Antibody capable of binding to thymic stromal lymphopoietin and use thereof

Abstract

To provide an antibody capable of binding to thymic stromal lymphopoietin and methods for its use.SOLUTION: Disclosed are an anti-TSLP antibody, comprising a murine antibody, chimeric antibody and humanized antibody of the light chain and heavy chain variable regions of the anti-TSLP antibody and antigen-binding fragments thereof, or a pharmaceutically acceptable salt or solvent compound thereof, and the use thereof as a medicament for treating asthma, especially the use thereof in the preparation of a drug for treating TSLP-positive diseases or conditions.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to the field of antibody agents. Specifically, the present disclosure relates to anti-TSLP antibody agents and uses thereof. [Background technology]

[0002] The statements in this specification are merely intended to provide background information related to the present disclosure and do not necessarily constitute prior art.

[0003] Asthma is a serious chronic inflammatory airway disease. There are approximately 334 million asthma patients worldwide, including approximately 30 million in China, with a much higher mortality rate than in developed countries. Environmental degradation and air pollution can increase the number of people affected by the disease, causing significant damage to human life and health.

[0004] Thymic stromal lymphocyte growth factor (TSLP) is an epithelial cell-derived cytokine produced in response to proinflammatory stimuli, and promotes allergic inflammation mainly through its action on dendritic cells and mast cells. TSLP is a type of interleukin-7 (IL-7)-like cytokine, and was first discovered in the conditioned medium of mouse thymic stromal cells. TSLP is mainly expressed in lung, skin, and intestinal epithelial cells. TSLP consists of four α-helices and two loops AB and CD. There are three pairs of disulfide bonds consisting of six cysteines in the molecule, two N-glycosylation sites, and the molecular weight is about 15 to 20 kD. The TSLP receptor is a complex consisting of two parts, one is TSLPR and the other is IL7Rα. TSLP first binds to TSLPR with relatively low affinity, then recruits the binding of IL7Rα with high affinity, and finally activates signal pathways such as stat5, leading to DC maturation and T cell differentiation.

[0005] Myeloid dendritic cells (mDCs) are the main effector cells of TSLP. TSLP acts on immature mDCs, which secrete cytokines IL-8, eotaxin-2, TARC, and MDC, while highly expressing OX40L. In the absence of IL-12, OX40L binds to natural CD4+ T cells, leading to their differentiation into Th2 cells. Th2 cells then secrete Th2 cytokines, such as IL-5, IL-4, IL-9, and TNF, to induce Th2 inflammatory responses in the body. In addition, TSLP induces DC cells to produce the cytokine IL-8, which in turn recruits neutrophils, resulting in neutrophilic innate immune inflammation. TSLP also induces DCs to produce eotaxin-2, which recruits eosinophils and, acting together with IL5, can rapidly transition the body into an inflammatory state of eosinophil infiltration. TSLP also acts on mast cells and natural killer cells, mediating innate inflammation by inducing the production of IL-4, IL-6, IgE, etc. In summary, TSLP simultaneously induces innate inflammation and Th2 inflammation, which increases tissue mucus, alters the airway leading to tracheal stenosis, and aggravates cellular fibrosis. Inflammation gradually develops into the three major allergic diseases, namely, asthma, allergic dermatitis, and allergic rhinitis. Therefore, inhibiting TSLP is potentially an effective strategy for treating diseases such as asthma and allergic dermatitis.

[0006] Currently, anti-TSLP antibodies are disclosed in Patent Documents 1, 2, 3, 4, and 5. However, the corresponding antibodies are not commercially available. Therefore, there is a continuing need to develop pharmaceutical agents that are effective in treating TSLP-related diseases. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2008 / 155365 [Patent Document 2] International Publication No. 2009 / 035577 [Patent Document 3] International Publication No. 2011 / 056772 [Patent Document 4] International Publication No. 2016 / 142426 [Patent Document 5] International Publication No. 2017 / 004149 Summary of the Invention [Problem to be solved by the invention]

[0008] The present disclosure provides anti-TSLP antibodies. [Means for solving the problem]

[0009] In some embodiments, the anti-TSLP antibody as described above comprises an antibody heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:47, respectively, and the light chain variable region comprises LCDR1 and LCDR2 as set forth in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3 as set forth in SEQ ID NO:48 or 55; The sequence of SEQ ID NO:47 is EDYDYDGYAMDX1, the sequence of SEQ ID NO:48 is QQWSSX2RT, and the sequence of SEQ ID NO:55 is QQSDX3X4RX5, where X1 is H or Y, X2 is N or D, X3 is N or S, X4 is V or G, and X5 is G or E; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:76, SEQ ID NO:24 and SEQ ID NO:25, respectively; The sequence of SEQ ID NO:76 is RASESVDX6SGLSFMH, wherein X6 is selected from N, S or Q; or iii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:96 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:118 and SEQ ID NO:31, respectively; The sequence of SEQ ID NO:96 is VIDPGX7X8DTNYNE, and the sequence of SEQ ID NO:118 is X9VX 10 X 11 X 12 X 13 T, X7 is selected from N, Q and V, X8 is G or V, X9 is Y or E, 10 is selected from S, D and E, and X 11 is selected from N, Q, D and E, and X 12 is selected from H, Y, D and E; X 13 is E or Y; or iv) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:32, SEQ ID NO:33 and SEQ ID NO:34, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:35, SEQ ID NO:36 and SEQ ID NO:37, respectively.

[0010] In some embodiments, the anti-TSLP antibody described above comprises a heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:16, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:19, respectively; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:46, respectively; or iii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:53, respectively; or iv) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:54, respectively; or v) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:23, SEQ ID NO:24 and SEQ ID NO:25, respectively; or vi) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:70, SEQ ID NO:24 and SEQ ID NO:25, respectively; or vii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:71, SEQ ID NO:24 and SEQ ID NO:25, respectively; or viii) the heavy chain variable region comprises HCDR1 and HCDR3 as depicted in SEQ ID NO:26 and SEQ ID NO:28, respectively, and an HCDR2 as depicted in SEQ ID NO:27, 93, 94 or 95, and the light chain variable region comprises LCDR1 and LCDR3 as depicted in SEQ ID NO:29 and SEQ ID NO:31, respectively, and an LCDR2 as depicted in SEQ ID NO:30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117.

[0011] In some embodiments, the anti-TSLP antibody described above comprises a heavy chain variable region and a light chain variable region, a) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or b) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:93 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or c) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:94 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or d) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:95 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or e) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:29, SEQ ID NO:108 and SEQ ID NO:31, respectively; or f) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:109 and SEQ ID NO:31, respectively; or g) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:110 and SEQ ID NO:31, respectively; or h) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:29, SEQ ID NO:111 and SEQ ID NO:31, respectively; or i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:112 and SEQ ID NO:31, respectively; or j) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:113 and SEQ ID NO:31, respectively; or k) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:114 and SEQ ID NO:31, respectively; or l) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:115 and SEQ ID NO:31, respectively; or m) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, 116 and 31, respectively; or n) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:29, SEQ ID NO:117 and SEQ ID NO:31, respectively.

[0012] In some embodiments of the anti-TSLP antibody described above, the anti-TSLP antibody is a murine antibody, a chimeric antibody, or a humanized antibody.

[0013] In some embodiments of the anti-TSLP antibodies described above, the anti-TSLP antibodies comprise framework regions derived from a human antibody, or comprise a light chain variable region and / or a heavy chain variable region selected from those set forth in (a), (b), (c), or (d) below: a) the heavy chain variable region comprises HCDR1 and HCDR2 as depicted in SEQ ID NO:14 and SEQ ID NO:15, respectively, and HCDR3 as depicted in SEQ ID NO:16 or 45, the framework regions of which comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 38K, 48I, 67A, 69L, 71V and 73K; and / or the light chain variable region comprises LCDR1 and LCDR2 as depicted in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3 as depicted in EQ ID NO:19, SEQ ID NO:46, SEQ ID NO:53 or SEQ ID NO:54, the framework regions of which comprise up to 10 amino acid back mutations, preferably the back mutations are selected from one or more of 46P, 47W, 58V and 70S; b) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the framework regions comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 2A, 27F, 38K, 39H, 48I, 67A, 69L, 71V and 76R; and / or the light chain variable region comprises LCDR2 and LCDR3 as depicted in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 as depicted in SEQ ID NO:23, SEQ ID NO:70 or SEQ ID NO:71, and the framework regions comprise up to 10 amino acid back mutations, preferably the back mutations are selected from one or more of 1D, 4L, 43P, 48L and 58I; c) the heavy chain variable region comprises HCDR1 and HCDR3 as shown in SEQ ID NO:26 and SEQ ID NO:28, respectively, and HCDR2 as shown in SEQ ID NO:27, SEQ ID NO:93, SEQ ID NO:94 or SEQ ID NO:95, the framework regions of which comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR; and / or the light chain variable region comprises LCDR1 and LCDR3 as shown in SEQ ID NO:29 and SEQ ID NO:31, respectively, and HCDR2 as shown in SEQ ID NO:27, SEQ ID NO:93, SEQ ID NO:94 or SEQ ID NO:95, the framework regions of which comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR; or, comprising an LCDR2 as set forth in SEQ ID NO:30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117, the framework regions of which comprise up to 10 back mutations, preferably the back mutations being selected from one or more of 1S, 43S, 67Y and 73F; or d) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:32, SEQ ID NO:33 and SEQ ID NO:34, respectively, and the framework regions comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 8K, 48I, 66K, 67A, 69L, 71V, 73K and 78A; and / or the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:35, SEQ ID NO:36 and SEQ ID NO:37, respectively, and the framework regions comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 43S, 45Q, 48V, 66V and 70Q.

[0014] In some embodiments, the anti-TSLP antibody described above comprises a heavy chain variable region and a light chain variable region, i) the heavy chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO:6, 42, 43, 44 or 50, and the light chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO:7, 38, 39, 40, 41, 49, 51 or 52; or ii) the heavy chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO:8, 62, 63, 64, 65, 66, 67, 68 or 69, and the light chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO:9, 56, 57, 58, 59, 60, 61, 72, 73, 74 or 75; or iii) the heavy chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 10, 85, 86, 87, 88, 89, 90, 91, 92 or 97 and the light chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO: 11, 77, 78, 79, 80, 81, 82, 83, 84, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119; or, iv) the heavy chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO:12, 126, 127, 128, 129, 130, 131 or 132, and the light chain variable region has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO:13, 120, 121, 122, 123, 124 or 125.

[0015] In some embodiments of the anti-TSLP antibodies described above, the anti-TSLP antibody is a humanized antibody, which comprises a framework region derived from a human antibody or a framework region variant thereof, wherein the framework region variant has up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid back mutations in the light chain framework region and / or the heavy chain framework region of the human antibody, respectively.

[0016] In some embodiments of the anti-TSLP antibodies described above, the framework region variant comprises a back mutation selected from those set forth in (a), (b), (c), or (d) below: a) one or more amino acid backmutations selected from the group consisting of 46P, 47W, 58V, 70S and 71Y in the framework region of the light chain variable region as depicted in SEQ ID NO: 38, 49, 51 or 52, and / or one or more amino acid backmutations selected from the group consisting of 38K, 48I, 67A, 69L, 71V and 73K in the framework region of the heavy chain variable region as depicted in SEQ ID NO: 42 or 50; b) one or more amino acid backmutations selected from the group consisting of 1D, 4L, 43P, 48L and 58I in the framework region of the light chain variable region as depicted in SEQ ID NO: 56, 59, 72, 73, 74 or 75, and / or one or more amino acid backmutations selected from the group consisting of 2A, 27F, 38K, 39H, 48I, 67A, 69L, 71V and 76R in the framework region of the heavy chain variable region as depicted in SEQ ID NO: 62; c) one or more amino acid back-mutations selected from the group consisting of 1S, 43S, 67Y and 73F in the framework region of the light chain variable region as set forth in SEQ ID NO: 77, 81, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119, and / or one or more amino acid back-mutations selected from the group consisting of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR in the framework region of the heavy chain variable region as set forth in SEQ ID NO: 85, 90, 91, 92 or 97; or d) one or more amino acid backmutations selected from the group consisting of 43S, 45Q, 48V, 66V and 70Q in the framework region of the light chain variable region as shown in SEQ ID NO:120, and / or one or more amino acid backmutations selected from the group consisting of 38K, 48I, 66K, 67A, 69L, 71V, 73K and 78A in the framework region of the heavy chain variable region as shown in SEQ ID NO:126.

[0017] In some embodiments, the anti-TSLP antibody described above comprises a heavy chain variable region and a light chain variable region, i) the amino acid sequence of the heavy chain variable region is as set forth in SEQ ID NO:6, 42, 43, 44 or 50 and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO:7, 38, 39, 40, 41, 49, 51 or 52; or ii) the amino acid sequence of the heavy chain variable region is as set forth in SEQ ID NO:8, 62, 63, 64, 65, 66, 67, 68 or 69 and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO:9, 56, 57, 58, 59, 60, 61, 72, 73, 74 or 75; or iii) the amino acid sequence of the heavy chain variable region is as set forth in SEQ ID NO: 10, 85, 86, 87, 88, 89, 90, 91, 92 or 97 and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO: 11, 77, 78, 79, 80, 81, 82, 83, 84, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119; or iv) the amino acid sequence of the heavy chain variable region is as set forth in SEQ ID NO:12, 126, 127, 128, 129, 130, 131 or 132, and the amino acid sequence of the light chain variable region is as set forth in SEQ ID NO:13, 120, 121, 122, 123, 124 or 125.

[0018] In some embodiments, the anti-TSLP antibodies described above comprise the heavy and light chain variable regions shown below: (a) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:6 and the sequence of the light chain variable region is as set forth in SEQ ID NO:7; (b) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:42, 43 or 44 and the sequence of the light chain variable region is as set forth in SEQ ID NO:39, 40 or 41; (c) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:43 and the sequence of the light chain variable region is as set forth in SEQ ID NO:38; (d) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:50 and the sequence of the light chain variable region is as set forth in SEQ ID NO:49, 51 or 52; (e) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:8 and the sequence of the light chain variable region is as set forth in SEQ ID NO:9; (f) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:62, 63, 64 or 65 and the sequence of the light chain variable region is as set forth in SEQ ID NO:56, 57 or 58; (g) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:64, 66, 67, 68 or 69 and the sequence of the light chain variable region is as set forth in SEQ ID NO:59, 60 or 61; (h) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:64 and the sequence of the light chain variable region is as set forth in SEQ ID NO:72 or 73; (i) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:69 and the sequence of the light chain variable region is as set forth in SEQ ID NO:74; (j) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:10 and the sequence of the light chain variable region is as set forth in SEQ ID NO:11; (k) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:85 and the sequence of the light chain variable region is as set forth in SEQ ID NO:77, 78, 102 or 104; (l) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:86 or 88 and the sequence of the light chain variable region is as set forth in SEQ ID NO:77 or 78; (m) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:87 and the sequence of the light chain variable region is as set forth in SEQ ID NO:77, 78, 79, 81, 82, 83, 84, 98, 99, 100, 101, 103, 105, 106 or 107; (n) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:89 and the sequence of the light chain variable region is as set forth in SEQ ID NO:79, 81, 82, 83 or 84; (o) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:90, 91 or 92 and the sequence of the light chain variable region is as set forth in SEQ ID NO:78; (p) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:97 and the sequence of the light chain variable region is as set forth in SEQ ID NO:119; (q) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:12 and the sequence of the light chain variable region is as set forth in SEQ ID NO:13; (r) the sequence of the heavy chain variable region is as set forth in SEQ ID NO: 127, 128, 129, 130, 131 or 132 and the sequence of the light chain variable region is as set forth in SEQ ID NO: 120, 121, 123, 124 or 125; or (s) the sequence of the heavy chain variable region is as set forth in SEQ ID NO:132 and the sequence of the light chain variable region is as set forth in SEQ ID NO:125.

[0019] In some embodiments of the anti-TSLP antibodies described above, the light and heavy chain variable regions of the antibodies are as follows:

[0020] [Table 1]

[0021] [Table 2]

[0022] [Table 3]

[0023] [Table 4]

[0024] In some embodiments of the anti-TSLP antibodies described above, the antibody further comprises an antibody constant region. Preferably, the heavy chain constant region of the antibody constant region is selected from the group consisting of human IgG1, IgG2, IgG3 and IgG4 constant regions and conventional variants thereof, and the light chain constant region of the antibody constant region is selected from the group consisting of human antibody kappa and lambda chain constant regions and conventional variants thereof, and more preferably, the antibody comprises the heavy chain constant region shown in SEQ ID NO:133 and the light chain constant region shown in SEQ ID NO:134.

[0025] In some embodiments, the above-mentioned anti-TSLP antibodies comprise the heavy and light chains shown below: (a) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:135, or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:136, or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto; (b) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:137, or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:138, or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto; (c) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:139, or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:140, or has at least 90% sequence identity thereto; or (d) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:141 or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:142 or has at least 90%, 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0026] In some embodiments, the above-mentioned anti-TSLP antibodies comprise the heavy and light chains shown below: (a) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:135 and the amino acid sequence of the light chain is as set forth in SEQ ID NO:136; (b) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:137 and the amino acid sequence of the light chain is as set forth in SEQ ID NO:138; (c) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:139 and the amino acid sequence of the light chain is as set forth in SEQ ID NO:140; or (d) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:141 and the amino acid sequence of the light chain is as set forth in SEQ ID NO:142.

[0027] In some embodiments, the antibody binds to human TSLP competitively with an anti-TSLP antibody or antigen-binding fragment thereof described above.

[0028] In another embodiment, the disclosure also provides a nucleic acid molecule encoding an anti-TSLP antibody as described above.

[0029] In another aspect, the present disclosure also provides an expression vector comprising a nucleic acid molecule as described above.

[0030] In another aspect, the present disclosure also provides a host cell comprising a nucleic acid molecule or an expression vector as described above, preferably, the cell is a bacterial cell, a fungal cell, an insect cell or a mammalian cell.

[0031] In some embodiments, the disclosure provides methods for preparing TSLP antibodies as described above.

[0032] In some embodiments, the disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of an anti-TSLP antibody as described above, a nucleic acid molecule as described above, or a host cell as described above, and one or more pharma- ceutically acceptable carriers, diluents, buffers, or excipients. Preferably, the therapeutically effective amount is 0.1-3000 mg or 1-1000 mg of an anti-TSLP antibody as described above in a unit dose of the composition.

[0033] In some embodiments, the disclosure provides methods for in vitro or ex vivo immunodetection or determination of TSLP, comprising using an anti-TSLP antibody as described above.

[0034] In some embodiments, the disclosure provides for the use of an anti-TSLP antibody as described above in preparing a reagent for the immunodetection of human TSLP.

[0035] In some embodiments, the disclosure provides an anti-TSLP antibody, as described above, for use in the immunodetection or determination of TSLP.

[0036] In some embodiments, the disclosure provides a kit comprising an anti-TSLP antibody as described above.

[0037] In some embodiments, the disclosure provides for the use of an anti-TSLP antibody as described above, a nucleic acid molecule as described above, a host cell as described above, or a pharmaceutical composition as described above in the preparation of a medicament for treating a TSLP-associated disorder, including asthma, idiopathic pulmonary fibrosis, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic sinusitis, urticaria, Netherton syndrome, eosinophilic esophagitis, food allergies, allergic diarrhea, eosinophilic gastroenteritis, allergic bronchopulmonary aspergillosis, and / or pulmonary aspergillosis. These include, but are not limited to, chronic pruritus, cancer, breast cancer, colon cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, chronic obstructive pulmonary disease, systemic sclerosis, multiple sclerosis, keloid disease, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, hypereosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, fibrosis due to radiation, and fibrosis due to wound healing.

