. Changes in the number of molecules bound to the biosensor tip cause shifts in the interference pattern that can be measured in real time. A non-limiting example of a bilayer interferometry device is the Octet® system (Pall ForteBio LLC). see, for example, Abdiche et al., 2008, Anal. Biochem. 377:209-277.

[00100] In some embodiments, the anti-IL31 antibody binds to canine IL31, feline IL31, or equine IL31 with a dissociation constant (Kd) of less than 5 x 10 ^-6 M, less than 1 x 10 ^-6 M, less than 5 x 10 ^–7 M, less than 1× ^10–7 M, less than 5× ^{10–8 M} , less than 1×10–8 M, less than 5× ^10–9 M, less than 1× ^10–9 M, less less than 5× ^10–10 M, less than 1× ^10–10 M, less than 5× ^10–11 M, less ^than 1×10–11 M, less than 5× ^10–12 M, or less than 1×10 ^{– 12} M according to the results of measurements by the bilayer interferometry method. In some embodiments, the anti-IL31 antibody binds to canine IL31, feline IL31, or equine IL31 with a Kd of 5x10 ^-6 M to 1x10 ^-6 M, 5x10 ^-6 M to 5x10 ^-7 M, from 5× ^10–6 M to 1× ^10–7 M, from 5× ^10–6 M to 5× ^10–8 M, from 5× ^10–6 M to 1× ^10–8 M, from 5×10 ^{– 6} M to 5× ^10–9 M, 5× ^10–6 M to 1× ^10–9 M, 5× ^10–6 M to 5× ^10–10 M, 5× ^10–6 M to 1× ^10–10 M, from 5× ^10–6 M to 5× ^10–11 M, from 5× ^10–6 M to 1× ^10–11 M, from 5× ^10–6 M to 5× ^10–12 M, from 5× ^10–6 M to 1× ^10–12 M, from 1× ^10–6 M to 5× ^10–7 M, from 1× ^10–6 M to 1× ^10–7 M, from 1×10 ^{– 6} M to 5 ^{× 10–8} M, 1× ^10–6 M to 1×10–8 M, 1× ^10–6 M to 5× ^10–9 M, 1× ^10–6 M to 1× ^10–9 M, from 1× ^10–6 M to 5× ^10–10 M, from 1× ^10–6 M to 1× ^10–10 M, from 1× ^10–6 M to 5× ^10–11 M, from 1× ^10–6 M to 1× ^10–11 M, from 1× ^10–6 M to 5× ^10–12 M, from 1× ^10–6 M to 1× ^10–12 M, from 5×10 ^{– 7} M to 1× ^10–7 M, 5× ^10–7 M to 5× ^10–8 M, 5× ^10–7 M to 1× ^10–8 M, 5× ^10–7 M to 5× ^10–9 M, from 5× ^10–7 M to 1× ^10–9 M, from 5× ^10–7 M to 5× ^10–10 M, from 5× ^10–7 M to 1× ^10–10 M, from 5× ^10–7 M to 5× ^10–11 M, from 5× ^10–7 M to 1× ^10–11 M, from 5× ^10–7 M to 5× ^10–12 M, from 5×10 ^{– 7} M to 1× ^10–12 M, 1× ^10–7 M to 5× ^10–8 M, 1× ^10–7 M to 1× ^10–8 M, 1× ^10–7 M to 5× ^10–9 M, from 1× ^10–7 M to 1× ^10–9 M, from 1× ^10–7 M to 5× ^10–10 M, from 1× ^10–7 M to 1× ^10–10 M, from 1× ^10–7 M to 5× ^10–11 M, from 1× ^10–7 M to 1× ^10–11 M, from 1× ^10–7 M to 5× ^10–12 M, from 1×10 ^{– 7} M to 1 ^{× 10–12} M, 5× ^10–8 M to 1×10–8 M, 5× ^10–8 M to 5× ^10–9 M, 5× ^10–8 M to 1× ^10–9 M, from 5× ^10–8 M to 5× ^10–10 M, from 5× ^10–8 M to 1× ^10–10 M, from 5× ^10–8 M to 5× ^10–11 M, from 5× ^10–8 M to 1× ^10–11 M, from 5× ^10–8 M to 5× ^10–12 M, from 5× ^10–8 M to 1× ^10–12 M, from 1×10 ^{– 8} M to 5× ^10–9 M, 1× ^{10–8 M} to 1×10–9 M, 1× ^10–8 M to 5× ^10–10 M, 1× ^10–8 M to 1× ^10–10 M, from 1× ^10–8 M to 5× ^10–11 M, from 1× ^10–8 M to 1× ^10–11 M, from 1× ^10–8 M to 5× ^10–12 M, from 1× ^10–8 M to 1× ^10–12 M, from 5× ^10–9 M to 1× ^{10–9 M, from 5×10–9 M} to 5× ^10–10 M, from 5×10 ^{– 9} M to 1× ^10–10 M, 5× ^10–9 M to 5× ^10–11 M, 5× ^10–9 M to 1× ^10–11 M, 5× ^10–9 M to 5× ^10–12 M, from 5× ^10–9 M to 1× ^10–12 M, from 1× ^10–9 M to 5× ^10–10 M, from 1× ^10–9 M to 1× ^10–10 M, from 1× ^10–9 M to 5× ^10–11 M, from 1× ^10–9 M to 1× ^10–11 M, from 1× ^10–9 M to 5× ^10–12 M, from 1×10 ^{– 9} M to 1× ^10–12 M, 5× ^10–10 M ^{to 1×10–10 M, 5×10–10 M to 5×10–11 M, 1×10–10 M to 5} × ^10–11 M, from 1× ^10–10 M to 1× ^10–11 M, from 1× ^10–10 M to 5× ^10–12 M, from 1× ^10–10 M to 1× ^10–12 M, from 5×10 ^–11 M to 1×10 ^–12 M, from 5×10 ^–11 M to 5×10 ^–12 M, from 5×10 ^–11 M to 1×10 ^–12 M, from 1×10 ^{– 11} M to 5×10 ^–12 M or from 1×10 ^–11 M to 1×10 ^–12 M, according to the results of measurement by the bilayer interferometry method. In some embodiments, the anti-IL31 antibody binds to canine IL31, feline IL31, or equine IL31 as determined by immunoblot analysis.

[00101] In some embodiments, the anti-IL31 antibody does not bind to human IL31 as determined by immunoblotting and/or bilayer interferometry analysis.

[00102] In some embodiments, an anti-IL31 antibody is provided that competes with an anti-IL31 antibody described herein (such as M14, M18, M19, or M87) for binding to IL31. In some embodiments, an antibody can be made or used that competes for binding with any of the antibodies described herein. In some embodiments, an anti-IL31 antibody is provided that competes with the M14 monoclonal antibody for binding to canine IL31 or feline IL31.

[00103] "Variant" means a biologically active polypeptide having at least about 50% amino acid sequence identity with a native sequence polypeptide after sequence alignment and, if necessary, gaps (gaps) introduced to achieve the maximum percentage of sequence identity and without considering any conservative substitutions as part of sequence identity. Such variants include, for example, polypeptides in which one or more amino acid residues are added, removed at the N or C terminus of the polypeptide.

[00104] In some embodiments, a variant has at least about 50% amino acid sequence identity, at least about 60% amino acid sequence identity, at least about 65% amino acid sequence identity, at least about 70% amino acid sequence identity, at least about 75% amino acid sequence identity, at least about 80% amino acid sequence identity, at least about 85% amino acid sequence identity, at least about 90% amino acid sequence identity, at least about 95% amino acid sequence identity with the polypeptide native sequence.

[00105] As used herein, "percentage (%) amino acid sequence identity" and "homology" with respect to the sequence of a peptide, polypeptide, or antibody is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence after alignment of sequences and introduction of gaps, if necessary to achieve the maximum percentage of sequence identity, and without considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in a variety of ways that are known to those skilled in the art, for example, using open source software such as BLAST, BLAST-2, ALIGN or MEGALINE ^™ (DNASTAR) software. Suitable parameters for measuring alignment can be determined by those skilled in the art, including any algorithms necessary to achieve maximum alignment over the entire length of the compared sequences.

[00106] An amino acid substitution may include, without limitation, substitution of one amino acid in a polypeptide with another amino acid. Examples of substitutions are shown in Table 2. Amino acid substitutions can be introduced into an antibody of interest and products can be screened for desired activity, eg, retained/improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[00107] Table 2

original balance Substitution examples Ala (A) Val; Leu; ile Arg(R) Lys; gln; Asn Asn(N) gln; His; asp; Lys; Arg Asp(D) Glu; Asn Cys(C) Ser; Ala Gln(Q) asn; Glu Glu(E) asp; Gln Gly (G) Ala His(H) asn; gln; Lys; Arg Ile (I) Leu; Val; met; Ala; Ph; Norleucine Leu(L) norleucine; Ile; Val; met; Ala; Phe Lys (K) Arg; gln; Asn Met(M) Leu; Ph; ile Phe(F) trp; Leu; Val; Ile; Ala; Tyr Pro (P) Ala Ser(S) Thr Thr(T) Val; Ser TRP(W) Tyr; Phe Tyr (Y) trp; Ph; Thr; Ser Val(V) Ile; Leu; met; Ph; Ala; norleucine

[00108] Amino acids can be grouped according to the general properties of the side chains:

(1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile;

(2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

(3) acidic: Asp, Glu;

(4) main: His, Lys, Arg;

(5) residues that affect chain orientation: Gly, Pro;

(6) aromatic: Trp, Tyr, Phe.

[00109] Non-conservative substitutions will entail replacing a member of one of these classes with another class.

[00110] In some embodiments, the anti-IL31 antibody comprises a heavy chain and a light chain, where:

A. the heavy chain contains a CDR-H1 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:1; a CDR-H2 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:2, SEQ ID NO: 62, SEQ ID NO:89 or SEQ ID NO:87; and a CDR-H3 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:3, and

b. the light chain contains a CDR-L1 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:63; a CDR-L2 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:9; and a CDR-L3 sequence having at least 85% sequence identity, at least 90% sequence identity, at least 95% sequence identity, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:10.

[00111] In some embodiments, the anti-IL31 antibody comprises a heavy chain and a light chain, where:

A. (i) the variable light chain sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:24; (ii) the variable heavy chain sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:25; or (iii) a variable light chain sequence as defined in (i) and a variable heavy chain sequence as defined in (ii); or

b. (i) the light chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:16; (ii) the heavy chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:15; or (iii) the variable light chain sequence is as defined in (i) and the variable heavy chain sequence is as defined in (ii); or

With. (i) the light chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:32; (ii) the heavy chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:33; or (iii) the variable light chain sequence is as defined in (i) and the variable heavy chain sequence is as defined in (ii); or

d. (i) the light chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:64; (ii) the heavy chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:67; or (iii) the variable light chain sequence is as defined in (i) and the variable heavy chain sequence is as defined in (ii); or

e. (i) the light chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:65; (ii) the heavy chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:68; or (iii) the variable light chain sequence is as defined in (i) and the variable heavy chain sequence is as defined in (ii); or

f. (i) the light chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:66; (ii) the heavy chain variable sequence has at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:69; or (iii) the variable light chain sequence is as defined in (i) and the variable heavy chain sequence is as defined in (ii).

[00112] The term "vector" is used to describe a polynucleotide that can be engineered to contain a cloned polynucleotide or polynucleotides that can propagate in a host cell. The vector may include one or more of the following: an origin of replication, one or more regulatory sequences (such as, for example, promoters or enhancers) that regulate the expression of the polypeptide of interest, or one or more selectable marker genes (such as, for example, genes antibiotic resistance and genes that can be used in colorimetric assays, such as β-galactosidase). The term "expression vector" refers to a vector that is used to express a polypeptide of interest in a host cell.

[00113] "Host cell" refers to a cell that can be or has been a recipient of a vector or an isolated polynucleotide. The host cells may be prokaryotic cells or eukaryotic cells. Exemplary eukaryotic cells include mammalian cells such as primate or non-primate cells; fungal cells such as yeast; plant cells; and insect cells. Non-limiting exemplary mammalian cells include, without limitation, NS0 cells, PER.C6® cells (Crucell), 293 cells, and CHO cells and their derivatives such as 293-6E, DG44, CHO-S, and CHO-K cells. Host cells include the progeny of a single host cell, and the progeny may not necessarily be completely identical (in morphology or genomic DNA complementarity) to the original parent cell due to natural, accidental or intentional mutation. The host cell includes cells transfected in vivo with a polynucleotide(s) encoding(s) the amino acid sequence(s) provided herein.

[00114] As used herein, the term "isolated" refers to a molecule that has been separated from at least some of the components with which it is commonly found in nature or produced. For example, a polypeptide is said to be "isolated" when it has been separated from at least some of the components of the cell in which it was produced. If the polypeptide is secreted by a cell, then the polypeptide is considered "isolated" upon expression, the physical separation of the supernatant containing the polypeptide from the cell that produces it. Similarly, a polynucleotide is said to be "isolated" when it is not part of a larger polynucleotide (such as, for example, genomic DNA or mitochondrial DNA, in the case of a DNA polynucleotide) in which it is normally found in nature, or is separated from at least some components of the cell in which it was produced, for example, in the case of an RNA polynucleotide. Thus, a DNA polynucleotide that is contained in a vector within a host cell may be referred to as "isolated". In some embodiments, the anti-IL31 antibody is purified by chromatography, such as size exclusion chromatography, ion exchange chromatography, protein A column chromatography, hydrophobic interaction chromatography, and CHT chromatography.

[00115] The term "kind of companion animal" refers to an animal suitable for being a companion animal. In some embodiments, the companion animal species is small mammals such as canines, felines, dogs, cats, horses, rabbits, ferrets, guinea pigs, rodents, and the like. In some embodiments, the companion animal species is a farm animal such as a horse, cow, pig, and the like.

[00116] The term "IL31 signaling function" refers to any subsequent activity, or combination thereof, that occurs when IL31 binds to its receptor or receptor complex. In some embodiments, IL31 signaling includes activation of Janus kinase (Jak) 1 or Jak 2 signaling molecules. In some embodiments, IL31 signaling includes phosphorylation of STAT-3 or STAT-5 proteins. In some embodiments, the IL31 signaling function comprises activation of the MAP kinase ERK1/2 signaling pathway. In some embodiments, IL31 signaling function includes activation of the PI3K/AKT signaling pathway. In some embodiments, IL31 signaling function includes activation of the Jak1/2 signaling pathway.

[00117] "Phosphorylation of STAT" means the modification of the STAT protein after expression by phosphorylation. For example, "STAT-3 phosphorylation" refers to STAT-3 phosphorylation, and "STAT-5 phosphorylation" refers to STAT-5 phosphorylation. In some embodiments, STAT-3 phosphorylation is measured using an immunoblot assay. For example, cells (eg, DH82 canine monocytic cells) are seeded in a 96-well cell culture plate at a density of 1 x 10 ⁵ cells per well in growth media (eg, MEM, Life Technologies®) containing 15% heat-inactivated fetal bovine serum. , 2 mmol/L GlutaMax, 1 mmol/L sodium pyruvate, and 10 nM/mL canine interferon-c (R&D Systems, Minneapolis, MN, USA) for 24 hours at 37°C in the presence of an anti-IL31 antibody described herein. description. Cell lysate immunoblotting analysis using anti-phospho STAT-3 and anti-STAT-3 antibodies (R&D Systems) was used to determine the concentrations of phosphorylated STAT-3 and non-phosphorylated STAT-3 relative to each other and compared with beta-actin control. Methods for qualitative or quantitative determination of the concentration of proteins using immunoblot known to experts in this field. In some embodiments, methods are used to qualitatively determine the relative concentration by visual inspection of the immunoblot. In some embodiments, methods are used to quantify the concentrations of phosphorylated STAT 3 and non-phosphorylated STAT 3 using immunoblot digital imaging, determining band intensities, and using a linear standard curve of known STAT-3 protein concentrations to back-calculate the concentrations of phosphorylated or non-phosphorylated STAT-3 in a sample.

