Antibodies capable of binding to thymic interstitial lymphocyte growth factor and their use

Abstract

This invention discloses an antibody capable of binding to thymic interstitial lymphocyte growth factor and a method for using the same antibody. [Solution] Disclosed are mouse antibodies, chimeric antibodies and humanized antibodies of the light chain variable region and heavy chain variable region of anti-TSLP antibodies, as well as anti-TSLP antibodies comprising their antigen-binding fragments, or pharmaceutically acceptable salts or solvent compounds thereof, and their use as an asthma treatment, in particular in the preparation of drugs for treating TSLP-positive diseases or symptoms.

Description

[Technical Field]

[0001] This disclosure relates to the field of antibody agents. Specifically, this disclosure relates to anti-TSLP antibody agents and their use. [Background technology]

[0002] The descriptions herein are for the sole purpose of providing background information relating to this disclosure and do not necessarily constitute prior art.

[0003] Asthma is a serious chronic inflammatory airway disease. There are approximately 334 million people with asthma worldwide, and about 30 million in China, with a much higher mortality rate than in developed countries. As environmental degradation and air pollution worsen, the number of people suffering from this disease will increase, potentially posing a significant threat to human life and health.

[0004] Thymus interstitial lymphocyte growth factor (TSLP) is an epithelial cell-derived cytokine produced in response to pro-inflammatory stimuli, primarily promoting allergic inflammation through its action on dendritic and mast cells. TSLP is a type of interleukin-7 (IL-7)-like cytokine and was first discovered in conditioned medium for 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. The molecule contains three pairs of disulfide bonds made up of six cysteine ​​units and two N-glycosylation sites, with a molecular weight of approximately 15-20 kD. The TSLP receptor is a two-part complex consisting of TSLPR and IL7Rα. TSLP first binds to TSLPR with relatively low affinity, then recruits IL7Rα with high affinity, and finally activates signaling pathways such as stat5, leading to DC maturation and T cell differentiation.

[0005] Myeloid dendritic cells (mDC) are the main effector cells of TSLP. TSLP acts on immature mDCs that secrete the 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 secrete Th2 cytokines such as IL-5, IL-4, IL-9, and TNF, inducing Th2 inflammatory responses in the body. Furthermore, TSLP induces DC cells to produce the cytokine IL-8, which sequentially mobilizes neutrophils, resulting in neutrophilic innate immune inflammation. TSLP also induces DCs to produce eotaxin-2, which mobilizes eosinophils and, acting together with IL5, can rapidly shift 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 causes innate inflammation and Th2 inflammation, thereby increasing tissue mucus, changing the airway and causing tracheal stenosis, and exacerbating cell fibrosis. The inflammation gradually develops into 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, it is necessary to continue the development of pharmaceuticals effective for the treatment of TSLP-related diseases.

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Patent Document 2

Patent Document 3

[0008] This disclosure provides an anti-TSLP antibody. [Means for solving the problem]

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

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

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

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

[0014] In some embodiments, the above-described anti-TSLP antibody 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 with the heavy chain variable region shown in 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 with the light chain variable region shown in 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 with the heavy chain variable region shown in 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 with the light chain variable region shown in 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 with the heavy chain variable region shown in 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 with the light chain variable region shown in 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 with the heavy chain variable region shown in 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 with the light chain variable region shown in amino acid sequence SEQ ID NO: 13, 120, 121, 122, 123, 124, or 125.

[0015] In some embodiments of the anti-TSLP antibody described above, the anti-TSLP antibody is a humanized antibody which comprises a framework region or a framework region variant derived from a human antibody, the framework region variant having 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 heavy chain framework region of the human antibody, respectively.

[0016] In some embodiments of the anti-TSLP antibody described above, the framework region variant includes a reverse mutation selected from those described in (a), (b), (c), or (d) below: a) One or more amino acid reverse mutations selected from the group consisting of 46P, 47W, 58V, 70S, and 71Y, located within the framework region of the light chain variable region shown in SEQ ID NO: 38, 49, 51, or 52, and / or one or more amino acid reverse mutations selected from the group consisting of 38K, 48I, 67A, 69L, 71V, and 73K, located within the framework region of the heavy chain variable region shown in SEQ ID NO: 42 or 50; b) One or more amino acid reverse mutations selected from the group consisting of 1D, 4L, 43P, 48L, and 58I, located within the framework region of the light chain variable region shown in SEQ ID NO: 56, 59, 72, 73, 74, or 75, and / or one or more amino acid reverse mutations selected from the group consisting of 2A, 27F, 38K, 39H, 48I, 67A, 69L, 71V, and 76R, located within the framework region of the heavy chain variable region shown in SEQ ID NO: 62; c) One or more amino acid reverse mutations selected from the group consisting of 1S, 43S, 67Y, and 73F included in the framework region of the light chain variable region shown 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 reverse mutations selected from the group consisting of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I, and 82bR included in the framework region of the heavy chain variable region shown in SEQ ID NO: 85, 90, 91, 92, or 97; or d) One or more amino acid reverse mutations selected from the group consisting of 43S, 45Q, 48V, 66V, and 70Q, which are included in the framework region of the light chain variable region shown in SEQ ID NO:120, and / or one or more amino acid reverse mutations selected from the group consisting of 38K, 48I, 66K, 67A, 69L, 71V, 73K, and 78A, which are included in the framework region of the heavy chain variable region shown in SEQ ID NO:126.

[0017] In some embodiments, the above-described anti-TSLP antibody 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 shown in SEQ ID NO: 6, 42, 43, 44 or 50, and the amino acid sequence of the light chain variable region is as shown 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 shown 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 shown 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 shown 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 shown 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 sequences of the heavy chain variable region are as shown in SEQ ID NO: 12, 126, 127, 128, 129, 130, 131 or 132, and the amino acid sequences of the light chain variable region are as shown in SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125.

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

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

[0020] [Table 1]

[0021] [Table 2]

[0022] [Table 3]

[0023] [Table 4]

[0024] In some embodiments of the anti-TSLP antibody 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 their conventional variants, and the light chain constant region of the antibody constant region is selected from the group consisting of human antibody κ and λ chain constant regions and their conventional variants. More preferably, the antibody comprises a heavy chain constant region shown in SEQ ID NO:133 and a light chain constant region shown in SEQ ID NO:134.

[0025] In some embodiments, the above-described anti-TSLP antibody comprises the following heavy and light chains: (a) The amino acid sequence of the heavy chain is as shown 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 shown 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 shown 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 shown 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 shown 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 shown 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 shown 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 shown 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-described anti-TSLP antibody comprises the following heavy and light chains: (a) The amino acid sequence of the heavy chain is as shown in SEQ ID NO:135, and the amino acid sequence of the light chain is as shown in SEQ ID NO:136; (b) The amino acid sequence of the heavy chain is as shown in SEQ ID NO:137, and the amino acid sequence of the light chain is as shown in SEQ ID NO:138; (c) The amino acid sequence of the heavy chain is as shown in SEQ ID NO:139, and the amino acid sequence of the light chain is as shown in SEQ ID NO:140; or, (d) The amino acid sequence of the heavy chain is as shown in SEQ ID NO:141, and the amino acid sequence of the light chain is as shown in SEQ ID NO:142.

[0027] In some embodiments, the antibody binds competitively to human TSLP with the aforementioned anti-TSLP antibody or its antigen-binding fragment.

[0028] In another embodiment, the disclosure also provides nucleic acid molecules encoding anti-TSLP antibodies as described above.

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

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

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

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

[0033] In some embodiments, the present disclosure provides a method for immunodetection or determination of TSLP in vitro or ex vivo, which includes the step of using an anti-TSLP antibody as described above.

[0034] In some embodiments, the disclosure provides the use of such anti-TSLP antibodies in preparing reagents for immunodetection of human TSLP.

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

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

[0037] In some embodiments, the Disclosure provides the use of the above-described anti-TSLP antibodies, nucleic acid molecules, host cells, or pharmaceutical compositions in preparing a drug for the treatment of TSLP-related diseases, including asthma, idiopathic pulmonary fibrosis, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic sinusitis, urticaria, Netherton syndrome, eosinophilic esophagitis, food allergies, allergic diarrhea, eosinophilic gastroenteritis, and allergic bronchopulmonary aspergillosis. This includes, but is not limited to, fibrosis due to chronic fungal sinusitis, chronic pruritus, cancer, breast cancer, colorectal cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, chronic obstructive pulmonary disease, systemic sclerosis, multiple sclerosis, keloids, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, eosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, radiation fibrosis, and fibrosis due to wound healing.

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

[0039] In some embodiments, the present disclosure provides anti-TSLP antibodies for use as pharmaceutical agents, wherein the anti-TSLP antibodies are for use in the treatment of TSLP-related diseases, 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 allergic fungal infections. This includes, but is not limited to, sinusitis, chronic pruritus, cancer, breast cancer, colorectal cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, chronic obstructive pulmonary disease, systemic sclerosis, multiple sclerosis, keloids, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, eosinophilic syndrome, eosinophilic granulomatosis with polyangiitis, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, radiation fibrosis, and fibrosis due to wound healing. [Brief explanation of the drawing]

[0040] [Figure 1] This figure shows the results of inhibiting the binding activity of TSLP to the TSLP receptor by an antibody. [Figure 2] This figure shows the results of inhibiting the binding activity of TSLP to the cell surface TSLP receptor by an antibody. [Figure 3] This figure shows the results of inhibiting the proliferative activity of BaF3 cells induced by TSLP with an antibody. [Figure 4A] This figure shows the results of antibody activity inhibiting the production of the chemokine TARC induced by TSLP. [Figure 4B] This figure shows the results of antibody activity inhibiting the production of the chemokine OPG induced by TSLP. [Figure 5A] This figure shows the results of antibody activity inhibiting the production of the Th2 cytokine IL-13. [Figure 5B] This figure shows the results of antibody activity inhibiting the production of the Th2 cytokine IL-4. [Figure 5C]This figure shows the results of antibody activity inhibiting the production of the Th2 cytokine TNF-α. [Figure 5D] This figure shows the results of antibody activity inhibiting the production of the Th2 cytokine IL-5. [Modes for carrying out the invention]

[0041] term

[0042] To facilitate understanding of this disclosure, certain technical and scientific terms are defined below. Unless expressly defined herein, all other technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art.