[0038] In some embodiments, the disclosure provides a method for treating a TSLP-associated disease, comprising administering to a subject a therapeutically effective amount of an anti-TSLP antibody as described above, a nucleic acid molecule as described above, a host cell as described above, or a pharmaceutical composition as described above, wherein the TSLP-associated disease includes asthma, idiopathic pulmonary fibrosis, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic sinusitis, urticaria, Netherton syndrome, eosinophilic esophagitis, food allergies, allergic diarrhea, eosinophilic gastroenteritis, allergic bronchitis, and the like. These include, but are not limited to, pulmonary aspergillosis, allergic fungal sinusitis, chronic pruritus, cancer, breast cancer, colon cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, chronic obstructive pulmonary disease, systemic sclerosis, multiple sclerosis, keloid disease, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, hypereosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, fibrosis due to radiation, and fibrosis due to wound healing.

[0039] In some embodiments, the disclosure provides an anti-TSLP antibody for use as a medicament, wherein the anti-TSLP antibody is for use in the treatment of a TSLP-associated disease, including asthma, idiopathic pulmonary fibrosis, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic sinusitis, urticaria, Netherton syndrome, eosinophilic esophagitis, food allergy, allergic diarrhea, eosinophilic gastroenteritis, allergic bronchopulmonary aspergillosis, allergic fungal These include, but are not limited to, sinusitis, chronic pruritus, cancer, breast cancer, colon cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, chronic obstructive pulmonary disease, systemic sclerosis, multiple sclerosis, keloid disease, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, hypereosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, fibrosis due to radiation, and fibrosis due to wound healing. [Brief description of the drawings]

[0040] [Figure 1] FIG. 1 shows the results of antibody inhibition of the binding activity of TSLP to the TSLP receptor. [Diagram 2] FIG. 1 shows the results of antibody inhibition of the binding activity of TSLP to the cell surface TSLP receptor. [Diagram 3] FIG. 1 shows the results of antibody inhibition of TSLP-induced proliferation activity of BaF3 cells. [Figure 4A] FIG. 1 shows the results of antibody activity inhibiting TSLP-induced production of the chemokine TARC. [Figure 4B] FIG. 1 shows the results of antibody activity inhibiting TSLP-induced production of the chemokine OPG. [Figure 5A] FIG. 1 shows the results of antibody activity inhibiting the production of the Th2 cytokine IL-13. [Figure 5B] FIG. 1 shows the results of antibody activity inhibiting the production of Th2 cytokine IL-4. [Figure 5C]FIG. 1 shows the results of antibody activity inhibiting the production of Th2 cytokine TNF-α. [Figure 5D] FIG. 1 shows the results of antibody activity inhibiting the production of Th2 cytokine IL-5. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0041] term

[0042] In order that the present disclosure may be more readily understood, certain technical and scientific terms are specifically defined below. Unless expressly defined herein, all other technical and scientific terms used herein shall have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0043] The three-letter and one-letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, 3558 (1968).

[0044] The term "thymic stromal lymphocyte growth factor (TSLP)" is a type I cytokine with four α-helical bundles, also known as an epithelial cell-derived cytokine, produced in response to proinflammatory stimuli. It is closely related to interleukin-7 (IL-7) and is a key factor in initiating allergic reactions by stimulating dendritic cells (DCs) and regulating the immune response of the human body. The term "TSLP" includes variants, isoforms, homologs, orthologs, and paralogs of TSLP.

[0045] As used herein, "antibody" refers to an immunoglobulin, and generally, an intact antibody is a tetrapeptide chain structure in which two identical heavy chains and two identical light chains are linked by interchain disulfide bonds. The immunoglobulin heavy chain constant regions exhibit different amino acid compositions and sequences, and therefore different antigenicity. Thus, immunoglobulins can be classified into five types or named immunoglobulin isotypes, namely IgM, IgD, IgG, IgA and IgE, with the corresponding heavy chains being μ, δ, γ, α and ε chains, respectively. Igs of the same type can be further divided into different subclasses depending on the difference in amino acid composition of the hinge region and the number and position of the heavy chain disulfide bonds. For example, IgG can be divided into IgG1, IgG2, IgG3 and IgG4. The light chains are divided into κ or λ chains depending on the difference in the constant region. Each of the five types of Igs can have κ or λ chains.

[0046] Approximately 110 amino acid sequences near the N-terminus of the heavy and light chains of an antibody are highly variable and known as the variable region (Fv region), while the remaining amino acid sequences near the C-terminus are relatively stable and are the constant region. The variable region contains three hypervariable regions (HVRs) and four framework regions (FRs) with relatively conservative sequences. The three hypervariable regions determine the specificity of the antibody and are also known as complementarity determining regions (CDRs). The light chain variable region (VL) and heavy chain variable region (VH) are composed of three CDR regions and four FR regions, respectively. The order from amino terminus to carboxy terminus is FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDR regions of the light chain are referred to as LCDR1, LCDR2 and LCDR3, while the three CDR regions of the heavy chain are referred to as HCDR1, HCDR2 and HCDR3.

[0047] The antibodies of the present disclosure include murine antibodies, chimeric antibodies, and humanized antibodies.

[0048] The term "mouse antibody" in this disclosure refers to an anti-human TSLP monoclonal antibody prepared according to the knowledge and techniques of those skilled in the art. During preparation, a subject is injected with TSLP antigen, and then hybridomas expressing antibodies with the desired sequence or functional properties are isolated. In a preferred embodiment of the present disclosure, the mouse anti-TSLP antibody or antigen-binding fragment thereof may further comprise a light chain constant region of a mouse κ chain, λ chain or a mutant thereof, or may further comprise a heavy chain constant region of a mouse IgG1, IgG2, IgG3 or a mutant thereof.

[0049] The term "chimeric antibody" refers to an antibody formed by fusing the variable region of a mouse antibody with the constant region of a human antibody, which can mitigate the immune response induced by mouse antibodies. To establish a chimeric antibody, it is necessary to first establish a hybridoma secreting a mouse-specific monoclonal antibody, then clone the variable region gene from the mouse hybridoma cell, and optionally clone the constant region gene of a human antibody, combine the mouse variable region gene with the human constant region gene to form a chimeric gene that is inserted into an expression vector, and finally express the chimeric antibody molecule in a eukaryotic or prokaryotic system. In a preferred embodiment of the present disclosure, the light chain of the TSLP chimeric antibody further comprises a light chain constant region derived from a human kappa, lambda chain or a mutant thereof. The heavy chain of the TSLP chimeric antibody further comprises a heavy chain constant region of human IgG1, IgG2, IgG3, IgG4 or a variant thereof, preferably a heavy chain constant region of human IgG1, IgG2 or IgG4, or an IgG1, IgG2 or IgG4 variant having an amino acid mutation (e.g., an L234A or L235A mutation, and / or an S228P mutation).

[0050] The term "humanized antibody", also known as CDR-grafted antibody, refers to an antibody produced by grafting mouse CDR sequences into the framework of a human antibody variable region, i.e., an antibody produced with a different type of human germline antibody framework sequence. Since it carries a large amount of mouse protein components, it can overcome the heterogeneous reaction caused by chimeric antibodies. Such framework sequences can be obtained from public DNA databases containing germline antibody gene sequences and published literature. For example, germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database (Internet www.mrccpe.com.ac.uk / vbase) and in Kabat, EA et al., Sequences of Proteins of Immunological Interest, 5th Edition, 1991. To avoid a decrease in activity caused by a concomitant decrease in immunogenicity, minimal back mutations or reverse mutations can be made in the human antibody variable region framework sequence to maintain activity. The humanized antibody of the present disclosure also includes humanized antibodies that are subjected to CDR affinity maturation by yeast display.

[0051] CDR grafting may result in a decrease in the affinity of the produced antibody or antigen-binding fragment thereof for the antigen due to changes in framework residues in contact with the antigen. Such interactions may be the result of somatic hypermutation. Thus, grafting of such donor framework amino acids into the framework of the humanized antibody may still be necessary. By examining the sequence and structure of the animal monoclonal antibody variable region, it is possible to identify the amino acid residues of the non-human antibody or antigen-binding fragment thereof that are involved in antigen binding. Residues of the CDR donor framework that differ from the germline may be considered relevant. If the closest germline cannot be determined, the sequence may be compared to a consensus sequence of subclass or animal antibody sequences with a high percentage of similarity. Rare framework residues are believed to be the result of somatic hypermutation and therefore play an important role in binding.

[0052] In one embodiment of the present disclosure, the antibody or antigen-binding fragment thereof may further comprise a light chain constant region of a human or mouse kappa chain, lambda chain or a mutant thereof, and may further comprise a heavy chain constant region of a human or mouse IgG1, IgG2, IgG3, IgG4 or a mutant thereof, preferably a heavy chain constant region of human IgG1, IgG2, IgG4, or an IgG1, IgG2, IgG4 variant having amino acid mutations (e.g., L234A / L235A mutations, S228P mutations, YTE mutations).

[0053] The "conventional variants" of the human antibody heavy chain constant region and the human antibody light chain constant region described in this disclosure refer to variants of the heavy or light chain constant region disclosed in the prior art that do not change the structure and function of the antibody variable region. Exemplary variants include IgG1, IgG2, IgG3, or IgG4 heavy chain constant region variants with site-specific modifications and amino acid substitutions in the heavy chain constant region. Specific substitutions include YTE mutations, L234A and / or L235A mutations, S228P mutations, and / or mutations to obtain knob-in-hole structures (having knob-Fc and hole-Fc combinations in the antibody heavy chain) that are well known in the art. These mutations have been identified to provide new properties to the antibody without changing the function of the antibody variable region.

[0054] "Human antibody (HuMAb)", "human-derived antibody", "fully human antibody" and "fully human antibody" may be used interchangeably and are antibodies derived from humans or genetically modified organisms that have been "engineered" to produce specific human antibodies in response to antigenic stimulation, and may be produced by any method known in the art. In some techniques, elements of human heavy and light chain loci are introduced into cell lines of organisms derived from embryonic stem cell lines in which the endogenous heavy and light chain loci have been targeted disrupted. The transgenic organisms are capable of synthesizing human antibodies specific to human antigens, and the organisms can be used to generate human antibody-secreting hybridomas. Human antibodies may also be antibodies in which the heavy and light chains are encoded by nucleotide sequences derived from one or more human DNA sources. Fully human antibodies may also be constructed by genetic or chromosomal transfection methods and phage display techniques, or by in vitro activated B cells, all of which are well known in the art.

[0055] The terms "full-length antibody," "intact antibody," "complete antibody," and "whole antibody" are used interchangeably herein and refer to an antibody in a substantially intact form, as distinguished from an antigen-binding fragment, as defined below. These terms specifically refer to antibodies whose light and heavy chains include constant regions. "Antibody" in the present disclosure includes "full-length antibodies" and antigen-binding fragments thereof.

[0056] In some embodiments, the full length antibodies of the present disclosure include antibodies formed by linking a light chain variable region to a light chain constant region and linking a heavy chain variable region to a heavy chain constant region as shown in the light and heavy chain combinations in the following Tables 1 to 4. Those skilled in the art can select light chain constant regions and heavy chain constant regions from different antibodies according to actual needs, for example, light chain constant regions and heavy chain constant regions from a human antibody.

[0057] The term "antigen-binding fragment" or "functional fragment" of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., TSLP). It has been found that fragments of a full-length antibody can be used to perform the antigen-binding function of the antibody. Examples of binding fragments encompassed by the term "antigen-binding fragment" of an antibody include the following: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment containing two Fab fragments linked by a disulfide bond in the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VH and VL domains of a single arm of an antibody; (V) a dsFv, which is a stable antigen-binding fragment formed by an interchain disulfide bond between the VH and VL; and (vi) diabodies, bispecific antibodies and multispecific antibodies, including fragments such as scFv, dsFv, Fab, etc. Furthermore, the two domains of the Fv fragment, VL and VH, are encoded by separate genes, but can be produced as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (called a single chain Fv (scFv)) by recombinantly linking them with a synthetic linker (see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci USA 85:5879-5883). Such single chain antibodies are also included in the term "antigen-binding portion" of an antibody. These antibody fragments are obtained using conventional techniques known to those of skill in the art and screened for utility in the same manner as intact antibodies. Antigen-binding portions may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact immunoglobulins. The antibodies may be of different isotypes, for example IgG (eg, IgG1, IgG2, IgG3 or IgG4 subtypes), IgA1, IgA2, IgD, IgE or IgM antibodies.

[0058] Fab is an antibody fragment with a molecular weight of approximately 50,000, which is obtained by treating an IgG antibody molecule with papain (which cleaves the amino acid residue at position 224 of the H chain). Of these fragments, approximately half of the N-terminal H chain and the entire L chain are linked by disulfide bonds and have antigen-binding activity.

[0059] F(ab')2 is an antibody fragment having a molecular weight of approximately 100,000, which has antigen-binding activity and contains two Fab regions bonded at the hinge position, and which can be obtained by digesting the downstream portions of the two disulfide bonds in the hinge region of IgG with the enzyme pepsin.

[0060] Fab' has a molecular weight of about 50,000 and is an antibody fragment having antigen-binding activity obtained by cleaving the disulfide bond in the hinge region of F(ab')2. The Fab' of the present disclosure can be produced by treating the F(ab')2 of the present invention, which specifically recognizes TSLP and binds to the amino acid sequence of its extracellular domain or its three-dimensional structure, with a reducing agent, such as dithiothreitol.

[0061] Fab' can also be produced by inserting DNA encoding the Fab' fragment of an antibody into a prokaryotic or eukaryotic expression vector and introducing the vector into a prokaryote or eukaryote to express Fab'.

[0062] The term "single chain antibody", "single chain Fv" or "scFv" refers to a molecule comprising an antibody heavy chain variable domain (or region; VH) and an antibody light chain variable domain (or region; VL) connected by a linker. Such scFv molecules have the general structure NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. A suitable linker in the prior art consists of a repeating GGGGS amino acid sequence or a variant thereof, for example a variant having 1 to 4 repeats (Holliger et al., (1993), Proc. Natl. Acad. Sci. USA 90:6444-6448). Other linkers that can be used in the present disclosure include those described in Alfthan et al. (1995), Protein Eng. 8:725-731; Choi et al. (2001), Eur. J. Immunol. 31:94-106; Hu et al. (1996), Cancer Res. 56:3055-3061; Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56; and Roovers et al. (2001), Cancer Immunol.

[0063] A diabody is an antibody fragment in which scFv or Fab are dimerized, and has bivalent antigen-binding activity, in which the two antigens may be the same or different.

[0064] Bispecific and multispecific antibodies refer to antibodies that can bind two or more antigens or antigenic determinants simultaneously, including scFv or Fab fragments that are capable of binding TSLP.

[0065] The diabody of the present disclosure can be produced by the following steps: obtain coding of the VH and VL of a monoclonal antibody of the present disclosure that specifically recognizes human TSLP and binds to the amino acid sequence of the extracellular domain or its three-dimensional structure, construct DNA encoding the scFv, ensure that the amino acid sequence of the peptide linker is 8 residues or less in length, insert the DNA into a prokaryotic or eukaryotic expression vector, and introduce the expression vector into a prokaryote or eukaryote to express the diabody.

[0066] dsFv is obtained by linking a VH polypeptide and a VL polypeptide, and one amino acid residue of each of the VH polypeptide and the VL polypeptide is replaced with a cysteine ​​residue by a disulfide bond between the cysteine ​​residues. The amino acid residue to be replaced with a cysteine ​​residue can be selected according to a known method (Protein Engineering, Vol. 7, p. 697 (1994)) based on the three-dimensional structure prediction of the antibody.

[0067] The full-length antibody or antigen-binding fragment of the present disclosure can be produced by the following steps: obtaining cDNA encoding the VH and VL of a monoclonal antibody of the present disclosure that specifically recognizes human TSLP and binds to the amino acid sequence of its extracellular domain or its three-dimensional structure; constructing DNA encoding the full-length antibody or antigen-binding fragment; inserting the DNA into a prokaryotic or eukaryotic expression vector; and introducing the expression vector into a prokaryote or eukaryote for expression.

[0068] The term "amino acid difference" or "amino acid mutation" refers to the presence of an amino acid change or mutation in a variant protein or polypeptide compared to an original protein or polypeptide, including the occurrence of an insertion, deletion or substitution of one, two, three or more amino acids relative to the original protein or polypeptide.

[0069] The term "antibody framework" or "FR region" refers to that part of a VL or VH variable domain that serves as a scaffold for the antigen binding loops (CDRs) of this variable domain. Essentially, it is a variable domain without the CDRs.

[0070] The term "complementarity determining region", "CDR" or "hypervariable region" refers to the six hypervariable regions in the variable domain of an antibody that mainly contribute to antigen binding. Generally, each heavy chain variable region has three CDRs (HCDR1, HCDR2, HCDR3), and each light chain variable region has three CDRs (LCDR1, LCDR2, LCDR3). Any of a variety of well-known schemes can be used to determine the amino acid sequence boundaries of the CDRs, including the "Kabat" numbering convention (Kabat et al. (1991), "Sequences of Proteins of Immunological Interest", 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD), the "Chothia" numbering convention (Al-Lazikani et al. (1997) JMB 273:927-948), and the ImmunoGenTics (IMGT) numbering convention (Lefranc MP, Immunologist, 7, 132-136 (1999); Lefranc, MP et al., Dev. Comp. Immunol., 27, 55-77 (2003)). For example, in the case of a traditional format, the amino acid residue numbers of the CDRs of the heavy chain variable domain (VH) are 31-35 (HCDR1), 50-65 (HCDR2) and 95-102 (HCDR3) according to the Kabat rules, and the amino acid residue numbers of the CDRs of the light chain variable domain (VL) are 24-4 (LCDR1), 50-56 (LCDR2) and 89-97 (LCDR3). According to the Chothia rules, the amino acid residue numbers of the CDRs of the VH are 26-32 (HCDR1), 52-56 (HCDR2) and 95-102 (HCDR3), and the amino acid residue numbers of the VL are 26-32 (LCDR1), 50-52 (LCDR2) and 91-96 (LCDR3). Combining the Kabat and Chothia CDR definitions, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH, and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL.According to the IMGT rules, the amino acid residue numbers of the CDRs in VH are approximately 26-35 (CDR1), 51-57 (CDR2), and 93-102 (CDR3), and the amino acid residue numbers of the CDRs in VL are approximately 27-32 (CDR1), 50-52 (CDR2), and 89-97 (CDR3). According to the IMGT rules, the CDR regions of an antibody can be determined by the IMGT / DomainGap Align program.

[0071] The term "epitope" or "antigenic determinant" refers to a site on an antigen (e.g., a specific site on a TSLP molecule) to which an immunoglobulin or antibody specifically binds. An epitope typically includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 consecutive or non-consecutive amino acids in a unique spatial conformation. See, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, edited by GE Morris (1996).

[0072] The terms "specific binding," "selective binding," "selectively binds," and "specifically binds" refer to antibody binding to an epitope on a given antigen. Typically, an antibody binds to an epitope on a given antigen, e.g., at about 10 -9 Less than M, about 10 -10 Less than M, about 10 -11 Less than M, about 10 -12 Approximately 10, such as less than M or less -8 It binds with an affinity (KD) of less than M.

[0073] The term "KD" refers to the dissociation equilibrium constant of a particular antibody-antigen interaction. Generally, the antibodies of the present disclosure have a dissociation equilibrium constant of about 10 -7 Less than M, e.g., about 10 -8 Less than M or about 10 -9 In the present disclosure, antibodies bind to TSLP with an affinity (KD) of M, and the KD value is determined by measuring the affinity of the antibody for cell surface antigens by FACS or Biacore methods.