[00118] "Reduce" or "inhibit" means to reduce, reduce, or block an activity, function, or amount compared to a reference. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 20% or more. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 50% or more. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 75%, 85%, 90%, 95%, or more. In some embodiments, the above amount is inhibited or reduced over a period of time relative to a control dose (such as placebo) over the same period of time. The term "reference" as used in this application refers to any sample, standard or level that is used for comparison purposes. The reference can be obtained from a healthy sample or a disease-free sample. In some examples, the reference is obtained from a disease free or untreated sample from a companion animal. In some examples, the reference is obtained from one or more healthy animals of a particular species, which are not subject to testing or treatment.

[00119] The term "substantially reduced" as used herein refers to a sufficiently large decrease in a numerical value relative to a reference numerical value, such that a person skilled in the art would consider the difference between the two values to be statistically significant in the context of the biological characteristic measured by these values. In some embodiments, substantially reduced numerical values are reduced from the control value by more than about any of: 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60% , 70%, 80%, 90% or 100%.

[00120] In some embodiments, the anti-IL31 antibody can reduce IL31 signaling in companion animal species by at least 10%, at least 15%, at least 20%, at least 30%, by at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or 100% compared to IL31 signaling in the absence of antibody, as measured by the reduction in STAT-3 phosphorylation. In some embodiments, the reduction in IL31 signaling or the reduction in STAT-3 phosphorylation is 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 10% to 60%, 10% to 70%, 10% to 80%, 10% to 90%, from 10% to 100%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% % to 50%, 15% to 60%, 15% to 70%, 15% to 80%, 15% to 90%, 15% to 100%, 20% to 25%, 20% up to 30%, from 20% to 35%, from 20% to 40%, from 20% to 45%, from 20% to 50%, from 20% to 60%, from 20% to 70%, from 20% to 80%, 20% to 90%, 20% to 100%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50 %, 25% to 60%, 25% to 70%, 25% to 80%, 25% to 90%, 25% to 100%, 30% to 35%, 30% to 40% , 30% to 45%, 30% to 50%, 30% to 60%, 30% to 70%, 30% to 80%, 30% to 90%, 30% to 100%, 35% to 40%, 35% to 45%, 35% to 50%, 35% to 60%, 35% to 70%, 35% to 80%, 35% to 90%, from 35% to 100%, 40% to 45%, 40% to 50%, 40% to 60%, 40% to 70%, 40% to 80%, 40% to 90%, 40% % to 100%, 45% to 50%, 45% to 60%, 45% to 70%, 45% to 80%, 45% to 90%, 45% to 100%, 50% up to 60%, 50% to 70%, 50% to 80%, 50% to 90%, 50% to 100%, 60% to 70%, 60% to 80%, 60% to 90%, 60% to 100%, 70% to 80%, 70% to 90%, 70% to 100%, 80% to 90%, 80% to 100% or 90% to 100 %.

Pharmaceutical compositions

[00121] The terms "pharmaceutical composition" and "pharmaceutical composition" refer to a preparation that is in a form that provides effective biological activity of the active(s) ingredient(s), and that does not contain additional components that are unacceptably toxic to the subject, to which the composition is introduced.

[00122] A "pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid, or liquid excipient, diluent, encapsulating material, adjuvant formulation, or carrier conventional in the art for use with a therapeutic agent, which together constitute a "pharmaceutical composition" intended for administration. subject. The pharmaceutically acceptable carrier is non-toxic to recipients at the dosages and concentrations employed and is compatible with the other ingredients of the formulation. The pharmaceutically acceptable carrier is suitable for the composition used. Examples of pharmaceutically acceptable carriers include alumina; aluminum stearate; lecithin; serum proteins such as human serum albumin, albumin of a dog or other animal; buffers such as phosphate, citrate, tromethamine or HEPES buffers; glycine; sorbic acid; potassium sorbate; mixtures of partial glycerides of saturated vegetable fatty acids; water; salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide or magnesium trisilicate; polyvinylpyrrolidone, cellulose-based substances; polyethylene glycol; sucrose; mannitol; or amino acids, including, without limitation, arginine.

[00123] The pharmaceutical composition may be stored in lyophilized form. Thus, in some embodiments, the implementation of the method of obtaining includes the stage of lyophilization. Then, prior to administration to a dog, cat or horse, the lyophilized composition may be reconstituted, typically as an aqueous composition suitable for parenteral administration. In other embodiments, in particular where the antibody is highly stable to thermal and oxidative denaturation, the pharmaceutical composition may be stored as a liquid, i. in the form of an aqueous composition which can be administered to a dog, cat or horse directly or after appropriate dilution. The lyophilized composition may be reconstituted with sterile water for injection (WFI). Antibacterial agents (eg, bacteriostatic agents such as benzyl alcohol) may be included. Thus, the invention provides pharmaceutical compositions in solid or liquid form.

[00124] The pH value of the pharmaceutical compositions when administered can range from about 5 to about 8. The compositions of the invention are sterile when intended for therapeutic use. Sterility can be achieved by any of several methods known in the art, including filtration through sterile filter membranes (eg, 0.2 micron membranes). Sterility can be maintained with or without antibacterial agents.

[00125] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition has a pH of 5.0 to 6.2, 5.0 to 6.0, or 5.3 to 5.7. In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition has a pH of 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1 or 6.2.

[00126] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition comprises L-histidine, sodium chloride, and polysorbate 80.

[00127] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition comprises sodium chloride at a concentration of 80 nM to 200 nM, 100 nM to 180 nM, 100 nM to 175 nM, 110 nM to 170 nM, 120 nM to 160 nM, 120 nM to 150 nM, 130 nM to 150 nM, 130 nM to 160 nM, 100 nM, 80 nM, 110 nM, 120 nM, 130 nM, 140 nM, 150 nM, 160 nM, 170 nM , 180 nM or 200 nM.

[00128] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition comprises polysorbate 80 at a concentration of 0.005 mg/mL to 0.5 mg/mL, 0.01 mg/mL to 0.1 mg/mL, 0.1 mg/ml to 0.5 mg/ml, 0.005 mg/ml to 0.01 mg/ml, 0.1 mg/ml, 0.2 mg/ml, 0.3 mg/ml, 0.4 mg/ml ml, 0.05 mg/ml, 0.06 mg/ml, 0.07 mg/ml, 0.08 mg/ml, 0.09 mg/ml or 0.1 mg/ml.

[00129] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition comprises L-histidine at a concentration of 5 mM to 100 mM, 10 mM to 50 mM, 20 mM to 30 mM, 10 mM to 30 mM, 20 mM up to 80 mM, 30 mM to 70 mM, 40 mM to 60 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 40 or 50 mM.

[00130] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains m-cresol or benzyl alcohol. In some embodiments, the m-cresol concentration is about 0.2%, about 0.1% to about 0.3%, about 0.08% to about 0.25%, or about 0.05% to about 0 .25%. In some embodiments, the concentration of benzyl alcohol is about 1%, about 0.5% to about 2%, about 0.2% to about 2.5%, about 1% to about 5%, about 0.5 % to about 5% or from about 1% to about 3%.

[00131] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains a sugar. In some embodiments, the sugar is sucrose, trehalose, D-mannitol, maltose, and/or sorbitol. In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains sugar at a concentration of 0.5% to 20%, 1% to 10%, 1% to 5%, 1% to 3%, 0.5%, 1 %, 2%, 3%, 4%, 5% or 10%.

[00132] In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains an antibacterial agent. In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains m-cresol or methyl paraben. In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains 0.2% m-cresol. In some embodiments, the pharmaceutically acceptable carrier or pharmaceutical composition contains 0.9% methylparaben.

The use of antibodies and pharmaceutical compositions

[00133] Antibodies or pharmaceutical compositions containing the antibodies of the invention may be useful in the treatment of an IL-31-induced condition. As used herein, the term "IL31-induced condition" means a disease associated with, caused by, or characterized by elevated levels or altered concentration gradients of IL31. Such IL31-induced conditions include, without limitation, pruritus or allergic disease. In some embodiments, the IL31-induced condition is atopic dermatitis, pruritus, asthma, psoriasis, scleroderma, or eczema. The IL31-induced condition may be present in a companion animal, including, but not limited to, dogs, cats, or horses.

[00134] Used in the present description, the term "treatment" is an approach to obtain useful or desired clinical results. The term "treatment" as used herein encompasses any administration or use of a therapeutic agent for the treatment of a disease in a mammal, including companion animals. For the purposes of this disclosure, useful or desirable clinical results include, without limitation, any one or more of the following: relief of one or more symptoms, reduction in the severity of the disease, prevention or delay in the spread of the disease, prevention or delay in the recurrence of the disease, delay or slowing in the progression of the disease, relief disease state, inhibition of a disease or progression of a disease, inhibition or slowing of a disease or its progression, arrest of its development and remission (partial or complete). The term "treatment" also includes the reduction of the pathological consequences of a proliferative disease. The methods provided herein include any one or more of these aspects of treatment. In accordance with the foregoing, the term "treatment" does not require one hundred percent elimination of all aspects of the disorder.

[00135] In some embodiments, an anti-IL31 antibody, or pharmaceutical compositions containing it, can be used in accordance with the methods described herein for the treatment of IL31-induced conditions. In some embodiments, the anti-IL31 antibody or pharmaceutical compositions are administered to a companion animal, such as a dog, cat, or horse, for the treatment of an IL31-induced condition.

[00136] A "therapeutically effective amount" of a substance/molecule, agonist, or antagonist may vary depending on such factors as the type of disease being treated, the condition of the disease, the severity and course of the disease, the type of therapeutic prescription of any prior therapy, medical history, response to prior treatment, discretion of the treating veterinarian, age, sex and weight of the animal and the ability of the substance/molecule, agonist or antagonist to elicit the desired response in the animal. A therapeutically effective amount is also the amount at which the therapeutically beneficial effects outweigh any toxic or detrimental effects of the substance/molecule, agonist or antagonist. A therapeutically effective amount may be delivered in one or more administrations. A therapeutically effective amount refers to an amount effective at dosages and for periods of time necessary to achieve the desired therapeutic or prophylactic result.

[00137] In some embodiments, the anti-IL31 antibody or pharmaceutical composition comprising the anti-IL31 antibody is administered parenterally, subcutaneously, by intravenous infusion, or by intramuscular injection. In some embodiments, the anti-IL31 antibody or pharmaceutical composition containing the anti-IL31 antibody is administered as a bolus injection or by continuous infusion over a period of time. In some embodiments, the anti-IL31 antibody or pharmaceutical composition comprising the anti-IL31 antibody is administered intramuscularly, intraperitoneally, intracerebrospinally, subcutaneously, intraarterially, intrasynovially, intrathecally, or by inhalation.

[00138] The anti-IL31 antibodies described herein can be administered in an amount of 0.1 mg/kg body weight to 100 mg/kg body weight per dose. In some embodiments, anti-IL31 antibodies can be administered in an amount of 0.5 mg/kg body weight to 50 mg/kg body weight per dose. In some embodiments, anti-IL31 antibodies can be administered in an amount of 1 mg/kg body weight to 10 mg/kg body weight per dose. In some embodiments, anti-IL31 antibodies can be administered in an amount of 0.5 mg/kg body weight to 100 mg/kg body weight, 1 mg/kg body weight to 100 mg/kg body weight, 5 mg/kg body weight body weight up to 100 mg/kg body weight, from 10 mg/kg body weight to 100 mg/kg body weight, from 20 mg/kg body weight to 100 mg/kg body weight, from 50 mg/kg body weight to 100 mg/day kg of body weight, from 1 mg/kg of body weight to 10 mg/kg of body weight, from 5 mg/kg of body weight to 10 mg/kg of body weight, from 0.5 mg/kg of body weight to 10 mg/kg of body weight or from 5 mg/kg body weight to 50 mg/kg body weight.

[00139] An anti-IL3 antibody, or a pharmaceutical composition comprising an anti-IL31 antibody, can be administered to a companion animal once or in multiple administrations. For example, an anti-IL31 antibody or a pharmaceutical composition containing an anti-IL31 antibody can be administered at least once, more than once, at least two times, at least three times, at least four times, or at least five times. once.

[00140] In some embodiments, the dose is administered once a week for at least two or three consecutive weeks, and in some embodiments, this treatment cycle is repeated two or more times, optionally alternating with one or more weeks without treatment. In other embodiments, a therapeutically effective dose is administered once daily for two to five consecutive days, and in some embodiments, this treatment cycle is repeated two or more times, optionally with one or more days or weeks without treatment.

[00141] The introduction "in combination with" one or more additional therapeutic agents includes simultaneous (parallel) and sequential administration in any order. The term "simultaneously" is used herein to refer to the administration of two or more therapeutic agents, when the administrations at least partially overlap in time, or when the administration of one therapeutic agent occurs within a short period of time relative to the administration of the other therapeutic agent. For example, two or more therapeutic agents are administered at intervals of no more than about a certain number of minutes. The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents, when the administration of one or more agents continues after the administration of one or more other agents has ceased, or when the administration of one or more agents begins before the administration of one or more other agents. For example, the administration of two or more therapeutic agents occurs with a time difference exceeding a certain number of minutes. Used in the present description, the term "in combination with" refers to the use of one method of treatment in addition to another method of treatment. Thus, "in combination with" refers to animals receiving one treatment before, during, or after receiving another treatment.

[00142] In some embodiments, the method includes administering an anti-IL31 antibody or a pharmaceutical composition comprising an anti-IL31 antibody in combination with a Jak inhibitor, PI3K inhibitor, AKT inhibitor, or MAPK inhibitor. In some embodiments, the method includes administering an anti-IL31 antibody or a pharmaceutical composition comprising an anti-IL31 antibody in combination with an anti-IL17 antibody, an anti-TNFα antibody, an anti-CD20 antibody, an anti-CD19 antibody, an anti-CD25 antibody, an anti- -IL4 antibody, anti-IL13 antibody, anti-IL23 antibody, anti-IgE antibody, anti-CD11α antibody, anti-IL6R antibody, anti-α4-integrin antibody, anti-IL12 antibody, anti-IL1β antibody, or anti-BlyS antibody.

[00143] Provided herein are methods for exposing an anti-IL31 antibody or a pharmaceutical composition containing an anti-IL31 antibody to a cell under conditions favorable for the antibody to bind to IL31. In some embodiments, the cell is exposed to the antibody or pharmaceutical composition ex vivo . In some embodiments, the cell is exposed to the antibody or pharmaceutical composition in vivo . In some embodiments, the cell is exposed to an anti-IL31 antibody or pharmaceutical composition under conditions favorable for the antibody to bind to intracellular IL31. In some embodiments, the cell is exposed to an anti-IL31 antibody or pharmaceutical composition under conditions favorable for the antibody to bind to extracellular IL31. In some embodiments, a cell may be exposed in vivo to an anti-IL31 antibody or pharmaceutical composition when administered by any one or more of the methods described herein, including, without limitation, intraperitoneal, intramuscular, intravenous injection to a subject. In some embodiments, a cell may be exposed ex vivo to an anti-IL31 antibody or pharmaceutical composition by exposing the cell to a culture medium containing the antibody or pharmaceutical composition. In some embodiments, prior to exposing the cell to the culture medium containing the antibody or pharmaceutical composition, the permeability of the cell membrane may be affected by any number of methods known to those skilled in the art (such as electroporation of the cells or exposing the cells to a solution containing calcium chloride).