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

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

[0045] As used herein, “antibody” refers to 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 constant regions of immunoglobulin heavy chains exhibit different amino acid compositions and sequences, and therefore exhibit different antigenicity. Thus, immunoglobulins can be classified into five types, or named immunoglobulin isotypes, namely IgM, IgD, IgG, IgA, and IgE, with corresponding heavy chains being μ, δ, γ, α, and ε chains, respectively. Ig of the same type can be further divided into different subclasses depending on the differences in the amino acid composition of the hinge region and the number and position of disulfide bonds in the heavy chain. For example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. The light chains can be divided into κ chains or λ chains depending on the differences in the constant region. Each of the five types of Ig can have either a κ chain or a λ chain.

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

[0047] The antibodies disclosed herein include mouse antibodies, chimeric antibodies, and humanized antibodies.

[0048] In this disclosure, the term “mouse antibody” refers to an anti-human TSLP monoclonal antibody prepared in accordance with the knowledge and art of those skilled in the art. During preparation, a subject is injected with the TSLP antigen, and then a hybridoma expressing an antibody having a desired sequence or functional property is isolated. In preferred embodiments of this disclosure, the mouse anti-TSLP antibody or its antigen-binding fragment may further comprise the light chain constant region of the mouse κ chain, λ chain or a variant thereof, or the heavy chain constant region of mouse IgG1, IgG2, IgG3 or a variant 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 the mouse antibody. Establishing a chimeric antibody requires first establishing a hybridoma that secretes a mouse-specific monoclonal antibody, then cloning the variable region gene from the mouse hybridoma cells, and, if necessary, cloning the constant region gene of a human antibody. The mouse variable region gene is then combined with the human constant region gene to form a chimeric gene for insertion into an expression vector, and finally, the chimeric antibody molecule is expressed in a eukaryotic or prokaryotic system. In a preferred embodiment of this disclosure, the light chain of the TSLP chimeric antibody further comprises a light chain constant region derived from a human κ, λ chain or a variant thereof. The heavy chain of the TSLP chimeric antibody further comprises the heavy chain constant region of human IgG1, IgG2, IgG3, IgG4 or a variant thereof, preferably the heavy chain constant region of human IgG1, IgG2, or IgG4, or a variant of IgG1, IgG2, or IgG4, having an amino acid mutation (e.g., L234A or L235A mutation, and / or S228P mutation).

[0050] The term "humanized antibody," also known as a CDR-transplanted antibody, refers to an antibody produced by transplanting a mouse CDR sequence into a human antibody variable region framework; that is, an antibody produced with a different type of human germline antibody framework sequence. Because it carries a large amount of mouse protein components, it can overcome the heterogeneous reactions caused by chimeric antibodies. Such framework sequences can be obtained from publicly available DNA databases and literature containing germline antibody gene sequences. For example, germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database (www.mrccpe.com.ac.uk / vbase) and in Kabat, EA et al., *Sequences of Proteins of Immunological Interest*, 5th edition, 1991. To avoid the decrease in activity simultaneously caused by decreased immunogenicity, minimal reverse or reverse mutations can be introduced into the human antibody variable region framework sequence to maintain activity. The humanized antibodies in this disclosure also include humanized antibodies in which CDR affinity maturation is performed by yeast display.

[0051] CDR transplantation can lead to a decrease in the affinity of the produced antibody or its antigen-binding fragment to the antigen due to changes in the framework residues in contact with the antigen. Such interactions may result from hypermutations in somatic cells. Therefore, transplantation of such donor framework amino acids into the framework of humanized antibodies may still be necessary. By examining the sequence and structure of the variable region of animal monoclonal antibodies, amino acid residues of non-human antibodies or their antigen-binding fragments that are involved in antigen binding can be identified. Residues of CDR donor frameworks that differ from germline can be considered relevant. If the closest germline cannot be determined, the sequence can be compared to a consensus sequence of a subclass or animal antibody sequence with a high degree of similarity. Rare framework residues are thought to be the result of hypermutations in somatic cells and therefore are thought to play a crucial role in binding.

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

[0053] The "conventional variants" of the human antibody heavy chain constant region and human antibody light chain constant region described in this disclosure refer to variants of the heavy chain constant region or light chain constant region disclosed in the prior art that do not alter the structure and function of the antibody variable region. Exemplary variants include heavy chain constant region variants of IgG1, IgG2, IgG3, or IgG4 that have site-directed 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 the knob-in-hole structure known in the art (giving the antibody heavy chain a combination of knob-Fc and hole-Fc). These mutations have been shown to impart new properties to the antibody without altering the function of the antibody variable region.

[0054] "Human antibodies (HuMAb)," "human-derived antibodies," "whole-human antibodies," and "fully human antibodies" are interchangeable terms and can be derived from humans or from genetically modified organisms "engineered" to produce specific human antibodies in response to antigen stimulation, and can 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 endogenous heavy and light chain loci have been targeted and disrupted. Transgenic organisms can synthesize human antibodies specific to human antigens, and human antibody-secreting hybridomas can be constructed using these organisms. Human antibodies can also be antibodies whose heavy and light chains are encoded by nucleotide sequences derived from one or more human DNA sources. Fully human antibodies can also be constructed by gene or chromosome transfection methods and phage display techniques, or by activated B cells in vitro, 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, distinct from the antigen-binding fragment defined below. Specifically, these terms refer to an antibody whose light and heavy chains contain a constant region. “Antibody” as used in this disclosure includes “full-length antibody” and its antigen-binding fragment.

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

[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 is known that fragments of full-length antibodies can be used to perform the antigen-binding function of the antibody. Examples of binding fragments included in the term "antigen-binding fragment" of an antibody include: (i) a monovalent fragment consisting of Fab fragments, VL, VH, CL, and CH1 domains; (ii) a bivalent fragment containing two Fab fragments linked by a disulfide bond in the hinge region, F(ab')2 fragments; (iii) an Fd fragment consisting of a VH domain and a CH1 domain; (iv) an Fv fragment consisting of the VH and VL domains of a single arm of the antibody; (v) dsFv, a stable antigen-binding fragment formed by an interchain disulfide bond between VH and VL; and (vi) diabodies, bispecific antibodies, and multispecific antibodies containing fragments such as scFv, dsFv, and Fab. Furthermore, although the two domains VL and VH of the Fv fragment are encoded by separate genes, they can be linked together using a synthetic linker via recombinant DNA techniques to produce a single protein chain in which the VL and VH regions pair up to form a monovalent molecule (referred to as single-chain Fv (scFv)) (see, for example, Bird et al. (1988) Science vol. 242: pp. 423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci USA vol. 85: pp. 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 skilled in the art and are screened for utility, just like intact antibodies. The antigen-binding portion may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact immunoglobulins. The antibody may be of a different isotype, for example, IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtype), IgA1, IgA2, IgD, IgE, or IgM antibody.

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

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

[0060] Fab' has a molecular weight of approximately 50,000 and is an antibody fragment having light-binding activity obtained by cleaving a disulfide bond in the hinge region of F(ab')2. Fab' of this disclosure can be prepared by treating 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] Furthermore, Fab' can be produced by inserting the DNA encoding the Fab' fragment of an antibody into a prokaryotic or eukaryotic expression vector, and then introducing the vector into a prokaryote or eukaryote to express Fab'.

[0062] The terms "single-chain antibody," "single-chain Fv," or "scFv" refer to molecules containing an antibody heavy chain variable domain (or region; VH) and an antibody light chain variable domain (or region; VL) linked by a linker. Such scFv molecules have the general structure of NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Appropriate linkers in the prior art consist of repeating GGGGS amino acid sequences or their variants, for example, variants having 1 to 4 repeats (Holliger et al., (1993), Proc. Natl. Acad. Sci. USA vol. 90: pp. 6444-6448). Other linkers available for use in this disclosure include those described in Alfthan et al. (1995), Protein Eng. Vol. 8: pp. 725-731, Choi et al. (2001), Eur. J. Immunol. Vol. 31: pp. 94-106, Hu et al. (1996), Cancer Res. Vol. 56: pp. 3055-3061, Kipriyanov et al. (1999), J. Mol. Biol. Vol. 293: pp. 41-56, and Roovers et al. (2001), Cancer Immunol.

[0063] A Diabody is an antibody fragment formed by the dimerization of scFv or Fab, and is an antibody fragment having bivalent antigen-binding activity. In this bivalent antigen-binding activity, the two antigens may be the same or different.

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

[0065] The diabody of this disclosure can be fabricated by the following steps: Obtain the VH and VL coding of the monoclonal antibody of this disclosure that specifically recognizes human TSLP and binds to the amino acid sequence or three-dimensional structure of the extracellular region; construct the DNA encoding scFv; adjust the length of the amino acid sequence of the peptide linker to 8 residues or less; 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 conjugating VH polypeptide and VL polypeptide, in which one amino acid residue in each of the VH polypeptide and VL polypeptide is replaced by a cysteine ​​residue via a disulfide bond between cysteine ​​residues. The amino acid residue to be replaced by the cysteine ​​residue can be selected based on the three-dimensional structure prediction of the antibody, according to a known method (Protein Engineering, Vol. 7, p. 697 (1994)).

[0067] The full-length antibodies or antigen-binding fragments of the present disclosure can be produced by the following steps: obtaining cDNA encoding VH and VL of the 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 terms "amino acid difference" or "amino acid mutation" refer to the presence of changes or mutations in amino acids in a mutant protein or polypeptide compared to the original protein or polypeptide, including the occurrence of one, two, three or more amino acid insertions, deletions, or substitutions relative to the original protein or polypeptide.

[0069] The terms "antibody framework" or "FR region" refer to a portion of the variable domain of the VL or VH that functions as a scaffold for the antigen-binding loop (CDR) of this variable domain. Essentially, it is a variable domain without a CDR.