[0074] The term "competition," when used in the context of antigen-binding proteins (e.g., neutralizing antigen-binding proteins or antibodies) competing for the same epitope, means that competition occurs between the antigen-binding proteins, as measured by an assay in which the antigen-binding protein being tested (e.g., an antibody or an immunologically functional fragment thereof) prevents or inhibits (e.g., reduces) the specific binding of a reference antigen-binding protein (e.g., a ligand or reference antibody) to a generic antigen (e.g., the TSLP antigen or a fragment thereof). Numerous types of competitive binding assays are available to determine whether an antigen-binding protein competes with another protein. These assays include, for example, solid-phase direct or indirect radioimmunoassays (RIA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competition assays (see Stahli et al., 1983, Methods in Enzymology 9:242-253), solid-phase direct biotin-avidin EIA (see Kirkland et al., 1986, J. Immunol. 137:3614-3619), solid-phase direct labeling assays, solid-phase direct labeling sandwich assays (see Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press), solid-phase direct labeling RIA using an I-125 label (see Morel et al., 1988, Molec. Immunol. 25:7-15), solid-phase direct biotin-avidin EIA (Cheung et al., 1990, Virology 176:546-552) and direct labeling RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:77-82). Generally, the assay involves binding purified antigen bound to a solid surface or cells using either an unlabeled test antigen binding protein and a labeled reference antigen binding protein. Competitive inhibition is measured by measuring the amount of label bound to the solid surface or cells in the presence of the antigen binding protein being tested. Generally, the antigen binding protein being tested is present in excess.Antigen binding proteins identified by competitive assays (competing antigen binding proteins) include: antigen binding proteins that bind to the same epitope as a reference antigen binding protein, and antigen binding proteins that bind to an adjacent epitope sufficiently close to the binding epitope of the reference antigen binding protein such that these two epitopes sterically interfere with each other's binding. Generally, when a competing antigen binding protein is present in excess, it inhibits (or reduces) specific binding between the reference antigen binding protein and a common antigen by at least 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 65-70%, 70-75% or 75% or more. In some cases, binding is inhibited by at least 80-85%, 85-90%, 90-95%, 95-97% or 97% or more.

[0075] The term "nucleic acid molecule" as used herein refers to DNA molecules and RNA molecules. Nucleic acid molecules may be single-stranded or double-stranded, but are preferably double-stranded DNA, single-stranded mRNA, or modified mRNA. A nucleic acid is "operably linked" when it is placed in a functional relationship with another nucleic acid sequence. For example, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the coding sequence.

[0076] "Identity" of an amino acid sequence refers to the percentage of amino acid residues that are identical between a first sequence and a second sequence when the amino acid sequences are aligned (with gaps introduced if necessary) to maximize sequence identity, and conservative substitutions are not considered part of the sequence identity. For the purpose of determining the percentage of identity of an amino acid sequence, alignment can be achieved using various methods within the skill of the art, for example, publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for measuring alignment, including any algorithms required to achieve maximum alignment over the entire length of the sequences being compared.

[0077] The term "expression vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. In one embodiment, the vector is a "plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. In another embodiment, the vector is a viral vector into which additional DNA segments can be ligated into the viral genome. The vectors disclosed herein can replicate autonomously in a host cell into which they are introduced (e.g., bacterial vectors with a bacterial origin of replication and episomal mammalian vectors, etc.) or can be integrated into the genome of the host cell after introduction into the host cell and replicate along with the host genome (e.g., non-episomal mammalian vectors, etc.).

[0078] Methods for generating and purifying antibodies and antigen-binding fragments are well known in the art and can be found, for example, in Antibody Laboratory Techniques Guide, Cold Spring Harbor Laboratory Press, Chapters 5-8 and 15. For example, mice can be immunized with human TSLP or a fragment thereof, and the resulting antibodies can be refolded, purified, and amino acid sequenced using conventional methods. Antigen-binding fragments can also be prepared using conventional methods. The antibodies or antigen-binding fragments of the present disclosure are engineered to add one or more human FR regions to the non-human CDR regions. Germline sequences of human FRs can be obtained from the ImmunoGeneTics (IMGT) website, http: / / imgt.cines.fr, by comparing the IMGT human antibody variable region germline gene database with MOE software, or from Immunoglobulin Facts Book, 2001 ISBN012441351.

[0079] The term "host cell" refers to a cell into which an expression vector has been introduced. Host cells may include bacterial, microbial, plant or animal cells. Bacteria that are easily transformed include members of the enterobacteriaceae, such as strains of Escherichia coli or Salmonella, the Bacillaceae, such as Bacillus subtilis, Pneumococcus, Streptococcus, and Haemophilus influenzae. Suitable microorganisms include Saccharomyces cerevisiae and Pichia pastoris. Suitable animal host cell lines include CHO (Chinese Hamster Ovary) cells and NSO cells.

[0080] The engineered antibodies or antigen-binding fragments in the present disclosure can be prepared and purified by conventional methods. For example, cDNA sequences encoding the heavy and light chains can be cloned and recombined into a GS expression vector. The recombinant immunoglobulin expression vector can be stably transfected into CHO cells. As a more recommended prior art, mammalian expression systems can result in glycosylation of the antibody, especially at the highly conserved N-terminal part of the Fc region. Stable clones can be obtained by expressing an antibody that specifically binds to human TSLP. Positive clones are grown in serum-free medium in a bioreactor to produce the antibody. The medium into which the antibody is secreted can be purified by conventional techniques. For example, it can be purified using a Sepharose FF column of A or G equilibrated with the conditioning buffer. Non-specifically bound components are washed away, the bound antibody is eluted by a pH gradient method, and the antibody fragments are detected and recovered by SDS-PAGE. The antibody can be filtered and concentrated by conventional methods. Soluble mixtures and polymers can also be removed by conventional methods, such as molecular sieves and ion exchange. The resulting product must be immediately frozen, such as at -70°C, or lyophilized.

[0081] "Administering" or "treating", when applied to an animal, human, experimental subject, cell, tissue, organ, or bodily fluid, refers to contacting an exogenous pharmaceutical, therapeutic, diagnostic, or composition with an animal, human, subject, cell, tissue, organ, or bodily fluid. "Administering" and "treating" can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Treatment of a cell includes contacting a reagent with the cell, and contacting a reagent with a fluid in contact with the cell. "Administering" and "treating" also refer to the treatment of a cell with another cell, for example, with a reagent, diagnostic, binding composition, in vitro and ex vivo. "Treatment", when applied to a human, veterinary, or experimental subject, refers to therapeutic procedures, preventative measures, research, and diagnostic applications.

[0082] "Treating" refers to administering a therapeutic agent, such as a composition comprising any one of the binding compounds of the present disclosure, internally or externally to a patient having one or more disease symptoms for which the therapeutic agent is known to have a therapeutic effect. Typically, the therapeutic agent is administered in an amount effective to alleviate one or more disease symptoms, induce regression of the symptoms, or inhibit the onset of the symptoms to any clinically measurable extent in the patient or population being treated. The amount of a therapeutic agent effective to alleviate a particular disease symptom (also referred to as a "therapeutically effective amount") may vary depending on the condition, age and weight of the patient, and the ability of the agent to produce the desired therapeutic effect in the patient. Whether the symptoms of the disease have been alleviated can be evaluated by any clinical testing method commonly used by physicians or other medical professionals to assess the severity or progression of the symptoms. Embodiments of the present disclosure (e.g., methods of treatment or products) should alleviate the intended target disease symptoms in a statistically significant number of patients as measured by statistical testing methods known in the art (e.g., Student's test, chi-square test, Mann and Whitney U-test, Kruskal-Wallis test (H-test), Jonckheere-Terpstra-test and Wilcoxon-test).

[0083] "Conservative modification" or "conservative exchange or substitution" refers to the replacement of amino acids in a protein with other amino acids having similar properties (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, etc.), and these changes can often be made without altering the biological activity of the protein. Those skilled in the art recognize that single amino acid substitutions in non-essential regions of a polypeptide generally do not substantially alter biological activity (see Watson et al., (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224, 4th ed.). Furthermore, substitution of amino acids with similar structure or function is unlikely to disrupt biological activity. Exemplary conservative substitutions are listed in the table "Exemplary Amino Acid Conservative Substitutions" below.

[0084] [Table 5]

[0085] "Effective amount" refers to the amount of a drug, compound or pharmaceutical composition required to obtain any one or more beneficial or desired therapeutic results. In the case of prophylactic use, beneficial or desired results include eliminating or reducing the risk, reducing the severity or delaying the onset of disease, including the biochemical, histological and / or behavioral symptoms of the disease, their complications and intermediate pathological phenotypes manifested during its development. In the case of therapeutic use, beneficial or desired results include reducing the incidence of various target antigen-related disorders of the present disclosure, or improving one or more symptoms of the disorder, reducing the dose of other drugs required to treat the disorder, improving the therapeutic efficacy of another drug, and clinical results such as slowing the progression of a patient's disorder associated with the target antigen of the present disclosure.

[0086] "Exogenous" refers to a substance that is produced outside an organism, cell, or body, depending on the situation. "Endogenous" refers to a substance that is produced within a cell, organism, or body, depending on the situation.

[0087] "Homology" refers to the similarity of sequences between two polynucleotide sequences or between two polypeptides. If the positions of the two sequences being compared are occupied by the same base or amino acid monomer subunit, for example, if each position of the two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percentage of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared, multiplied by 100. For example, in an optimal sequence alignment, if 6 out of 10 positions of the two sequences are matching or homologous, then the two sequences are 60% homologous, and if 95 out of 100 positions of the two sequences are matching or homologous, then the two sequences are 95% homologous. In general, when two sequences are aligned, the comparison is made to maximize the percentage of homology. For example, the comparison can be performed by the BLAST algorithm, with the parameters of the algorithm being selected to give the maximum match between each sequence over the entire length of each reference sequence. The following references relate to the BLAST algorithms commonly used for sequence analysis: BLAST ALGORITHMS: Altschul, SF et al. (1990) J. Mol. Biol. 215: 403-410; Gish, W. et al. (1993) Nature Genet. 3: 266-272; Madden, TL et al. (1996) Meth. Enzymol. 266: 131-141; Altschul, SF et al. (1997) Nucleic Acids Res. 25: 3389-3402; Zhang, J. et al. (1997) Genome Res. 7: 649-656. Other conventional BLAST algorithms, such as those provided by NCBI BLAST, are also familiar to those skilled in the art.

[0088] As used herein, the terms "cell", "cell line" and "cell culture" can be used interchangeably, and all such designations include progeny. Thus, the terms "transformant" and "transformed cell" include the primary test cell and the culture derived therefrom, regardless of the number of passages. It should also be understood that not all progeny are completely identical in terms of DNA content due to intentional or unintentional mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. When referring to something different, it will be clear from the context.

[0089] As used herein, "polymerase chain reaction" or "PCR" refers to a procedure or technique for amplifying specific portions of minute amounts of nucleic acid, RNA and / or DNA, as described, for example, in U.S. Pat. No. 4,683,195. Generally, sequence information must be obtained from the ends or outside of the target region so that oligonucleotide primers can be designed, which are identical or similar in sequence to the corresponding strands of the template to be amplified. The 5' terminal nucleotides of the two primers can be identical to the ends of the material to be amplified. PCR can be used to amplify specific RNA sequences, specific DNA sequences from total genomic DNA, and cDNA sequences transcribed from total cellular RNA, phage or plasmid sequences, etc. See Mullis et al. (1987) Cold Spring Harbor, Symp. Ouant. Biol. 51:263; Erlich ed., (1989) PCR TECHNOLOGY (Stockton Press, NY). As used herein, PCR is considered to be one example, but not the only example, of a nucleic acid polymerase reaction method for amplifying a nucleic acid test sample, which involves using known nucleic acids as primers and a nucleic acid polymerase to amplify or generate specific portions of a nucleic acid.

[0090] "Isolated" refers to a purified state, in which case it is meant that the specified molecule is substantially free of other biological molecules, such as nucleic acids, proteins, lipids, carbohydrates, or other substances, such as cell debris and growth medium. In general, the term "isolated" is not intended to mean complete absence of these substances, or absence of water, buffers, or salts, unless present in amounts that would significantly interfere with the experimental or therapeutic use of the compounds described herein.

[0091] "Optional" or "optionally" means that the described event or circumstance may occur, but does not necessarily occur. The description includes cases where the event or circumstance occurs and cases where it does not occur.

[0092] "Pharmaceutical composition" refers to a mixture containing one or more compounds described herein or their physiologically / pharmaceutical acceptable salts or prodrugs, and other chemical components, such as physiologically / pharmaceutical acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby facilitating absorption of the active ingredient and exerting its biological activity.

[0093] The term "pharmaceutical acceptable carrier" refers to any inert substance suitable for use in a formulation for delivery of an antibody or antigen-binding fragment. Carriers can be anti-adhesive agents, binders, coating agents, disintegrants, fillers or diluents, preservatives (such as antioxidants, antibacterial agents or antifungal agents), sweeteners, absorption retardants, wetting agents, emulsifiers, buffers, etc. Examples of suitable pharmaceutical acceptable carriers include water, ethanol, polyols (such as glycerol, propanediol, polyethylene glycol, etc.), dextrose, vegetable oils (such as olive oil), saline, buffers, and isotonicity agents, such as sugars, polyols, sorbitol, and sodium chloride.

[0094] Furthermore, the present disclosure includes a pharmaceutical agent for treating a TSLP-associated disease, comprising an anti-TSLP antibody or antigen-binding fragment thereof of the present disclosure as an active ingredient.

[0095] The TSLP-related disease in the present disclosure is not particularly limited as long as it is a disease related to TSLP.For example, the therapeutic response induced by the molecule of the present disclosure can be achieved by binding to human TSLP, then blocking the binding of TSLP to its receptor, or killing the cells that overexpress TSLP.

[0096] Furthermore, the present disclosure relates to methods for immunodetecting or measuring a target antigen (e.g., TSLP), reagents for immunodetecting or measuring a target antigen (e.g., TSLP), methods for immunodetecting or measuring cells expressing a target antigen (e.g., TSLP), and diagnostic agents for diagnosing diseases associated with target antigen (e.g., TSLP)-positive cells, which contain the antibody or antibody fragment of the present disclosure as an active ingredient, specifically recognize the target antigen (e.g., human TSLP), and bind to the amino acid sequence of the extracellular domain or its three-dimensional structure.

[0097] In the present disclosure, the method used to detect or measure the amount of a target antigen (eg, TSLP) can be any known method, including, for example, immunodetection or measurement methods.

[0098] Immunodetection or measurement methods are methods that use labeled antigens or antibodies to detect or measure the amount of antibodies or antigens. Examples of immunodetection or measurement methods include radioimmunoassay (RIA), enzyme immunoassay (EIA or ELISA), fluorescent immunoassay (FIA), luminescence immunoassay, Western blotting, physicochemical methods, etc.

[0099] The above-mentioned TSLP-related diseases can be diagnosed by detecting or measuring cells that express TSLP using the antibodies or antibody fragments of the present disclosure.

[0100] To detect cells expressing a polypeptide, known immunodetection methods can be used, preferably immunoprecipitation, fluorescent cell staining, immunohistochemical staining, etc. Also, fluorescent antibody staining using the FMAT8100HTS system (Applied Biosystem) can be used.

[0101] In the present disclosure, the in vivo sample used to detect or measure a target antigen (e.g., TSLP) is not particularly limited as long as it is likely to contain cells expressing the target antigen (e.g., TSLP), and examples that can be used include histiocytes, blood, plasma, serum, pancreatic juice, urine, feces, tissue fluid, or culture fluid.

[0102] According to the required diagnostic method, the diagnostic agent containing the monoclonal antibody or its antibody fragment of the present disclosure may also contain a reagent for carrying out an antigen-antibody reaction or a reagent for detecting the reaction. The reagent used for carrying out the antigen-antibody reaction includes buffers, salts, etc. The reagent used for detection includes reagents commonly used in immunodetection or measurement methods, such as a labeled secondary antibody that recognizes the monoclonal antibody, its antibody fragment, or its conjugate, and a substrate corresponding to the label.