[00144] In some embodiments, the implementation of the binding leads to a decrease in the signaling function of IL31 in the cell. In some embodiments, an anti-IL31 antibody can reduce IL31 signaling in a cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or 100% compared to IL31 signaling in the absence of antibody, as measured by the reduction in STAT-3 phosphorylation. In some embodiments, the reduction in IL31 signaling or the reduction in STAT-3 phosphorylation is 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% and 35%. , 10% to 40%, 10% to 45%, 10% to 50%, 10% to 60%, 10% to 70%, 10% to 80%, 10% to 90%, 10% to 100%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, from 15% to 50%, 15% to 60%, 15% to 70%, 15% to 80%, 15% to 90%, 15% to 100%, 20% to 25% from 20% up to 30%, from 20% to 35%, from 20% to 40%, from 20% to 45%, from 20% to 50%, from 20% to 60%, from 20% to 70%, from 20% to 80%, 20% to 90%, 20% to 100%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50 %, 25% to 60%, 25% to 70%, 25% to 80%, 25% to 90%, 25% to 100%, 30% to 35%, 30% to 40% , 30% to 45%, 30% to 50%, 30% to 60%, 30% to 70%, 30% to 80%, 30% to 90%, 30% to 100%, 35% to 40%, 35% and 45%, 35% to 50%, 35% to 60%, 35% to 70%, 35% to 80%, 35% to 90%, from 35% to 100%, 40% to 45%, 40% to 50%, 40% to 60%, 40% to 70%, 40% to 80%, 40% to 90%, 40% % to 100%, 45% to 50%, 45% to 60%, 45% to 70%, 45% to 80%, 45% to 90%, 45% to 100%, 50% up to 60%, 50% to 70% 50% to 80%, 50% to 90%, 50% to 100%, 60% to 70%, 60% to 80%, 60% to 90 %, 60% to 100%, 70% to 80%, 70% to 90%, 70% to 100%, 80% to 90%, 80% to 100% or 90% to 100% .

[00145] This application provides methods for using anti-IL31 antibodies, polypeptides, and polynucleotides to detect, diagnose, and monitor an IL31-induced condition. Provided herein are methods for determining whether a companion animal will respond to anti-IL31 antibody therapy. In some embodiments, the method includes determining, using an anti-IL31 antibody, whether the animal has cells that express IL31. In some embodiments, the detection method includes contacting a sample with an antibody, polypeptide, or polynucleotide and determining if the level of binding differs from that of a reference or reference sample (such as a control). In some embodiments, the method may be useful in determining whether the antibodies or polypeptides described herein are suitable for treating an animal subject.

[00146] In some embodiments, the sample is a biological sample. The term "biological sample" means the amount of a substance obtained from a living being or a formerly living being. In some embodiments, the biological sample is a cell or cell/tissue lysate. In some embodiments, the biological sample includes, without limitation, blood (eg, whole blood), plasma, serum, urine, synovial fluid, and epithelial cells.

[00147] In some embodiments, the cells or cell/tissue lysate are contacted with an anti-IL31 antibody and the binding between the antibody and the cell is determined. When test cells show binding activity compared to a control cell of the same tissue type, this may indicate that the subject will benefit from anti-IL31 antibody treatment. In some embodiments, the test cells are taken from the tissue of a companion animal.

[00148] Various methods known in the art can be used to detect antibody-antigen specific binding. Typical immunoassays that can be performed include polarization fluorescence immunoassay (FPIA), fluorescence immunoassay (FIA), enzyme immunoassay (EIA), nephelometric inhibition immunoassay (NIA), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay (RIA). An indicator fragment or group of tags can be attached to the antibodies of interest and chosen to meet the requirements for various applications of the method, which are often determined by the availability of assay equipment and compatible immunoassay procedures. Suitable labels include, without limitation, radionuclides (eg, ¹²⁵ I, ¹³¹ I, ³⁵ S, ³ H, or ³² P), enzymes (eg, alkaline phosphatase, horseradish peroxidase, luciferase, or β-lactosidase), fluorescent moieties, or proteins (eg. , fluorescein, rhodamine, phycoerythrin, GFP, or BFP) or luminescent moieties (eg, Qdot ^™ nanoparticles available from Quantum Dot Corporation, Palo Alto, CA). General methods used in the implementation of various types of immunoassays mentioned above are known to specialists in this field.

[00149] For diagnostic purposes, a polypeptide, including antibodies, can be labeled with a detectable moiety, including, without limitation, radioisotopes, fluorescent labels, and various enzyme-substrate labels known in the art. Methods for conjugating labels to antibodies are known in the art. In some embodiments, anti-IL31 antibodies do not need to be labeled and their presence can be detected by a second labeled antibody that binds to the first anti-IL31 antibody. In some embodiments, the anti-IL31 antibody can be used in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal antibodies: A Manual of Techniques, pp. 147–158 (CRC Press, Inc. 1987). Anti-IL31 antibodies and polypeptides can also be used in in vivo diagnostic assays such as in vivo imaging. Typically, the antibody or polypeptide is labeled with a radionuclide (such as ¹¹¹ In, ⁹⁹ Tc, ¹⁴ C, ¹³¹ I, ¹²⁵ I, ³ H, or any other radionuclide label, including those described herein) to allow localization cells or tissues of interest using immunoscintigraphy. The antibody can also be used as a staining reagent for pathology using techniques well known in the art.

[00150] In some embodiments, the first antibody is used for diagnosis and the second antibody is used as a therapeutic. In some embodiments, the first and second antibodies are different. In some embodiments, the first and second antibodies may simultaneously bind to an antigen by binding to separate epitopes.

[00151] The following examples illustrate specific aspects of the invention and are not intended to limit the scope of the present invention in any way.

EXAMPLES

Example 1 Identification of mouse monoclonal antibodies that bind to canine IL31.

[00152] The canine IL31 gene encoding the IL31 protein (SEQ ID NO:22) was synthesized with a poly-His tag at the C-terminus and cloned into a mammalian expression vector. A plasmid carrying the canine IL31 gene was transfected into 293 cells.

[00153] The supernatant containing canine IL31 protein was collected and filtered. Canine IL31 was affinity purified on a Ni–NTA column (CaptivA® Protein A Affinity Resin, Repligen).

[00154] Mouse monoclonal antibodies were identified by standard immunization using canine IL31 produced by 293 cells as the immunogen. Two different adjuvants were used during immunization (Antibody Solutions, Sunnyvale, CA) and monoclonal antibodies were generated by the standard hybridoma method. To screen for clones producing antibodies that bind to IL31, developed enzyme-linked immunosorbent assay (ELISA). Canine IL31 was first biotinylated and then injected into streptavidin coated wells. Then, immunized serum was added to the wells, followed by washing and detection with HRP-conjugated anti-mouse antibodies. The presence of canine IL31-binding antibody gave a positive signal. Of the thousands of clones tested by ELISA, the 170 clones with the highest binding affinity were selected for further testing by biosensor analysis (Forte Bio Octet) based on signal intensity. Biotinylated canine IL31 was immobilized on the sensor tip, and hybridoma clone supernatants containing antibodies to canine IL31 were selected for slow dissociation rate (dissociation rate between antibody and ligand). The binding affinity of the top 19 candidates was measured at a single concentration and expressed as the equilibrium dissociation constant (Kd) after antibody concentration was measured by protein A binding based titer analysis on the Biosensor Octet. The Kd values of the top 19 candidates were less than 10 nM.

[00155] In addition, each of the 170 clones with high binding activity, according to the results of ELISA, was tested for neutralizing activity. To assess the potency of the best candidates to reduce canine IL31-mediated pSTAT signaling, the cell function assay described in Example 4 below was performed using canine DH82 cells. Four best clones (M14, M18, M19 and M87) were selected for further study. It is noteworthy that most of the clones with high binding affinity identified by ELISA turned out to be non-functional.

Example 2. Identification of DNA sequences encoding VH and VL monoclonal antibodies.

[00156] Hybridoma cells producing M14, M18, M19 and M87 were pelleted by centrifugation. RNA was extracted and oligonucleotide primers were used to amplify mouse immunoglobulin (Ig) variable domains to obtain cDNA by standard methods. The variable heavy chain (VH) and variable light chain (VL) of each of the four clones were sequenced and analyzed by sequence alignment (FIG. 1; SEQ ID NOS:36-43). Notably, three of the four active antibodies (M14, M18, and M19) had six CDRs that were identical except that CDR-L1 M18 had isoleucine at position 14 (SEQ ID NO:63), while both M14 and M19 had methionine at position 14 (SEQ ID NO:8), as well as the fact that CDR-H2 M18 had tyrosine at position 9 (SEQ ID NO:62 and SEQ ID NO:87), while as in M14 and M19, aspartic acid was present at position 14 (SEQ ID NO:2 and SEQ ID NO:89). The sequence similarity of the CDRs suggests that M14, M18 and M19 share a common epitope.

Example 3: Expression and purification of mouse-canine chimeric and caninized IL31-mAb M14 from CHO cells

[00157] The DNA sequences encoding the chimeric antibody were designed to fuse mouse VH (SEQ ID NO:25) and mouse VL (SEQ ID NO:24) M14 with a canine constant heavy chain and a canine constant light chain. Nucleotide sequences were chemically synthesized and inserted into an expression vector suitable for transfection into a CHO host cell. After transfection into CHO cells, either the light chain protein or the heavy chain protein, or both, is secreted from the cell. For example, chimeric M14, which uses canine IgG-B, was purified using one-step protein A column chromatography.

[00158] M14 mouse VH and VL were caninized by searching for and selecting the appropriate germline canine antibody sequences as a template for CDR grafting followed by protein modeling. Caninized IgG-B M14 (SEQ ID NO:18 and SEQ ID NO:21) was readily expressed and was subjected to one-step purification on a protein A column or other chromatographic methods such as ion exchange column chromatography, hydrophobic interaction column chromatography, mixed column chromatography. mode, for example on CHT, or column chromatography with multimodal mode, such as CaptoMMC. Low pH steps or other virus inactivation and removal steps may be used. The purified protein was mixed with excipients and sterilized by filtration to prepare the pharmaceutical composition of the invention. The pharmaceutical composition was administered to a dog with atopic dermatitis in an amount sufficient to bind to inhibit IL31.

[00159] Vectors were then used to pilot transfect into CHO–S cells with the ^{FreestyleMaxTM} transfection reagent (Life Technologies). The supernatant was collected by clarifying the conditioned medium. The protein was purified using one-step protein A chromatography and used for further analysis.

Example 4 Demonstration of IL31 Binding Activity

[00160] This example shows that the antibodies of the invention, exemplified by chimeric M14 (SEQ ID NO:26 and SEQ ID NO:27) and caninized M14 (SEQ ID NO:18 and SEQ ID NO:21), bind to canine IL31 with the kinetics required for therapeutic activity.

[00161] The binding assay was performed using an Octet biosensor as follows. Briefly, canine IL31 was biotinylated at primary amine groups using EZ-Link ^™ NHS-LC-biotin (Thermo Scientific, cat. no.: 21336) or at glycan groups using EZ-Link ^™ biotin-LC-hydrazide (ThermoFisher Scientific, Item No.: 21340), according to the manufacturer's instructions. Free unreacted biotin was removed from biotinylated IL31 by extensive dialysis. Biotinylated canine IL31 was immobilized on a streptavidin coated sensor tip. Association at four different concentrations (400, 200, 66.6 and 33 nM) of chimeric and caninized M14 antibody and human and canine IL31 (amine-conjugated biotin) and association of 100 nM caninized M14 antibody and canine IL31 (glycan-conjugated-biotin) tracked for ninety seconds. Dissociation was monitored for 600 seconds. The curve containing only the buffer was subtracted to correct for any deviation. Data were fitted to a 2:1 binding model using ^ForteBioTM data analysis software to determine k _on , k _off and Kd. Buffer for dilutions and all binding steps: 20 mM phosphate, 150 mM NaCl, pH 7.2.

[00162] Canine IL31 with a C-terminal poly-His-tag was expressed and purified from CHO-S cells. Human IL31 was obtained from Sino Biological biosensors and streptavidin was obtained from ForteBio (cat. #18-509). The binding kinetics were as follows: in the case of the canine IL31 ligand (amine-conjugated biotin), the Kd(M) value of chimeric M14 was <1.0×10 ^–11 (Fig. 2), and the Kd(M) value of caninized M14 was <1. 0×10 ^-11 (Fig. 3). In the case of the canine IL31 ligand (glycan-conjugated biotin), the Kd(M) value of caninized M14 was <1.0× ^10–12 and the k _off (1/s) value was <1.0× ^10–7 .

[00163] Chimeric M14 and caninized M14 did not give a visible binding signal to human IL31. Thus, Kd was not measured.

Example 5 Demonstration that M14 inhibits canine IL31 signaling.

[00164] Upon binding to the IL-31 receptor, IL-31 induced Janus kinase (Jak) 1 and Jak2 signaling molecules are activated. In turn, activated Jak stimulate phosphorylation of downstream signaling molecules STAT-3 and STAT-5. Detection of anti-IL31 activity from the fraction of cell-free culture medium purified with protein A was performed using the analysis of anti-phospho-Stat3 by immunoblotting (Gonzales et. Al. Vet Dermatol 2013; 24:48-e12). Briefly, DH82 canine monocytic cells (American Type Culture Collection, Manassas, VA, USA) were plated in 96-well flat-bottomed cell culture plates at a seeding density of 1×10 ⁵ cells/well in MEM (Life Technologies) medium containing 15% heat-inactivated fetal bovine serum, 2 mmol/l GlutaMax, 1 mmol/l sodium pyruvate and 10 ng/ml canine interferon-c (R&D Systems, Minneapolis, Minnesota, USA) for 24 h at 37°C. In this experiment, the concentration of canine IL31-Fc was 5 ng/ml (8 nM). Anti-phospho-STAT-3 and anti-STAT-3 antibodies were purchased from R&D Systems. Antibodies to beta-actin were purchased from Sigma-Aldrich. As shown in Figure 4, canine IL31 signaling decreases (as evidenced by a decrease in STAT-3 phosphorylation) as the cell-targeting concentration of caninized M14 increases (lane 1: anti-IL31 antibody absent; lane 2: 3.3 nM; lane 3: 6.6 nM, lane 4: 9.9 nM and lane 5: 13.2 nM).

Example 6 Identification of an epitope of canine IL31 binding to M14.

[00165] To identify the epitope of canine IL31 recognized by M14, a variety of recombinant GST molecules-canine IL31 fragment were generated and the proteins were expressed intracellularly in E. coli . After the transfer of GST-recombinant proteins to the membrane, chimeric M14 was used to probe the membrane. A positive signal occurred if the IL31 fragment contained an epitope.

[00166] Figure 5, in combination with Figure 6, demonstrates that M14 can recognize a minimal fragment (SEQ ID NO:23).

Example 7 Demonstration of cross-reactivity of M14 with feline IL31.