[0070] The terms "complementarity-determining region," "CDR," or "hypervariable region" refer to the region within the antibody's variable domain that primarily contributes to antigen binding, among the six hypervariable regions. Generally, each heavy chain variable region has three CDRs (HCDR1, HCDR2, HCDR3), and each light chain variable region has three CDRs (LCDR1, LCDR2, LCDR3). The amino acid sequence boundaries of CDRs can be determined using any of the various well-known schemes, including the "Kabat" numbering system (Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD), the "Chothia" numbering system (Al-Lazikani et al., (1997) JMB Vol. 273: pp. 927-948), and the ImmunoGenTics (IMGT) numbering system (Lefranc MP, Immunologist, Vol. 7, pp. 132-136 (1999); Lefranc, MP et al., Dev. Comp. Immunol., Vol. 27, pp. 55-77 (2003)). For example, in the traditional format, according to Kabat's rules, the amino acid residue numbers of the CDRs in the heavy chain variable domain (VH) are 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3), and the amino acid residue numbers of the CDRs in the light chain variable domain (VL) are 24-4 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). According to Chothia's rules, the amino acid residue numbers of the CDRs in 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 definitions of CDR, the CDR consists 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 CDRs in VH are approximately 26 - 35 (CDR1), 51 - 57 (CDR2), and 93 - 102 (CDR3), and the amino acid residue numbers of CDRs in VL are approximately 27 - 32 (CDR1), 50 - 52 (CDR2), and 89 - 97 (CDR3). According to the IMGT rules, the CDR regions of antibodies can be determined by the IMGT / DomainGap Align program.

[0071] The term "epitope" or "antigenic determinant" refers to the site on an antigen to which an immunoglobulin or antibody specifically binds (e.g., a specific site on the TSLP molecule). An epitope usually contains 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, for example, "Epitope Mapping Protocols in Methods in Molecular Biology", Volume 66, edited by G.E. 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. Usually, an antibody binds with an affinity (KD) of less than about 10 -9 M, less than about 10 -10 M, less than about 10 -11 M, less than about 10 -12 M, or less, such as less than about 10 -8 M.

[0073] The term "KD" refers to the dissociation equilibrium constant of a specific antibody - antigen interaction. Generally, the antibodies of the present disclosure bind to TSLP with an affinity (KD) of less than about 10 -7 M, for example, less than about 10 -8 M or less than about 10 -9 M. In the present disclosure, however, the affinity of an antibody for a cell - surface antigen is measured by FACS or the Biacore method to determine the KD value.

[0074] The term "competition," when used in the context of antigen-binding proteins competing for the same epitope (e.g., neutralizing antigen-binding proteins or neutralizing antibodies), means that competition occurs between antigen-binding proteins, which is measured by the following assay: in this assay, 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 common antigen (e.g., TSLP antigen or a fragment thereof). Numerous types of competitive binding assays can be used to determine whether an antigen-binding protein competes with other proteins. These assays include, for example, solid-phase direct or indirect radioimmunoassays (RIAs), solid-phase direct or indirect enzyme immunoassays (EIAs), sandwich competition assays (see Stahli et al., 1983, Methods in Enzymology, Vol. 9: pp. 242-253), solid-phase direct biotin-avidin EIA (see Kirkland et al., 1986, J.Immunol., Vol. 137: pp. 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 I-125 labels (see Morel et al., 1988, Molec.Immunol., Vol. 25: pp. 7-15), and solid-phase direct biotin-avidin EIA (see Cheung et al., 1990, Virology). This includes direct labeling RIAs (see Vol. 176: pp. 546-552) and direct labeling RIAs (see Moldenhauer et al., 1990, Scand. J. Immunol. Vol. 32: pp. 77-82). Generally, assays involve binding purified antigens bound to a solid surface or cell using either an unlabeled test antigen-binding protein or a labeled reference antigen-binding protein. Competitive inhibition is measured by determining the amount of label bound to the solid surface or cell 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 (competitive antigen-binding proteins) include: antigen-binding proteins that bind to the same epitope as the reference antigen-binding protein, and antigen-binding proteins that bind to an adjacent epitope that is sufficiently close to the binding epitope of the reference antigen-binding protein. These two epitopes sterically interfere with each other's binding. Generally, an excess of competing antigen-binding proteins inhibits (or reduces) specific binding between the reference antigen-binding protein and the 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] As used herein, the term “nucleic acid molecule” 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. When nucleic acids are placed in a functional relationship with another nucleic acid sequence, they are “operably ligated.” For example, if a promoter or enhancer affects the transcription of a coding sequence, the promoter or enhancer is operably ligated to the coding sequence.

[0076] "Sequence identity" of amino acids 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 to maximize sequence identity (with gaps introduced as necessary). Conservative substitutions are not considered part of sequence identity. For the purpose of determining the percentage of amino acid sequence identity, alignment can be achieved using various methods within the technical scope of this art, such as publicly available computer software like BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR) software. Those skilled in the art can determine parameters suitable for measuring alignment, including any algorithm 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 is ligated. In one embodiment, the vector is a “plasmid,” which refers to a circular double-stranded DNA loop to which additional DNA segments can be ligated. In another embodiment, the vector is a viral vector to which additional DNA segments can be ligated into a viral genome. The vectors disclosed herein can replicate autonomously in the introduced host cell (e.g., bacterial vectors with a bacterial origin of replication and episomal mammalian vectors), or, after being introduced into a host cell, can be integrated into the host cell's genome and replicate together with the host genome (e.g., non-episomal mammalian vectors).

[0078] Methods for generating and purifying antibodies and antigen-binding fragments are well known in the art and can be found, for example, in the Antibody Experiment 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 regenerated, purified, and sequenced using conventional methods. Antigen-binding fragments can also be prepared using conventional methods. The antibodies or antigen-binding fragments of this disclosure are genetically engineered to add one or more human FR regions to a non-human CDR region. The germline sequences of human FR 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 FactsBook, 2001, ISBN 012441351.

[0079] The term "host cell" refers to the cell into which the expression vector has been introduced. Host cells can include bacteria, microorganisms, plant, or animal cells. Easily transformable bacteria include members of the Enterobacteriaceae family, such as strains of Escherichia coli or Salmonella; Bacillaceae family, such as Bacillus subtilis; Pneumococcus; Streptococcus; and members of Haemophilus influenzae. Suitable microorganisms include Saccharomyces cerevisiae and Pichia pastoris. Suitable animal host cell lines include CHO (Chinese hamster ovary strain) cells and NSO cells.

[0080] The manipulated antibodies or antigen-binding fragments in this disclosure can be prepared and purified by conventional methods. For example, cDNA sequences encoding heavy and light chains can be cloned and recombinant into a GS expression vector. The recombinant immunoglobulin expression vector can be stably transfected into CHO cells. As a more preferred prior art, mammalian expression systems can induce antibody glycosylation, particularly at the highly conserved N-terminus of the Fc region. Stable clones can be obtained by expressing antibodies that specifically bind to human TSLP. Positive clones are grown in serum-free medium in a bioreactor to produce antibodies. The medium from which the antibodies are secreted can be purified by conventional methods. For example, it can be purified using a Sepharose FF column of type A or G equilibrated with a prepared buffer. Nonspecifically bound components are washed away, the bound antibodies are eluted by a pH gradient, and antibody fragments are detected and recovered by SDS-PAGE. The antibodies can be filtered and concentrated by conventional methods. Soluble mixtures and polymers can also be removed by conventional methods, such as molecular sieving or ion exchange. The resulting product needs to be immediately frozen at -70°C or similar temperature, or freeze-dried.

[0081] "Administer" or "treat" means, when applied to animals, humans, research subjects, cells, tissues, organs, or body fluids, bringing an exogenous drug, therapeutic agent, diagnostic agent, or composition into contact with an animal, human, research subject, cell, tissue, organ, or body fluid. "Administer" and "treat" can also refer, for example, to treatment, pharmacokinetics, diagnostics, research, and experimental methods. Cell therapy includes bringing a reagent into contact with cells and bringing a reagent into contact with the fluid in which the cells are in contact. "Administer" and "treat" can also refer, for example, to the treatment of cells by another cell in vitro and ex vivo, with a reagent, diagnostic, or conjugated composition. "Treat" means, when applied to humans, animals (veterinary), or research subjects, to therapeutic procedures, preventive measures, research, and diagnostic applications.

[0082] "To treat" means to administer, internally or externally, a therapeutic agent, such as a composition containing any one of the conjugate compounds of this disclosure, to a patient having one or more disease symptoms for which the therapeutic agent is known to be therapeutic. Typically, the therapeutic agent is administered in an amount effective to alleviate, induce regression of, or suppress the onset of one or more disease symptoms in the patient or population being treated to any clinically measurable degree. The amount of therapeutic agent effective to alleviate a particular disease symptom (also referred to as the "therapeutic dose") may vary depending on the patient's condition, age and weight, and the ability of the drug to produce the desired therapeutic effect in the patient. Whether the symptoms of the disease have been alleviated can be assessed by any clinical trial method commonly used by a physician or other healthcare professional to assess the severity or progression of the symptoms. Embodiments of the present disclosure (e.g., a treatment method or product) should alleviate a target disease symptom 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] "Isomorphic modification" or "isomorphic exchange or substitution" refers to the substitution of an amino acid in a protein with another amino acid that has similar properties (e.g., charge, side chain size, hydrophobic / hydrophilicity, skeletal conformation and stiffness), and these modifications can often be made without altering the biological activity of the protein. Those skilled in the art generally recognize that a single amino acid substitution in a non-essential region of a polypeptide does not substantially alter its biological activity (see Watson et al., Molecular Biology of the Gene (1987), The Benjamin / Cummings Pub. Co., p. 224, 4th edition). Furthermore, substitutions of amino acids with similar structure or function are unlikely to interfere with biological activity. Exemplary isomorphic substitutions are listed in the table below, "Exemplary Amino Acid Isomorphic Substitutions."

[0084] [Table 5]

[0085] “Effective dose” means the amount of drug, compound, or pharmaceutical composition required to obtain any one or more beneficial or desired therapeutic outcome. In the case of prophylactic use, beneficial or desired outcomes include the elimination or reduction of risk, a reduction in the severity or delay of disease onset, including the biochemical, histological, and / or behavioral symptoms of the disease, their complications, and intermediate pathological phenotypes that appear during their development. In the case of therapeutic use, beneficial or desired outcomes include clinical outcomes such as a reduction in the incidence of various target antigen-associated disorders of this disclosure, improvement of one or more symptoms of the disorder, a reduction in the dose of other drugs required to treat the disorder, an improvement in the therapeutic effect of another drug, and a delay in the progression of a patient's disorder associated with the target antigen of this disclosure.