[0103] In the above specification, the details of one or more embodiments of the present disclosure are presented. Although any methods and materials similar or identical to those described herein can be used to carry out or test the present invention, the preferred methods and materials are described below. Other features, objects and advantages of the present disclosure will become clear from the specification and claims. In the specification and claims, the singular form includes plural referents unless the context clearly dictates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the common meanings understood by those skilled in the art to which the present invention belongs. All patents and publications cited in this specification are incorporated by reference. The following examples are presented to more comprehensively describe the preferred embodiments of the present invention. These examples should not be interpreted in any way as limiting the scope of the present invention, which is defined by the claims. The present invention includes, for example, the following embodiments: [Embodiment 1] An anti-TSLP antibody comprising a heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:47, respectively, and the light chain variable region comprises LCDR1 and LCDR2 as set forth in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3 as set forth in SEQ ID NO:48 or 55; The sequence of SEQ ID NO: 47 is EDYDYDGYAMDX1, the sequence of SEQ ID NO: 48 is QQWSSX2RT, and the sequence of SEQ ID NO: 55 is QQSDX3X4RX5; X1 is H or Y, X2 is N or D, X3 is N or S, X4 is V or G, and X5 is G or E; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:76, SEQ ID NO:24 and SEQ ID NO:25, respectively; The sequence of SEQ ID NO:76 is RASESVDX6SGLSFMH, wherein X6 is selected from N, S and Q; or iii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:96 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:118 and SEQ ID NO:31, respectively; The sequence of SEQ ID NO:96 is VIDPGX7X8DTNYNE and the sequence of SEQ ID NO:118 is X9VX 10 X 11 X 12 X 13 T, X7 is selected from N, Q and V, X8 is G or V, X9 is Y or E, 10 is selected from S, D and E, and X 11 is selected from N, Q, D and E, and X 12 is selected from H, Y, D and E, and X 13 is E or Y; or iv) An anti-TSLP antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:32, SEQ ID NO:33 and SEQ ID NO:34, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:35, SEQ ID NO:36 and SEQ ID NO:37, respectively. [Embodiment 2] A heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1 and HCDR2 as set forth in SEQ ID NO:14 and SEQ ID NO:15, respectively, and an HCDR3 as set forth in SEQ ID NO:16 or 45, and the light chain variable region comprises LCDR1 and LCDR2 as set forth in SEQ ID NO:17 and SEQ ID NO:18, respectively, and an LCDR3 as set forth in SEQ ID NO:19, 46, 53 or 54; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR2 and LCDR3 as set forth in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 as set forth in SEQ ID NO:23, 70 or 71; or iii) the heavy chain variable region comprises HCDR1 and HCDR3 as set forth in SEQ ID NO:26 and SEQ ID NO:28, respectively, and HCDR2 as set forth in SEQ ID NO:27, 93, 94 or 95, and the light chain variable region comprises LCDR1 and LCDR3 as set forth in SEQ ID NO:29 and SEQ ID NO:31, respectively, and LCDR2 as set forth in SEQ ID NO:30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117; The anti-TSLP antibody of embodiment 1. [Embodiment 3] A heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:54, respectively; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:53, respectively; or iii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:70, SEQ ID NO:24 and SEQ ID NO:25, respectively; or iv) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:71, SEQ ID NO:24 and SEQ ID NO:25, respectively; or v) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:94 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:113 and SEQ ID NO:31, respectively; or vi) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:94 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or vii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:16, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:19, respectively; or viii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:46, respectively; or ix) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:23, SEQ ID NO:24 and SEQ ID NO:25, respectively; or x) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or xi) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:93 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or xii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:95 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or xiii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1 and LCDR3 as set forth in SEQ ID NO:29 and SEQ ID NO:31, respectively, and LCDR2 as set forth in SEQ ID NO:108, 109, 110, 111, 112, 113, 114, 115, 116 or 117; The anti-TSLP antibody of embodiment 2. [Embodiment 4] The anti-TSLP antibody according to any one of embodiments 1 to 3, wherein the anti-TSLP antibody is a mouse antibody, a chimeric antibody, or a humanized antibody. [Embodiment 5] The anti-TSLP antibody of embodiment 4, wherein the anti-TSLP antibody comprises a framework region derived from a human antibody, and the anti-TSLP antibody comprises a light chain variable region and / or a heavy chain variable region selected from those described in (a), (b), (c), or (d) below. a) said heavy chain variable region comprises HCDR1 and HCDR2 as depicted in SEQ ID NO:14 and SEQ ID NO:15, respectively, and HCDR3 as depicted in SEQ ID NO:16 or 45, the framework regions of which comprise up to 10 back mutations, preferably selected from one or more of 38K, 48I, 67A, 69L, 71V and 73K, and / or said light chain variable region comprises LCDR1 and LCDR2 as depicted in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3 as depicted in SEQ ID NO:19, 46, 53 or 54, the framework regions of which comprise up to 10 amino acid back mutations, preferably selected from one or more of 46P, 47W, 58V, 70S and 71Y; b) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, whose framework regions comprise up to 10 back mutations, preferably selected from one or more of 2A, 27F, 38K, 39H, 48I, 67A, 69L, 71V and 76R, and / or the light chain variable region comprises LCDR2 and LCDR3 as depicted in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 as depicted in SEQ ID NO:23, 70 or 71, whose framework regions comprise up to 10 amino acid back mutations, preferably selected from one or more of 1D, 4L, 43P, 48L and 58I; c) the heavy chain variable region comprises HCDR1 and HCDR3 as shown in SEQ ID NO:26 and SEQ ID NO:28, respectively, and HCDR2 as shown in SEQ ID NO:27, 93, 94 or 95, the framework regions of which comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR, and / or the light chain variable region comprises LCDR1 and LCDR3 as shown in SEQ ID NO:29 and SEQ ID NO:31, respectively, and HCDR2 as shown in SEQ ID NO:27, 93, 94 or 95, the framework regions of which comprise up to 10 back mutations, preferably the back mutations are selected from one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR, or, comprising an LCDR2 as set forth in SEQ ID NO:30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117, the framework regions of which comprise up to 10 backmutations, preferably the backmutations being selected from one or more of 1S, 43S, 67Y and 73F; or d) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:32, SEQ ID NO:33 and SEQ ID NO:34, respectively, and the framework regions comprise up to 10 back mutations, preferably selected from one or more of 38K, 48I, 66K, 67A, 69L, 71V, 73K and 78A, and / or the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO:35, SEQ ID NO:36 and SEQ ID NO:37, respectively, and the framework regions comprise up to 10 back mutations, preferably selected from one or more of 43S, 45Q, 48V, 66V and 70Q. [Embodiment 6] The present invention comprises a heavy chain variable region and a light chain variable region, i) the heavy chain variable region has at least 90% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO:6, 42, 43, 44 or 50, and the light chain variable region has at least 90% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO:7, 38, 39, 40, 41, 49, 51 or 52; or ii) the heavy chain variable region has at least 90% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO:8, 62, 63, 64, 65, 66, 67, 68 or 69, and the light chain variable region has at least 90% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO:9, 56, 57, 58, 59, 60, 61, 72, 73, 74 or 75; or iii) the heavy chain variable region has at least 90% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 10, 85, 86, 87, 88, 89, 90, 91, 92 or 97, and the light chain variable region has at least 90% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO: 11, 77, 78, 79, 80, 81, 82, 83, 84, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119; or iv) the heavy chain variable region has at least 90% sequence identity to the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 12, 126, 127, 128, 129, 130, 131 or 132, and the light chain variable region has at least 90% sequence identity to the light chain variable region set forth in the amino acid sequence SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125; The anti-TSLP antibody of embodiment 4. [Embodiment 7] The present invention comprises a heavy chain variable region and a light chain variable region, i) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:6, 42, 43, 44 or 50, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO:7, 38, 39, 40, 41, 49, 51 or 52; or ii) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO:8, 62, 63, 64, 65, 66, 67, 68 or 69 and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO:9, 56, 57, 58, 59, 60, 61, 72, 73, 74 or 75; or iii) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 10, 85, 86, 87, 88, 89, 90, 91, 92 or 97 and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 11, 77, 78, 79, 80, 81, 82, 83, 84, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119; or iv) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 12, 126, 127, 128, 129, 130, 131 or 132, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125; The anti-TSLP antibody of embodiment 6. [Embodiment 8] The anti-TSLP antibody of embodiment 7, comprising the heavy chain variable region and light chain variable region shown below. a) the sequence of the heavy chain variable region is set forth in SEQ ID NO:50 and the sequence of the light chain variable region is set forth in SEQ ID NO:52; or b) the sequence of the heavy chain variable region is set forth in SEQ ID NO:50 and the sequence of the light chain variable region is set forth in SEQ ID NO:51; or c) the sequence of the heavy chain variable region is set forth in SEQ ID NO:69 and the sequence of the light chain variable region is set forth in SEQ ID NO:74; or d) the sequence of the heavy chain variable region is set forth in SEQ ID NO:64 and the sequence of the light chain variable region is set forth in SEQ ID NO:73; or e) the sequence of the heavy chain variable region is set forth in SEQ ID NO:97 and the sequence of the light chain variable region is set forth in SEQ ID NO:119; or f) the sequence of the heavy chain variable region is set forth in SEQ ID NO:91 and the sequence of the light chain variable region is set forth in SEQ ID NO:78; or g) the sequence of the heavy chain variable region is shown in SEQ ID NO:132 and the sequence of the light chain variable region is shown in SEQ ID NO:125. [Embodiment 9] The anti-TSLP antibody of any one of embodiments 1 to 8, wherein the antibody further comprises an antibody heavy chain constant region and a light chain constant region, preferably, the heavy chain constant region is selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 constant regions and conventional mutants thereof, and the light chain constant region is selected from the group consisting of human antibody κ chain and λ chain constant regions and conventional mutants thereof, more preferably, the antibody comprises the heavy chain constant region shown in SEQ ID NO: 133 and the light chain constant region shown in SEQ ID NO: 134. [Embodiment 10] The anti-TSLP antibody of embodiment 9, wherein the antibody comprises the heavy and light chains shown below. a) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:135 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:136 or has at least 90% sequence identity thereto; or b) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:137 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:138 or has at least 90% sequence identity thereto; or c) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:139 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:140 or has at least 90% sequence identity thereto; or d) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO:141 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO:142 or has at least 90% sequence identity thereto. [Embodiment 11] An isolated anti-TSLP antibody that binds to human TSLP competitively with the anti-TSLP antibody of any one of Embodiments 1 to 10. [Embodiment 12] A nucleic acid molecule encoding the anti-TSLP antibody of any one of Embodiments 1 to 11. [Embodiment 13] A host cell comprising the nucleic acid molecule described in embodiment 12. [Embodiment 14] A pharmaceutical composition comprising a therapeutically effective amount of an anti-TSLP antibody described in any one of embodiments 1 to 11, a nucleic acid molecule described in embodiment 12, or a host cell described in embodiment 13, and one or more pharma- ceutically acceptable carriers, diluents, buffers, or excipients. [Embodiment 15] A method for immunodetecting or determining TSLP in vitro or ex vivo, comprising a step of using an anti-TSLP antibody described in any one of embodiments 1 to 11. [Embodiment 16] A kit comprising an anti-TSLP antibody described in any one of embodiments 1 to 11. [Embodiment 17] A method for treating a TSLP-related disease, comprising administering to a subject a therapeutically effective amount of the anti-TSLP antibody of any one of Embodiments 1 to 11, the nucleic acid molecule of Embodiment 12, the host cell of Embodiment 13, or the pharmaceutical composition of Embodiment 14. Preferably, the TSLP-related disease is asthma, idiopathic pulmonary fibrosis, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic sinusitis, urticaria, Netherton syndrome, eosinophilic esophagitis, food allergy, allergic diarrhea, eosinophilic gastroenteritis, allergic bronchopulmonary aspergillosis, allergic fungal sinusitis, chronic pruritus, cancer, breast cancer, colon cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, or chronic obstructive pulmonary disease. The method is selected from the group consisting of systemic sclerosis, multiple sclerosis, keloidosis, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, hypereosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, inflammatory bowel disease, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, fibrosis due to radiation, and fibrosis due to wound healing.

[0104] Working Example

[0105] The following examples are included to further illustrate the present disclosure, but these examples are not intended to limit the scope of the disclosure.

[0106] Experimental methods for which conditions are not specified in the examples or test examples of this disclosure generally follow conventional conditions or conditions recommended by the manufacturers of raw materials or commercial products. See Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor; Current Protocols Molecular Biology, Ausubel et al., Greene Publishing Associates, Wiley Interscience, NY. Reagents for which manufacturers are not specified are commercially available conventional reagents.

[0107] Example 1. Expression of TSLP and TSLP receptors

[0108] Sequences encoding His-tagged human TSLP and cyno TSLP, human IgG1-Fc-tagged human TSLP and cyno TSLP, and the TSLP receptor extracellular domain sequence were loaded into the phr vector to construct expression plasmids, which were then transfected into HEK293. The specific transfection process was as follows: The day before, HEK293E cells were transfected in Freestyle expression medium (containing 1% FBS) at 0.8 × 10 6The cells were seeded at 100μg / ml, placed on a 37℃ thermostatic shaker (120 rpm), and cultured for 24 hours. After 24 hours, the transfection plasmid and transfection reagent PEI were sterilized with a 0.22μm filter. Then, the transfection plasmid was adjusted to 100μg / 100ml cells, with a mass ratio of PEI (1mg / ml) to plasmid of 3:1. Taking the transfection of 200ml of HEK293E cells as an example, 10ml of Opti-MEM and 200μg of plasmid were taken, mixed well, and left to stand for 5 minutes; another 10ml of Opti-MEM and 600μg of PEI were taken, mixed well, and left to stand for 5 minutes. The plasmid and PEI were thoroughly mixed and left to stand for 15 minutes. It is better not to exceed 20 minutes. The mixture of plasmid and PEI was slowly added to 200 ml of HEK293E cells and placed on a shaker at 8% CO2, 120 rpm, 37°C for cultivation. On the third day of transfection, the culture was supplemented with 10% of the medium. Until the sixth day of transfection, samples were taken and centrifuged at 4500 rpm for 10 minutes to recover the cell supernatant. The supernatant was filtered and purified to obtain recombinant TSLP and TSLP receptor proteins according to Example 2. The purified proteins were used in the experiments in each of the following examples. The relevant sequences are as follows:

[0109] 1. Amino acid sequence of his-tagged human TSLP antigen (huTSLP-his) [ka] Note: The underlined part is the signal peptide sequence, and the shaded part is the Flag-His6-tag. SEQ ID NO:1

[0110] 2. Amino acid sequence of Fc-tagged human TSLP antigen (huTSLP-Fc) [ka] Note: The underlined portion is the signal peptide sequence, and the shaded portion is the linker human Fc tag. SEQ ID NO:2

[0111] 3. Amino acid sequence of his-tagged cynoTSLP antigen (cynoTSLP-his) [ka] Note: The underlined portion is the signal peptide sequence, the shaded portion is the Flag-His6-tag. SEQ ID NO:3

[0112] 4. Amino acid sequence of Fc-tagged cynoTSLP antigen (cynoTSLP-Fc) [ka] Note: The underlined portion is the signal peptide sequence, and the shaded portion is the linker human Fc tag. SEQ ID NO:4

[0113] 5. Amino acid sequence of Fc-tagged human TSLP receptor extracellular domain (human-TSLPR-Fc-ECD) [ka] Note: The underlined part is the human TSLPR extracellular domain, the shaded part is the linker-human Fc-tag. SEQ ID NO:5

[0114] Example 2. Purification of TSLP and TSLP Receptor (TSLPR) Recombinant Proteins

[0115] 2.1 Purification of various His-tagged TSLP recombinant proteins

[0116] The cell expression supernatant sample was centrifuged at high speed to remove impurities and filtered. The nickel column was equilibrated with PBS solution and washed with 10 column volumes. The filtered supernatant sample was applied to the column. 280The column was washed with PBS solution containing 30 mM imidazole until the readings dropped to baseline. The target protein was then eluted with PBS solution containing 300 mM imidazole and the elution peak was collected. The protein was concentrated and exchanged into PBS, and after being identified as correct by LC-MS, it was aliquoted for use. His-tagged human TSLP and cyno TSLP were obtained.

[0117] 2.2 Purification of various human Fc-tagged TSLP and human TSLP receptor extracellular domain recombinant proteins

[0118] The cell expression supernatant samples were centrifuged at high speed to remove impurities. The recombinant antibody expression supernatant was purified on a Protein A column. The column was washed with PBS until the A280 readings dropped to baseline. The target protein was eluted with 100 mM acetate buffer pH 3.5 and neutralized with 1 M Tris-HCl pH 8.0. The resulting protein was concentrated, exchanged into fresh solution, and identified as correct by electrophoresis and LC-MS, after which it was aliquoted for use.

[0119] Example 3. Construction and identification of recombinant TSLP receptor and IL7Rα receptor cell lines

[0120] To screen for antibodies that can block TSLP from binding to the TSLP receptor, we constructed CHO-K1 and BaF3 cell lines that simultaneously express both human TSLP receptor and human IL7Rα (TSLPR / IL7Rα). The target gene TSLPR / IL7Rα was packaged using lentivirus and cloned into the target cell line to form a stable high-expressing cell line. First, human TSLPR and human IL7Rα genes were cloned into plasmids pCDH-CMV-MCS-EF1-puro and pCDH-CMV-MCS-EF1-Neo (SBI, CD500B-1), respectively. Then, using the lentivirus infection method, human TSLPR was inserted into CHO-K1 and BaF3 cell lines and cultured under the selection pressure of 10 μg / ml puromycin (Gibco, US) for 3 weeks. A second round of infection was then performed. The human IL7Rα gene was inserted into the cell lines and screened with 1mg / ml G418 (Gibco,US) and 10μg / ml puromycin for 2-3 weeks.Finally, CHO-K1 and BaF3 monoclonal cell lines that simultaneously expressed high levels of TSLPR and IL7Rα were screened by flow sorting.

[0121] Example 4. Preparation and screening of anti-human TSLP antibodies

[0122] Anti-human TSLP monoclonal antibodies were generated by immunizing 6- to 8-week-old laboratory SJL white mice (Beijing Charles River Laboratory Animal Technology Co.,Ltd. Animal Production Permit Number: SCXK(Beijing)2012-0001). Feeding environment: SPF level. Mice were kept in a laboratory environment with a 12 / 12-h light / dark cycle, a temperature of 20-25°C, and a humidity of 40-60% for 1 week after purchase. Mice adapted to the environment were immunized with recombinant proteins huTSLP-Fc (25 μg), huTSLP-his (12.5 μg), cyno TSLP-his (12.5 μg), and TiterMax, alum, or CpG adjuvant. After 4-5 immunizations, mice with high antibody titers in the serum and a tendency for titers to reach a plateau were selected and sacrificed. Spleen cells were collected and fused with myeloma cells. Splenic lymphocytes were fused with myeloma cells Sp2 / 0 cells (ATCC® CRL-8287™) to obtain hybridoma cells using an optimized PEG-mediated fusion step.

[0123] In the first screening, we performed ELISA binding assays of human and cyno TSLP, assays to inhibit the binding of human TSLP to its receptor TSLPR, and experiments to inhibit the proliferation of BaF3 cells induced by TSLP. After transferring hybridoma cells to 24-well plates, the supernatants were rescreened. Hybridoma clones were obtained by subcloning the selected positive clones twice and used for antibody production, and purification was performed by affinity methods.

[0124] After screening, we obtained monoclonal hybridoma cell lines NO:3, NO:119, NO:179 and NO:199 with good activity, and harvested hybridoma cells in logarithmic growth phase. RNA was extracted with NucleoZol (MN) and reverse transcription was performed (PrimeScript™ Reverse Transcriptase, Takara, Cat. No. 2680A). cDNA obtained by reverse transcription was amplified by PCR using Mouse Ig-Primer Set (Novagen, TB326 Rev.B 0503) and sent to a sequencing company for sequencing. Mouse anti-TSLP antibodies were obtained after sequencing: mab3, mab119, mab179 and mab199 sequences, the amino acid sequences of their variable regions are as follows: > mab3 mouse heavy chain variable region sequence: [ka] SEQ ID NO:6 >mab3 mouse light chain variable region sequence: [ka] SEQ ID NO:7 >mab119 mouse heavy chain variable region sequence: [ka] SEQ ID NO:8 >mab119 mouse light chain variable region sequence: [ka] SEQ ID NO:9 >mab179 mouse heavy chain variable region sequence: [ka] SEQ ID NO:10 >mab179 mouse light chain variable region sequence: [ka] SEQ ID NO:11 >mab199 mouse heavy chain variable region sequence: [ka] SEQ ID NO:12 >mab199 mouse light chain variable region sequence: [ka] SEQ ID NO:13

[0125] The amino acid sequences of the CDR regions obtained according to the Kabat numbering convention are shown in the following table:

[0126] [Table 6]

[0127] Chimeric antibodies were formed by linking the light and heavy chain variable regions of the above-mentioned mouse antibodies with the light and heavy chain constant regions of human antibodies (such as the kappa constant region shown in SEQ ID NO:134, the IgG1-YTE constant region shown in SEQ ID NO:133, etc.). The chimeric antibody corresponding to clone mab3 was named Ch3, and similarly for the other antibodies.

[0128] Example 5. Humanized design of anti-human TSLP monoclonal antibody

[0129] To reduce the immunogenicity of mouse antibodies, the screened mab3, mab119, mab179 and mab199 antibodies with excellent in vivo and in vitro activity were humanized. Humanization of mouse monoclonal antibodies was performed according to the methods published in many documents in the art. Briefly, human antibody constant domains were used to replace the parent (mouse antibody) constant domains, human germline antibody sequences were selected according to the homology between mouse and human antibodies, and CDR grafting was performed. Then, based on the three-dimensional structure of the mouse antibody, amino acid residues in VL and VH were back-mutated, and the constant regions of the mouse antibody were replaced with human constant regions to obtain the final humanized molecule.

[0130] 5.1 Selection and backmutation of the human FR region of mab3

[0131] (1) Selection and backmutation of human FR regions

[0132] For mab3, the humanized VH template is IGHV1-3 * 01+IGHJ6 * 01, and the humanized VL template is IGKV3-20+IGKJ4 * The CDRs of mab3 were grafted into the human template, and the resulting variable region sequences after grafting were as follows: hu3 VL-CDR after transplant: [ka] SEQ ID NO:38 hu3 VH-CDR after transplant: [ka] SEQ ID NO:42

[0133] The backmutation designs of the mab3 humanized antibody are shown in the table below:

[0134] [Table 7] Note: Grafted refers to the grafting of mouse antibody CDRs to human germline FR region sequences. L46P refers to the mutation of L at position 46 back to P according to the Kabat numbering convention.

[0135] The sequences of the variable regions of the mab3 humanized antibody are as follows: > hu3VL1 (hu3 VL-CDR port) [ka] SEQ ID NO:38 [ka] [ka] NOTE: Single underlines represent CDR regions and double underlines represent backmutation sites.

[0136] The above light and heavy chain variable regions were combined with human germline light and heavy chain constant region sequences to finally form complete light and heavy chain sequences, resulting in an antibody with a full-length sequence. Illustratively, for the mab3 humanized antibody in the present disclosure, the heavy chain constant region is the IgG1-YTE constant region shown in SEQ ID NO:133, and the light chain constant region is the kappa chain constant region shown in SEQ ID NO:134, but they can also be replaced with other constant regions known in the art.