[00167] To evaluate whether the M14 antibody recognizes feline IL31 (SEQ ID NO:28) or equine IL31 (SEQ ID NO:29), each protein was fused to human Fc and expressed in 293 mammalian cells. Partially purified proteins were blotted and probed with M14 antibody. The immunoblot in Figure 1 demonstrates that M14 binds to feline IL31. Western blot analysis did not reveal binding between M14 and equine IL31. However, bilayer interferometry analysis showed that the M14 antibody binds to equine IL31, but with less affinity. Preliminary measurement of Kd with biotinylated equine IL31 immobilized on the sensor indicated that the affinity (Kd) was in the range of about 10 to 50 nM.

Example 8 Felineized M14

[00168] The M14 variable light chain was felinized as (SEQ ID NO:32) and the M14 variable heavy chain was felinized as (SEQ ID NO:33). First, mouse heavy chain and light chain variable sequences were used to search for suitable feline VH and VL variants. Suitable feline reading frames were selected for CDR inoculation. They were further optimized using structural modeling. The felinized VH and VL were fused to the feline IgG heavy chain constant domains (CH1, CH2 and CH3) and the feline light chain constant domain (CL1).

[00169] Feline chimeric M14 antibody (SEQ ID NO:30 and SEQ ID NO:31) or feline M14 antibody (SEQ ID NO:34 and SEQ ID NO:35) can be administered to cats for the treatment of an IL31-induced condition.

Example 9 Identification of canine IL31 binding epitope to M14.

[00170] To further identify the amino acid residues of the canine IL31 epitope recognized by M14, a variety of recombinant GST molecules—a fragment of the canine IL31 epitope carrying alanine mutations were expressed in E. coli . In FIG. 8-12 shows immunoblots of detailed epitope mapping of GST-canine IL31 fusion proteins probed with anti-canine IL31-mAb (M14) or caninized M14 (upper panels) and anti-GST antibody control (lower panels). The epitope fragment tested in each lane is indicated below the immunoblot. The fragment name indicates the amino acid range of mature canine IL31 tested (SEQ ID NO:44) and the position of the alanine mutation, if any. A positive signal arose when the IL31 fragment contained a wild-type epitope, while a negative signal arose when the IL31 fragment contained an alanine substitution at a residue important for antibody–ligand interaction.

[00171] The results of epitope mapping are shown in Table 3 below. The results show that P12, S13, D14 and K17 of mature canine IL31 (SEQ ID NO:44) are involved in binding of the M14 antibody, and that R16 and I18 are partially involved in M14 recognition. Overall, the results of this study suggest that the IL31 polypeptide motif that binds the CDR of the M14 antibody is: PSDX ₁ X ₂ KI, where X ₁ and X ₂ are variable (SEQ ID NO:45).

[00172] Table 3

Amino acid replacement Does substitution prevent binding? P12A Yes S13A Yes D14A Yes V15A No R16A Partially K17A Yes I18A Partially I19A No L20A No

Example 10: M14 binds specifically to IL31 of other species containing the PSDX ₁ X ₂ KI motif in the epitope

[00173] The IL31 epitope motif recognized by M14 (PSDX ₁ X ₂ KI; SEQ ID NO:45) is absent from human (SEQ ID NO:46) or mouse (SEQ ID NO:61) IL31 amino acid sequences. However, this motif has been identified in several other species, including felids:

Felis catus (XP_011286140.1; SEQ ID NO:28)

Odobenus rosmarus divergens (XP_004395998.1; SEQ ID NO:47)

Papio Anubis (XP_003907358.1; SEQ ID NO:49)

Ursus maritimus (XP_008687166.1; SEQ ID NO:51)

Leptonychotes weddellii (XP_006746595.1; SEQ ID NO:52)

Panthera tigris altaica (XP_007079636.1; SEQ ID NO:53)

Acinonyx jubatus (XP_014919275.1; SEQ ID NO:54)

Macaca flavicularis (EHH66805.1; SEQ ID NO:55)

Macaca mulatta (EHH21279.1; SEQ ID NO:56)

Mandrillus leucophaeus (XP_011819882.1; SEQ ID NO:57)

Chlorocebus sabaeus (XP_008003211.1; SEQ ID NO:58)

Cercocebus atys (XP_011926625.1; SEQ ID NO:59)

Rhinopithecus roxellana (XP_010366647.1; SEQ ID NO:60)

[00174] Walrus IL31 (SEQ ID NO:47) and Olive baboon IL31 (SEQ ID NO:49) have a PSDX ₁ X ₂ KI epitope (SEQ ID NO:45) that can be recognized by the M14 antibody. To facilitate protein purification, a C-terminal His-tag was added to walrus IL31 (SEQ ID NO:48) and olive baboon IL31 (SEQ ID NO:50). Western blot analysis confirmed that M14 binds to walrus IL31 (Fig. 13). The M14 antibody or derivatives thereof can be used as therapeutic and diagnostic agents for IL31-induced disease in any of the species listed above.

Example 11 Thermal Stability of Caninized M14 Antibody.

[00175] The thermal stability of the caninized M14 antibody was compared with the commercially available Zoetis' ^CYTOPOINTTM anti-IL31 antibody over a wide pH range using Differential Scanning Fluorescence (DSF). The melting point (Tm) of each antibody at various pH values is shown in Table 4 below. Both CYTOPOINT ^™ and caninized M14 antibodies were buffer exchanged with the assay buffer shown in Table 4 using a PD Minitrap ^™ G-25 column. (GE Healthcare). The Tm of each antibody was evaluated using the same buffer and protein concentration. Caninized M14 antibody showed improved thermal stability compared to CYTOPOINT ^TM over a wide pH range. In addition, under stress conditions, such as keeping at 55°C for 2 days at an antibody concentration of 0.22 mg/ml, CYTOPOINT ^TM precipitated, while in the case of caninized M14 antibody, no precipitation was observed.

[00176] Table 4

Tested pH Analysis Buffer Melting point of canned M14 (Tm, °C) Melting point of ^CYTOPOINTTM (Tm, °C) 3 0.1M NaAc 55.44 49.21 4 0.1M NaAc 61.68 57.54 5 0.1M NaAc 66.84 61.71 6 0.1M _NaPO4 68.20 62.84 7 0.1M _NaPO4 67.25 61.76 7.2 2xPBS 66.71 60.98 8 0.1M _NaPO4 65.08 60.08 9 0.1M TrisHCl 64.25 59.25

Example 12 Buffer formulations for caninized M14 antibodies.

[00177] The thermal stability of the caninized M14 antibody was analyzed in various buffer formulations. Buffers containing sodium phosphate, sodium acetate, or L-histidine were considered. Other compositional variables included: various pH values (pH 5.2, 5.5, 6.0, 6.5 and 7.0), various concentrations of sodium chloride (50 mM and 140 mM), various polysorbates (polysorbate 20 and polysorbate 80) and various antibacterial agents (m-cresol and methylparaben). The melting point (Tm) of the caninized M14 antibody in each buffer was measured by differential scanning fluorescence (DSF) at 20 to 95°C. Table 5 shows the Tm values for the caninized M14 antibody in the various buffers tested.

[00178] Table 5

Description of composition Buffer Composition Melting temperature
(Tm °C) A1 20 mM sodium phosphate
140 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 6.0 67.6 A2 A1+
m-cresol (0.2%) 65.0 A3 A1+
Methylparaben 0.9 mg/ml 67.3 A4 20 mM sodium phosphate
140 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 6.0 66.9 A5 A4+
0.2% m-cresol 64.4 A6 A4+
Methylparaben 0.9 mg/ml 67.3 A7 20 mM sodium phosphate
50 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 6.0 66.3 A8 A7+
0.2% m-cresol No peak* A9 A7+
Methylparaben 0.9 mg/ml 66.1 A10 20 mM sodium phosphate
50 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 6.0 65.8 A11 A10+
0.2% m-cresol 63.6 A12 A10+
Methylparaben 0.9 mg/ml 65.5 B1 20 mM sodium phosphate
140 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 7.0 68.0 B2 B1+
m-cresol (0.2%) 66.2 B3 B1+
Methylparaben 0.9 mg/ml 68.1 B4 20 mM sodium phosphate
140 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 7.0 68.1 B5 B4+
0.2% m-cresol 64.6 B6 B4+
Methylparaben 0.9 mg/ml 67.2 B7 20 mM sodium phosphate
50 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 7.0 67.6 B8 A7+
0.2% m-cresol 64.7 B9 B7+
Methylparaben 0.9 mg/ml 66.9 B10 20 mM sodium phosphate
50 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 7.0 67.3 B11 B10+
0.2% m-cresol 64.8 B12 B10+
Methylparaben 0.9 mg/ml 67.4 C1 20 mM sodium acetate
140 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 5.2 67.9 C2 C1+
m-cresol (0.2%) 65.4 C3 C1+
Methylparaben 0.9 mg/ml 67.6 C4 20 mM sodium acetate
140 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 5.2 67.7 C5 C4+
0.2% m-cresol 64.7 C6 C4+
Methylparaben 0.9 mg/ml 67.7 C7 20 mM sodium acetate
50 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 5.2 66.6 C8 C7+
0.2% m-cresol No peak* C9 C7+
Methylparaben 0.9 mg/ml 66.9 C10 20 mM sodium acetate
50 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 5.2 68.0 C11 C10+
0.2% m-cresol 64.7 C12 C10+
Methylparaben 0.9 mg/ml 67.7 D1 20 mM L-histidine
140 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 5.5 69.5 D2 D1+
m-cresol (0.2%) 68.1 D3 D1+
Methylparaben 0.9 mg/ml 69.6 D4 20 mM L-histidine
140 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 5.5 69.6 D5 D4+
0.2% m-cresol 67.2 D6 D4+
Methylparaben 0.9 mg/ml 69.1 D7 20 mM L-histidine
50 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 5.5 68.0 D8 D7+
0.2% m-cresol 66.2 D9 D7+
Methylparaben 0.9 mg/ml 67.7 D10 20 mM L-histidine
50 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 5.5 68.1 D11 D10+
0.2% m-cresol 66.6 D12 D10+
Methylparaben 0.9 mg/ml 68.1 E1 20 mM L-histidine
140 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 6.5 68.2 E2 E1+
m-cresol (0.2%) No peak* E3 E1+
Methylparaben 0.9 mg/ml 67.9 E4 20 mM L-histidine
140 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 6.5 67.9 E5 E4+
0.2% m-cresol No peak* E6 E4+
Methylparaben 0.9 mg/ml 67.6 E7 20 mM L-histidine
50 mM sodium chloride
Polysorbate 80 (0.05 mg/ml)
pH 6.5 67.3 E8 E7+
0.2% m-cresol No peak* E9 E7+
Methylparaben 0.9 mg/ml 67.5 E10 20 mM L-histidine
50 mM sodium chloride
Polysorbate 20 (0.05 mg/ml)
pH 6.5 67.2 E11 E10+
0.2% m-cresol 64.6 E12 E10+
Methylparaben 0.9 mg/ml 66.9

* The absence of a peak indicates that no distinct melting point was observed.

[00179] DSF also tested the Tm of formulations D1, D2, D3, D4, D5, and D6 in the presence of one of the following sugars (1%): sucrose, trehalose, D-mannitol, maltose, and sorbitol.

[00180] Formulas with and without sugars were also tested under stress conditions: 1 day at 40°C, then 1 day at 45°C, then 4 days at 55°C. SEC HPLC was used to identify and quantify the monomeric and aggregated forms of antibodies in various compositions after exposure to stressful conditions. The sample was loaded onto a SHODEX ^TM KW803 column (8 mm x 300 mm) and attached to a KW–G guard column. An Agilent 1100 Chromatography System was used with 2X PBS (270 mM NaCl, 5.4 mM KCl, 8.6 mM Na ₂ PO ₄ ), pH 7.2 as running buffer at a constant flow rate of 0.5 ml/min. The column was calibrated with a BIORAD gel filtration standard (cat. no. 151-1901) consisting of thyroglobulin, bovine γ-globulin, chicken ovalbumin, horse myoglobin, and vitamin B12 (molecular weight 1350-670,000). The amount of monomeric antibody remaining in solution was determined by measuring the UV absorbance at 214 or 280 nm and calculating the peak area under the curve.

[00181] According to DSF and HPLC analysis, formulations containing L-histidine, sodium chloride, polysorbate 80 and having a pH of 5.0 to 6.2, such as formulations D1, D2, D3, D4, D6, D7, D10 and D12 were found to be more preferable. For example, formulations preferred for single dosing are: 20 mM L-histidine; 140 mM sodium chloride; polysorbate 80 (0.05 mg/ml); pH 5.5. Formulations preferred for multiple administration (in the presence of preservatives) are as follows:

1. 20 mM L-histidine; 140 mM sodium chloride; Polysorbate 80 (0.05 mg/ml); sucrose (1–3%); m-cresol (0.2%); pH 5.5; And

2. 20 mM L-histidine; 140 mM sodium chloride; Polysorbate 80 (0.05 mg/ml); trehalose (1–3%); methylparaben (0.9%); pH 5.5.

--->

SEQUENCE LISTS

<110> KINDRED BIOSIENCES, INC.

<120> ANTI-IL31 ANTIBODIES FOR VETERINARY USE

<130> 01157-0007-01PCT

<140> PCT/US18/17623

<141> 2018-02-09

<150> US 62/463,543

<151> 2017-02-24

<150> PCT/US2017/023788

<151> 2017-03-23

<160> 89

<170>PatentIn version 3.5

<210> 1

<211> 9

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(9)

<223> Variable heavy chain CDR-H1 amino acid sequence

clones M14, M18 and M19 mouse antibodies

<400> 1

Gly Asp Ser Ile Thr Ser Gly Tyr Trp

15

<210> 2

<211> 16

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(16)

<223> Variable heavy chain CDR-H2 amino acid sequence

M14 and M19 mouse antibody clones

<400> 2

Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 3

<211> 12

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(12)

<223> Variable heavy chain CDR-H3 amino acid sequence

clones M14, M18 and M19 mouse antibodies

<400> 3

Ala Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr

1 5 10

<210> 4

<211> 25

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(25)

<223> Amino acid sequence of HC-FR1 framework region

heavy chain variable region of mouse antibody clone M14

<400> 4

Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Thr

20 25

<210> 5

<211> 15

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(15)

<223> Amino acid sequence of HC-FR2 framework region

heavy chain variable region of mouse antibody clone M14

<400> 5

Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met Gly

1 5 10 15

<210> 6

<211> 30

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(30)

<223> Amino acid sequence of HC-FR3 framework region

heavy chain variable region of mouse antibody clone M14

<400> 6

Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu Gln

1 5 10 15

Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

20 25 30

<210> 7

<211> 11

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(11)

<223> Amino acid sequence of HC-FR4 framework region

heavy chain variable region of mouse antibody clone M14

<400> 7

Trp Gly Glyn Gly Thr Ser Val Thr Val Ser Ser

1 5 10

<210> 8

<211> 15

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(15)

<223> Variable heavy chain CDR-L1 amino acid sequence

M14 and M19 mouse antibody clones

<400> 8

Arg Ala Ser Glu Ser Val Asp Thr Tyr Gly Asn Ser Phe Met His

1 5 10 15

<210> 9

<211> 7

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(7)

<223> Variable heavy chain CDR-L2 amino acid sequence

clones M14, M18 and M19 mouse antibodies

<400> 9

Arg Ala Ser Asn Leu Glu Ser

15

<210> 10

<211> 9

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(9)

<223> Variable heavy chain CDR-L3 amino acid sequence

clones M14, M18 and M19 mouse antibodies

<400> 10

Gln Gln Ser Tyr Glu Asp Pro Trp Thr

15

<210> 11

<211> 23

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(23)

<223> Amino acid sequence of LC-FR1 framework region

light chain variable region of mouse antibody clone M14

<400> 11

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys

20

<210> 12

<211> 15

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(15)