[0086] "Exogenous" refers to substances produced outside of living organisms, cells, or the human body, depending on the circumstances. "Endogenous" refers to substances produced within cells, living organisms, or the human body, depending on the circumstances.

[0087] "Homologousity" refers to the similarity between two polynucleotide sequences or two polypeptides. For example, if the positions of two sequences being compared are occupied by the same base or amino acid monomer subunit, such as when each position in two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The homology rate 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 and multiplied by 100. For example, in optimal sequence alignment, if 6 out of 10 positions of two sequences are matching or homologous, the two sequences are 60% homologous; if 95 out of 100 positions of two sequences are matching or homologous, the two sequences are 95% homologous. Generally, when aligning two sequences, the comparison is performed in a way that maximizes the homology rate. For example, the comparison can be performed using the BLAST algorithm, and the algorithm's parameters are selected to give the greatest match between each sequence across the entire length of each reference sequence. The following references pertain to the BLAST algorithm, which is commonly used in sequence analysis: BLAST ALGORITHMS: Altschul, SF et al., (1990) J.Mol.Biol. 215: pp. 403-410; Gish, W. et al., (1993) Nature Genet. 3: pp. 266-272; Madden, TL et al., (1996) Meth.Enzymol. 266: pp. 131-141; Altschul, SF et al., (1997) Nucleic Acids Res. 25: pp. 3389-3402; Zhang, J. et al., (1997) Genome Res. 7: pp. 649-656. Other conventional BLAST algorithms, such as those provided by NCBI BLAST, are also well known to those skilled in the art.

[0088] The terms “cells,” “cell lines,” and “cell cultures” used herein are interchangeable, and all such terms include offspring. Therefore, the terms “transformed organisms” and “transformed cells” include primary test cells and cultures derived therefrom, regardless of the number of passages. It should also be understood that, due to intentional or unintentional mutations, not all offspring will be exactly the same in terms of DNA content. This includes mutant offspring that have the same function or biological activity as those screened in the initially transformed cells. Where something else is being referred to, 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 trace amounts of nucleic acids, RNA, and / or DNA, as described, for example, in U.S. Patent 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, and these primers have sequences identical or similar to those of the strand corresponding to the template being amplified. The 5' terminal nucleotides of two primers may be identical to those of the ends of the material being amplified. PCR can be used to amplify specific RNA sequences, specific DNA sequences from whole-genomic DNA, and cDNA sequences transcribed from whole-cellular RNA, phage, or plasmid sequences. See Mullis et al. (1987) Cold Spring Harbor, Symp. Ouant. Biol. Vol. 51: 263; Erlich ed., (1989) PCR TECHNOLOGY (Stockton Press, NY). As used herein, PCR is considered an example, but not the only, of nucleic acid polymerase reaction methods for amplifying nucleic acid test samples. This method involves using known nucleic acids as primers and nucleic acid polymerases to amplify or generate specific sites of nucleic acids.

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

[0091] "Optional" or "at the discretion of the user" means that the events or environments described below may occur, but are not necessarily guaranteed to occur. This description includes cases where the events or environments occur and cases where they do not.

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

[0093] The term "pharmaceutically acceptable carrier" refers to any inert substance suitable for use in formulations for the delivery of antibodies or antigen-binding fragments. Carriers can be anti-adhesion agents, binders, coatings, disintegrants, fillers or diluents, preservatives (such as antioxidants, antibacterial or antifungal agents), sweeteners, absorption retarders, wetting agents, emulsifiers, buffers, etc. Examples of suitable pharmaceutically acceptable carriers include water, ethanol, polyols (e.g., glycerol, propanediol, polyethylene glycol, etc.), dextrose, vegetable oils (e.g., olive oil), saline, buffers, and isotonic agents, such as sugars, polyols, sorbitol, and sodium chloride.

[0094] Furthermore, this disclosure includes agents for treating TSLP-related diseases, comprising the anti-TSLP antibody or its antigen-binding fragment as an active ingredient.

[0095] The TSLP-related diseases described herein are not particularly limited, as long as they are diseases associated with TSLP. For example, therapeutic responses induced by the molecules of this disclosure can be achieved by binding to human TSLP and then blocking the binding of TSLP to its receptor, or by killing cells that overexpress TSLP.

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

[0097] In this disclosure, the method used to detect or measure the amount of the target antigen (e.g., TSLP) may be any known method, including, for example, immunodetection and measurement methods.

[0098] Immunodetection or measurement methods are methods for detecting or measuring the amount of an antibody or antigen using a labeled antigen or antibody. Examples of immunodetection or measurement methods include radioimmunoassay (RIA), enzyme immunoassay (EIA or ELISA), fluorescence immunoassay (FIA), luminescence immunoassay, Western blotting, and physicochemical methods.

[0099] The aforementioned TSLP-related diseases can be diagnosed by detecting or measuring cells expressing TSLP using the antibodies or antibody fragments of this disclosure.

[0100] To detect cells expressing polypeptides, known immunodetection methods can be used, preferably immunoprecipitation, fluorescent cell staining, or immunohistochemical staining. Fluorescent antibody staining using the FMAT8100HTS system (Applied Biosystems) can also be used.

[0101] In this disclosure, the in vivo sample used for the detection or measurement of a target antigen (e.g., TSLP) is not particularly limited insofar as it may contain cells expressing the target antigen (e.g., TSLP), and may include, for example, histiocytes, blood, plasma, serum, pancreatic juice, urine, feces, tissue fluid, or culture medium.

[0102] Depending on the required diagnostic method, a diagnostic agent comprising the monoclonal antibody or an antibody fragment of the present disclosure may also include reagents for carrying out an antigen-antibody reaction or reagents for detecting that reaction. Reagents used to carry out an antigen-antibody reaction include buffers, salts, etc. Reagents used for detection include reagents commonly used in immunoassay or assay methods, such as a monoclonal antibody, a labeled secondary antibody that recognizes its antibody fragment or conjugate, and a substrate corresponding to the label.