[0137] The sequences of the heavy and light chain variable regions of the resulting mab3 humanized antibody are shown in the table below.

[0138] [Table 8]

[0139] The binding activity of the mab3 humanized antibody to human TSLP was detected by ELISA, and it was confirmed that the mab3 humanized antibody has an extremely high binding ability to human TSLP.

[0140] (2) Point mutations in the hu3 antibody

[0141] The detection revealed that there were hot spots in the MDH sequence of HCDR3 and the NTR sequence of LCDR3 of the mab3 humanized antibody. Therefore, the corresponding hot spots were mutated. The sequences of the CDR regions of the mab3 humanized antibody obtained after mutation are as follows:

[0142] [Table 9] NOTE: The positions of the mutation sites in Table 9 are numbered according to the natural order of the variable region sequence.

[0143] It can be concluded that the CDR sequences of the mab3 humanized antibody are as follows:

[0144] [Table 10] where X1 is selected from H or Y, and X2 is selected from N or D.

[0145] Exemplarily, the CDRs, heavy and light chain variable regions of the humanized antibody hu3-11 obtained after mutation are as follows:

[0146] [Table 11]

[0147] > hu3-11 light chain variable region (hu3VL4-N93D) [ka] SEQ ID NO:49 > hu3-11 heavy chain variable region (hu3VH2-H110Y) [ka] SEQ ID NO:50

[0148] The hotspot mutated light and heavy chain variable regions were recombined with human germline light and heavy chain constant region sequences to form complete light and heavy chain sequences, resulting in an antibody with full-length sequence.

[0149] The binding activity of the antibody obtained after the mutation to human TSLP was detected using ELISA. The results showed that the affinity activity of hu3-11 to human TSLP was still high, and the hotspot mutations on HCDR3 and LCDR3 of mab3 humanized antibody did not affect the activity of the antibody.

[0150] (3) Affinity maturation of hu3-11 antibody

[0151] The hu3-11 molecule was affinity matured. The affinity maturation process is as follows.

[0152] Yeast library construction: Degenerate primers were designed and the designed amino acid mutations were introduced into the antibody hu3-11scFv mutant library by PCR. The size of each library was approximately 10 9 The diversity of the constructed yeast library was verified by sequencing.

[0153] In the first round of screening, approximately 5 × 10 10The cells and biotinylated TSLP-Fc protein (1-10 μg / ml) were incubated in 50 ml of 0.1% bovine serum albumin (BSA)-containing phosphate-buffered saline (PBSA) for 1 h at room temperature. The mixture was then washed with 0.1% PBSA to remove unbound antibody fragments. Then, 100 μl of streptomycin beads (Milenyi Biotec, Auburn, CA) were added to the hu3-11-scFv antibody mutant library bound to biotinylated TSLP-Fc and loaded onto the AutoMACS system for sorting. Cells with high affinity to TSLP-Fc were collected from the antibody library and induced at 250 rpm and 20 °C for 18 h. The resulting enriched library was subjected to a second round of screening against biotinylated recombinant TSLP-Fc protein.

[0154] For the third and fourth rounds of screening, the library cells from the previous round were incubated with biotinylated recombinant TSLP-Fc protein (0.1-1 μg / ml) and 10 μg / ml mouse anti-cMyc (9E10, Sigma) antibody in 0.1% PBSA for 1 h at room temperature. The mixture was washed with 0.1% PBSA to remove unbound antibody fragments. Goat anti-mouse-Alexa488 (A-11001, Life Technologies) and streptavidin-PE (S-866, Life Technologies) were added and incubated at 4°C for 1 h. The mixture was washed with 0.1% PBSA to remove unbound antibody fragments. Finally, antibodies with high affinity were screened by FACS screening (BD FACSAriaTM FUSION).

[0155] The hu3-11-scFv mutant library was subjected to two rounds of MACS screening and two rounds of FACS screening using biotinylated TSLP-Fc antigen. Approximately 400 yeast single clones were then selected for culture and expression induction. The binding of the yeast single clones to the TSLP-Fc antigen was detected using FACS, and yeast single clones with high affinity were selected and sequence verified. The sequenced clones were compared and analyzed. After removing redundant sequences, the non-redundant sequences were converted into full-length antibodies for mammalian cell expression.

[0156] The sequences of the light chain variable regions obtained by affinity maturation are as follows: [ka]

[0157] The resulting light chain variable region was recombined with the heavy chain variable region of mab3 humanized antibody to obtain new mab3 humanized antibodies. For example, huVL5 and huVL6 were combined with hu3VH2-H110Y, respectively, to obtain new antibody molecules hu3-12 and hu3-13. Details are as follows:

[0158] [Table 12]

[0159] The CDR sequences of the mab humanized antibody obtained after affinity maturation are shown below.

[0160] [Table 13]

[0161] The new antibody mab3 humanized antibody was subjected to ELISA to detect its binding activity to human TSLP. The results showed that hu3-12 and hu3-13 still had high binding ability to human TSLP. It was shown that changing LCDR3 did not affect the activity of the hu3 series antibodies.

[0162] In summary, the CDR sequences of the mab3 humanized antibody are as follows:

[0163] [Table 14] Here, X1 is H or Y, X3 is N or S, X4 is V or G, and X5 is G or E.

[0164] The antibody heavy and light chain variable region combinations of the mab3 humanized antibody after hotspot mutation and affinity maturation are shown in the table below.

[0165] [Table 15]

[0166] 5.2 Selection and backmutation of the human FR region of mab119

[0167] For mab119, IGHV1-69 was used as the VH template. * 02 and HJ6 * 01 was selected as the template for VL, and IGKV4-1 * 01 and IGKJ2 * 01, and IGKV3-11 * 01 and IGKJ2 * 01 was selected. The CDR regions of the mouse antibody were grafted into the selected humanized template, and the FR regions were backmutated to obtain different light and heavy chain variable regions. The variable region sequences obtained by CDR grafting are as follows: [ka]

[0168] The back mutations of the mab119 humanized antibody are shown in the table below.

[0169] [Table 16] Note: For example, M4L denotes that M at position 4 is backmutated to L according to the Kabat numbering convention. Grafted denotes that the mouse antibody CDRs are grafted onto human germline FR region sequences.

[0170] The specific sequences of the variable regions of the mab119 humanized antibody are as follows: >hu119VL1 (Grafted (IGKV4-1*01)) [ka] >hu119VL2 [ka] SEQ ID NO:57 >hu119VL3 [ka] SEQ ID NO:58 >hu119VL4 (Grafted, IGKV3-11*01) [ka] SEQ ID NO:59 >hu119VL5 [ka] SEQ ID NO:60 >hu119VL6 [ka] SEQ ID NO:61 [ka] [ka] [ka] NOTE: Single underlines represent variable regions, while double underlines represent back mutations.

[0171] The above light and heavy chain variable regions were combined with human germline light and heavy chain constant region sequences to finally form complete light and heavy chain sequences, thus obtaining an antibody with a full-length sequence. Illustratively, for the mab119 humanized antibody in the present disclosure, the heavy chain constant region is the IgG1-YTE constant region shown in SEQ ID NO:133, and the light chain constant region is the kappa chain constant region shown in SEQ ID NO:134, but they can also be replaced with other constant regions known in the art.

[0172] The heavy and light chain variable regions of the mab119 humanized antibody are shown in Table 17.

[0173] [Table 17]

[0174] The binding activity of the humanized antibody to human TSLP was detected by ELISA, and the results showed that the mab119 humanized antibody was able to specifically bind to human TSLP.

[0175] (2) hu119 mutation

[0176] Since hot spots were detected in the LCDR1 DNS sequence of the mab119 humanized antibody, the corresponding sites were mutated to N31S or N31Q. The LCDR1 sequences obtained after mutation are as follows:

[0177] [Table 18] NOTE: The mutation site positions in Table 19 are numbered according to natural order.

[0178] As an example, the mutant sequences of hu119VL2 and hu119VL6 obtained after mutation are as follows: [ka] SEQ ID NO:73 [ka] NOTE: Single underlines represent variable regions, while double underlines represent back mutations.

[0179] The resulting hu119VL2 and hu119VL6 mutants were combined with hu119VH to generate new humanized hu119 antibodies. As examples, hu119VL2-N31S, hu119VL2-N31Q were each combined with hu119VH3 to generate antibodies hu119-28 and hu119-29, and hu119VL3-N31S was combined with hu119VH8 to generate antibody hu119-30. Exemplary combinations of variable regions of mutant antibodies are as follows:

[0180] [Table 19]

[0181] The affinity of the antibodies obtained after the mutations to human TSLP was detected by ELISA. The results showed that the hu119-28 and hu119-29 antibodies still had relatively high affinity to human TSLP, indicating that the N31S and N31Q mutations in LCDR2 did not affect the anti-TSLP antibody activity.

[0182] In summary, the sequences of the CDRs of the mab119 humanized antibody are as follows:

[0183] [Table 20] X6 is selected from N, S, and Q.

[0184] Humanization of mab179

[0185] (1) Template Selection and Backmutation for Humanization of the Mab179 Mouse Antibody

[0186] In the case of mab179, IGHV1-69 was used as the VH template. * 02 and IGHJ6 * 01 was selected as the template for VL, and IGKV4-1 * 01 and IGKJ2 * 01, or IGKV2-29 * 02 and IGKJ2 * 01 was selected. The CDR regions of the mouse antibody were grafted into the selected humanized template, and the FR regions were backmutated to obtain light and heavy chain variable regions with different sequences. The humanized variable region sequences and backmutations are as follows: [ka] [ka]

[0187] [Table 21] NOTE: For example, P43S denotes a backmutation of P at position 43 to S according to the Kabat numbering convention. Grafted denotes that the mouse antibody CDRs have been grafted onto human germline FR region sequences.

[0188] The variable regions of the mab179 humanized antibody are shown below. >hu179VL1 (Graft (IGKV4-1 * 01) [ka] [ka] [ka] SEQ ID NO:82 [ka] [ka] [ka] Note: Single underlined regions represent CDRs and double underlined regions represent backmutation sites.

[0189] The above light and heavy chain variable regions were combined with human germline light and heavy chain constant region sequences to finally form complete light and heavy chain sequences, thus obtaining an antibody with a full-length sequence. For example, for the mab199 humanized antibody in the present disclosure, the heavy chain constant region is the IgG1-YTE constant region shown in SEQ ID NO:133, and the light chain constant region is the kappa chain constant region shown in SEQ ID NO:134, but they can also be replaced with other constant regions known in the art.

[0190] [Table 22]

[0191] The affinity of the mab179 humanized antibody to human TSLP was detected by ELISA, and the results showed that the mab179 humanized antibody has very good affinity to human TSLP.

[0192] (2) Mutation of the hu179 antibody

[0193] The detection revealed the existence of hot spots on the HCDR2 and LCDR2 sequences of the mab179 humanized antibody, and the corresponding hot spots were then mutated to eliminate the risk of molecular modification.

[0194] In one embodiment, an amino acid mutation is made to GNG in HCDR2 of hu179VH1, and the sequence of hu179VH1 after mutation is as follows: hu179VH1-N55Q [ka] SEQ ID NO:90 [ka] [ka] Note: The single underlined regions represent the CDRs and the double underlined regions represent the backmutation sites.

[0195] The sequence of the HCDR2 region of the mab179 humanized antibody obtained after mutation is as follows:

[0196] [Table 23] NOTE: The mutation site positions in Table 24 are numbered according to natural order.

[0197] The CDR regions of the mab179 humanized antibody can be taken from above and are shown as follows:

[0198] [Table 24] X7 is selected from N, Q, and V, and X8 is selected from G or V.

[0199] The hu179VH1 mutants obtained after mutation were combined with the humanized hu179VL to obtain new mab179 humanized antibodies. Exemplary antibodies of the combination of hu179VH1 mutants and hu179VL2 are as follows:

[0200] [Table 25]

[0201] The affinity of the antibody with human TSLP obtained after the mutation was detected by ELISA. The results showed that the antibody after HCDR2 mutation still maintained a relatively high affinity with human TSLP. This indicates that the point mutations N55Q, N55V, and G56V in HCDR2 of the mab179 humanized antibody do not fundamentally affect the affinity activity of the antibody with TSLP.

[0202] Using the same method, we mutated hu179VH2, hu179VH3, hu179VH4, and hu179VH5 with N55Q, N55V, and G56V point mutations (numbered in natural order), respectively, and recombined the heavy and light chain variable regions obtained by the mutations to obtain a new humanized mab179. As an example, the mutated sequence of hu179VH3 is shown below. [ka] Note: Single underlines represent CDRs and double underlines represent backmutation sites.

[0203] In some other examples, the LCDR2 of the mab179 humanized antibody was mutated. For example, the sequence of hu179VL2 after mutation is shown below: [ka] [ka] [ka] [ka] Note: Single underlines represent CDRs and double underlines represent backmutation sites.

[0204] The sequence of the mab179 humanized antibody LCDR2 obtained after mutation is as follows:

[0205] LCDR2 mutants of the mab179 humanized antibody [Table 26]

[0206] From the above, the general formula of LCDR2 of the mab179 humanized antibody is X9VX 10 X 11 X 12 X 13 X9 is selected from Y or E; 10 is selected from S, D or E, and X 11 is selected from N, Q, D, or E, and X 12 is selected from H, Y, D or E, and X 13 is selected from E or Y. The CDR regions of the mab179 humanized antibody are as shown in the table below.

[0207] [Table 27] X7 is selected from N, Q, and V, X8 is selected from G and V, X9 is selected from Y and E, and X 10 is selected from S, D, and E, and X 11 is selected from N, Q, D, E, and X 12 is selected from H, Y, D, and E, and X 13 was selected from E and Y.

[0208] The hu179VL2 mutant obtained after mutation was combined with the humanized hu179 heavy chain variable region to obtain a new mab179 humanized antibody. As an example, the hu179VL2 mutant was combined with hu179VH1 and hu179VH3, and the combination of CDRs and heavy and light chain variable regions of the resulting mab179 humanized antibody is shown below.

[0209] [Table 28] Here, X5 is selected from Y, E, X6 is selected from S, D, E, X7 is selected from N, Q, D, E, X8 is selected from H, Y, D, E, and X9 is selected from E, Y.

[0210] [Table 29]

[0211] The affinity of the mab179 humanized antibody obtained after LCDR2 mutation to human TSLP was detected by ELISA. The results showed that the antibody obtained after hot spot site mutation in LCDR2 still had a relatively good affinity to human TSLP. This indicates that the hot spot site mutation in LCDR2 does not affect the binding activity of the mab179 humanized antibody.

[0212] Following the same method, the following mutations were made in LCDR2 of hu179VL3, hu179VL4, hu179VL5, hu179VL6, hu179VL7 and hu179VL8: N53Q, N53D, N53S, H54Y, Y50E, S52D, S52E, N53E, H54D, H54E, Y55E. The mutated light and heavy chain variable regions were combined to form a new mab humanized antibody. In one embodiment, the sequence of mutated hu179VL8 is shown as follows: [ka]

[0213] The mutagenic results of hu179VL8-N53E and hu179VH3-N55V were combined to obtain a new antibody molecule, hu179-33, whose CDR sequences are as follows:

[0214] CDR region of hu179-33 antibody [Table 30]

[0215] The binding activity of the resulting antibodies after mutation to human TSLP was detected by Biacore. Exemplary binding activities of the antibodies are shown below.

[0216] [Table 31]

[0217] As a result, it was found that the antibody hu179-33 has a relatively high specific binding activity to human TSLP. This indicates that the point mutations in the hot spots of both HCDR2 and LCDR2 do not affect the affinity of the mab179 humanized antibody to human TSLP. In the mab179 humanized antibody molecule, the mutations N55Q, N55V, and G56V made on HCDR2 and the mutations N53Q, N53D, N53S, H54Y, Y50E, S52D, S52E, N53E, H54D, H54E, and Y55E made on LCDR2 do not affect the binding of the antibody to human TSLP, i.e., the activity of the anti-TSLP antibody.

[0218] 5.4 Selection and backmutation of human FR regions for the mab199 antibody

[0219] For mab199, IGHV1-46 was used as the VH template. * 01 and HJ6 * 01 was selected, and IGKV1-39 was used as the VL template. * 01 and IGKJ4 * 01 was selected. The CDR regions of the mouse antibody were grafted into the selected humanized template, and the FR regions were backmutated to obtain light and heavy chain variable regions of different sequences. The backmutations are shown in Table 32.

[0220] [Table 32] Note: For example, I48V denotes that the I at position 48 is backmutated to a V according to the Kabat numbering convention. "Grafted" denotes that the mouse antibody CDRs are grafted onto human germline FR region sequences.

[0221] The variable regions of the mab199 humanized antibody are shown below. [ka] [ka] [ka] [ka] Note: Single underlines represent CDRs and double underlines represent backmutation sites.

[0222] The above light and heavy chain variable regions were combined with human germline light and heavy chain constant region sequences to finally form complete light and heavy chain sequences, thus obtaining an antibody with a full-length sequence. For the mab199 humanized antibody, unless otherwise specified in this disclosure, the light chain constant region is the constant region shown in SEQ ID NO:134, and the heavy chain constant region is the constant region shown in SEQ ID NO:133.

[0223] The resulting mab199 humanized antibody is shown below.

[0224] [Table 33]

[0225] The activity of the humanized antibody mab199, which inhibits the binding of TSLP to the TSLP receptor, was detected using the ELISA method. The detection results are as follows.

[0226] [Table 34]

[0227] As a result, it was found that the mab199 humanized antibody still had relatively high activity in inhibiting the binding of TSLP to the TSLP receptor.

[0228] 5.5 Antibody constant regions

[0229] The heavy chain constant region of the humanized and chimeric antibodies can be selected from the group consisting of the constant regions of IgG1, IgG2, IgG4 and variants thereof. Exemplarily, the IgG1-YTE constant region is used in the present disclosure, the sequence of which is as shown in SEQ ID NO:133. The light chain constant region can be selected from the light chain constant region of human kappa chain, lambda chain or variants thereof. Exemplarily, the human kappa chain constant region is used in the present disclosure, the sequence of which is as shown in SEQ ID NO:134. >IgG1-YTE heavy chain constant region: [ka] NOTE: Underlines refer to the designed M252Y, S254T, T256E mutations. > Kappa light chain constant region: [ka]

[0230] The humanized heavy and light chain variable regions in this disclosure were recombined with the constant regions described above to obtain full-length heavy and light chain sequences. As an example, the antibody sequences are as follows: hu3-13 antibody heavy chain: [ka] hu3-13 antibody light chain: [ka] hu119-30 antibody heavy chain [ka] hu119-30 antibody light chain [ka] hu179-33 antibody heavy chain [ka] hu179-33 antibody light chain [ka] hu199-36 antibody heavy chain [ka] hu199-36 antibody light chain: [ka] Note: The underlined parts represent the CDRs and the italicized parts represent the constant regions.

[0231] AMG157 was used as a positive control in the present disclosure and its sequence is as shown in SEQ ID NO:143 and SEQ ID NO:144. AMG157 heavy chain sequence [ka] AMG157 light chain sequence [ka]

[0232] Furthermore, when testing antibody activity, the present disclosure also constructed cell lines with human TSLP receptor and human IL7Rα, whose sequences are as follows: The full length amino acid sequence of the human TSLP receptor: [ka] Note: The underlined portion indicates the signal peptide. SEQ ID NO:145 Full-length amino acid sequence of human IL7Rα (Uniprot ID: P16871) [ka] Note: The underlined portion indicates the signal peptide. SEQ ID NO:146

[0233] The antibodies of the disclosure can be cloned, expressed, and purified using conventional gene cloning and recombinant expression methods.