<223> Amino acid sequence of LC-FR2 framework region

light chain variable region of mouse antibody clone M14

<400> 12

Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 13

<211> 32

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(32)

<223> Amino acid sequence of LC-FR3 framework region

light chain variable region of mouse antibody clone M14

<400> 13

Gly Ile Pro Ala Arg Phe Gly Gly Ser Gly Ser Arg Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Asp Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys

20 25 30

<210> 14

<211> 10

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(10)

<223> Amino acid sequence of LC-FR4 framework region

light chain variable region of mouse antibody clone M14

<400> 14

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 15

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized amino acid sequence

variable heavy chain clone M14 mouse antibody

<400> 15

Glu Val Gln Leu Val Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 16

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized amino acid sequence

variable light chain of mouse antibody clone M14

<400> 16

Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala

50 55 60

Arg Phe Gly Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Gln Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 17

<211> 448

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized heavy chain sequence from

clone M14 mouse antibody and canine IgG-A

<400> 17

Glu Val Gln Leu Val Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Ser Thr Ser Gly Ser Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Ser Gly Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ser Leu Thr Ser Gly Val His Thr Phe Pro Ser Val Leu Gln

165 170 175

Ser Ser Gly Leu His Ser Leu Ser Ser Met Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Pro Ser Glu Thr Phe Thr Cys Asn Val Val His Pro Ala Ser

195 200 205

Asn Thr Lys Val Asp Lys Pro Val Phe Asn Glu Cys Arg Cys Thr Asp

210 215 220

Thr Pro Cys Pro Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile

225 230 235 240

Phe Pro Pro Lys Pro Lys Asp Ile Leu Arg Ile Thr Arg Thr Pro Glu

245 250 255

Val Thr Cys Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln

260 265 270

Ile Ser Trp Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln

275 280 285

Ser Arg Glu Gln Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu

290 295 300

Pro Ile Glu His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg

305 310 315 320

Val Asn His Ile Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys

325 330 335

Ala Arg Gly Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser

340 345 350

Pro Lys Glu Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile

355 360 365

Lys Asp Phe Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly

370 375 380

Gln Gln Glu Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp

385 390 395 400

Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Thr

420 425 430

Leu Gln Asn His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys

435 440 445

<210> 18

<211> 453

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized heavy chain sequence from

clone M14 mouse antibody and canine IgG-B

<400> 18

Glu Val Gln Leu Val Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Ser Thr Ser Gly Ser Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Ser Gly Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ser Leu Thr Ser Gly Val His Thr Phe Pro Ser Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Met Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Pro Ser Glu Thr Phe Thr Cys Asn Val Ala His Pro Ala Ser

195 200 205

Lys Thr Lys Val Asp Lys Pro Val Pro Lys Arg Glu Asn Gly Arg Val

210 215 220

Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu

245 250 255

Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro

260 265 270

Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met

275 280 285

Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly

305 310 315 320

Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile

325 330 335

Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val

340 345 350

Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser

355 360 365

Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu

370 375 380

Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr

385 390 395 400

Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu

405 410 415

Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser

435 440 445

His Ser Pro Gly Lys

450

<210> 19

<211> 451

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized heavy chain sequence from

clone M14 mouse antibody and canine IgG-C

<400> 19

Glu Val Gln Leu Val Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Ser Gln Ser Gly Ser Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Ser Gly Tyr Ile Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Val Ser Leu Thr Ser Gly Val His Thr Phe Pro Ser Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Met Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Pro Ser Glu Thr Phe Thr Cys Asn Val Ala His Pro Ala Thr

195 200 205

Asn Thr Lys Val Asp Lys Pro Val Ala Lys Glu Cys Glu Cys Lys Cys

210 215 220

Asn Cys Asn Asn Cys Pro Cys Pro Gly Cys Gly Leu Leu Gly Gly Pro

225 230 235 240

Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Ile Leu Val Thr Ala

245 250 255

Arg Thr Pro Thr Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asn

260 265 270

Pro Glu Val Gln Ile Ser Trp Phe Val Asp Ser Lys Gln Val Gln Thr

275 280 285

Ala Asn Thr Gln Pro Arg Glu Glu Gln Ser Asn Gly Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Ser Gly Lys Gln

305 310 315 320

Phe Lys Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Glu

325 330 335

Ile Ile Ser Lys Thr Pro Gly Gln Ala His Gln Pro Asn Val Tyr Val

340 345 350

Leu Pro Pro Ser Arg Asp Glu Met Ser Lys Asn Thr Val Thr Leu Thr

355 360 365

Cys Leu Val Lys Asp Phe Phe Pro Glu Ile Asp Val Glu Trp Gln

370 375 380

Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Met Thr Pro Pro

385 390 395 400

Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val

405 410 415

Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Ile Ser Leu Ser His Ser

435 440 445

Pro Gly Lys

450

<210> 20

<211> 449

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized heavy chain sequence from

clone M14 mouse antibody and canine IgG-D

<400> 20

Glu Val Gln Leu Val Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Ser Thr Ser Gly Ser Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Ser Gly Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ser Leu Thr Ser Gly Val His Thr Phe Pro Ser Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Thr Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Pro Ser Glu Thr Phe Thr Cys Asn Val Val His Pro Ala Ser

195 200 205

Asn Thr Lys Val Asp Lys Pro Val Pro Lys Glu Ser Thr Cys Lys Cys

210 215 220

Ile Ser Pro Cys Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Ile Phe Pro Pro Lys Pro Lys Asp Ile Leu Arg Ile Thr Arg Thr Pro

245 250 255

Glu Ile Thr Cys Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val

260 265 270

Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr

275 280 285

Gln Pro Arg Glu Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Pro Ile Glu His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys

305 310 315 320

Arg Val Asn His Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser

325 330 335

Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro

340 345 350

Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu

355 360 365

Ile Lys Asp Phe Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn

370 375 380

Gly Gln Pro Glu Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu

385 390 395 400

Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys

405 410 415

Ser Arg Trp Gln Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu

420 425 430

Ala Leu Gln Asn His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly

435 440 445

Lys

<210> 21

<211> 221

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: caninized light chain sequence from

mouse antibody clone M14 and canine light chain constant region

<400> 21

Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala

50 55 60

Arg Phe Gly Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Gln Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Asn Asp Ala Gln Pro Ala Val Tyr Leu Phe Gln Pro Ser Pro Asp Gln

115 120 125

Leu His Thr Gly Ser Ala Ser Val Val Cys Leu Leu Asn Ser Phe Tyr

130 135 140

Pro Lys Asp Ile Asn Val Lys Trp Lys Val Asp Gly Val Ile Gln Asp

145 150 155 160

Thr Gly Ile Gln Glu Ser Val Thr Glu Gln Asp Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Met Ser Ser Thr Glu Tyr Leu Ser His

180 185 190

Glu Leu Tyr Ser Cys Glu Ile Thr His Lys Ser Leu Pro Ser Thr Leu

195 200 205

Ile Lys Ser Phe Gln Arg Ser Glu Cys Gln Arg Val Asp

210 215 220

<210> 22

<211> 159

<212> Protein

<213> Unknown

<220>

<223> Canidae sp.

<220>

<221> misc_feature

<222> (1)..(159)

<223> Canine IL31 amino acid sequence

<400> 22

Met Leu Ser His Thr Gly Pro Ser Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Ser Met Glu Thr Leu Leu Ser Ser His Met Ala Pro Thr His Gln Leu

20 25 30

Pro Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser

35 40 45

Arg Gly Leu Leu Glu Asp Tyr Gln Lys Lys Glu Thr Gly Val Pro Glu

50 55 60

Ser Asn Arg Thr Leu Leu Leu Cys Leu Thr Ser Asp Ser Gln Pro Pro

65 70 75 80

Arg Leu Asn Ser Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Ile Ile Asp Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln His Glu Pro Glu Thr Glu Ile Ser Val Pro Ala Asp

115 120 125

Thr Phe Glu Cys Lys Ser Phe Ile Leu Thr Ile Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Glu Ser Val Phe Lys Ser Leu Asn Ser Gly Pro Gln

145 150 155

<210> 23

<211> 17

<212> Protein

<213> Unknown

<220>

<223> Canidae sp.

<220>

<221> misc_feature

<222> (1)..(17)

<223> Canine IL31 epitope

<400> 23

Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser Arg

1 5 10 15

gly

<210> 24

<211> 111

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(111)

<223> Variable light chain amino acid sequence of clone M14

mouse antibody

<400> 24

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala

50 55 60

Arg Phe Gly Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 25

<211> 118

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> misc_feature

<222> (1)..(118)

<223> Variable heavy chain amino acid sequence of clone M14

mouse antibody

<400> 25

Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Ser Val Thr Val Ser Ser

115

<210> 26

<211> 221

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: chimeric variable light chain of clone M14 murine

antibodies and canine light chain constant region

<400> 26

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala

50 55 60

Arg Phe Gly Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Asn Asp Ala Gln Pro Ala Val Tyr Leu Phe Gln Pro Ser Pro Asp Gln

115 120 125

Leu His Thr Gly Ser Ala Ser Val Val Cys Leu Leu Asn Ser Phe Tyr

130 135 140

Pro Lys Asp Ile Asn Val Lys Trp Lys Val Asp Gly Val Ile Gln Asp

145 150 155 160

Thr Gly Ile Gln Glu Ser Val Thr Glu Gln Asp Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Met Ser Ser Thr Glu Tyr Leu Ser His

180 185 190

Glu Leu Tyr Ser Cys Glu Ile Thr His Lys Ser Leu Pro Ser Thr Leu

195 200 205

Ile Lys Ser Phe Gln Arg Ser Glu Cys Gln Arg Val Asp

210 215 220

<210> 27

<211> 453

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: chimeric variable heavy chain of clone M14 murine

antibodies and canine IgG-B

<400> 27

Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Ser Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Ser Thr Ser Gly Ser Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Ser Gly Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ser Leu Thr Ser Gly Val His Thr Phe Pro Ser Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Met Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Pro Ser Glu Thr Phe Thr Cys Asn Val Ala His Pro Ala Ser

195 200 205

Lys Thr Lys Val Asp Lys Pro Val Pro Lys Arg Glu Asn Gly Arg Val

210 215 220

Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu

245 250 255

Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro

260 265 270

Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met

275 280 285

Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly

305 310 315 320

Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile

325 330 335

Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val

340 345 350

Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser

355 360 365

Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu

370 375 380

Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr

385 390 395 400

Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu

405 410 415

Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser

435 440 445

His Ser Pro Gly Lys

450

<210> 28

<211> 159

<212> Protein

<213> Felis catus

<220>

<221> misc_feature

<222> (1)..(159)

<223> Amino acid sequence of feline IL31 NCBI ref:

XP_011286140.1 [felis catus]

<400> 28

Met Leu Ser His Ala Gly Pro Ala Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Cys Met Glu Thr Leu Leu Pro Ser His Met Ala Pro Ala His Arg Leu

20 25 30

Gln Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Arg Pro Met Ser

35 40 45

Lys Gly Leu Leu Gln Asp Tyr Leu Lys Lys Glu Ile Gly Leu Pro Glu

50 55 60

Ser Asn His Ser Ser Leu Pro Cys Leu Ser Ser Asp Ser Gln Leu Pro

65 70 75 80

His Ile Asn Gly Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Thr Ile Asp Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln Arg Glu Pro Glu Ala Lys Val Ser Met Pro Ala Asp

115 120 125

Asn Phe Glu Arg Lys Asn Phe Ile Leu Ala Val Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Glu His Val Leu Gln Ser Leu Asn Ser Gly Pro Gln

145 150 155

<210> 29

<211> 209

<212> Protein

<213> Equus caballus

<220>

<221> misc_feature

<222> (1)..(209)

<223> Equine IL31 amino acid sequence

<400> 29

Met Val Ser His Ile Gly Ser Thr Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Thr Leu Met Phe Ser His Thr Gly Pro Ile Tyr Gln Leu

20 25 30

Gln Pro Lys Glu Ile Gln Ala Ile Ile Val Glu Leu Gln Asn Leu Ser

35 40 45

Lys Lys Leu Leu Asp Asp Tyr Leu Asn Lys Glu Lys Gly Val Gln Lys

50 55 60

Phe Asp Ser Asp Leu Pro Ser Cys Phe Thr Ser Asp Ser Gln Ala Pro

65 70 75 80

Gly Asn Ile Asn Ser Ser Ala Ile Leu Pro Tyr Phe Lys Ala Ile Ser

85 90 95

Pro Ser Leu Asn Asn Asp Lys Ser Leu Tyr Ile Ile Glu Gln Leu Asp

100 105 110

Lys Leu Asn Phe Gln Asn Ala Pro Glu Thr Glu Val Ser Met Pro Thr

115 120 125

Asp Asn Phe Glu Arg Lys Arg Phe Ile Leu Thr Ile Leu Arg Trp Phe

130 135 140

Ser Asn Cys Leu Glu Leu Ala Met Lys Thr Leu Thr Thr Ala Glu Gln

145 150 155 160

Ala Leu Pro Pro Leu Asp Pro Ser Thr Pro His Ala Gly Ala Val Ala

165 170 175

Leu Thr His His Gln Gln Asp Arg Thr Ala Leu Asp Arg Ala Val Phe

180 185 190

Pro Phe Val Trp Ala Ala Pro Arg Gly Gly Glu Val Gly Asp Gly Gly

195 200 205

His

<210> 30

<211> 221

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: chimeric variable light chain of clone M14 murine

antibodies and feline light chain constant region

<400> 30

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Ile Pro Ala

50 55 60

Arg Phe Gly Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asp

65 70 75 80

Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Ser Asp Ala Gln Pro Ser Val Phe Leu Phe Gln Pro Ser Leu Asp Glu

115 120 125

Leu His Thr Gly Ser Ala Ser Ile Val Cys Ile Leu Asn Asp Phe Tyr

130 135 140

Pro Lys Glu Val Asn Val Lys Trp Lys Val Asp Gly Val Val Gln Asn

145 150 155 160

Lys Gly Ile Gln Glu Ser Thr Thr Glu Gln Asn Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Met Ser Ser Thr Glu Tyr Gln Ser

180 185 190

His Glu Lys Phe Ser Cys Glu Val Thr His Lys Ser Leu Ala Ser Thr

195 200 205

Leu Val Lys Ser Phe Asn Arg Ser Glu Cys Gln Arg Glu

210 215 220

<210> 31

<211> 453

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: chimeric variable heavy chain of clone M14 murine

antibodies and feline heavy chain constant region

<400> 31

Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met

35 40 45

Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro Ser Leu Lys

50 55 60

Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu

65 70 75 80

Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Ser Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Thr Thr Ser Gly Ala Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Leu Gly Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ala Ser Gly Leu Tyr Ser Leu Ser Ser Met Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Leu Ser Asp Thr Phe Thr Cys Asn Val Ala His Pro Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Thr Val Arg Lys Thr Asp His Pro Pro Gly

210 215 220

Pro Lys Pro Cys Asp Cys Pro Lys Cys Pro Pro Pro Glu Met Leu Gly

225 230 235 240

Gly Pro Ser Ile Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Ser

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Leu Val Val Asp Leu Gly Pro

260 265 270

Asp Asp Ser Asp Val Gln Ile Thr Trp Phe Val Asp Asn Thr Gln Val

275 280 285

Tyr Thr Ala Lys Thr Ser Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Pro Ile Leu His Gln Asp Trp Leu Lys Gly