[0103] The above-mentioned specification provides details of one or more embodiments of the present disclosure. The present invention can be carried out or tested using any methods and materials similar to or identical to those described herein, but preferred methods and materials are described below. Other features, purposes and advantages of the present disclosure will become apparent in the specification and claims. In the specification and claims, unless the context clearly indicates otherwise, singular nouns include plural subjects. Unless otherwise defined, all technical and scientific terms used herein have the general meanings understood by those skilled in the art in which the present invention pertains. All patents and publications cited herein are incorporated by reference. The following examples are provided to more comprehensively illustrate preferred embodiments of the present invention. These examples should not be construed as limiting the scope of the present invention in any way, and the scope of the present invention 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 includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:14, SEQ ID NO:15, and SEQ ID NO:47, respectively, and the light chain variable region includes LCDR1 and LCDR2, indicated in SEQ ID NO:17 and SEQ ID NO:18, respectively, and LCDR3, indicated in SEQ ID NO:48 or 55. The sequence for SEQ ID NO:47 is EDYDYDGYAMDX1, the sequence for SEQ ID NO:48 is QQWSSX2RT, and the sequence for 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, X5 is G or E; or ii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:76, SEQ ID NO:24, and SEQ ID NO:25, respectively. The sequence for SEQ ID NO:76 is RASESVDX6SGLSFMH, where X6 is selected from N, S, and Q; or iii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:26, SEQ ID NO:96, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:29, SEQ ID NO:118, and SEQ ID NO:31, respectively. The sequence for SEQ ID NO:96 is VIDPGX7X8DTNYNE, and the sequence for SEQ ID NO:118 is X9VX 10 X 11 X 12 X 13 T is selected from N, Q, and V, X8 is G or V, X9 is Y or E, X 10 is selected from S, D, and E, 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 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 shown in SEQ ID NO:35, SEQ ID NO:36, and SEQ ID NO:37, respectively. [Embodiment 2] Includes a heavy chain variable region and a light chain variable region, i) The heavy chain variable region includes HCDR1 and HCDR2 shown in SEQ ID NO: 14 and SEQ ID NO: 15, respectively, and HCDR3 shown in SEQ ID NO: 16 or 45, and the light chain variable region includes LCDR1 and LCDR2 shown in SEQ ID NO: 17 and SEQ ID NO: 18, respectively, and LCDR3 shown in SEQ ID NO: 19, 46, 53 or 54; or ii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3 shown in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and the light chain variable region includes LCDR2 and LCDR3 shown in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 shown in SEQ ID NO:23, 70, or 71; or, iii) The heavy chain variable region includes HCDR1 and HCDR3 shown in SEQ ID NO: 26 and SEQ ID NO: 28, respectively, and HCDR2 shown in SEQ ID NO: 27, 93, 94, or 95, and the light chain variable region includes LCDR1 and LCDR3 shown in SEQ ID NO: 29 and SEQ ID NO: 31, respectively, and LCDR2 shown in SEQ ID NO: 30, 108, 109, 110, 111, 112, 113, 114, 115, 116, or 117, The anti-TSLP antibody described in Embodiment 1. [Embodiment 3] Includes a heavy chain variable region and a light chain variable region, i) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:14, SEQ ID NO:15, and SEQ ID NO:45, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:54, respectively; or, ii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:14, SEQ ID NO:15, and SEQ ID NO:45, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:53, respectively; or, iii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:70, SEQ ID NO:24, and SEQ ID NO:25, respectively; or, iv) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:71, SEQ ID NO:24, and SEQ ID NO:25, respectively; or, v) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:26, SEQ ID NO:94, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:29, SEQ ID NO:113, and SEQ ID NO:31, respectively; or, vi) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:26, SEQ ID NO:94, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, respectively; or, vii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:14, SEQ ID NO:15, and SEQ ID NO:16, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19, respectively; or, viii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:14, SEQ ID NO:15, and SEQ ID NO:45, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:46, respectively; or, ix) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:23, SEQ ID NO:24, and SEQ ID NO:25, respectively; or, x) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:26, SEQ ID NO:27, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, respectively; or, xi) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:26, SEQ ID NO:93, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, respectively; or, xii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, indicated in SEQ ID NO:26, SEQ ID NO:95, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, indicated in SEQ ID NO:29, SEQ ID NO:30, and SEQ ID NO:31, respectively; or, xiii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3 shown in SEQ ID NO:26, SEQ ID NO:27, and SEQ ID NO:28, respectively, and the light chain variable region includes LCDR1 and LCDR3 shown in SEQ ID NO:29 and SEQ ID NO:31, respectively, and LCDR2 shown in SEQ ID NO:108, 109, 110, 111, 112, 113, 114, 115, 116, or 117, The anti-TSLP antibody described in 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 according to 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) The heavy chain variable region includes HCDR1 and HCDR2 shown in SEQ ID NO: 14 and SEQ ID NO: 15, respectively, and HCDR3 shown in SEQ ID NO: 16 or 45, and the framework region includes up to 10 reverse mutations, preferably one or more of 38K, 48I, 67A, 69L, 71V, and 73K, and / or the light chain variable region includes LCDR1 and LCDR2 shown in SEQ ID NO: 17 and SEQ ID NO: 18, respectively, and LCDR3 shown in SEQ ID NO: 19, 46, 53, or 54, and the framework region includes up to 10 amino acid reverse mutations, preferably one or more of 46P, 47W, 58V, 70S, and 71Y; b) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3 shown in SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22, respectively, and its framework region includes up to 10 reverse 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 includes LCDR2 and LCDR3 shown in SEQ ID NO:24 and SEQ ID NO:25, respectively, and LCDR1 shown in SEQ ID NO:23, 70, or 71, and its framework region includes up to 10 amino acid reverse mutations, preferably selected from one or more of 1D, 4L, 43P, 48L, and 58I; c) The heavy chain variable region includes HCDR1 and HCDR3 shown in EQ ID NO:26 and SEQ ID NO:28, respectively, and HCDR2 shown in SEQ ID NO:27, 93, 94, or 95, and the framework region includes up to 10 revert mutations, preferably one or more of 27Y, 28A, 38K, 48I, 66K, 67A, 69L, 80I, and 82bR, and / or the light chain variable region includes LCDR1 and LCDR3 shown in SEQ ID NO:29 and SEQ ID NO:31, respectively, and SEQ ID The LCDR2 includes the one shown in NO:30, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117, and its framework region includes up to 10 revert mutations, preferably the revert mutations are selected from one or more of 1S, 43S, 67Y and 73F; or, d) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3 shown in SEQ ID NO:32, SEQ ID NO:33, and SEQ ID NO:34, respectively, and its framework region includes up to 10 revert mutations, preferably selected from one or more of 38K, 48I, 66K, 67A, 69L, 71V, 73K, and 78A, and / or the light chain variable region includes LCDR1, LCDR2, and LCDR3 shown in SEQ ID NO:35, SEQ ID NO:36, and SEQ ID NO:37, respectively, and its framework region includes up to 10 revert mutations, preferably selected from one or more of 43S, 45Q, 48V, 66V, and 70Q. [Embodiment 6] Includes 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in amino acid sequence SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125, The anti-TSLP antibody described in Embodiment 4. [Embodiment 7] Includes a heavy chain variable region and a light chain variable region, i) The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 6, 42, 43, 44 or 50, and the amino acid sequence of the light chain variable region is shown 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 shown 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 shown 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 shown 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 shown 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 shown 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 shown in SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125, The anti-TSLP antibody described in Embodiment 6. [Embodiment 8] The anti-TSLP antibody according to 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 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. [Embodiment 9] The anti-TSLP antibody according to 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 their conventional variants, and the light chain constant region is selected from the group consisting of human antibody κ chain and λ chain constant regions and their conventional variants, and more preferably the antibody comprises a heavy chain constant region shown in SEQ ID NO:133 and a light chain constant region shown in SEQ ID NO:134. [Embodiment 10] The anti-TSLP antibody according to Embodiment 9, wherein the antibody comprises the heavy chain and light chain shown below. a) The amino acid sequence of the heavy chain is as shown in SEQ ID NO:135, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown 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 shown in SEQ ID NO:137, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown 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 shown in SEQ ID NO:139, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown 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 shown in SEQ ID NO:141, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown in SEQ ID NO:142, or has at least 90% sequence identity thereto. [Embodiment 11] An isolated anti-TSLP antibody that competitively binds to human TSLP with the anti-TSLP antibody described in any of Embodiments 1 to 10. [Embodiment 12] A nucleic acid molecule encoding an anti-TSLP antibody as described in any one of Embodiments 1 to 11. [Embodiment 13] A host cell containing the nucleic acid molecule described in Embodiment 12. [Embodiment 14] A pharmaceutical composition comprising a therapeutically effective amount of an anti-TSLP antibody according to any of Embodiments 1 to 11, a nucleic acid molecule according to Embodiment 12, or a host cell according to Embodiment 13, and one or more pharmaceutically acceptable carriers, diluents, buffers, or excipients. [Embodiment 15] A method for immunodetecting or determining TSLP in vitro or ex vivo, comprising the step of using an anti-TSLP antibody described in any of Embodiments 1 to 11. [Embodiment 16] A kit comprising the anti-TSLP antibody described in any of Embodiments 1 to 11. [Embodiment 17] A method for treating a TSLP-related disease, comprising administering a therapeutically effective amount of an anti-TSLP antibody according to any of Embodiments 1 to 11, a nucleic acid molecule according to Embodiment 12, a host cell according to Embodiment 13, or a pharmaceutical composition according to Embodiment 14, wherein the TSLP-related disease is preferably 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, colorectal cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, or chronic obstructive pulmonary disease. A method selected from the group consisting of systemic sclerosis, multiple sclerosis, keloids, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, eosinophilia syndrome, eosinophilic granulomatosis with polyangiitis, inflammatory bowel disease, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, radiation-induced fibrosis, and fibrosis due to wound healing.

[0104] Examples

[0105] The following embodiments are incorporated to further illustrate the present disclosure, but they do not limit the scope of the present disclosure.

[0106] Experimental methods in the examples or test cases of this disclosure where conditions are not specified generally follow conventional conditions or conditions recommended by the manufacturer of the raw materials or 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 whose manufacturers are not specified are commercially available conventional reagents.

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

[0108] Expression plasmids were constructed by loading His-tagged human TSLP and cyno-TSLP, human IgG1-Fc-tagged human TSLP and cyno-TSLP, and sequences encoding the TSLP receptor extracellular domain sequence into a phr vector, and then transfecting HEK293 cells. The specific transfection procedure was as follows: The day before, HEK293E cells were placed in Freestyle expression medium (containing 1% FBS) at a rate of 0.8 × 10⁶. 6Cells were seeded at 1 / ml and placed in a 37°C constant temperature stirrer (120 rpm) for 24 hours of incubation. After 24 hours, the transfection plasmid and transfection reagent PEI were sterilized through a 0.22 μm filter. The transfection plasmid was then prepared at 100 μg / 100 ml of cells, with a PEI (1 mg / ml) to plasmid mass ratio of 3:1. For example, when transfecting 200 ml of HEK293E cells, 10 ml of Opti-MEM and 200 μg of plasmid were taken, mixed well, and allowed to stand for 5 minutes; another 10 ml of Opti-MEM and 600 μg of PEI were taken, mixed well, and allowed to stand for 5 minutes. The plasmid and PEI were thoroughly mixed and allowed to stand for 15 minutes. It is best not to exceed 20 minutes. The plasmid and PEI mixture was slowly added to 200 ml of HEK293E cells, and the culture was placed on a stirrer at 8% CO2, 120 rpm, and 37°C. On day 3 of transfection, the culture was supplemented with 10% of the culture medium. Samples were collected up to day 6 of transfection, centrifuged at 4500 rpm for 10 minutes, and the cell supernatant was collected. 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 of 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 portion is the signal peptide sequence, and the shaded portion 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 italicized 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, and the italicized 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 italicized portion is the Linker Human Fc tag. SEQ ID NO:4

[0113] 5. Amino acid sequence of the Fc-tagged human TSLP receptor extracellular domain (human-TSLPR-Fc-ECD) [ka] Note: The underlined portion is the human-TSLPR extracellular domain, and the italicized portion 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 then filtered. The nickel column was equilibrated with PBS solution and washed with 10 times the column volume. The filtered supernatant sample was applied to the column. A 280The column was washed with PBS solution containing 30 mM imidazole until the readings decreased to baseline. The target protein was then eluted with PBS solution containing 300 mM imidazole, and the eluted peaks were collected. The proteins were concentrated and transferred to PBS, and after being identified as correct by LC-MS, aliquots were obtained. 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 sample was centrifuged at high speed to remove impurities. The recombinant antibody expression supernatant was purified using a Protein A column. The column was washed with PBS until the A280 reading decreased 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, replaced with fresh solution, identified as correct by electrophoresis and LC-MS, and then 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 the human TSLP receptor and human IL7Rα (TSLPR / IL7Rα). We packaged the target genes TSLPR / IL7Rα using lentivirus and cloned them into target cell lines to form stable, high-expression cell lines. First, the 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. Next, using lentiviral infection, human TSLPR was inserted into CHO-K1 and BaF3 cell lines and cultured for 3 weeks under selective pressure of 10 μg / ml puromycin (Gibco, US). A second infection was then performed. Human IL7Rα genes were inserted into cell lines, and screening was performed for 2-3 weeks with 1 mg / ml G418 (Gibco, US) and 10 μg / ml puromycin. Finally, CHO-K1 and BaF3 monoclonal cell lines that simultaneously highly expressed TSLPR and IL7Rα were screened using flow sorting.

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

[0122] Anti-human TSLP monoclonal antibodies were produced by immunizing 6-8 week old experimental SJL white mice (Beijing Charles River Laboratory Animal Technology Co., Ltd., Animal Production License No.: SCXK (Beijing) 2012-0001). Feeding environment: SPF level. Mice were stored in a laboratory environment for one week after purchase under a 12 / 12 hour light / dark cycle, at a temperature of 20-25°C and a humidity of 40-60%. 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 serum antibody titers and a tendency to reach a titer plateau were selected and sacrificed. Splenocytes were collected and fused with myeloma cells. Splenocytes and myeloma cells (Sp2 / 0 cells, ATCC® CRL-8287®) were fused, and hybridoma cells were obtained using an optimized PEG-mediated fusion step.