[0234] Test Example:

[0235] Biological evaluation of in vitro activity

[0236] Test Example 1: ELISA detection of binding of anti-TSLP antibodies to human TSLP

[0237] Human TSLP-his (SEQ ID NO:1) was diluted to 1 μg / ml in PBS (Shanghai BasalMedia, B320) buffer at pH 7.4, added to a 96-well microtiter plate (Corning, CLS3590-100EA) at 100 μg / well, and incubated at 4°C overnight. The liquid was discarded, and 200 μl / well of inhibitor solution containing 5% skim milk powder (Bright Dairy skim milk powder) diluted in PBS was added, and the plate was incubated for 2 hours in a 37°C incubator to perform inhibition. After inhibition, the inhibitor solution was discarded, and the plate was washed three times with PBST buffer (PBS containing 0.1% Tween-20, pH 7.4). The test antibody and the positive antibody AMG157 diluted to different concentrations with the sample diluent were added at 100 μl / well, and incubated for 1 hour in a 37°C incubator. After the end of the incubation, the plate was washed three times with PBST. HRP-labeled goat anti-mouse secondary antibody (Jackson Immuno Research, 115-035-003) diluted with sample diluent was added at 100 μl / well and incubated at 37°C for 1 hour. After washing the plate six times with PBST, 50 μl / well TMB chromogenic substrate (KPL, 52-00-03) was added and incubated at room temperature for 10-15 minutes, and 50 μl / well 1 M H2SO4 was added to stop the reaction. Absorbance was read at 450 nm using a NOVOStar microplate reader. The EC50 value for TSLP antibody binding to TSLP was calculated, and the results are shown in the table below.

[0238] [Table 35]

[0239] The results showed that the antibodies in the present disclosure have very good binding activity with human TSLP.

[0240] Test Example 2: Biacore detection of the affinity of anti-TSLP humanized antibodies to TSLP of different species

[0241] The affinity of the tested humanized TSLP antibodies to human and cynoTSLP was detected by a Biacore T200 (GE) instrument.

[0242] The tested molecules were affinity captured with a Protein A biosensor chip (catalog number 29127556, GE). Then, antigens (huTSLP-his and cynoTSLP-his prepared in Example 1) were flowed over the chip surface, and the reaction signals were detected in real time using Biacore T200 to obtain binding and dissociation curves. After the dissociation of each experimental cycle was completed, the biosensor chip was washed and regenerated with glycine-HCl regeneration solution (pH 1.5, catalog number BR-1003-54, GE). The data was fitted to the (1:1) Langmuir model using BIAevaluation version 4.1, GE software, and affinity values ​​were obtained as shown in the table below.

[0243] [Table 36]

[0244] The results demonstrated that the anti-TSLP antibodies of the present disclosure have relatively high affinity for human TSLP and are capable of binding to cynoTSLP.

[0245] Test Example 3: ELISA-based experiment of anti-TSLP antibodies that inhibit the binding of TSLP to the TSLP receptor

[0246] The TSLP receptor has two subunits, TSLPR and IL7R, of which TSLPR is a specific receptor for TSLP and IL7R is a common receptor for TSLP and IL7. TSLP first binds to TSLPR and then to IL7R. This example was used to determine whether TSLP antibodies could inhibit the binding of TSLP to the extracellular domain of recombinantly expressed TSLPR receptor protein.

[0247] ELISA plates were coated with human-TSLPR-Fc-ECD (2 μg / ml, SEQ ID NO:5) and incubated overnight at 4°C. After discarding the liquid, 200 μl / well of inhibition solution containing 5% nonfat dry milk diluted in PBS was added and incubated in a 37°C incubator for 2 hours to perform inhibition. After inhibition was completed, the inhibition solution was discarded and the plate was washed three times with PBST buffer (PBS containing 0.05% Tween-20, pH 7.4). Biotin-labeled huTSLP-Fc antigen was prepared at 3 nM, and the tested antibodies were serially diluted starting from 200 nM. Antigen and antibody were mixed 1:1, placed at 37°C for 15 minutes, and added to the microtiter plate at 100 μl per well and placed at 37°C for 1 hour. The plate was washed three times with PBST. Streptavidin-peroxidase polymer diluted 1:4000 in sample diluent was added at 100 μl / well and incubated at 37°C for 1 hour. After washing the plate five times with PBST, 100 μl / well TMB chromogenic substrate (KPL, 52-00-03) was added and incubated at room temperature for 3-10 minutes, and the reaction was stopped by adding 100 μl / well 1M H2SO4. Absorbance values ​​were read at 450 nm using a NOVOStar microplate reader. IC50 values ​​of TSLP antibodies that inhibited binding of TSLP to TSLPR were calculated and the results are shown in Table 37 and Figure 1.

[0248] [Table 37]

[0249] The results showed that all of the antibodies disclosed herein were capable of potently inhibiting the binding of TSLP to its receptor, TSLPR.

[0250] Test Example 5: FACS-based experiment to inhibit binding of TSLP antibodies to the TSLP receptor

[0251] This example was used to determine whether anti-TSLP antibodies could inhibit the binding of TSLP to the TSLPR / IL7R receptor on the surface of the CHOK1 cell line.

[0252] In the detailed method, CHOK1-TSLPR / IL7R was cultured in DME / F12 containing 10% FBS, 1 mg / ml G418 and 10 μg / ml puromycin. Well-established CHOK1-TSLPR / IL7R cells were centrifuged (1000 rpm, 5 min) and washed once with 2% FBS in PBS. Cells were counted and diluted to 1 × 10 6 The cells were adjusted to a cell concentration of 100 / ml. 50 μl of cells were added to a round-bottom 96-well plate. The antibodies to be tested were diluted in PBS containing 2% BSA, with an initial concentration of 20 nM, a gradient of 8, and a ratio of 1:4. Biotin-labeled TSLP-Fc antigen was prepared at 2 nM. Antigen and antibody were mixed 1:1 and placed at 37°C for 15 min. 50 μl of the mixture was added per well to the 96-well plate plated with cells and incubated at 4°C for 1 h. After incubation, the plate was centrifuged (800g, 5 min) at 4°C and the supernatant was discarded. The plate was washed twice with 200 μl of pre-chilled PBS by centrifugation. PE-SA secondary antibody diluted 1:1000 was added and incubated at 4°C for 40 min in the dark. The plate was then centrifuged (800g, 5 min) at 4°C and the supernatant was discarded. 200 μl pre-chilled PBS was added to rupture the cells, which were then washed by centrifugation at 4° C. three times. 100 μl PBS was added, and the plate was loaded into the machine for reading. The IC50 values ​​of TSLP antibodies that inhibited the binding of TSLP to TSLPR / IL7R were calculated according to the values ​​of the fluorescent signal. The results are shown in Table 38.

[0253] [Table 38]

[0254] The results showed that all of the antibodies disclosed herein could relatively strongly inhibit the binding of TSLP to cell surface TSLPR / IL7R.

[0255] Test Example 6: Inhibition of TSLP-induced chemokine production by anti-TSLP antibodies

[0256] TSLP can induce naive myeloid dendritic cells (mDCs) to mature and secrete the chemokines thymus activation-regulated chemokine (TARC) and osteoprotegerin (OPG), which can further mediate innate and adaptive immune inflammatory responses. This example demonstrated that the resulting antibodies can inhibit TSLP-induced chemokine production by mDCs, thereby inhibiting the development of innate and adaptive inflammatory responses.

[0257] Naïve bone marrow tissue mDCs were isolated and purified from human peripheral blood mononuclear cells (PBMCs) using magnetic bead sorting (CD1c (BDCA-1) + dendritic cell isolation kit, Miltenyi Biotec). The obtained mDCs were seeded into 96-well cell culture plates. Serially diluted antibody samples and human TSLP (huTSLP-his, final concentration 50 ng / ml) were pre-incubated for approximately 45 min (37 °C) and then added to each cell culture well containing mDCs, respectively, to stimulate mDCs in vitro. The plates were placed in an incubator and cultured for 48 h. The cell culture supernatants were collected and appropriately diluted, and then the chemokine content was detected using the ELISA method. TARC was detected using the human CCL17 / TARC Quantikine ELISA Kit from R&D, and OPG content was detected using the human CCL22 / MDC Quantikine ELISA Kit (R&D). The results are shown in Figure 4A to Figure 4B.

[0258] The results showed that all of the antibodies in the present disclosure could significantly inhibit TSLP-induced TARC and OPG chemokine production, indicating that the antibodies in the present disclosure could block the development of innate and adaptive inflammatory responses.

[0259] Test Example 7. Inhibition of proliferation of BaF3-TLSPR / IL7R cells induced by native TSLP by anti-TSLP antibodies

[0260] BaF3-hTSLPR / hIL7R cells can proliferate under the stimulation of native TSLP. When the antibody binds to native TSLP, the stimulatory effect of TSLP on BaF3-hTSLPR / hIL7R cells is reduced.

[0261] NHLF cells (BeNa Culture Collection BNCC340764) and HLF1 cells (BeNa Culture Collection BNCC337730) were cultured until the cells grew to 80% and the supernatant was discarded. Human lung fibroblasts, NHLF (BeNa Culture Collection BNCC340764) and HLF1 (BeNa Culture Collection BNCC337730), were stimulated with 10ng / ml human IL1-β (Sino Biological GMP-10139-HNAE), 20ng / ml IL13 (R&D 213-ILB-005), and 20ng / ml TNF-α (PEPROTECH 300-01A) for 72 hours to induce the production of native TSLP. After stimulation, the cell supernatant was collected and centrifuged at 4500 rpm for 5 minutes to remove cell debris. The supernatant was collected, concentrated approximately 10-fold using a concentration column, and filtered for further use.

[0262] BaF3-hTSLPR / hIL17R cells were cultured in RPMI1640 with 10% FBS (10 ng / mL mIL3, R&D 213-ILB-005) at 1 × 10 4The cells were adjusted to a density of 1000 cells / ml and cultured in a 37°C, 5% CO2 incubator until logarithmic growth phase. The cells were harvested, centrifuged at 800 rpm / min for 5 min, and the supernatant was discarded. The cells were washed three times with PBS to remove cytokines that stimulate proliferation in culture. The cells were resuspended in RPMI1640 medium containing 4% FBS and plated at 4000 cells / 50 μl / well in a 96-well plate and cultured in an incubator for 2 h. The antibody tested was native TSLP at a 10-fold ratio, serially diluted with an initial antibody concentration of 100 nM, resulting in three dilution gradients of 100 nM, 10 nM, and 1 nM. 50 μl / well of the diluted antibody / antigen mixture was added to the cells at final antibody concentrations of 50 nM, 5 nM, and 0.5 nM. The plates were incubated in a 37°C, 5% CO2 incubator for 72 h. Then 30 μL of CellTiter-Glo (Promega) was added to each well, incubated at room temperature in the dark for 10 minutes, and detected using a Cytation5 cell imager with the luminescence program. The results are shown in the table below.

[0263] [Table 39]

[0264] As a result, it was found that all of the antibodies obtained in the present disclosure were able to significantly suppress the proliferation-stimulating activity of natural TSLP, BaF3, and in particular, hu179-33, which has an activity 100-fold or more higher than that of AMG157.

[0265] Test Example 8: Experiments with anti-TSLP antibodies that inhibit TSLP-induced proliferation of BaF3 cells overexpressing TSLPR / IL7R

[0266] TSLP binds to TSLPR / IL7R on the surface of BaF3, thereby promoting the proliferation of BaF3. This example was used to determine whether the antibodies of the present disclosure can block the activity of TSLP to induce proliferation of BaF3.

[0267] Specifically, BaF3 cells overexpressing TSLPR / IL7R were cultured in RPMI1640 containing 10% FBS and 2 ng / mL rhIL3 (MultiSciences, catalog no. 96-AF-300-03-20) at 37°C in a 5% CO2 incubator at a concentration of 1 × 10 6 The cells were cultured at a cell density not exceeding 1000 cells / ml. When the antibody was detected, the logarithmic growth phase cells were washed with PBS three times and centrifuged at 800 rpm for 5 minutes. The cell density was adjusted to 8000 cells / well / 90 μl using RPMI1640 (2% FBS, recombinant human TSLP-Fc: 40 ng / ml). 10 μl of serially diluted antibodies to be tested were added to the 96-well plate and cultured for 2 days. 30 μl of cell titer was added and mixed for detection. The IC50 was calculated according to the readings. The results are shown in Table 40 and Figure 3.

[0268] [Table 40]

[0269] The results showed that all of the antibodies of the present disclosure had a relatively strong ability to inhibit TSLP-mediated proliferation of BaF3 cells.

[0270] Test Example 9: Humanized anti-TSLP antibody inhibits TSLP-induced natural CD4 + Inhibition of T cell differentiation into Th2 cells

[0271] TSLP can induce maturation of primary bone marrow mDC cells. Mature mDC cells highly express OX40 ligand, which binds naturally to CD4. + It can bind OX40 on the surface of T cells, thereby forming natural CD4 + It induces T to differentiate into Th2 cells, produces factors related to immune responses such as IL4 / IL5 / IL13, and induces Th2 inflammatory reactions in the body. This test example was used to detect whether the antibody obtained in the present disclosure can block the differentiation of Th2 cells induced by TSLP.

[0272] Natural bone marrow DCs were isolated and purified from human peripheral blood mononuclear cells (PBMCs) using magnetic bead sorting (CD1c (BDCA-1)+ dendritic cell isolation kit, Miltenyi Biotec). The obtained mDCs were seeded into 96-well cell culture plates. Serially diluted antibody samples and recombinantly expressed human TSLP (huTSLP-his, final concentration 50 ng / ml) were pre-incubated for approximately 45 min (37°C) and then added to each cell culture well containing mDCs, respectively, and cultured at 37°C for 24 h. Mature mDCs after stimulation were collected and washed twice with PBS. CD4 + CD45RA + Natural T cells were extracted from PBMCs by magnetic bead separation (Miltenyi, Biotec). Natural T cells obtained by separation were mixed with mature mDCs, seeded in a 96-well cell culture plate at a ratio of 5:1, and co-cultured for 6 days. The cells were collected and seeded in a 96-well plate precoated with anti-CD3 (10 μg / ml), and anti-CD28 (1 μg / mL) was added to stimulate the differentiated T cells again. The cells were cultured for 24 hours, and finally the cell culture supernatant was collected. Th2-related cytokines secreted by the cells in the supernatant were detected by ELISA. IL-4 and IL-5 cytokines were detected with an ELISA kit from R&D, and TNF-α and IL-13 were detected with an ELISA kit from NeoBioscience. The results are shown in Figures 5A-5D.

[0273] The results showed that the antibodies obtained in the present disclosure could significantly suppress the production of Th2 cytokines IL4, IL5, IL13 and TNF-α, and demonstrated that the antibodies obtained in the present disclosure could inhibit TSLP-induced differentiation of Th2 cells.