305 310 315 320

Lys Glu Phe Lys Cys Lys Val Asn Ser Lys Ser Leu Pro Ser Pro Ile

325 330 335

Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln Pro His Glu Pro Gln Val

340 345 350

Tyr Val Leu Pro Pro Ala Gln Glu Glu Leu Ser Arg Asn Lys Val Ser

355 360 365

Val Thr Cys Leu Ile Lys Ser Phe His Pro Pro Asp Ile Ala Val Glu

370 375 380

Trp Glu Ile Thr Gly Gln Pro Glu Pro Glu Asn Asn Tyr Arg Thr Thr

385 390 395 400

Pro Pro Gln Leu Asp Ser Asp Gly Thr Tyr Phe Val Tyr Ser Lys Leu

405 410 415

Ser Val Asp Arg Ser His Trp Gln Arg Gly Asn Thr Tyr Thr Cys Ser

420 425 430

Val Ser His Glu Ala Leu His Ser His His Thr Gln Lys Ser Leu Thr

435 440 445

Gln Ser Pro Gly Lys

450

<210> 32

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: feline variable light chain sequence

clone M14 mouse antibody

<400> 32

Glu Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Pro Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 33

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: felinized variable sequence

mouse antibody clone M14 heavy chain

<400> 33

Asp Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gln Trp Val

35 40 45

Ala Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Leu Asn Asn Leu Lys Ala Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 34

<211> 221

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: felinized clone light chain sequence

M14 mouse antibody

<400> 34

Glu Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Pro Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Thr Tyr

20 25 30

Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro

35 40 45

Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr

85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Ser Asp Ala Gln Pro Ser Val Phe Leu Phe Gln Pro Ser Leu Asp Glu

115 120 125

Leu His Thr Gly Ser Ala Ser Ile Val Cys Ile Leu Asn Asp Phe Tyr

130 135 140

Pro Lys Glu Val Asn Val Lys Trp Lys Val Asp Gly Val Val Gln Asn

145 150 155 160

Lys Gly Ile Gln Glu Ser Thr Thr Glu Gln Asn Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Met Ser Ser Thr Glu Tyr Gln Ser

180 185 190

His Glu Lys Phe Ser Cys Glu Val Thr His Lys Ser Leu Ala Ser Thr

195 200 205

Leu Val Lys Ser Phe Asn Arg Ser Glu Cys Gln Arg Glu

210 215 220

<210> 35

<211> 453

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic: felinized clone heavy chain sequence

M14 mouse antibody

<400> 35

Asp Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr Gly Asp Ser Ile Thr Ser Gly

20 25 30

Tyr Trp Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gln Trp Val

35 40 45

Ala Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Ala Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Leu Asn Asn Leu Lys Ala Glu Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Thr Ala Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Cys Gly Thr Thr Ser Gly Ala Thr Val Ala Leu Ala

130 135 140

Cys Leu Val Leu Gly Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ala Ser Gly Leu Tyr Ser Leu Ser Ser Met Val Thr Val Pro Ser Ser

180 185 190

Arg Trp Leu Ser Asp Thr Phe Thr Cys Asn Val Ala His Pro Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Thr Val Arg Lys Thr Asp His Pro Pro Gly

210 215 220

Pro Lys Pro Cys Asp Cys Pro Lys Cys Pro Pro Pro Glu Met Leu Gly

225 230 235 240

Gly Pro Ser Ile Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Ser

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Leu Val Val Asp Leu Gly Pro

260 265 270

Asp Asp Ser Asp Val Gln Ile Thr Trp Phe Val Asp Asn Thr Gln Val

275 280 285

Tyr Thr Ala Lys Thr Ser Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Pro Ile Leu His Gln Asp Trp Leu Lys Gly

305 310 315 320

Lys Glu Phe Lys Cys Lys Val Asn Ser Lys Ser Leu Pro Ser Pro Ile

325 330 335

Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln Pro His Glu Pro Gln Val

340 345 350

Tyr Val Leu Pro Pro Ala Gln Glu Glu Leu Ser Arg Asn Lys Val Ser

355 360 365

Val Thr Cys Leu Ile Lys Ser Phe His Pro Pro Asp Ile Ala Val Glu

370 375 380

Trp Glu Ile Thr Gly Gln Pro Glu Pro Glu Asn Asn Tyr Arg Thr Thr

385 390 395 400

Pro Pro Gln Leu Asp Ser Asp Gly Thr Tyr Phe Val Tyr Ser Lys Leu

405 410 415

Ser Val Asp Arg Ser His Trp Gln Arg Gly Asn Thr Tyr Thr Cys Ser

420 425 430

Val Ser His Glu Ala Leu His Ser His His Thr Gln Lys Ser Leu Thr

435 440 445

Gln Ser Pro Gly Lys

450

<210> 36

<211> 238

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(238)

<223> Light chain amino acid sequence of clone M14 murine

antibodies

<400> 36

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

35 40 45

Val Asp Thr Tyr Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Ser

50 55 60

Gly Gln Ser Pro Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser

65 70 75 80

Gly Ile Pro Ala Arg Phe Gly Gly Ser Gly Ser Arg Thr Asp Phe Thr

85 90 95

Leu Thr Ile Asp Pro Val Glu Ala Asp Asp Val Ala Thr Tyr Tyr Cys

100 105 110

Gln Gln Ser Tyr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro

130 135 140

Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu

145 150 155 160

Asn Asn Phe Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly

165 170 175

Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser

180 185 190

Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp

195 200 205

Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr

210 215 220

Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys

225 230 235

<210> 37

<211> 238

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(238)

<223> Light chain amino acid sequence of clone M18 murine

antibodies

<400> 37

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Pro Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

35 40 45

Val Asp Thr Tyr Gly Asn Ser Phe Ile His Trp Tyr Gln Gln Lys Pro

50 55 60

Gly Gln Ser Pro Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser

65 70 75 80

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr

85 90 95

Leu Thr Ile Asn Pro Val Glu Thr Asp Asp Val Ala Thr Tyr Tyr Cys

100 105 110

Gln Gln Ser Tyr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro

130 135 140

Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu

145 150 155 160

Asn Asn Phe Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly

165 170 175

Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser

180 185 190

Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp

195 200 205

Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr

210 215 220

Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys

225 230 235

<210> 38

<211> 238

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(238)

<223> Light chain amino acid sequence of clone M19 murine

antibodies

<400> 38

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

35 40 45

Val Asp Thr Tyr Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro

50 55 60

Gly Gln Pro Pro Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser

65 70 75 80

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr

85 90 95

Leu Thr Ile Asn Pro Val Glu Ala Asp Asp Ile Ala Thr Tyr Tyr Cys

100 105 110

Gln Gln Ser Tyr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro

130 135 140

Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu

145 150 155 160

Asn Asn Phe Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly

165 170 175

Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser

180 185 190

Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp

195 200 205

Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr

210 215 220

Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys

225 230 235

<210> 39

<211> 238

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(238)

<223> Light chain amino acid sequence of clone M87 murine

antibodies

<400> 39

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Tyr Arg Ala Ser Lys Ser

35 40 45

Val Ser Thr Ser Gly Tyr Ser Tyr Met His Trp Asn Gln Gln Lys Pro

50 55 60

Gly Gln Pro Pro Arg Leu Leu Ile Tyr Leu Val Ser Asn Leu Glu Ser

65 70 75 80

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

85 90 95

Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys

100 105 110

Gln His Ile Arg Glu Leu Thr Arg Ser Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro

130 135 140

Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu

145 150 155 160

Asn Asn Phe Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly

165 170 175

Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser

180 185 190

Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp

195 200 205

Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr

210 215 220

Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys

225 230 235

<210> 40

<211> 152

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(152)

<223> Variable and hinge region amino acid sequence

mouse antibody clone M14 heavy chain

<400> 40

Met Ala Val Leu Gly Leu Leu Leu Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys

20 25 30

Pro Ser Gln Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile

35 40 45

Thr Ser Gly Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu

50 55 60

Glu Tyr Met Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro

65 70 75 80

Ser Leu Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln

85 90 95

Tyr Tyr Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr

100 105 110

Tyr Cys Ala Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser

130 135 140

Val Tyr Pro Leu Ala Pro Gly Ser

145 150

<210> 41

<211> 152

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(152)

<223> Variable and hinge region amino acid sequence

mouse antibody clone M18 heavy chain

<400> 41

Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys

20 25 30

Pro Ser Gln Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile

35 40 45

Thr Ser Gly Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu

50 55 60

Glu Tyr Met Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro

65 70 75 80

Ser Leu Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln

85 90 95

Tyr Tyr Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr

100 105 110

Tyr Cys Ala Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser

130 135 140

Val Tyr Pro Leu Ala Pro Gly Ser

145 150

<210> 42

<211> 152

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(152)

<223> Variable and hinge region amino acid sequence

mouse antibody clone M19 heavy chain

<400> 42

Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys

20 25 30

Pro Ser Gln Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile

35 40 45

Thr Ser Gly Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Glu Leu

50 55 60

Glu Tyr Met Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Tyr Tyr Asn Pro

65 70 75 80

Ser Leu Lys Ser Arg Phe Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln

85 90 95

Tyr Tyr Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr

100 105 110

Tyr Cys Ala Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser

130 135 140

Val Tyr Pro Leu Ala Pro Gly Ser

145 150

<210> 43

<211> 154

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(154)

<223> Variable and hinge region amino acid sequence

mouse antibody clone M87 heavy chain

<400> 43

Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Gln

20 25 30

Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe

35 40 45

Thr Asp Tyr Tyr Met Asn Trp Val Arg Gln Pro Pro Gly Lys Ala Leu

50 55 60

Glu Trp Leu Gly Phe Ile Arg Asn Lys Ala Asn Gly Tyr Thr Thr Glu

65 70 75 80

Tyr Ser Ala Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser

85 90 95

Gln Ser Ile Leu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Ser

100 105 110

Ala Thr Tyr Tyr Cys Ala Arg Asp Tyr Tyr Gly Ser Cys Phe Asp Tyr

115 120 125

Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Lys Thr Thr Pro

130 135 140

Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser

145 150

<210> 44

<211> 137

<212> Protein

<213> Unknown

<220>

<223> Canidae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(137)

<223> Mature canine IL31 amino acid sequence

<400> 44

Ser Ser His Met Ala Pro Thr His Gln Leu Pro Pro Ser Asp Val Arg

1 5 10 15

Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser Arg Gly Leu Leu Glu Asp

20 25 30

Tyr Gln Lys Lys Glu Thr Gly Val Pro Glu Ser Asn Arg Thr Leu Leu

35 40 45

Leu Cys Leu Thr Ser Asp Ser Gln Pro Arg Leu Asn Ser Ser Ala

50 55 60

Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro Leu Ser Asp Lys Asn Ile

65 70 75 80

Ile Asp Lys Ile Ile Glu Gln Leu Asp Lys Leu Lys Phe Gln His Glu

85 90 95

Pro Glu Thr Glu Ile Ser Val Pro Ala Asp Thr Phe Glu Cys Lys Ser

100 105 110

Phe Ile Leu Thr Ile Leu Gln Gln Phe Ser Ala Cys Leu Glu Ser Val

115 120 125

Phe Lys Ser Leu Asn Ser Gly Pro Gln

130 135

<210> 45

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> IL31 epitope

<220>

<221> MISC_FEATURE

<222> (4)..(4)

<223> X is any amino acid; or X is hydrophobic

amino acid, or X is selected from A, V, I and L, or X is selected from V and I

<220>

<221> MISC_FEATURE

<222> (5)..(5)

<223> X is any amino acid; or X is hydrophobic

amino acid, or X is selected from A, R, K, Q and N; or X is selected from R and Q

<400> 45

Pro Ser Asp Xaa Xaa Lys Ile

15

<210> 46

<211> 164

<212> Protein

<213> Homo sapiens

<220>

<221> MISC_FEATURE

<222> (1)..(164)

<223> Human IL31 precursor amino acid sequence

NCBI ref: NP_001014358.1

<400> 46

Met Ala Ser His Ser Gly Pro Ser Thr Ser Val Leu Phe Leu Phe Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Ala Ser His Thr Leu Pro Val Arg Leu Leu

20 25 30

Arg Pro Ser Asp Asp Val Gln Lys Ile Val Glu Glu Leu Gln Ser Leu

35 40 45

Ser Lys Met Leu Leu Lys Asp Val Glu Glu Glu Lys Gly Val Leu Val

50 55 60

Ser Gln Asn Tyr Thr Leu Pro Cys Leu Ser Pro Asp Ala Gln Pro Pro

65 70 75 80

Asn Asn Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg

85 90 95

Gln Leu Asp Asn Lys Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp

100 105 110

Lys Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr

115 120 125

Asp Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe

130 135 140

Ser Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln

145 150 155 160

Gln Ala Thr Thr

<210> 47

<211> 177

<212> Protein

<213> Odobenus rosmarus divergens

<220>

<221> MISC_FEATURE

<222> (1)..(177)

<223> Predicted amino acid sequence of walrus IL31 NCBI ref:

XP_004395998.1 [odobeus rosmarus divergens]

<400> 47

Met Leu Ser His Ala Gly Pro Ala Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Phe Met Gly Thr Ser Leu Thr Ser Gln Thr Ala Pro Ile His Gln Leu

20 25 30

His Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser

35 40 45

Lys Gly Leu Leu Glu Asp Tyr Leu Lys Lys Glu Met Gly Val Pro Glu

50 55 60

Ser Asn His Phe Leu Leu Pro Cys Leu Thr Ser Asp Ser Gln Pro Pro

65 70 75 80

Arg Ile Asn Ser Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Thr Ile Asn Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln His Glu Pro Glu Thr Glu Val Ser Val Pro Ala Asp

115 120 125

Thr Phe Glu Ser Lys Ser Phe Ile Leu Ala Ile Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Asp His Val Phe Lys Ser Leu Asn Pro Gly Pro Gln Gln

145 150 155 160

Val Met Gln Gly His Leu Ile Glu Pro Ile Pro Ser Gly Thr Ala Asp

165 170 175

Val

<210> 48

<211> 187

<212> Protein

<213> Odobenus rosmarus divergens

<220>

<221> MISC_FEATURE

<222> (1)..(187)

<223> Walrus_IL31_His6

<400> 48

Met Leu Ser His Ala Gly Pro Ala Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Phe Met Gly Thr Ser Leu Thr Ser Gln Thr Ala Pro Ile His Gln Leu

20 25 30

His Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser

35 40 45

Lys Gly Leu Leu Glu Asp Tyr Leu Lys Lys Glu Met Gly Val Pro Glu

50 55 60

Ser Asn His Phe Leu Leu Pro Cys Leu Thr Ser Asp Ser Gln Pro Pro

65 70 75 80

Arg Ile Asn Ser Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Thr Ile Asn Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln His Glu Pro Glu Thr Glu Val Ser Val Pro Ala Asp

115 120 125

Thr Phe Glu Ser Lys Ser Phe Ile Leu Ala Ile Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Asp His Val Phe Lys Ser Leu Asn Pro Gly Pro Gln Gln

145 150 155 160

Val Met Gln Gly His Leu Ile Glu Pro Ile Pro Ser Gly Thr Ala Asp

165 170 175

Val Gly Ser Gly Ser His His His His His His

180 185

<210> 49

<211> 163

<212> Protein

<213> Papio anubis

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> The predicted amino acid sequence of IL31 olive

baboon NCBI ref: XP_003907358.1 [papio anubis]

<400> 49

Met Ala Ser His Ser Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Gln Pro Ser Asp Ile Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Thr Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Glu

85 90 95

Leu Asp Asn Lys Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr

<210> 50

<211> 173

<212> Protein

<213> Papio anubis

<220>

<221> MISC_FEATURE

<222> (1)..(173)

<223> Olive baboon_IL31_His6

<400> 50

Met Ala Ser His Ser Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Gln Pro Ser Asp Ile Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Thr Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Glu

85 90 95

Leu Asp Asn Lys Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr Gly Ser Gly Ser His His His His His His