[0123] Initial screening involved ELISA binding assays for human and cyno-TSLP, assays to inhibit human TSLP binding to its receptor TSLPR, and experiments to inhibit TSLP-induced BaF3 cell proliferation. After transferring hybridoma cells to 24-well plates, the supernatant was re-screened. Hybridoma clones were obtained by subcloning selected positive clones twice, used for antibody production, and purified by affinity assays.

[0124] After screening, monoclonal hybridoma cell lines NO:3, NO:119, NO:179, and NO:199 with good activity were obtained, and hybridoma cells in the logarithmic growth phase were collected. RNA was extracted with NucleoZol (MN) and reverse transcription was performed (PrimeScript® reverse transcriptase, Takara, catalog no. 2680A). The cDNA obtained by reverse transcription was amplified by PCR using a 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: the sequences of mab3, mab119, mab179, and mab199, and 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 region obtained according to Kabat's numbering rules are shown in the table below:

[0126] [Table 6]

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

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

[0129] To reduce the immunogenicity of mouse antibodies, screened mab3, mab119, mab179, and mab199 antibodies with excellent in vivo and in vitro activity were humanized. Humanization of mouse monoclonal antibodies was carried out according to methods published in many literatures in the art. Briefly, the constant domain of the parent (mouse antibody) was replaced with the constant domain of the human antibody, and human germline antibody sequences were selected according to the homology between the mouse antibody and the human antibody, followed by CDR grafting. Next, based on the three-dimensional structure of the mouse antibody, the VL and VH amino acid residues were reverse-mutated, and the constant region of the mouse antibody was replaced with the human constant region to obtain the final humanized molecule.

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

[0131] (1) Selection of the human FR region and reverse mutation

[0132] In the case of mab3, the humanized VH template is IGHV1-3 * 01+IGHJ6 * The 01 is the humanized VL template, and the IGKV3-20+IGKJ4 * The result was 01. The CDR of mab3 was transplanted into a human template, and the variable region sequence obtained after transplantation was as follows: hu3 VL-CDR after transplant: [ka] SEQ ID NO:38 hu3 VH-CDR after transplant: [ka] SEQ ID NO:42

[0133] The reverse mutation design for the mab3 humanized antibody is shown in the table below:

[0134] [Table 7] Note: "Grafted" indicates that the mouse antibody CDR has been transplanted into the human germline FR region sequence. L46P indicates that, according to Kabat's numbering rules, the L at position 46 has been mutated back to P.

[0135] The sequence of the variable region of the mab3 humanized antibody is as follows: > hu3VL1 (hu3 VL-CDR port) [ka] SEQ ID NO:38 [ka] [ka] Note: A single underline indicates a CDR region, and a double underline indicates a revert mutation site.

[0136] The above-mentioned light and heavy chain variable regions were combined with human germline light and heavy chain constant region sequences to ultimately form complete light and heavy chain sequences and obtain an antibody having the full-length sequence. Exemplarily, in the case of the mab3 humanized antibody in this 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 κ chain constant region shown in SEQ ID NO:134, but these can also be replaced with other constant regions known in the art.

[0137] The sequences of the heavy chain and light chain variable regions of the obtained MAB3 humanized antibody are shown in the table below.

[0138] [Table 8]

[0139] The binding activity of the mab3 humanized antibody against human TSLP was detected by ELISA, confirming that the mab3 humanized antibody has excellent binding ability to human TSLP.

[0140] (2) Point mutation to hu3 antibody

[0141] Detection revealed the presence of hotspots in the MDH sequence of HCDR3 and the NTR sequence of LCDR3 of the mab3 humanized antibody. Therefore, the corresponding hotspots were mutated. The sequence of the CDR region of the resulting mab3 humanized antibody is as follows:

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

[0143] We can conclude that the CDR sequence of the mab3 humanized antibody is as follows:

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

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

[0146] [Table 11]

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

[0148] The light and heavy chain variable regions after hotspot mutations were recombined with the light and heavy chain constant region sequences of human germline cells to form complete light and heavy chain sequences, and antibodies with full-length sequences were obtained.

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

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

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

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

[0153] In the first screening, approximately 5 × 10⁶ mutants were selected from the hu3-11-scFv mutant library. 10Cells and biotinylated TSLP-Fc protein (1-10 μg / ml) were cultured in 50 ml of 0.1% bovine serum albumin (BSA)-containing phosphate-buffered saline (PBSA) at room temperature for 1 hour. Next, the mixture was washed with 0.1% PBSA to remove unbound antibody fragments. Then, 100 μl of streptomycin beads (Milenyi Biotec, Auburn, CA) were added to a library of hu3-11-scFv antibody mutants bound to biotinylated TSLP-Fc, loaded into an AutoMACS system, and sorted. Cells with high affinity for TSLP-Fc were collected from the antibody library and induced at 250 rpm, 20°C for 18 hours. The resulting enriched library was screened a second time against biotinylated recombinant TSLP-Fc protein.

[0154] For the third and fourth screenings, library cells from the previous screening were cultured 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 at room temperature for 1 hour. 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 the cells were cultured at 4°C for 1 hour. The mixture was washed with 0.1% PBSA to remove unbound antibody fragments. Finally, antibodies with high affinity were screened using FACS screening (BD FACSAria™ FUSION).

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

[0156] The sequence of the light chain variable region obtained by affinity maturation is as follows.

Chemical formula

[0157] The obtained light chain variable region was recombined with the heavy chain variable region of the mab3 humanized antibody to obtain a new mab3 humanized antibody. For example, huVL5 and huVL6 were combined with hu3VH2-H110Y respectively to obtain new antibody molecules hu3-12 and hu3-13. The 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] ELISA was performed on the obtained new antibody mab3 humanized antibody to detect the binding activity to human TSLP. As a result, it was found that hu3-12 and hu3-13 still have a high binding ability to human TSLP. It was shown that changing LCDR3 does not affect the activity of the hu3 series of antibodies.

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

[0163]

Table 14

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

[0165] [Table 15]

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

[0167] For mab119, IGHV1-69 was used as the VH template. * 02 and HJ6 * Select 01 and use IGKV4-1 as the VL mold. * 01 and IGKJ2 * 01, and IGKV3-11 * 01 and IGKJ2 * 01 was selected. The CDR region of the mouse antibody was transplanted into the selected humanized template, and the FR region was reverse-mutated to obtain different light and heavy chain variable regions. The variable region sequences obtained by CDR transplantation are as follows. [ka]

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

[0169] [Table 16] Note: For example, M4L indicates that, according to Kabat's numbering rules, the M at position 4 is reverse-mutated to L. Grafted indicates that the mouse antibody CDR has been transplanted into the human germline FR region sequence.

[0170] The specific sequence of the variable region of the mab119 humanized antibody is as follows: >hu119VL1 (Grafted (IGKV4-1*01))

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0171] The above-mentioned light chain and heavy chain variable regions were combined with human germline light chain and heavy chain constant region sequences to ultimately form complete light chain and heavy chain sequences, thus obtaining an antibody having a full-length sequence. Exemplarily, in the case of the mab119 humanized antibody in this 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 κ chain constant region shown in SEQ ID NO:134, but these can also be replaced with other constant regions known in the art.

[0172] Table 17 shows the heavy and light chain variable regions of the MAB119 humanized antibody.

[0173] [Table 17]

[0174] ELISA detection of the binding activity of humanized antibodies to human TSLP revealed that the mab119 humanized antibody can specifically bind to human TSLP.

[0175] (2) Mutation of hu119

[0176] A hotspot was detected in the LCDR1DNS sequence of the mab119 humanized antibody, so the corresponding site was mutated to N31S or N31Q. The resulting LCDR1 sequence after mutation is as follows:

[0177] [Table 18] Note: The locations of the mutation sites in Table 19 are numbered according to their natural order.

[0178] For example, the mutant sequences of hu119VL2 and hu119VL6 obtained after mutation are as follows: [ka] SEQ ID NO:73 [ka] Note: A single underline indicates a variable region, and a double underline indicates a revertant mutation.

[0179] The obtained hu119VL2 and hu119VL6 mutants were combined with hu119VH to obtain new humanized hu119 antibodies. For example, hu119VL2-N31S and hu119VL2-N31Q were combined with hu119VH3, respectively, to obtain antibodies hu119-28 and hu119-29, and hu119VL3-N31S was combined with hu119VH8 to obtain antibody hu119-30. Exemplary combinations of the variable regions of the mutant antibodies are as follows:

[0180] [Table 19]

[0181] The affinity of antibodies obtained after mutation to human TSLP was detected using ELISA. As a result, 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 anti-TSLP antibody activity.

[0182] In summary, the CDR sequence of the mab119 humanized antibody is as follows:

[0183] [Table 20] X6 can be selected from N, S, or Q.

[0184] 5.3. Humanization of mab179

[0185] (1) Template selection and reverse mutation for humanization of mab179 mouse antibodies

[0186] For mab179, IGHV1-69 is used as the VH mold. * 02 and IGHJ6 * Select 01 and use IGKV4-1 as the VL mold. * 01 and IGKJ2 * 01, or IGKV2-29 * 02 and IGKJ2 * 01 was selected. The CDR region of the mouse antibody was transplanted into the selected humanized template, and the FR region was reverse-mutated to obtain light and heavy chain variable regions with different sequences. The humanized variable region sequences and reverse mutations are as follows. [ka] [ka]

[0187] [Table 21] Note: For example, P43S indicates a reversion mutation from P to S at position 43, according to Kabat's numbering rules. Grafted indicates that the mouse antibody CDR has been grafted onto a human germline FR region sequence.

[0188] The variable region of the MAB179 humanized antibody is shown below. >hu179VL1 (Graft (IGKV4-1 * 01)) [ka] [ka] [ka] SEQ ID NO:82 [ka] [ka] [ka] Note: A single underlined section represents a CDR, and a double underlined section represents a revert mutation site.

[0189] The above-mentioned light chain and heavy chain variable regions were combined with human germline light chain and heavy chain constant region sequences to ultimately form complete light chain and heavy chain sequences, thus obtaining antibodies with full-length sequences. For example, in the case of the mab199 humanized antibody in this 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 κ chain constant region shown in SEQ ID NO:134, but these 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 for human TSLP.

[0192] (2) Mutation of the hu179 antibody

[0193] Detection revealed the presence of hotspots on the HCDR2 and LCDR2 sequences of the mab179 humanized antibody. Therefore, the risk of molecular modification was eliminated by mutating the corresponding hotspots.