[0274] [Sequence table] SEQUENCE LISTING <110> JIANGSU HENGRUI MEDICINE CO., LTD. SHANGHAI HENGRUI PHARMACEUTICAL CO., LTD. <120> ANTIBODY CAPABLE OF BINDING TO THYMIC STROMAL LYMPHOPOIETIN AND USE THEREOF <130> PA25-027 <160> 146 <170> SIPOSequenceListing 1.0 <210> 1 <211> 176 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of his-tagged human TSLP antigen (huTSLP-his) <400> 1 Met Phe Pro Phe Ala Leu Leu Tyr Val Leu Ser Val Ser Phe Arg Lys 1 5 10 15 Ile Phe Ile Leu Gln Leu Val Gly Leu Val Leu Thr Tyr Asp Phe Thr 20 25 30 Asn Cys Asp Phe Glu Lys Ile Lys Ala Ala Tyr Leu Ser Thr Ile Ser 35 40 45 Lys Asp Leu Ile Thr Tyr Met Ser Gly Thr Lys Ser Thr Glu Phe Asn 50 55 60 Asn Thr Val Ser Cys Ser Asn Arg Pro His Cys Leu Thr Glu Ile Gln 65 70 75 80 Ser Leu Thr Phe Asn Pro Thr Ala Gly Cys Ala Ser Leu Ala Lys Glu 85 90 95 Met Phe Ala Met Lys Thr Lys Ala Ala Leu Ala Ile Trp Cys Pro Gly 100 105 110 Tyr Ser Glu Thr Gln Ile Asn Ala Thr Gln Ala Met Lys Lys Ala Arg 115 120 125 Lys Ser Lys Val Thr Thr Asn Lys Cys Leu Glu Gln Val Ser Gln Leu 130 135 140 Gln Gly Leu Trp Arg Arg Phe Asn Arg Pro Leu Leu Lys Gln Gln Gly 145 150 155 160 Ser Ser Asp Tyr Lys Asp Asp Asp Asp Lys His His His His His His 165 170 175 <210> 2 <211> 398 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of Fc-tagged human TSLP antigen (huTSLP-Fc) <400> 2 Met Phe Pro Phe Ala Leu Leu Tyr Val Leu Ser Val Ser Phe Arg Lys 1 5 10 15 Ile Phe Ile Leu Gln Leu Val Gly Leu Val Leu Thr Tyr Asp Phe Thr 20 25 30 Asn Cys Asp Phe Glu Lys Ile Lys Ala Ala Tyr Leu Ser Thr Ile Ser 35 40 45 Lys Asp Leu Ile Thr Tyr Met Ser Gly Thr Lys Ser Thr Glu Phe Asn 50 55 60 Asn Thr Val Ser Cys Ser Asn Arg Pro His Cys Leu Thr Glu Ile Gln 65 70 75 80 Ser Leu Thr Phe Asn Pro Thr Ala Gly Cys Ala Ser Leu Ala Lys Glu 85 90 95 Met Phe Ala Met Lys Thr Lys Ala Ala Leu Ala Ile Trp Cys Pro Gly 100 105 110 Tyr Ser Glu Thr Gln Ile Asn Ala Thr Gln Ala Met Lys Lys Ala Arg 115 120 125 Lys Ser Lys Val Thr Thr Asn Lys Cys Leu Glu Gln Val Ser Gln Leu 130 135 140 Gln Gly Leu Trp Arg Arg Phe Asn Arg Pro Leu Leu Lys Gln Gln Asp 145 150 155 160 Ile Glu Gly Arg Met Asp Glu Pro Lys Ser Ser Asp Lys Thr His Thr 165 170 175 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 180 185 190 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 195 200 205 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 210 215 220 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 225 230 235 240 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 245 250 255 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 260 265 270 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 275 280 285 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 290 295 300 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 305 310 315 320 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 325 330 335 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 340 345 350 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 355 360 365 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 370 375 380 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 385 390 395 <210> 3 <211> 168 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of his-tagged cyno TSLP antigen (cynoTSLP-his) <400> 3 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Tyr Asp Phe Thr Asn Cys Asp Phe Gln Lys Ile Glu 20 25 30 Ala Asp Tyr Leu Arg Thr Ile Ser Lys Asp Leu Ile Thr Tyr Met Ser 35 40 45 Gly Thr Lys Ser Thr Asp Phe Asn Asn Thr Val Ser Cys Ser Asn Arg 50 55 60 Pro His Cys Leu Thr Glu Ile Gln Ser Leu Thr Phe Asn Pro Thr Pro 65 70 75 80 Arg Cys Ala Ser Leu Ala Lys Glu Met Phe Ala Arg Lys Thr Lys Ala 85 90 95 Thr Leu Ala Leu Trp Cys Pro Gly Tyr Ser Glu Thr Gln Ile Asn Ala 100 105 110 Thr Gln Ala Met Lys Lys Ala Arg Lys Ser Lys Val Thr Thr Asn Lys 115 120 125 Cys Leu Glu Gln Val Ser Gln Leu Leu Gly Leu Trp Arg Arg Phe Ile 130 135 140 Arg Thr Leu Leu Lys Gln Gln Gly Ser Ser Asp Tyr Lys Asp Asp Asp 145 150 155 160 Asp Lys His His His His His His 165 <210> 4 <211> 390 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of Fc-tagged cyno TSLP antigen (cyno TSLP-Fc) <400> 4 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Tyr Asp Phe Thr Asn Cys Asp Phe Gln Lys Ile Glu 20 25 30 Ala Asp Tyr Leu Arg Thr Ile Ser Lys Asp Leu Ile Thr Tyr Met Ser 35 40 45 Gly Thr Lys Ser Thr Asp Phe Asn Asn Thr Val Ser Cys Ser Asn Arg 50 55 60 Pro His Cys Leu Thr Glu Ile Gln Ser Leu Thr Phe Asn Pro Thr Pro 65 70 75 80 Arg Cys Ala Ser Leu Ala Lys Glu Met Phe Ala Arg Lys Thr Lys Ala 85 90 95 Thr Leu Ala Leu Trp Cys Pro Gly Tyr Ser Glu Thr Gln Ile Asn Ala 100 105 110 Thr Gln Ala Met Lys Lys Ala Arg Lys Ser Lys Val Thr Thr Asn Lys 115 120 125 Cys Leu Glu Gln Val Ser Gln Leu Leu Gly Leu Trp Arg Arg Phe Ile 130 135 140 Arg Thr Leu Leu Lys Gln Gln Asp Ile Glu Gly Arg Met Asp Glu Pro 145 150 155 160 Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 165 170 175 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 180 185 190 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 195 200 205 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 210 215 220 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 225 230 235 240 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 245 250 255 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 260 265 270 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 275 280 285 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 290 295 300 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 305 310 315 320 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 325 330 335 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 340 345 350 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 355 360 365 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 370 375 380 Ser Leu Ser Pro Gly Lys 385 390 <210> 5 <211> 446 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of Fc-tagged human TSLP receptor extracellular domain (human-TSLPR-Fc-ECD) <400> 5 Gly Ala Ala Glu Gly Val Gln Ile Gln Ile Ile Tyr Phe Asn Leu Glu 1 5 10 15 Thr Val Gln Val Thr Trp Asn Ala Ser Lys Tyr Ser Arg Thr Asn Leu 20 25 30 Thr Phe His Tyr Arg Phe Asn Gly Asp Glu Ala Tyr Asp Gln Cys Thr 35 40 45 Asn Tyr Leu Leu Gln Glu Gly His Thr Ser Gly Cys Leu Leu Asp Ala 50 55 60 Glu Gln Arg Asp Asp Ile Leu Tyr Phe Ser Ile Arg Asn Gly Thr His 65 70 75 80 Pro Val Phe Thr Ala Ser Arg Trp Met Val Tyr Tyr Leu Lys Pro Ser 85 90 95 Ser Pro Lys His Val Arg Phe Ser Trp His Gln Asp Ala Val Thr Val 100 105 110 Thr Cys Ser Asp Leu Ser Tyr Gly Asp Leu Leu Tyr Glu Val Gln Tyr 115 120 125 Arg Ser Pro Phe Asp Thr Glu Trp Gln Ser Lys Gln Glu Asn Thr Cys 130 135 140 Asn Val Thr Ile Glu Gly Leu Asp Ala Glu Lys Cys Tyr Ser Phe Trp 145 150 155 160 Val Arg Val Lys Ala Met Glu Asp Val Tyr Gly Pro Asp Thr Tyr Pro 165 170 175 Ser Asp Trp Ser Glu Val Thr Cys Trp Gln Arg Gly Glu Ile Arg Asp 180 185 190 Ala Cys Ala Glu Thr Pro Thr Pro Pro Lys Pro Lys Leu Ser Lys Asp 195 200 205 Ile Glu Gly Arg Met Asp Glu Pro Lys Ser Ser Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 6 <211> 121 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab3 heavy chain variable region sequence <400> 6 Glu Val Gln Leu Gln Gln Ser Gly Pro Val Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asp 20 25 30 Tyr Met Asn Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp His Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 7 <211> 105 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab3 light chain variable region sequence <400> 7 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Arg Thr Phe 85 90 95 Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 8 <211> 120 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab119 heavy chain variable region sequence <400> 8 Gln Ala Tyr Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Phe Ala Phe Thr Thr Tyr 20 25 30 Asn Met His Trp Val Lys His Thr Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Ser Arg Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 9 <211> 111 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab119 light chain variable region sequence <400> 9 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 Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Leu Asn 65 70 75 80 Pro Val Glu Thr Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110 <210> 10 <211> 119 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab179 heavy chain variable region sequence <400> 10 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Ile Glu Leu Ser Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Leu Thr Val Ser Ser 115 <210> 11 <211> 107 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab179 light chain variable region sequence <400> 11 Ser Ile Val Met Thr Gln Thr Pro Lys Phe Leu Leu Val Ser Ala Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 12 <211> 117 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab199 heavy chain variable region sequence <400> 12 Gln Val Gln Leu Gln Gln Ser Gly Pro Gln Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Leu Thr Val Ser Ser 115 <210> 13 <211> 107 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> Murine mab199 light chain variable region sequence <400> 13 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Val Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 14 <211> 5 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab3 HCDR1 <400> 14 Asp Asp Tyr Met Asn 1 5 <210> 15 <211> 17 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> mab3 HCDR2 <400> 15 Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 16 <211> 12 <212> PRT <213> Mus musculus <220> <221> PEPTIDE <223> mab HCDR3 <400> 16 Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp His 1 5 10 <210> 17 <211> 10 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab3 LCDR1 <400> 17 Arg Ala Ser Ser Ser Val Ser Tyr Met His 1 5 10 <210> 18 <211> 7 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab LCDR2 <400> 18 Ala Thr Ser Asn Leu Ala Ser 1 5 <210> 19 <211> 8 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab3 LCDR3 <400> 19 Gln Gln Trp Ser Ser Asn Arg Thr 1 5 <210> 20 <211> 5 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab119 HCDR1 <400> 20 Thr Tyr Asn Met His 1 5 <210> 21 <211> 17 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab119 HCDR2 <400> 21 Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 22 <211> 11 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab119 HCDR3 <400> 22 Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val 1 5 10 <210> 23 <211> 15 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab119 LCDR1 <400> 23 Arg Ala Ser Glu Ser Val Asp Asn Ser Gly Leu Ser Phe Met His 1 5 10 15 <210> 24 <211> 7 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab119 LCDR2 <400> 24 Arg Ala Ser Asn Leu Gly Ser 1 5 <210> 25 <211> 9 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab119 LCDR3 <400> 25 Gln Gln Ile Asn Thr Asp Pro Leu Thr 1 5 <210> 26 <211> 5 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab179 HCDR1 <400> 26 Asn Tyr Leu Ile Glu 1 5 <210> 27 <211> 17 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab179 HCDR2 <400> 27 Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe Lys 1 5 10 15 Gly <210> 28 <211> 10 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab179 HCDR3 <400> 28 Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr 1 5 10 <210> 29 <211> 11 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab179 LCDR1 <400> 29 Lys Ala Ser Gln Ser Val Ser Ser Asp Val Thr 1 5 10 <210> 30 <211> 7 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab179 LCDR2 <400> 30 Tyr Val Ser Asn His Tyr Thr 1 5 <210> 31 <211> 9 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab179 LCDR3 <400> 31 Gln Gln His His Arg Phe Pro Leu Thr 1 5 <210> 32 <211> 5 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab199 HCDR1 <400> 32 Thr Tyr Trp Met His 1 5 <210> 33 <211> 17 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab199 HCDR2 <400> 33 Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe Lys 1 5 10 15 Asp <210> 34 <211> 8 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab199 HCDR3 <400> 34 Thr Leu Asp Gly Tyr Tyr Asp Tyr 1 5 <210> 35 <211> 11 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab199 LCDR1 <400> 35 Arg Ala Ser Glu Asn Ile Tyr Ser Tyr Leu Ala 1 5 10 <210> 36 <211> 7 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab199 LCDR2 <400> 36 Phe Ala Lys Thr Leu Ala Glu 1 5 <210> 37 <211> 9 <212> PRT <213> Mus musculus <220> <221> DOMAIN <223> mab199 LCDR3 <400> 37 Gln His His Tyr Gly Thr Pro Trp Thr 1 5 <210> 38 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 VL-CDR grafted <400> 38 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Arg Thr Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 39 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VL2 <400> 39 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Arg Thr Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 40 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VL3 <400> 40 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Arg Thr Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 41 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VL4 <400> 41 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Arg Thr Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 42 <211> 121 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VH1 <400> 42 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asp 20 25 30 Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp His Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 43 <211> 121 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VH2 <400> 43 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asp 20 25 30 Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Leu Thr Val Asp Lys Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp His Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 44 <211> 121 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VH3 <400> 44 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asp 20 25 30 Tyr Met Asn Trp Val Lys Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile 35 40 45 Gly Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp His Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 45 <211> 12 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 HCDR3-H110Y <400> 45 Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp Tyr 1 5 10 <210> 46 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3LCDR3-N93D <400> 46 Gln Gln Trp Ser Ser Asp Arg Thr 1 5 <210> 47 <211> 12 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 HCDR3 (general formula) <220> <221> DOMAIN <222> (12)..(12) <223> Xaa is selected from His or Tyr. <400> 47 Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp Xaa 1 5 10 <210> 48 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 LCDR (general formula 1) <220> <221> DOMAIN <222> (6)..(6) <223> Xaa is selected from Asn or Asp. <400> 48 Gln Gln Trp Ser Ser Xaa Arg Thr 1 5 <210> 49 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Light chain variable sequence of hu3-11 <400> 49 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asp Arg Thr Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 50 <211> 121 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Heavy chain variable sequence of hu3-11 <400> 50 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asp 20 25 30 Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Leu Thr Val Asp Lys Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 51 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VL5 <400> 51 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Asp Asn Val Arg Gly Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 52 <211> 105 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3VL6 <400> 52 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Asp Ser Gly Arg Glu Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 53 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 LCDR3-V1 <400> 53 Gln Gln Ser Asp Asn Val Arg Gly 1 5 <210> 54 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 LCDR3-V2 <400> 54 Gln Gln Ser Asp Ser Gly Arg Glu 1 5 <210> 55 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3 LCDR3 (general formula 2) <220> <221> DOMAIN <222> (5)..(5) <223> Xaa is selected from Asn or Ser. <220> <221> DOMAIN <222> (6)..(6) <223> Xaa is selected from Val or Gly. <220> <221> DOMAIN <222> (8)..(8) <223> Xaa is selected from Gly or Glu. <400> 55 Gln Gln Ser Asp Xaa Xaa Arg Xaa 1 5 <210> 56 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119-VL CDR grafted(IGKV4-1*01) <400> 56 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Asp Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Gly Ser Gly Val Pro Asp 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 Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 57 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL2 <400> 57 Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Asp Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Gly Ser Gly Val Pro Asp 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 Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 58 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL3 <400> 58 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Asp Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Asp 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 Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 59 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL4 (Grafted, IGKV3-11*01) <400> 59 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 35 40 45 Arg Leu Leu Ile Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 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 Phe Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 60 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL5 <400> 60 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 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 Phe Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 61 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL6 <400> 61 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 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 Phe Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 62 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH1 (Grafted) <400> 62 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 63 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH2 <400> 63 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 64 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH3 <400> 64 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Lys His Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 65 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH4 <400> 65 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Lys His Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 66 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH5 <400> 66 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 67 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH6 <400> 67 Glu Ala Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 68 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH7 <400> 68 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Arg Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 69 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VH8 <400> 69 Glu Ala Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Arg Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 70 <211> 15 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119 LCDR1-N31S <400> 70 Arg Ala Ser Glu Ser Val Asp Ser Ser Gly Leu Ser Phe Met His 1 5 10 15 <210> 71 <211> 15 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119 LCDR1-N31Q <400> 71 Arg Ala Ser Glu Ser Val Asp Gln Ser Gly Leu Ser Phe Met His 1 5 10 15 <210> 72 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL2-N31S <400> 72 Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Asp Ser Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Gly Ser Gly Val Pro Asp 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 Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 73 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL2-N31Q <400> 73 Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Asp Gln Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Gly Ser Gly Val Pro Asp 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 Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 74 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL6-N31S <400> 74 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Ser Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 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 Phe Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 75 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119VL6-N31Q <400> 75 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Gln Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 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 Phe Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 76 <211> 15 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu119 LCDR1-general formula <220> <221> DOMAIN <222> (9)..(9) <223> Xaa is selected from Asn, Ser or Gln. <400> 76 Arg Ala Ser Glu Ser Val Asp Xaa Ser Gly Leu Ser Phe Met His 1 5 10 15 <210> 77 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL1 (Graft (IGKV4-1*01)) <400> 77 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 78 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2 <400> 78 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 79 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 VL3 <400> 79 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 80 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 VL4 <400> 80 Ser Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 81 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL5 (Grafted(IGKV2-29*02)) <400> 81 Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala 65 70 75 80 Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 82 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL6 <400> 82 Ser Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala 65 70 75 80 Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 83 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL7 <400> 83 Ser Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Lys Ile Ser Arg Val Glu Ala 65 70 75 80 Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 84 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL8 <400> 84 Ser Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala 65 70 75 80 Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 85 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH1 (Grafted) <400> 85 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 86 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH2 <400> 86 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 87 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH3 <400> 87 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Ile Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 88 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH4 <400> 88 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Ile Glu Leu Ser Arg Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 89 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH5 <400> 89 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Gly Asn Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 90 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH1- N55Q <400> 90 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asp Pro Gly Gln Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 91 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH1- N55V <400> 91 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asp Pro Gly Val Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 92 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH1- G56V <400> 92 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asp Pro Gly Asn Val Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 93 <211> 17 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 HCDR2-N55Q <400> 93 Val Ile Asp Pro Gly Gln Gly Asp Thr Asn Tyr Asn Glu Asn Phe Lys 1 5 10 15 Gly <210> 94 <211> 17 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 HCDR2-N55V <400> 94 Val Ile Asp Pro Gly Val Gly Asp Thr Asn Tyr Asn Glu Asn Phe Lys 1 5 10 15 Gly <210> 95 <211> 17 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 HCDR2-G56V <400> 95 Val Ile Asp Pro Gly Asn Val Asp Thr Asn Tyr Asn Glu Asn Phe Lys 1 5 10 15 Gly <210> 96 <211> 17 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 HCDR2 (general formula) <220> <221> DOMAIN <222> (6)..(6) <223> Xaa is selected from Asn, Gln or Val. <220> <221> DOMAIN <222> (7)..(7) <223> Xaa is selected from Gly or Val. <400> 96 Val Ile Asp Pro Gly Xaa Xaa Asp Thr Asn Tyr Asn Glu Asn Phe Lys 1 5 10 15 Gly <210> 97 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VH3-N55V <400> 97 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Gly Val Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Ile Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 98 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-Y50E <400> 98 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Glu Val Ser Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 99 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-S52D <400> 99 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Asp Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 100 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-S52E <400> 100 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Glu Asn His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 101 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-N53Q <400> 101 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Gln His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 102 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-N53D <400> 102 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asp His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 103 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-N53E <400> 103 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Glu His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 104 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-H54Y <400> 104 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn Tyr Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 105 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-H54D <400> 105 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn Asp Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 106 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-H54E <400> 106 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn Glu Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 107 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL2-Y55E <400> 107 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Asn His Glu Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 108 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-Y50E <400> 108 Glu Val Ser Asn His Tyr Thr 1 5 <210> 109 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-S52D <400> 109 Tyr Val Asp Asn His Tyr Thr 1 5 <210> 110 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-S52E <400> 110 Tyr Val Glu Asn His Tyr Thr 1 5 <210> 111 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179LCDR2-N53Q <400> 111 Tyr Val Ser Gln His Tyr Thr 1 5 <210> 112 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-N53D <400> 112 Tyr Val Ser Asp His Tyr Thr 1 5 <210> 113 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-N53E <400> 113 Tyr Val Ser Glu His Tyr Thr 1 5 <210> 114 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-H54Y <400> 114 Tyr Val Ser Asn Tyr Tyr Thr 1 5 <210> 115 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-H54D <400> 115 Tyr Val Ser Asn Asp Tyr Thr 1 5 <210> 116 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-H54E <400> 116 Tyr Val Ser Asn Glu Tyr Thr 1 5 <210> 117 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179 LCDR2-Y55E <400> 117 Tyr Val Ser Asn His Glu Thr 1 5 <210> 118 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> mab179 humanized antibody LCDR2 general formula <220> <221> DOMAIN <222> (1)..(1) <223> Xaa is selected from Try or Glu. <220> <221> DOMAIN <222> (3)..(3) <223> Xaa is selected from Ser, Asp or Glu. <220> <221> DOMAIN <222> (4)..(4) <223> Xaa is selected from Asn, Gln, Asp or Glu. <220> <221> DOMAIN <222> (5)..(5) <223> Xaa is selected from His, Tyr, Asp or Glu. <220> <221> DOMAIN <222> (6)..(6) <223> Xaa is selected from Glu or Tyr. <220> <221> DOMAIN <223> hu179VL8-N53E <400> 118 Xaa Val Xaa Xaa Xaa Xaa Thr 1 5 <210> 119 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179VL8-N53E <400> 119 Ser Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Glu His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala 65 70 75 80 Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 120 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VL1 (Grafted) <400> 120 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 121 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VL2 <400> 121 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 122 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VL3 <400> 122 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 123 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VL4 <400> 123 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 124 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VL5 <400> 124 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Val Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 125 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VL6 <400> 125 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Val Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 126 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH1 (Grafted) <400> 126 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 127 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH2 <400> 127 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 128 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH3 <400> 128 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 129 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH4 <400> 129 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 130 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH5 <400> 130 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 131 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH6 <400> 131 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Lys Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 132 <211> 117 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199VH7 <400> 132 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Lys Val Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser 115 <210> 133 <211> 330 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> IgG1-YTE heavy chain constant region <400> 133 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 134 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> kappa light chain constant region <400> 134 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 135 <211> 451 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> hu3-13 antibody heavy chain <400> 135 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Asp 20 25 30 Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Ile Ile Ser Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Leu Thr Val Asp Lys Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Asp Tyr Asp Gly Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr 245 250 255 Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 136 <211> 212 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu3-13 antibody light chain <400> 136 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Asp Ser Gly Arg Glu Phe 85 90 95 Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser 100 105 110 Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala 115 120 125 Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 130 135 140 Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 145 150 155 160 Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr 165 170 175 Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys 180 185 190 Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn 195 200 205 Arg Gly Glu Cys 210 <210> 137 <211> 450 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> hu119-30 antibody heavy chain <400> 137 Glu Ala Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Glu Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Arg Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Tyr Gly Glu Gly Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile 245 250 255 Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> 138 <211> 218 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> hu119-30 antibody light chain <400> 138 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Ser Ser 20 25 30 Gly Leu Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Leu Tyr Arg Ala Ser Asn Leu Gly Ser Gly Ile Pro Ala 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 Phe Ala Val Tyr Tyr Cys Gln Gln Ile Asn 85 90 95 Thr Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 145 150 155 160 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 139 <211> 449 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179-33 antibody heavy chain <400> 139 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Gly Val Gly Asp Thr Asn Tyr Asn Glu Asn Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Ile Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asn Thr Gly Thr Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr 245 250 255 Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 140 <211> 214 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu179-33 antibody light chain <400> 140 Ser Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Ser Asp 20 25 30 Val Thr Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Tyr Val Ser Glu His Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala 65 70 75 80 Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln His His Arg Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 141 <211> 447 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> hu199-36 antibody heavy chain <400> 141 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Thr Leu Ile Gln Lys Phe 50 55 60 Lys Asp Lys Val Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Thr Leu Asp Gly Tyr Tyr Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 142 <211> 214 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> hu199-36 antibody light chain <400> 142 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Phe Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Val Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Gly Thr Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 143 <211> 448 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> AMG157 heavy chain sequence <400> 143 Gln Met Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Thr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys His Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Thr Arg Asp Asn Ser Lys Asn Thr Leu Asn 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys 210 215 220 Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val 290 295 300 Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 144 <211> 214 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> AMG157 light chain sequence <400> 144 Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln 1 5 10 15 Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Leu Gly Ser Lys Ser Val 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Val Tyr 35 40 45 Asp Asp Ser Asp Arg Pro Ser Trp Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Gly Glu Ala Gly 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His 85 90 95 Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys 100 105 110 Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln 115 120 125 Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly 130 135 140 Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly 145 150 155 160 Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala 165 170 175 Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser 180 185 190 Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val 195 200 205 Ala Pro Thr Glu Cys Ser 210 <210> 145 <211> 371 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of human TSLP receptor full-length sequence <400> 145 Met Gly Arg Leu Val Leu Leu Trp Gly Ala Ala Val Phe Leu Leu Gly 1 5 10 15 Gly Trp Met Ala Leu Gly Gln Gly Gly Ala Ala Glu Gly Val Gln Ile 20 25 30 Gln Ile Ile Tyr Phe Asn Leu Glu Thr Val Gln Val Thr Trp Asn Ala 35 40 45 Ser Lys Tyr Ser Arg Thr Asn Leu Thr Phe His Tyr Arg Phe Asn Gly 50 55 60 Asp Glu Ala Tyr Asp Gln Cys Thr Asn Tyr Leu Leu Gln Glu Gly His 65 70 75 80 Thr Ser Gly Cys Leu Leu Asp Ala Glu Gln Arg Asp Asp Ile Leu Tyr 85 90 95 Phe Ser Ile Arg Asn Gly Thr His Pro Val Phe Thr Ala Ser Arg Trp 100 105 110 Met Val Tyr Tyr Leu Lys Pro Ser Ser Pro Lys His Val Arg Phe Ser 115 120 125 Trp His Gln Asp Ala Val Thr Val Thr Cys Ser Asp Leu Ser Tyr Gly 130 135 140 Asp Leu Leu Tyr Glu Val Gln Tyr Arg Ser Pro Phe Asp Thr Glu Trp 145 150 155 160 Gln Ser Lys Gln Glu Asn Thr Cys Asn Val Thr Ile Glu Gly Leu Asp 165 170 175 Ala Glu Lys Cys Tyr Ser Phe Trp Val Arg Val Lys Ala Met Glu Asp 180 185 190 Val Tyr Gly Pro Asp Thr Tyr Pro Ser Asp Trp Ser Glu Val Thr Cys 195 200 205 Trp Gln Arg Gly Glu Ile Arg Asp Ala Cys Ala Glu Thr Pro Thr Pro 210 215 220 Pro Lys Pro Lys Leu Ser Lys Phe Ile Leu Ile Ser Ser Leu Ala Ile 225 230 235 240 Leu Leu Met Val Ser Leu Leu Leu Leu Ser Leu Trp Lys Leu Trp Arg 245 250 255 Val Lys Lys Phe Leu Ile Pro Ser Val Pro Asp Pro Lys Ser Ile Phe 260 265 270 Pro Gly Leu Phe Glu Ile His Gln Gly Asn Phe Gln Glu Trp Ile Thr 275 280 285 Asp Thr Gln Asn Val Ala His Leu His Lys Met Ala Gly Ala Glu Gln 290 295 300 Glu Ser Gly Pro Glu Glu Pro Leu Val Val Gln Leu Ala Lys Thr Glu 305 310 315 320 Ala Glu Ser Pro Arg Met Leu Asp Pro Gln Thr Glu Glu Lys Glu Ala 325 330 335 Ser Gly Gly Ser Leu Gln Leu Pro His Gln Pro Leu Gln Gly Gly Asp 340 345 350 Val Val Thr Ile Gly Gly Phe Thr Phe Val Met Asn Asp Arg Ser Tyr 355 360 365 Val Ala Leu 370 <210> 146 <211> 459 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Amino acid sequence of human IL7R alpha full-length sequence <400> 146 Met Thr Ile Leu Gly Thr Thr Phe Gly Met Val Phe Ser Leu Leu Gln 1 5 10 15 Val Val Ser Gly Glu Ser Gly Tyr Ala Gln Asn Gly Asp Leu Glu Asp 20 25 30 Ala Glu Leu Asp Asp Tyr Ser Phe Ser Cys Tyr Ser Gln Leu Glu Val 35 40 45 Asn Gly Ser Gln His Ser Leu Thr Cys Ala Phe Glu Asp Pro Asp Val 50 55 60 Asn Thr Thr Asn Leu Glu Phe Glu Ile Cys Gly Ala Leu Val Glu Val 65 70 75 80 Lys Cys Leu Asn Phe Arg Lys Leu Gln Glu Ile Tyr Phe Ile Glu Thr 85 90 95 Lys Lys Phe Leu Leu Ile Gly Lys Ser Asn Ile Cys Val Lys Val Gly 100 105 110 Glu Lys Ser Leu Thr Cys Lys Lys Ile Asp Leu Thr Thr Ile Val Lys 115 120 125 Pro Glu Ala Pro Phe Asp Leu Ser Val Ile Tyr Arg Glu Gly Ala Asn 130 135 140 Asp Phe Val Val Thr Phe Asn Thr Ser His Leu Gln Lys Lys Tyr Val 145 150 155 160 Lys Val Leu Met His Asp Val Ala Tyr Arg Gln Glu Lys Asp Glu Asn 165 170 175 Lys Trp Thr His Val Asn Leu Ser Ser Thr Lys Leu Thr Leu Leu Gln 180 185 190 Arg Lys Leu Gln Pro Ala Ala Met Tyr Glu Ile Lys Val Arg Ser Ile 195 200 205 Pro Asp His Tyr Phe Lys Gly Phe Trp Ser Glu Trp Ser Pro Ser Tyr 210 215 220 Tyr Phe Arg Thr Pro Glu Ile Asn Asn Ser Ser Gly Glu Met Asp Pro 225 230 235 240 Ile Leu Leu Thr Ile Ser Ile Leu Ser Phe Phe Ser Val Ala Leu Leu 245 250 255 Val Ile Leu Ala Cys Val Leu Trp Lys Lys Arg Ile Lys Pro Ile Val 260 265 270 Trp Pro Ser Leu Pro Asp His Lys Lys Thr Leu Glu His Leu Cys Lys 275 280 285 Lys Pro Arg Lys Asn Leu Asn Val Ser Phe Asn Pro Glu Ser Phe Leu 290 295 300 Asp Cys Gln Ile His Arg Val Asp Asp Ile Gln Ala Arg Asp Glu Val 305 310 315 320 Glu Gly Phe Leu Gln Asp Thr Phe Pro Gln Gln Leu Glu Glu Ser Glu 325 330 335 Lys Gln Arg Leu Gly Gly Asp Val Gln Ser Pro Asn Cys Pro Ser Glu 340 345 350 Asp Val Val Ile Thr Pro Glu Ser Phe Gly Arg Asp Ser Ser Leu Thr 355 360 365 Cys Leu Ala Gly Asn Val Ser Ala Cys Asp Ala Pro Ile Leu Ser Ser 370 375 380 Ser Arg Ser Leu Asp Cys Arg Glu Ser Gly Lys Asn Gly Pro His Val 385 390 395 400 Tyr Gln Asp Leu Leu Leu Ser Leu Gly Thr Thr Asn Ser Thr Leu Pro 405 410 415 Pro Pro Phe Ser Leu Gln Ser Gly Ile Leu Thr Leu Asn Pro Val Ala 420 425 430 Gln Gly Gln Pro Ile Leu Thr Ser Leu Gly Ser Asn Gln Glu Glu Ala 435 440 445 Tyr Val Thr Met Ser Ser Phe Tyr Gln Asn Gln 450 455