165 170

<210> 51

<211> 159

<212> Protein

<213> Ursus maritimus

<220>

<221> MISC_FEATURE

<222> (1)..(159)

<223> Predicted polar bear IL31 amino acid sequence

NCBI ref: XP_008687166.1 [ursus maritimus]

<400> 51

Met Leu Ser Arg Ala Val Pro Ala Gly Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Tyr Met Glu Thr Leu Leu Thr Ser His Thr Ala Pro Thr His Arg Leu

20 25 30

Pro Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser

35 40 45

Lys Gly Leu Leu Glu Asp Tyr Leu Lys Lys Glu Thr Gly Leu Pro Glu

50 55 60

Ser Asn His Ser Leu Val Pro Cys Leu Thr Ser Asp Ser Glu Ala Pro

65 70 75 80

His Ile Asn Ser Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Val Ile Asp Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln His Glu Pro Glu Thr Glu Val Ser Val Pro Ala Asp

115 120 125

Thr Phe Glu Gly Lys Ser Phe Ile Leu Thr Ile Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Glu Arg Val Phe Lys Ser Leu Asn Pro Gly Ala Gln

145 150 155

<210> 52

<211> 185

<212> Protein

<213> Ursus maritimus

<220>

<221> MISC_FEATURE

<222> (1)..(185)

<223> Predicted amino acid sequence of Weddell seal IL31

NCBI ref: XP_006746595.1 [leptonychotes weddellii]

<400> 52

Met Leu Ser His Ala Gly Pro Ala Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Phe Met Glu Thr Ser Leu Thr Ser Gln Thr Val Pro Ile His Gln Leu

20 25 30

Gln Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Gln Pro Leu Ser

35 40 45

Lys Gly Leu Leu Glu Asp Tyr Leu Lys Lys Glu Met Gly Val Pro Glu

50 55 60

Ser Asn His Phe Leu Leu Pro Cys Leu Thr Ser Asp Ser Gln Pro Pro

65 70 75 80

Arg Ile Asn Ser Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Ile Ile Asn Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln His Glu Pro Glu Thr Glu Val Ser Val Pro Ala Asp

115 120 125

Thr Phe Glu Ser Lys Ser Phe Ile Leu Thr Ile Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Gly His Val Leu Lys Ser Leu Asn Pro Gly Pro Gln Gln

145 150 155 160

Val Met Gln Gly His Leu Ala Lys Pro Ile Pro Ser Gly Thr Ala Asp

165 170 175

Met Tyr Glu Thr Leu Arg His His His

180 185

<210> 53

<211> 159

<212> Protein

<213> Panthera tigris altaica

<220>

<221> MISC_FEATURE

<222> (1)..(159)

<223> Predicted siberian tiger IL31 amino acid sequence

NCBI ref: XP_007079636.1 [panther tigris altaica]

<400> 53

Met Leu Ser His Ala Gly Pro Ala Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Cys Met Glu Thr Leu Leu Pro Ser His Met Ala Pro Thr His Arg Leu

20 25 30

Gln Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Arg Pro Met Ser

35 40 45

Lys Gly Leu Leu Gln Asp Tyr Leu Lys Lys Glu Met Gly Leu Pro Glu

50 55 60

Ser Asn His Ser Ser Leu Pro Cys Leu Ser Ser Asp Ser Gln Leu Pro

65 70 75 80

His Ile Asn Gly Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Thr Ile Asp Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln Arg Glu Pro Glu Ala Glu Val Ser Met Pro Ala Asp

115 120 125

Asn Phe Glu Arg Lys Asn Phe Ile Leu Ala Val Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Glu His Val Leu Gln Ser Leu Asn Ser Gly Pro Gln

145 150 155

<210> 54

<211> 159

<212> Protein

<213> Acinonyx jubatus

<220>

<221> MISC_FEATURE

<222> (1)..(159)

<223> Predicted amino acid sequence of cheetah IL31 NCBI ref:

XP_014919275.1 [acinonyx jubatus]

<400> 54

Met Leu Ser His Ala Gly Pro Ala Arg Phe Ala Leu Phe Leu Leu Cys

1 5 10 15

Cys Met Glu Thr Leu Leu Pro Ser His Met Ala Pro Thr His Arg Leu

20 25 30

Gln Pro Ser Asp Val Arg Lys Ile Ile Leu Glu Leu Arg Pro Met Ser

35 40 45

Lys Gly Leu Leu Gln Asp Tyr Leu Lys Lys Glu Met Gly Leu Pro Glu

50 55 60

Ser Asn His Ser Ser Leu Pro Cys Leu Ser Ser Asp Ser Gln Leu Pro

65 70 75 80

His Ile Asn Gly Ser Ala Ile Leu Pro Tyr Phe Arg Ala Ile Arg Pro

85 90 95

Leu Ser Asp Lys Asn Thr Ile Asp Lys Ile Ile Glu Gln Leu Asp Lys

100 105 110

Leu Lys Phe Gln Arg Glu Pro Glu Ala Glu Val Ser Met Pro Ala Asp

115 120 125

Asn Phe Glu Arg Lys Asn Phe Ile Leu Ala Val Leu Gln Gln Phe Ser

130 135 140

Ala Cys Leu Glu His Val Leu Gln Ser Leu Asn Ser Gly Ser Gln

145 150 155

<210> 55

<211> 163

<212> Protein

<213> Macaca fascicularis

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> Predicted amino acid sequence of cynomolgus monkey IL31

GenBank ref: EHH66805.1 [macaca fascicularis]

<400> 55

Met Ala Ser His Ser Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Gln Pro Ser Asp Ile Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Thr Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Gln

85 90 95

Leu Asp Asn Lys Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr

<210> 56

<211> 158

<212> Protein

<213> Macaca mulatta

<220>

<221> MISC_FEATURE

<222> (1)..(158)

<223> Predicted amino acid sequence of IL31 rhesus monkey

(partial) GenBank ref: EHH21279.1 [macaca mulatta]

<400> 56

Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys Cys Leu Gly Gly Trp

1 5 10 15

Leu Thr Ser His Thr Leu Pro Val His Phe Leu Gln Pro Ser Asp Ile

20 25 30

Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser Lys Met Leu Leu Lys

35 40 45

Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser Gln Asn Tyr Thr Leu

50 55 60

Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn Ile Ile His Ser Pro

65 70 75 80

Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Gln Leu Asp Asn Lys Ser

85 90 95

Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys Leu Ile Phe Gln Asp

100 105 110

Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp Thr His Glu Cys Lys

115 120 125

Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser Glu Cys Met Asp Leu

130 135 140

Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln Ala Thr Thr

145 150 155

<210> 57

<211> 163

<212> Protein

<213> Mandrillus leucophaeus

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> Predicted amino acid sequence of IL31 drill NCBI ref:

XP_011819882.1 [mandrillus leucophaeus]

<400> 57

Met Ala Ser His Ser Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Gln Pro Ser Asp Ile Gln Lys Ile Val Lys Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Thr Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Gln

85 90 95

Leu Asp Asn Arg Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr

<210> 58

<211> 163

<212> Protein

<213> Chlorocebus sabaeus

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> Green monkey IL31 predicted amino acid sequence

NCBI ref: XP_008003211.1 [chlorocebus sabaeus]

<400> 58

Met Ala Ser His Ser Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Gln Pro Ser Asp Ile Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Lys Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Gln

85 90 95

Leu Asp Asn Lys Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr

<210> 59

<211> 163

<212> Protein

<213> Cercocebus atys

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> Predicted amino acid sequence of IL31 smoky

Albatross NCBI ref: XP_011926625.1 [cercocebus atys]

<400> 59

Met Ala Ser His Ser Gly Pro Ala Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Gln Pro Ser Asp Ile Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Lys Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Thr Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Gln

85 90 95

Leu Asp Asn Arg Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr

<210> 60

<211> 163

<212> Protein

<213> Rhinopithecus roxellana

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> Predicted amino acid sequence of IL31 golden

snub-nosed monkey NCBI ref: XP_010366647.1 [rhinopithecus roxellana]

<400> 60

Met Ala Ser His Ser Gly Pro Thr Thr Ser Val Leu Phe Leu Leu Cys

1 5 10 15

Cys Leu Gly Gly Trp Leu Thr Ser His Thr Leu Pro Val His Phe Leu

20 25 30

Arg Pro Ser Asp Ile Gln Lys Ile Val Glu Glu Leu Gln Ser Leu Ser

35 40 45

Lys Met Leu Leu Lys Asp Val Glu Glu Asp Lys Gly Val Leu Val Ser

50 55 60

Gln Asn Tyr Thr Leu Pro Cys Leu Thr Pro Asp Ala Gln Pro Pro Asn

65 70 75 80

Ile Ile His Ser Pro Ala Ile Arg Ala Tyr Leu Lys Thr Ile Arg Gln

85 90 95

Leu Asp Asn Lys Ser Val Ile Asp Glu Ile Ile Glu His Leu Asp Lys

100 105 110

Leu Ile Phe Gln Asp Ala Pro Glu Thr Asn Ile Ser Val Pro Thr Asp

115 120 125

Thr His Glu Cys Lys Arg Phe Ile Leu Thr Ile Ser Gln Gln Phe Ser

130 135 140

Glu Cys Met Asp Leu Ala Leu Lys Ser Leu Thr Ser Gly Ala Gln Gln

145 150 155 160

Ala Thr Thr

<210> 61

<211> 163

<212> Protein

<213> Mus musculus

<220>

<221> MISC_FEATURE

<222> (1)..(163)

<223> Amino acid sequence of mouse NCBI IL31 precursor

ref: NP_083870 [mus musculus]

<400> 61

Met Ile Phe His Thr Gly Thr Thr Lys Pro Thr Leu Val Leu Leu Cys

1 5 10 15

Cys Ile Gly Thr Trp Leu Ala Thr Cys Ser Leu Ser Phe Gly Ala Pro

20 25 30

Ile Ser Lys Glu Asp Leu Arg Thr Thr Ile Asp Leu Leu Lys Gln Glu

35 40 45

Ser Gln Asp Leu Tyr Asn Asn Tyr Ser Ile Lys Gln Ala Ser Gly Met

50 55 60

Ser Ala Asp Glu Ser Ile Gln Leu Pro Cys Phe Ser Leu Asp Arg Glu

65 70 75 80

Ala Leu Thr Asn Ile Ser Val Ile Ile Ala His Leu Glu Lys Val Lys

85 90 95

Val Leu Ser Glu Asn Thr Val Asp Thr Ser Trp Val Ile Arg Trp Leu

100 105 110

Thr Asn Ile Ser Cys Phe Asn Pro Leu Asn Leu Asn Ile Ser Val Pro

115 120 125

Gly Asn Thr Asp Glu Ser Tyr Asp Cys Lys Val Phe Val Leu Thr Val

130 135 140

Leu Lys Gln Phe Ser Asn Cys Met Ala Glu Leu Gln Ala Lys Asp Asn

145 150 155 160

Thr Thr Cys

<210> 62

<211> 16

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(16)

<223> Variable heavy chain CDR-H2 amino acid sequence

clone M18 mouse antibody

<400> 62

Tyr Ile Ser Tyr Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 63

<211> 15

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(15)

<223> Variable light chain CDR-L1 amino acid sequence

clone M18 mouse antibody

<400> 63

Arg Ala Ser Glu Ser Val Asp Thr Tyr Gly Asn Ser Phe Ile His

1 5 10 15

<210> 64

<211> 131

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(131)

<223> Variable light chain amino acid sequence of clone M18

mouse antibody

<400> 64

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Pro Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

35 40 45

Val Asp Thr Tyr Gly Asn Ser Phe Ile His Trp Tyr Gln Gln Lys Pro

50 55 60

Gly Gln Ser Pro Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser

65 70 75 80

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr

85 90 95

Leu Thr Ile Asn Pro Val Glu Thr Asp Asp Val Ala Thr Tyr Tyr Cys

100 105 110

Gln Gln Ser Tyr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys

130

<210> 65

<211> 131

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(131)

<223> Variable light chain amino acid sequence of clone M19

mouse antibody

<400> 65

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser

35 40 45

Val Asp Thr Tyr Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro

50 55 60

Gly Gln Pro Pro Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser

65 70 75 80

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr

85 90 95

Leu Thr Ile Asn Pro Val Glu Ala Asp Asp Ile Ala Thr Tyr Tyr Cys

100 105 110

Gln Gln Ser Tyr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys

130

<210> 66

<211> 131

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(131)

<223> Variable light chain amino acid sequence of clone M87

mouse antibody

<400> 66

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro

1 5 10 15

Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala

20 25 30

Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Tyr Arg Ala Ser Lys Ser

35 40 45

Val Ser Thr Ser Gly Tyr Ser Tyr Met His Trp Asn Gln Gln Lys Pro

50 55 60

Gly Gln Pro Pro Arg Leu Leu Ile Tyr Leu Val Ser Asn Leu Glu Ser

65 70 75 80

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

85 90 95

Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys

100 105 110

Gln His Ile Arg Glu Leu Thr Arg Ser Phe Gly Gly Gly Thr Lys Leu

115 120 125

Glu Ile Lys

130

<210> 67

<211> 137

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(137)

<223> Variable heavy chain amino acid sequence of clone M18

mouse antibody

<400> 67

Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys

20 25 30

Pro Ser Gln Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile

35 40 45

Thr Ser Gly Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu

50 55 60

Glu Tyr Met Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr Asn Pro

65 70 75 80

Ser Leu Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln

85 90 95

Tyr Tyr Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr

100 105 110

Tyr Cys Ala Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Ser Val Thr Val Ser Ser

130 135

<210> 68

<211> 137

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(137)

<223> Variable heavy chain amino acid sequence of clone M19

mouse antibody

<400> 68

Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Gln Leu Gln Glu Ser Gly Pro Ser Leu Val Lys

20 25 30

Pro Ser Gln Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Asp Ser Ile

35 40 45

Thr Ser Gly Tyr Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Glu Leu

50 55 60

Glu Tyr Met Gly Tyr Ile Ser Tyr Ser Gly Ile Thr Tyr Tyr Asn Pro

65 70 75 80

Ser Leu Lys Ser Arg Phe Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln

85 90 95

Tyr Tyr Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr

100 105 110

Tyr Cys Ala Arg Tyr Gly Asn Tyr Gly Tyr Ala Met Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Ser Val Thr Val Ser Ser

130 135

<210> 69

<211> 139

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(139)

<223> Variable heavy chain amino acid sequence of clone M87

mouse antibody

<400> 69

Met Ala Val Leu Gly Leu Leu Phe Cys Leu Val Thr Phe Pro Ser Cys

1 5 10 15

Val Leu Ser Glu Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Gln

20 25 30

Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe

35 40 45

Thr Asp Tyr Tyr Met Asn Trp Val Arg Gln Pro Pro Gly Lys Ala Leu

50 55 60

Glu Trp Leu Gly Phe Ile Arg Asn Lys Ala Asn Gly Tyr Thr Thr Glu

65 70 75 80

Tyr Ser Ala Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser

85 90 95

Gln Ser Ile Leu Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Ser

100 105 110

Ala Thr Tyr Tyr Cys Ala Arg Asp Tyr Tyr Gly Ser Cys Phe Asp Tyr

115 120 125

Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser

130 135

<210> 70

<211> 25

<212> Protein

<213> Artificial sequence

<220>

<223> Caninized HC-FR1 M14

<400> 70

Glu Val Gln Leu Val Glu Ser Gly Pro Ser Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr

20 25

<210> 71

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<223> Canned HC-FR2 M14

<400> 71

Asn Trp Ile Arg Lys Phe Pro Gly Asn Lys Leu Glu Tyr Met Gly

1 5 10 15

<210> 72

<211> 30

<212> Protein

<213> Artificial sequence

<220>

<223> Canned HC-FR3 M14

<400> 72

Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Tyr Leu Gln

1 5 10 15

Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

20 25 30

<210> 73

<211> 36

<212> Protein

<213> Artificial sequence

<220>

<223> Alternative caninized HC-FR3 M14

<400> 73

Asn Pro Ser Leu Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys

1 5 10 15

Asn Gln Tyr Tyr Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala

20 25 30

Thr Tyr Tyr Cys

35

<210> 74

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> Caninized HC-FR4 M14

<400> 74

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 75

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<223> Caninized LC-FR1 M14

<400> 75

Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys

20

<210> 76

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<223> Caninized LC-FR2 M14

<400> 76

Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 77

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<223> Caninized LC-FR3 M14

<400> 77

Gly Ile Pro Ala Arg Phe Gly Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Asp Pro Val Gln Ala Asp Asp Val Ala Thr Tyr Tyr Cys

20 25 30

<210> 78

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<223> Caninized LC-FR4 M14

<400> 78

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 79

<211> 25

<212> Protein

<213> Artificial sequence

<220>

<223> Felined HC-FR1 M14

<400> 79

Asp Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Thr Cys Ser Val Thr

20 25

<210> 80

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<223> Felined HC-FR2 M14

<400> 80

Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gln Trp Val Ala

1 5 10 15

<210> 81

<211> 37

<212> Protein

<213> Artificial sequence

<220>

<223> Felineized HC-FR3 M14

<400> 81

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys

1 5 10 15

Asn Thr Leu Tyr Leu Gln Leu Asn Asn Leu Lys Ala Glu Asp Thr Ala

20 25 30

Thr Tyr Tyr Cys Ala

35

<210> 82

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> Felined HC-FR4 M14

<400> 82

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 83

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<223> Feline LC-FR1 M14

<400> 83

Glu Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Pro Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys

20

<210> 84

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<223> Felined LC-FR2 M14

<400> 84

Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 85

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<223> Felined LC-FR3 M14

<400> 85

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Ser Leu Glu Pro Glu Asp Ala Ala Thr Tyr Tyr Cys

20 25 30

<210> 86

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<223> Feline LC-FR4 M14

<400> 86

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 87

<211> 10

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(10)

<223> Alternative amino acid sequence of CDR-H2 variable

mouse antibody clone M18 heavy chain

<400> 87

Tyr Ile Ser Tyr Ser Gly Ile Thr Tyr Tyr

1 5 10

<210> 88

<211> 7

<212> Protein

<213> Unknown

<220>

<223> Canidae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(7)

<223> Alternate canine IL31 epitope

<400> 88

Pro Ser Asp Val Arg Lys Ile

15

<210> 89

<211> 10

<212> Protein

<213> Unknown

<220>

<223> Murinae sp.

<220>

<221> MISC_FEATURE

<222> (1)..(10)

<223> Alternative amino acid sequence of CDR-H2 variable

heavy chain clones M14 and M19 mouse antibodies

<400> 89

Tyr Ile Ser Tyr Ser Gly Ile Thr Asp Tyr

1 5 10

<---

Claims (97)

1. An isolated antibody or an isolated antibody fragment that binds to canine IL31, containing: a) CDR-H1 containing the amino acid sequence of SEQ ID NO: 1; CDR-H2 containing the amino acid sequence of SEQ ID NO: 89; And a CDR-H3 containing the amino acid sequence of SEQ ID NO: 3, and b) CDR-L1 containing the amino acid sequence of SEQ ID NO: 8; CDR-L2 containing the amino acid sequence of SEQ ID NO: 9; And CDR-L3 containing the amino acid sequence of SEQ ID NO: 10. 2. An antibody or antibody fragment according to claim 1, wherein the antibody or antibody fragment binds to the amino acid sequence PSDX ₁ X ₂ KI (SEQ ID NO: 45), where X is any amino acid residue. 3. An antibody or antibody fragment according to claim 1 or 2, wherein the antibody or antibody fragment binds to canine IL31 with a dissociation constant (Kd) of less than 5 x 10 ^-6 M, less than 1 x 10 ^-6 M, less than 5 x 10 ^–7 M, less than 1× ^10–7 M, less than 5× ^10–8 M, less than 1×10–8 ^M , less than 5× ^10–9 M, less than 1× ^10–9 M, less than 5× ^10–10 M, less than 1× ^10–10 M, less than 5× ^10–11 M, less than 1× ^10–11 M, less than 5× ^10–12 M, or less than 1× ^10–12 M, as measured by bilayer interferometry. 4. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody or antibody fragment reduces IL31 signaling in the companion animal species as measured by reduction in STAT-3 phosphorylation. 5. An antibody or antibody fragment according to claim 4, wherein the companion animal species is canine, feline or equine. 6. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody or antibody fragment binds to feline IL31 or equine IL31 as determined by immunoblot analysis and/or bilayer interferometry. 7. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody or antibody fragment competes with the M14 monoclonal antibody for binding to canine IL31. 8. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody or antibody fragment competes with the M14 monoclonal antibody for binding to feline IL31. 9. An antibody or antibody fragment according to any of the preceding claims, wherein the antibody or antibody fragment does not bind to human IL31 as determined by immunoblot analysis and/or bilayer interferometry. 10. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody is a monoclonal antibody. 11. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody or antibody fragment is caninized, felinized, equinized, or chimeric. 12. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody is a chimeric antibody comprising a murine variable heavy chain framework or a murine variable light chain framework. 13. An antibody or antibody fragment according to any of the preceding claims, further comprising one or more of: (a) heavy chain variable region framework region 1 (HC-FR1) sequences of SEQ ID NO: 4, 70 or 79; (b) HC-FR2 sequence SEQ ID NO: 5, 71 or 80; (c) HC-FR3 sequence SEQ ID NO: 6, 73 or 81; (d) HC-FR4 sequence SEQ ID NO: 7, 74 or 82; (e) light chain variable region framework region 1 (LC-FR1) sequences of SEQ ID NO: 11, 75, or 83; (f) LC-FR2 sequence SEQ ID NO: 12, 76 or 84; (g) LC-FR3 sequence SEQ ID NO: 13, 77 or 85; or (h) LC-FR4 sequences SEQ ID NO: 14, 78 or 86. 14. An antibody or antibody fragment according to any one of the preceding paragraphs, containing: a) (i) a light chain variable region comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, or at least 98% identical to the amino acid sequence of SEQ ID NO: 24 ; (ii) a heavy chain variable region comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, or at least 98% identical to the amino acid sequence of SEQ ID NO: 25; or (iii) a light chain variable region as defined in (i) and a heavy chain variable region as defined in (ii); or b) (i) a light chain variable region comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, or at least 98% identical to the amino acid sequence of SEQ ID NO: 16 ; (ii) a heavy chain variable region comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, or at least 98% identical to the amino acid sequence of SEQ ID NO: 15; or (iii) a light chain variable region as defined in (i) and a heavy chain variable region as defined in (ii); or c)(i) a light chain variable region comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, or at least 98% identical to the amino acid sequence of SEQ ID NO: 32 ; (ii) a heavy chain variable region comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, or at least 98% identical to the amino acid sequence of SEQ ID NO: 33; or (iii) a light chain variable region as defined in (i) and a heavy chain variable region as defined in (ii). 15. An antibody or antibody fragment according to any one of the preceding paragraphs, containing the variable region of the light chain containing the amino acid sequence of SEQ ID NO: 24; SEQ ID NO: 16; or SEQ ID NO: 32. 16. The antibody or antibody fragment according to any one of the preceding paragraphs, containing the variable region of the heavy chain containing the amino acid sequence of SEQ ID NO: 25; SEQ ID NO: 15; or SEQ ID NO: 33. 17. An antibody or antibody fragment according to any one of the preceding claims, containing a light chain variable region comprising the amino acid sequence of SEQ ID NO:24 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25; a light chain variable region comprising the amino acid sequence of SEQ ID NO: 16 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15; or a light chain variable region containing the amino acid sequence of SEQ ID NO:32; and a heavy chain variable region containing the amino acid sequence of SEQ ID NO:33. 18. An antibody or antibody fragment according to any one of the preceding claims, comprising a heavy chain constant region or a light chain constant region derived from a companion animal. 19. An antibody or antibody fragment according to any of the preceding claims, containing (a) a canine heavy chain constant region selected from IgG-A, IgG-B, IgG-C and IgG-D constant regions; (b) a feline heavy chain constant region selected from IgG1, IgG2a and IgG2b constant regions; or (c) an equine heavy chain constant region selected from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 and IgG7 constant regions. 20. An antibody or antibody fragment according to any one of the preceding paragraphs, containing: a)(i) a light chain containing the amino acid sequence of SEQ ID NO: 26; (ii) a heavy chain containing the amino acid sequence of SEQ ID NO: 27; or (iii) a light chain as defined in (i) and a heavy chain as defined in (ii); or b)(i) a light chain containing the amino acid sequence of SEQ ID NO: 30; (ii) a heavy chain containing the amino acid sequence of SEQ ID NO: 31; or (iii) a light chain as defined in (i) and a heavy chain as defined in (ii); or c) (i) a light chain containing the amino acid sequence of SEQ ID NO: 34; (ii) a heavy chain containing the amino acid sequence of SEQ ID NO: 35; or (iii) a light chain as defined in (i) and a heavy chain as defined in (ii). 21. An antibody or antibody fragment according to any of the preceding claims, containing a light chain containing the amino acid sequence of SEQ ID NO: 21. 22. An antibody or antibody fragment according to any one of the preceding paragraphs, containing a heavy chain containing the amino acid sequence of SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; or SEQ ID NO: 20. 23. An antibody or antibody fragment according to any one of the preceding claims, wherein the antibody fragment is selected from Fv, scFv, Fab, Fab', F(ab') ₂ and Fab'-SH. 24. An antibody or antibody fragment according to any of the preceding claims, wherein the antibody or antibody fragment is bispecific, where the antibody or antibody fragment binds to IL31 and one or more antigens selected from IL17, TNFα, CD20, CD19, CD25, IL4, IL13, IL23, IgE, CD11α, IL6R, α4-integrin, IL12, IL1β or BlyS. 25. An isolated nucleic acid encoding an antibody or antibody fragment according to any one of claims 1-24. 26. A host cell for producing an anti-IL31 antibody or antibody fragment, wherein the cell contains the nucleic acid of claim 25. 27. A method for producing an antibody or antibody fragment, comprising culturing a host cell according to claim 26 and isolating the antibody or antibody fragment. 28. A pharmaceutical composition for the treatment of an IL31-mediated condition, wherein the composition comprises an antibody or antibody fragment according to any one of claims 1-24 and a pharmaceutically acceptable carrier. 29. Pharmaceutical composition according to claim 28, where the pharmaceutically acceptable carrier includes L-histidine, sodium chloride and polysorbate 80 and where the pharmaceutical composition has a pH of 5.0 to 6.2. 30. The pharmaceutical composition according to claim 29, wherein the pH of the pharmaceutical composition is 5.0 to 6.0, or 5.3 to 5.7, or equal to 5.5. 31. The pharmaceutical composition according to claim 29 or 30, wherein the pharmaceutical composition contains L-histidine at a concentration of 5 to 100 mM, 10 to 50 mM, 20 to 30 mM, 10 to 30 mM, or 20 mM. 32. A pharmaceutical composition according to any one of claims 29-31, wherein the pharmaceutical composition contains sodium chloride at a concentration of 80 to 200 mM, 100 to 175 mM, 120 to 150 mM, or 140 mM. 33. The pharmaceutical composition according to any one of claims 29-32, where the pharmaceutical composition contains polysorbate 80 at a concentration of 0.005 to 0.5 mg/ml, 0.01 to 0.1 mg/ml or equal to 0.05 mg/ml ml. 34. A pharmaceutical composition according to any one of claims 29-33, wherein the pharmaceutically acceptable carrier comprises at least one sugar. 35. The pharmaceutical composition according to claim 34, wherein the pharmaceutical composition contains sugar at a concentration of 0.5 to 20%, 1 to 10%, 1 to 5%, or 1 to 3%. 36. A pharmaceutical composition according to any one of claims 29-35, wherein the pharmaceutically acceptable carrier comprises sucrose, trehalose, D-mannitol, maltose and/or sorbitol. 37. A pharmaceutical composition according to any one of claims 29 to 36, wherein the pharmaceutical composition contains an antibacterial agent. 38. A pharmaceutical composition according to any one of claims 29-37, wherein the pharmaceutical composition contains m-cresol or methylparaben. 39. A pharmaceutical composition according to any one of claims 29-38, wherein the pharmaceutical composition contains 0.2% m-cresol and/or 0.9% methylparaben. 40. A method for treating an IL31-induced condition in a companion animal species, wherein the method comprises administering to a companion animal species a therapeutically effective amount of an antibody or antibody fragment according to any one of claims 1-24 or a pharmaceutical composition according to any one of claims 28-39. 41. The method of claim 40 wherein the companion animal species is a canine, feline or equine species. 42. The method of claim 40 or 41, wherein the IL31-induced condition is an itching or allergic condition. 43. The method of any one of claims 40-42, wherein the IL31-induced condition is selected from atopic dermatitis, pruritus, asthma, psoriasis, scleroderma, and eczema. 44. The method according to any one of claims 40-43, wherein the antibody or antibody fragment or pharmaceutical composition is administered parenterally. 45. The method according to any one of claims 40 to 44, wherein the antibody or antibody fragment or pharmaceutical composition is administered by the intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intraarterial, intrasynovial, intrathecal, or inhalation routes. 46. The method of any one of claims 40-45, wherein the method comprises administering the antibody or antibody fragment or pharmaceutical composition in combination with a Jak inhibitor, a PI3K inhibitor, an AKT inhibitor, or a MAPK inhibitor. 47. The method of any one of claims 40-46, wherein the method comprises administering the antibody or antibody fragment or pharmaceutical composition in combination with one or more antibodies selected from anti-IL17 antibody, anti-TNFα antibody, anti-CD20 antibody, anti- CD19 antibodies, anti-CD25 antibodies, anti-IL4 antibodies, anti-IL13 antibodies, anti-IL23 antibodies, anti-IgE antibodies, anti-CD11α antibodies, anti-IL6R antibodies, anti-α4-integrin antibodies, anti-IL12 antibodies, anti –IL1β antibodies and anti-BlyS antibodies. 48. A method for reducing the signaling function of IL31 in a cell, which includes exposing the cell to an antibody or antibody fragment according to any one of claims 1–24 or a pharmaceutical composition according to any one of claims 28–39 under conditions under which binding of the antibody or antibody fragment to extracellular IL31, thereby reducing binding to the IL31 receptor and/or reducing IL31 signaling in the cell. 49. The method of claim 48, wherein the cell is exposed to the antibody or antibody fragment or pharmaceutical composition ex vivo . 50. The method of claim 48, wherein the cell is exposed to the antibody or antibody fragment or pharmaceutical composition in vivo . 51. The method of any one of claims 48-50, wherein the cage is a dog cage, cat cage, or horse cage. 52. A method for detecting IL31 in a sample obtained from a companion animal species, comprising contacting the sample with an antibody or antibody fragment according to any one of claims 1-24 or a pharmaceutical composition according to any one of claims 28-39 under conditions that allow binding of the anti-IL31 antibody or antibody fragment, and detecting the formation of a complex of the antibody or antibody fragment and IL31 in the sample. 53. The method of claim 52, wherein the sample is a biological sample obtained from a dog, cat, or horse.

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