[0194] In one embodiment, an amino acid mutation is performed on the GNG of HCDR2 in hu179VH1, and the sequence of the mutated hu179VH1 is as follows: hu179VH1- N55Q [ka] SEQ ID NO:90 [ka] [ka] Note: A single underlined section represents a CDR, and a double underlined section represents a revert mutation site.

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

[0196] [Table 23] Note: The locations of the mutation sites in Table 24 are numbered according to their natural order.

[0197] The CDR region of the mab179 humanized antibody can be obtained from the above and is 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 mutant obtained after mutation was combined with the humanized hu179VL to obtain a new humanized mab179 antibody. An example antibody combining the hu179VH1 mutant and hu179VL2 is shown below.

[0200] [Table 25]

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

[0202] Using the same method, point mutations (numbered in natural order) of N55Q, N55V, and G56V were introduced into hu179VH2, hu179VH3, hu179VH4, and hu179VH5, respectively. The variable regions of the heavy and light chains obtained by these mutations were then recombined to obtain a new, humanized mab179. As an example, the mutant sequence of hu179VH3 is shown below. [ka] Note: A single underline indicates a CDR, and a double underline indicates a reverse mutation site.

[0203] In several other cases, the LCDR2 molecule of the mab179 humanized antibody underwent amino acid mutations. For example, the sequence of the mutated hu179VL2 molecule is shown below. [ka] [ka] [ka] [ka] Note: A single underline indicates a CDR, and a double underline indicates a revert mutation site.

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

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

[0206] From the above, the general formula for the LCDR2 of the mab179 humanized antibody is X9VX 10 X 11 X 12 X 13 T (SEQ ID NO: 118), X9 is selected from Y or E, X 10 is selected from S, D, or E, X 11 is selected from N, Q, D, or E, X 12 is selected from H, Y, D, or E, X 13 The CDR region of the mab179 humanized antibody is shown in the table below.

[0207] [Table 27] X7 is selected from N, Q, V, X8 is selected from G, V, X9 is selected from Y, E, X 10 The first is selected from S, D, and E, and X 11 The first is selected from N, Q, D, and E, and X 12 The first is selected from H, Y, D, E, and X 13 It 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 combination of the hu179VL2 mutant with hu179VH1 and hu179VH3, and the resulting CDRs and the heavy and light chain variable regions of the obtained mab179 humanized antibody are shown below.

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

[0210] [Table 29]

[0211] The affinity of the mab179 humanized antibody obtained after LCDR2 mutation to human TSLP was detected using ELISA. The results showed that the antibody obtained after hotspot site mutation to LCDR2 still exhibited relatively good affinity to human TSLP. This indicates that hotspot site mutation to LCDR2 does not affect the binding activity of the mab179 humanized antibody.

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

[0213] A new antibody molecule, hu179-33, was obtained by combining hu179VL8-N53E and hu179VH3-N55V, which were obtained through mutation. Its CDR sequence is as follows:

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

[0215] The binding activity of antibodies obtained after mutation into human TSLP was detected by Biacore. Exemplary antibody binding activities are shown below.

[0216] [Table 31]

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

[0218] 5.4 Selection of the human FR region and reverse mutation for mab199 antibodies

[0219] Regarding mab199, IGHV1-46 is used as the VH template. * 01 and HJ6 * Select 01, and use IGKV1-39 as the VL mold. * 01 and IGKJ4 * 01 was selected. The CDR region of the mouse antibody was transplanted into the selected humanized template, and the FR region was reverse-mutated to obtain light and heavy chain variable regions with different sequences. The reverse mutations are shown in Table 32.

[0220] [Table 32] Note: For example, I48V indicates that, according to Kabat's numbering rules, the I at position 48 is reverse-mutated to V. "Grafted" indicates that the mouse antibody CDR has been transplanted into a human germline FR region sequence.

[0221] The variable region of the mab199 humanized antibody is shown below. [ka] [ka] [ka] [ka] Note: A single underline indicates a CDR, and a double underline indicates a revert mutation site.

[0222] The above-mentioned light chain and heavy chain variable regions were combined with human germline light chain and heavy chain constant region sequences to ultimately form complete light chain and heavy chain sequences, thus obtaining antibodies with full-length sequences. With respect to the mab199 humanized antibody, unless otherwise explicitly stated 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 obtained MAB199 humanized antibody is shown below.

[0224] [Table 33]

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

[0226] [Table 34]

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

[0228] 5.5 Antibody constant region

[0229] The heavy chain constant regions of humanized antibodies and chimeric antibodies can be selected from the group consisting of the constant regions of IgG1, IgG2, IgG4, and their variants. Exemplarily, the IgG1-YTE constant region is used in this disclosure, and its sequence is shown in SEQ ID NO:133. The light chain constant region can be selected from the light chain constant regions of the human κ chain, λ chain, or their variants. Exemplarily, the human κ chain constant region is used in this disclosure, and its sequence is shown in SEQ ID NO:134. >IgG1-YTE heavy chain constant region: [ka] Note: Underlined terms refer to the designed M252Y, S254T, and T256E mutations. > Kappa light chain constant region: [ka]

[0230] The humanized heavy chain and light chain variable regions in this disclosure were rearranged with the constant regions described above to obtain the full-length sequences of the heavy chain and light chain. As an example, the antibody sequence is 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: Underlined portions represent CDR, and italicized portions represent the steady-state region.

[0231] AMG157 is used as a positive control in this disclosure, and its sequences are as shown in SEQ ID NO:143 and SEQ ID NO:144. Heavy chain arrangement of AMG157 [ka] AMG157 light chain arrangement [ka]

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

[0233] The antibodies described herein 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: ELSA 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 pH 7.4 PBS (Shanghai BasalMedia, B320) buffer and added at 100 μg / well to a 96-well microtiter plate (Corning, CLS3590-100EA), and incubated overnight at 4°C. The solution was discarded, and 200 μl / well of inhibition solution containing 5% skim milk powder (Bright Dairy skim milk powder) diluted in PBS was added. Inhibition was performed by incubation in a 37°C incubator for 2 hours. After inhibition, the inhibition solution was discarded, and the plate was washed three times with PBST buffer (PBS containing 0.1% Tween-20, pH 7.4). 100 μl / well of the positive antibody AMG157, diluted to different concentrations with the test antibody and sample diluent, was added and incubated in a 37°C incubator for 1 hour. After 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 of TMB chromogenic substrate (KPL, 52-00-03) was added and incubated at room temperature for 10-15 minutes. The reaction was stopped by adding 50 μl / well of 1 M H2SO4. Absorption values ​​were read at 450 nm using a NOVOStar microplate reader. The EC50 value when the TSLP antibody binds to TSLP was calculated, and the results are shown in the table below.

[0238] [Table 35]

[0239] The results showed that the antibody in this disclosure has very good binding activity to human TSLP.

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

[0241] The affinity of the tested humanized TSLP antibody for human and cynoTSLP was detected using a Biacore T200(GE) instrument.

[0242] The test molecule was affinity-captured using a Protein A biosensor chip (catalog number 29127556, GE). Then, antigens (huTSLP-his and cynoTSLP-his prepared in Example 1) were flowed onto the chip surface, and the reaction signal was detected in real time using a Biacore T200 to obtain binding and dissociation curves. After dissociation was complete in each experimental cycle, the biosensor chip was washed and regenerated with a glycine-hydrochloride regeneration solution (pH 1.5, catalog number BR-1003-54, GE). The data were matched to a (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 showed that the anti-TSLP antibody in this disclosure has a relatively high affinity for human TSLP and can bind to cynoTSLP.

[0245] Experiment Example 3: ELSA-based experiment of an anti-TSLP antibody that inhibits the binding of TSLP to the TSLP receptor.

[0246] The TSLP receptor has two subunits, TSLPR and IL7R. TSLPR is a receptor specific to TSLP, while IL7R is a receptor common to both TSLP and IL7. TSLP first binds to TSLPR, and then to IL7R. Using this example, we identified whether a TSLP antibody could inhibit the binding of TSLP to the extracellular domain of recombinant 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% skim milk powder diluted in PBS was added, and inhibition was performed by incubation in a 37°C incubator for 2 hours. After inhibition was complete, the inhibition solution was discarded, and the plates were 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 test antibody was sequentially diluted from 200 nM. The antigen and antibody were mixed 1:1, incubated at 37°C for 15 minutes, and added to a microtiter plate at 100 μl per well, and incubated at 37°C for 1 hour. The plates were washed three times with PBST. Streptoavidin-peroxidase polymer, diluted 1:4000 with sample diluent, was added at a rate of 100 μl / well and incubated at 37°C for 1 hour. After washing the plate five times with PBST, 100 μl / well of TMB chromogenic substrate (KPL, 52-00-03) was added and incubated at room temperature for 3-10 minutes. The reaction was stopped by adding 100 μl / well of 1M H2SO4. Absorption values ​​were read at 450 nm using a NOVOStar microplate reader. The IC50 values ​​of TSLP antibodies that inhibit the 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 antibodies in this disclosure can strongly inhibit the binding of TSLP to its receptor, TSLPR.

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

[0251] Using this example, we identified whether the anti-TSLP antibody could inhibit the binding of TSLP to the TSLPR / IL7R receptor on the surface of the CHOK1 cell line.

[0252] In the detailed procedure, CHOK1-TSLPR / IL7R cells were cultured in DME / F12 containing 10% FBS, 1 mg / ml G418, and 10 μg / ml puromycin. Good-condition CHOK1-TSLPR / IL7R cells were centrifuged (1000 rpm, 5 min) and washed once with 2% FBS in PBS. The cells were counted, and 1 × 10⁶ cells were identified. 6 The cell concentration was adjusted to 1 / ml. 50 μl of cells were added to a round-bottom 96-well plate. The test antibody was diluted in PBS solution containing 2% BSA to an initial concentration of 20 nM, gradient 8, and ratio of 1:4. Biotin-labeled TSLP-Fc antigen was prepared at 2 nM. The antigen and antibody were mixed 1:1 and incubated at 37°C for 15 minutes. The mixture was added to the 96-well plated with cells at a rate of 50 μl per well and incubated at 4°C for 1 hour. After incubation, the plate was centrifuged at 4°C (800 g, 5 min) and the supernatant was discarded. The plate was washed twice by centrifugation with 200 μl of pre-chilled PBS. A PE-SA secondary antibody diluted 1:1000 was added and incubated in the dark for 40 minutes at 4°C. The plate was then centrifuged at 4°C (800 g, 5 min) and the supernatant was discarded. 200 μl of pre-cooled PBS was added to rupture the cells, and these were washed by centrifugation three times at 4°C. 100 μl of PBS was added, and the plate was loaded into the machine for reading. The IC50 value of the TSLP antibody that inhibits the binding of TSLP to TSLPR / IL7R was calculated according to the fluorescence signal value. The results are shown in Table 38.