Claims

1. 1. An anti-TSLP antibody comprising a heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:47, respectively; the light chain variable region comprises LCDR1 and LCDR2 as set forth in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3 as set forth in SEQ ID NO:48 or 55; The sequence of SEQ ID NO: 47 is EDYDYDGYAMDX. １ , the sequence of SEQ ID NO: 48 is QQWSSX ２ RT, the sequence of SEQ ID NO: 55 is QQSDX ３ X ４ R.X. ５ and X １ is H or Y, X ２ is N or D, X ３ is N or S, X ４ is V or G, X ５ is G or E; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:76, SEQ ID NO:24 and SEQ ID NO:25, respectively; The sequence of SEQ ID NO: 76 is RASESVDX ６ SGLSFMH, X ６ is selected from N, S and Q; or iii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:96 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:118 and SEQ ID NO:31, respectively; The sequence of SEQ ID NO:96 is VIDPGX. ７ X ８ DTNYNE, and the sequence of SEQ ID NO: 118 is X ９ VX １０ X １１ X １２ X １３ T and X ７ is selected from N, Q and V; X ８ is G or V, and X ９ is Y or E, and X １０ is selected from S, D and E; X １１ is selected from N, Q, D and E; X １２ is selected from H, Y, D and E; X １３ is E or Y; or iv) an anti-TSLP antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:32, SEQ ID NO:33 and SEQ ID NO:34, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:35, SEQ ID NO:36 and SEQ ID NO:37, respectively.

2. comprising a heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1 and HCDR2 as set forth in SEQ ID NO:14 and SEQ ID NO:15, respectively, and HCDR3 as set forth in SEQ ID NO:16 or 45, and the light chain variable region comprises LCDR1 and LCDR2 as set forth in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3 as set forth in SEQ ID NO:19, 46, 53 or 54; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR2 and LCDR3 as set forth in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 as set forth in SEQ ID NO:23, 70 or 71; or iii) the anti-TSLP antibody of claim 1, wherein the heavy chain variable region comprises HCDR1 and HCDR3 as set forth in SEQ ID NO:26 and SEQ ID NO:28, respectively, and HCDR2 as set forth in SEQ ID NO:27, 93, 94 or 95, and the light chain variable region comprises LCDR1 and LCDR3 as set forth in SEQ ID NO:29 and SEQ ID NO:31, respectively, and LCDR2 as set forth in SEQ ID NO:30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117.

3. comprising a heavy chain variable region and a light chain variable region, i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:54, respectively; or ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:53, respectively; or iii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:70, SEQ ID NO:24 and SEQ ID NO:25, respectively; or iv) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:71, SEQ ID NO:24 and SEQ ID NO:25, respectively; or v) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:94 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:113 and SEQ ID NO:31, respectively; or vi) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:94 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or vii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:16, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:19, respectively; or viii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:45, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:17, SEQ ID NO:18 and SEQ ID NO:46, respectively; or ix) the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO:23, SEQ ID NO:24, and SEQ ID NO:25, respectively; or x) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or xi) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:93 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31, respectively; or xii) the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:95, and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, respectively; or xiii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:28, respectively, and the light chain variable region comprises LCDR1 and LCDR3 as set forth in SEQ ID NO:29 and SEQ ID NO:31, respectively, and LCDR2 as set forth in SEQ ID NO:108, 109, 110, 111, 112, 113, 114, 115, 116 or 117; The anti-TSLP antibody of claim 2.

4. The anti-TSLP antibody according to any one of claims 1 to 3, wherein the anti-TSLP antibody is a murine antibody, a chimeric antibody or a humanized antibody.

5. 5. The anti-TSLP antibody of claim 4, wherein the anti-TSLP antibody comprises a framework region derived from a human antibody, and the anti-TSLP antibody comprises a light chain variable region and / or a heavy chain variable region selected from those set forth in (a), (b), (c) or (d) below. a) the heavy chain variable region comprises HCDR1 and HCDR2 as depicted in SEQ ID NO: 14 and SEQ ID NO: 15, respectively, and HCDR3 as depicted in SEQ ID NO: 16 or 45, the framework regions of which comprise up to 10 back mutations, preferably selected from one or more of 38K, 48I, 67A, 69L, 71V and 73K, and / or the light chain variable region comprises LCDR1 and LCDR2 as depicted in SEQ ID NO: 17 and SEQ ID NO: 18, respectively, and LCDR3 as depicted in SEQ ID NO: 19, 46, 53 or 54, the framework regions of which comprise up to 10 amino acid back mutations, preferably selected from one or more of 46P, 47W, 58V, 70S and 71Y; b) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22, respectively, whose framework regions comprise up to 10 backmutations, preferably selected from one or more of 2A, 27F, 38K, 39H, 48I, 67A, 69L, 71V and 76R, and / or the light chain variable region comprises LCDR2 and LCDR3 as depicted in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 as depicted in SEQ ID NO:23, 70 or 71, whose framework regions comprise up to 10 amino acid backmutations, preferably selected from one or more of 1D, 4L, 43P, 48L and 58I; c) the heavy chain variable region comprises HCDR1 and HCDR3 as set forth in SEQ ID NO: 26 and SEQ ID NO: 28, respectively, and HCDR2 as set forth in SEQ ID NO: 27, 93, 94 or 95, the framework regions of which comprise up to 10 back mutations, preferably the back mutations being selected from one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR; and / or the light chain variable region comprises LCDR1 and LCDR3 as set forth in SEQ ID NO: 29 and SEQ ID NO: 31, respectively, and HCDR2 as set forth in SEQ ID NO: 27, 93, 94 or 95, the framework regions of which comprise up to 10 back mutations, preferably the back mutations being selected from one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I and 82bR. or, comprising an LCDR2 as set forth in SEQ ID NO: 30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117, the framework regions of which comprise up to 10 back mutations, preferably the back mutations being selected from one or more of 1S, 43S, 67Y and 73F; or d) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as depicted in SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34, respectively, whose framework regions comprise up to 10 back mutations, preferably selected from one or more of 38K, 48I, 66K, 67A, 69L, 71V, 73K and 78A, and / or the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as depicted in SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37, respectively, whose framework regions comprise up to 10 back mutations, preferably selected from one or more of 43S, 45Q, 48V, 66V and 70Q.

6. comprising a heavy chain variable region and a light chain variable region, i) the heavy chain variable region has at least 90% sequence identity with the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 6, 42, 43, 44 or 50, and the light chain variable region has at least 90% sequence identity with the light chain variable region set forth in the amino acid sequence SEQ ID NO: 7, 38, 39, 40, 41, 49, 51 or 52; or ii) the heavy chain variable region has at least 90% sequence identity with the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 8, 62, 63, 64, 65, 66, 67, 68 or 69, and the light chain variable region has at least 90% sequence identity with the light chain variable region set forth in the amino acid sequence SEQ ID NO: 9, 56, 57, 58, 59, 60, 61, 72, 73, 74 or 75; or iii) the heavy chain variable region has at least 90% sequence identity with the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 10, 85, 86, 87, 88, 89, 90, 91, 92 or 97, and the light chain variable region has at least 90% sequence identity with the light chain variable region set forth in the amino acid sequence SEQ ID NO: 11, 77, 78, 79, 80, 81, 82, 83, 84, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119; or iv) the heavy chain variable region has at least 90% sequence identity with the heavy chain variable region set forth in the amino acid sequence SEQ ID NO: 12, 126, 127, 128, 129, 130, 131 or 132, and the light chain variable region has at least 90% sequence identity with the light chain variable region set forth in the amino acid sequence SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125; The anti-TSLP antibody of claim 4.

7. comprising a heavy chain variable region and a light chain variable region, i) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 6, 42, 43, 44 or 50, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 7, 38, 39, 40, 41, 49, 51 or 52; or ii) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 8, 62, 63, 64, 65, 66, 67, 68 or 69, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 9, 56, 57, 58, 59, 60, 61, 72, 73, 74 or 75; or iii) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 10, 85, 86, 87, 88, 89, 90, 91, 92 or 97 and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 11, 77, 78, 79, 80, 81, 82, 83, 84, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107 or 119; or iv) the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 12, 126, 127, 128, 129, 130, 131 or 132, and the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125; The anti-TSLP antibody of claim 6.

8. 8. The anti-TSLP antibody of claim 7, comprising the heavy chain variable region and the light chain variable region as shown below. a) the sequence of the heavy chain variable region is shown in SEQ ID NO:50 and the sequence of the light chain variable region is shown in SEQ ID NO:52; or b) the sequence of the heavy chain variable region is shown in SEQ ID NO:50 and the sequence of the light chain variable region is shown in SEQ ID NO:51; or c) the sequence of the heavy chain variable region is shown in SEQ ID NO: 69 and the sequence of the light chain variable region is shown in SEQ ID NO: 74; or d) the sequence of the heavy chain variable region is shown in SEQ ID NO: 64 and the sequence of the light chain variable region is shown in SEQ ID NO: 73; or e) the sequence of the heavy chain variable region is shown in SEQ ID NO: 97 and the sequence of the light chain variable region is shown in SEQ ID NO: 119; or f) the sequence of the heavy chain variable region is shown in SEQ ID NO: 91 and the sequence of the light chain variable region is shown in SEQ ID NO: 78; or g) the sequence of the heavy chain variable region is shown in SEQ ID NO:132 and the sequence of the light chain variable region is shown in SEQ ID NO:

125.

9. 9. The anti-TSLP antibody of any one of claims 1 to 8, wherein said antibody further comprises an antibody heavy and light chain constant region, preferably said heavy chain constant region is selected from the group consisting of human IgG1, IgG2, IgG3 and IgG4 constant regions and conventional variants thereof, and said light chain constant region is selected from the group consisting of human antibody kappa and lambda chain constant regions and conventional variants thereof, more preferably said antibody comprises a heavy chain constant region as set forth in SEQ ID NO: 133, and a light chain constant region as set forth in SEQ ID NO:

134.

10. 10. The anti-TSLP antibody of claim 9, wherein the antibody comprises the heavy and light chains as set forth below: a) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO: 135 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO: 136 or has at least 90% sequence identity thereto; or b) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO: 137 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO: 138 or has at least 90% sequence identity thereto; or c) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO: 139 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO: 140 or has at least 90% sequence identity thereto; or d) the amino acid sequence of the heavy chain is as set forth in SEQ ID NO: 141 or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as set forth in SEQ ID NO: 142 or has at least 90% sequence identity thereto.

11. An isolated anti-TSLP antibody that binds to human TSLP competitively with the anti-TSLP antibody of any one of claims 1 to 10.

12. A nucleic acid molecule encoding an anti-TSLP antibody according to any one of claims 1 to 11.

13. A host cell comprising the nucleic acid molecule of claim 12.

14. A pharmaceutical composition comprising a therapeutically effective amount of an anti-TSLP antibody according to any one of claims 1 to 11, a nucleic acid molecule according to claim 12, or a host cell according to claim 13, and one or more pharma- ceutically acceptable carriers, diluents, buffers, or excipients.

15. A method for immunodetecting or determining TSLP in vitro or ex vivo, comprising the step of using an anti-TSLP antibody according to any one of claims 1 to 11.

16. A kit comprising an anti-TSLP antibody according to any one of claims 1 to 11.

17. A method of treating a TSLP-related disease comprising administering to a subject a therapeutically effective amount of an anti-TSLP antibody according to any one of claims 1 to 11, a nucleic acid molecule according to claim 12, a host cell according to claim 13, or a pharmaceutical composition according to claim 14, wherein preferably said TSLP-related disease is asthma, idiopathic pulmonary fibrosis, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic sinusitis, urticaria, Netherton syndrome, eosinophilic esophagitis, food allergy, allergic diarrhea, eosinophilic gastroenteritis, allergic bronchopulmonary aspergillosis, allergic fungal sinusitis, chronic pruritus, cancer, breast cancer, colon cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, chronic obstructive pulmonary disease. The method is selected from the group consisting of systemic sclerosis, multiple sclerosis, keloidosis, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, hypereosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, inflammatory bowel disease, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, fibrosis due to radiation, and fibrosis due to wound healing.

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