[0253] [Table 38]

[0254] The results showed that all of the antibodies in this disclosure can 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 mature naive bone marrow dendritic cells (mDCs) and induce the secretion of thymic activation-modulating chemokines (TARCs) and osteoprotegerin (OPGs), thereby further mediating innate and adaptive immune inflammatory responses. This study demonstrated that the resulting antibody inhibits TSLP-induced chemokine production by mDCs, thereby suppressing the development of innate and adaptive inflammatory responses.

[0257] Naive bone marrow tissue mDCs were isolated and purified from human peripheral blood mononuclear cells (PBMCs) using a magnetic bead sorting method (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 minutes (37°C) and then added to each cell culture well containing mDCs to stimulate the mDCs in vitro. The plates were placed in an incubator and cultured for 48 hours. The cell culture supernatant was collected, appropriately diluted, and chemokine content was detected using ELISA. TARC was detected using R&D's Human CCL17 / TARC Quantikine ELISA Kit, and OPG content was detected using R&D's Human CCL22 / MDC Quantikine ELISA Kit. The results are shown in Figures 4A-4B.

[0258] The results showed that all antibodies in this disclosure can significantly inhibit the production of TSLP-induced TARC and OPG chemokines, demonstrating that the antibodies in this disclosure can prevent the development of spontaneous and adaptive inflammatory responses.

[0259] Test Example 7. Inhibition of BaF3-TLSPR / IL7R cell proliferation induced by natural TSLP with an anti-TSLP antibody.

[0260] BaF3-hTSLPR / hIL7R cells can proliferate under stimulation with native TSLP. When an antibody binds to native TSLP, the stimulating 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 reached 80% growth, and the supernatant was discarded. Human lung fibroblasts, NHLF (BeNa Culture Collection BNCC340764) and HLF1 (BeNa Culture Collection BNCC337730) were stimulated for 72 hours with 10 ng / ml human IL1-β (Sino Biological GMP-10139-HNAE), 20 ng / ml IL13 (R&D 213-ILB-005), and 20 ng / ml TNF-α (PEPROTECH 300-01A) to induce the production of natural 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 then filtered for later use.

[0262] BaF3-hTSLPR / hIL17R cells were cultured in RPMI1640 with 10% FBS (10 ng / mL mIL3, R&D 213-ILB-005) and 1 × 10⁶ cells were cultured. 4Cells were adjusted to a density of cells / ml and cultured in a 37°C, 5% CO2 incubator until the logarithmic growth phase. Cells were harvested, centrifuged at 800 rpm / min for 5 minutes, and the supernatant was discarded. Cells were washed three times with PBS to remove cytokines that stimulate growth in the medium. Cells were resuspended in RPMI1640 medium containing 4% FBS, seeded at 4000 cells / 50 μl / well in a 96-well plate, and cultured in an incubator for 2 hours. The test antibody was natural TSLP used at a 10-fold ratio, serially diluted from an initial antibody concentration of 100 nM to 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. Plates were incubated in a 37°C, 5% CO2 incubator for 72 hours. Next, 30 μL of CellTiter-Glo(Promega) was added to each well, incubated in the dark at room temperature for 10 minutes, and detected using a Cytation5 cell imaging system with a luminescence program. The results are shown in the table below.

[0263] [Table 39]

[0264] As a result, it was found that all antibodies obtained in this disclosure were able to significantly suppress the activity that stimulates the proliferation of BaF3 in natural TSLP, particularly hu179-33, which has more than 100 times the activity of AMG157.

[0265] Experiment Example 8: Experiment on an anti-TSLP antibody that inhibits TSLP-induced proliferation in BaF3 cells overexpressing TSLPR / IL7R.

[0266] TSLP binds to TSLPR / IL7R on the surface of BaF3, thereby promoting BaF3 proliferation. Using this example, we identified whether the antibodies of this disclosure can block the activity of TSLP that induces BaF3 proliferation.

[0267] Specifically, BaF3 cells overexpressing TSLPR / IL7R were cultured in RPMI1640 containing 10% FBS and 2 ng / mL rhIL3 (MultiSciences, catalog number 96-AF-300-03-20), and incubated at 37°C in a 5% CO2 incubator for 1 × 10⁶ cells. 6 Cells were cultured at a cell density not exceeding cells / ml. Upon antibody detection, cells in the logarithmic growth phase were washed three times with PBS and centrifuged at 800 rpm for 5 minutes. Using RPMI1640 (2% FBS, recombinant human TSLP-Fc: 40 ng / ml), the cell density was adjusted to 8000 cells / well / 90 μl. 10 μl of serially diluted antibody to be tested was added to a 96-well plate and cultured for 2 days. 30 μl of cell titer was added and mixed for detection. 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 antibodies in this disclosure have a relatively strong ability to suppress TSLP-mediated proliferation of BaF3 cells.

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

[0271] TSLP can induce maturation of primary bone marrow tissue mDC cells. Mature mDC cells highly express the OX40 ligand, which is native to CD4 + OX40 can bind to the surface of T cells, thereby enabling the binding of natural CD4 + T cells are differentiated into Th2 cells, which produce immune response-related factors such as IL4 / IL5 / IL13, triggering a Th2 inflammatory response in the body. Using this example, we detected whether the antibodies obtained in this disclosure can inhibit Th2 cell differentiation induced by TSLP.

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

[0273] The results showed that the antibodies obtained in this disclosure can significantly suppress the production of Th2 cytokines IL4, I5, IL13, and TNF-α, and that the antibodies obtained in this disclosure can inhibit TSLP-induced differentiation of Th2 cells.

[0274] [Sequence List] 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> PA26-141 <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. An anti-TSLP antibody comprising a heavy chain variable region and a light chain variable region, i) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 47, respectively, and the light chain variable region includes LCDR1 and LCDR2 as shown in SEQ ID NO: 17 and SEQ ID NO: 18, respectively, and LCDR3 as shown in SEQ ID NO: 48 or 55, The sequence for SEQ ID NO: 47 is EDYDYDGYAMDX １ The sequence for SEQ ID NO: 48 is QQWSSX ２ The sequence for RT, SEQ ID NO: 55 is QQSDX. ３ X ４ RX 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 includes HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 22, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 76, SEQ ID NO: 24, and SEQ ID NO: 25, respectively. The sequence for SEQ ID NO: 76 is RASESVDX ６ SGLSFMH and X ６ is selected from N, S, and Q; or, iii) The heavy chain variable region includes HCDR1, HCDR2, and HCDR3 as indicated in SEQ ID NO: 26, SEQ ID NO: 96, and SEQ ID NO: 28, respectively, and the light chain variable region includes LCDR1, LCDR2, and LCDR3 as indicated in SEQ ID NO: 29, SEQ ID NO: 118, and SEQ ID NO: 31, respectively. The sequence for SEQ ID NO: 96 is VIDPGX ７ X ８ The sequence is DTNYNE, and SEQ ID NO: 118 is X ９ VX １０ X １１ X １２ X １３ T is X ７ is selected from N, Q, and V, X ８ is G or V, X ９ If Y or E, X １０ is selected from S, D, and E, 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) 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.

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

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

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

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

6. Including heavy chain variable regions and light chain variable regions, i) The heavy chain variable region has at least 90% sequence identity with the heavy chain variable region shown in 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in 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 shown in amino acid sequence SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125, The anti-TSLP antibody according to claim 4.

7. Including heavy chain variable regions and light chain variable regions, i) The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 6, 42, 43, 44 or 50, and the amino acid sequence of the light chain variable region is shown 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 shown 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 shown 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 shown 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 shown 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 shown 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 shown in SEQ ID NO: 13, 120, 121, 122, 123, 124 or 125, The anti-TSLP antibody according to claim 6.

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

125.

9. The anti-TSLP antibody according to any one of claims 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 their conventional variants, and the light chain constant region is selected from the group consisting of human antibody κ-chain and λ-chain constant regions and their conventional variants, and more preferably the antibody comprises a heavy chain constant region shown in SEQ ID NO: 133 and a light chain constant region shown in SEQ ID NO:

134.

10. The anti-TSLP antibody according to claim 9, wherein the antibody comprises the heavy chain and light chain shown below. a) The amino acid sequence of the heavy chain is as shown in SEQ ID NO: 135, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown 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 shown in SEQ ID NO: 137, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown 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 shown in SEQ ID NO: 139, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown 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 shown in SEQ ID NO: 141, or has at least 90% sequence identity thereto, and the amino acid sequence of the light chain is as shown in SEQ ID NO: 142, or has at least 90% sequence identity thereto.

11. An isolated anti-TSLP antibody that competitively binds to human TSLP with the anti-TSLP antibody described in 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 containing the nucleic acid molecule described in 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 pharmaceutically 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 the anti-TSLP antibody according to any one of claims 1 to 11.

17. A method for treating a TSLP-related disease, comprising administering 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 the TSLP-related disease is preferably 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, colorectal cancer, lung cancer, ovarian cancer, prostate cancer, rheumatoid arthritis, or chronic obstructive pulmonary disease. A method selected from the group consisting of systemic sclerosis, multiple sclerosis, keloids, ulcerative colitis, nasal polyps, chronic eosinophilic pneumonia, eosinophilic bronchitis, celiac disease, Churg-Strauss syndrome, eosinophilic myalgia syndrome, eosinophilia syndrome, eosinophilic granulomatosis with polyangiitis, inflammatory bowel disease, scleroderma, interstitial lung disease, fibrosis due to chronic hepatitis B or C, radiation-induced fibrosis, and fibrosis due to wound healing.

Images