SIRPγ variants and their fusion proteins

Abstract

SIRPγ variants and their fusion proteins are provided. Preparation and applications of peptides containing SIRPγ variants are also provided.

Description

Technical Field

[0001] This disclosure relates to SIRPγ variants, fusion proteins containing SIRPγ variants, bifunctional fusion proteins targeting PD-1 and CD47, and their use as medicines. Background Technology

[0002] The statements herein are provided only as background information in connection with this disclosure and do not necessarily constitute prior art.

[0003] CD47 is expressed or overexpressed in many tumor types, including acute myeloid leukemia, various subtypes of B-cell non-Hodgkin lymphoma, and many human solid tumor cells (Cell. 2009 Jul 23; 138(2):286-99; Res. 2011; 71:1374-84.). CD47 binds to signal regulatory protein α (SIRPα) on macrophages, resulting in a “don’t eat me” signal that prevents host cells from phagocytosing tumor cells, thus allowing them to be eliminated by the innate immune system (MolTher. 2017 Feb 1; 25(2):523-533). Data have shown that anti-CD47 antibodies help improve effective anti-tumor T-cell responses in immune-tolerant mice (Nat Med. 2015 Oct; 21(10):1209-15). Currently, there are many therapies targeting the CD47 / SIRPα interaction, including anti-CD47 antibodies, engineered SIRPα receptor proteins, anti-SIRPα antibodies, and bispecific antibodies.

[0004] Antibodies against CD47 can disrupt inhibitory signals of phagocytosis and can synergize with Fc-mediated prophagocytic signals to effectively eliminate tumor cells. However, CD47 is also expressed in normal cells, so therapeutic CD47 antibodies may be toxic (2017 Feb; 9(2): E168-E174). Currently, the novel humanized anti-CD47 monoclonal antibody Hu5F9 has caused transient and dose-dependent anemia in non-human primates with single doses of 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 10 mg / kg, and 30 mg / kg. The use of Hu5F9 at doses of 1 mg / kg, 3 mg / kg, or 10 mg / kg resulted in hemoglobin levels as low as below 10 g / dL between 5 and 7 days. The use of Hu5F9 at a dose of 30 mg / kg caused severe anemia in non-human primates, with hemoglobin levels below 8 g / dL (J Clin Oncol 2016; 34:abstr 3019).

[0005] The signal regulatory protein family (SIRP) plays a crucial role in regulating immune responses. This family comprises three type I transmembrane glycoproteins: SIRPα, SIRPβ, and SIRPγ. Both SIRPα and SIRPγ can bind to CD47, but SIRPγ's binding affinity to CD47 is approximately 10 times weaker than that of SIRPα. Studies have shown that the KD value for SIRPγ binding to CD47 is approximately 23 μM (The Journal of Immunology, 2004, 173:2562–2570.). SIRPγ is expressed on T cells and activated NK cells. The CD47-SIRPγ interaction participates in the contact between antigen-presenting cells and T cells, co-stimulating T cell activation and promoting T cell proliferation (Piccio et al., Blood 2005, 105, 2421-2427; BMC Struct Biol. 2013 Jul 4; 13:13.).

[0006] Patents related to the CD47 signal path include WO2016065329, WO2016109415, WO2014087248, WO2014093678, CN107849143A, CN108350048, etc. Summary of the Invention

[0007] This disclosure provides a variety of SIRPγ variants and fusion proteins comprising these SIRPγ variants, wherein the SIRPγ variants have higher CD47 affinity activity than the wild type and their fusion proteins have higher stability.

[0008] The SIRPγ variant disclosed herein utilizes the Disulfide Scan module of MOE (Molecular Operating Environment) software to design intrachain disulfide bonds through structural analysis of SIRPγ. Cysteine ​​mutations are introduced at appropriate positions in the SIRPγ sequence to form intrachain disulfide bonds, thereby increasing the stability of the SIRPγ variant sequence. Exemplarily, two cysteine ​​mutations are introduced to form a pair of disulfide bonds within the SIRPγ sequence. In some embodiments, cysteine ​​mutations are introduced at positions 14 and 115, and positions 8 and 107 of SIRPγ, respectively.

[0009] In some embodiments, the SIRPγ variant contains at least one amino acid mutation relative to the wild-type SIRPγ peptide, wherein the amino acid mutation site is selected from: Q8, L13, L14, N51, H56, N70, R77, S79, G107, M112, and G115.

[0010] In some embodiments, the SIRPγ variant contains at least one amino acid mutation relative to the wild-type SIRPγ peptide, wherein the amino acid mutation site is selected from: M6, Q8, L13, L14, K19, N51, Q52, K53, E54, H56, N70, M72, R77, S79, N101, G107, M112, and G115.

[0011] In some embodiments, the SIRPγ variant comprises at least three amino acid mutations selected from M6I, L13V, K19E, N51M or N51A, Q52S, K53G, E54R, H56Q, N70E, M72K, R77K, S79Q, N101D, and M112V.

[0012] In some embodiments, the SIRPγ variant, relative to the wild-type SIRPγ peptide shown in SEQ ID NO: 14, contains the following amino acid mutations:

[0013] i) L14C and G115C; or

[0014] ii) Q8C and G107C.

[0015] In some embodiments, the SIRPγ variant binds CD47 with an affinity up to 10 times greater than that of wild-type SIRPγ (which has the sequence of SEQ ID NO: 14). In some embodiments, the SIRPγ variant binds CD47 with an affinity up to 100 times greater than that of wild-type SIRPγ. In some embodiments, the SIRPγ variant binds CD47 with an affinity up to 1000 times greater than that of wild-type SIRPγ. In some embodiments, the SIRPγ variant binds CD47 with an affinity less than 1 × 10⁻⁶. -8 M, less than 5×10 - 9 M, less than 2×10 -9 M, less than 1×10 -9 M, less than 6×10 -10 M or less than 5×10 -10 The KD value of M binds to CD47; this affinity can be detected by the BIAcore method, as detailed in Test Example 1. In some embodiments, the SIRPγ variant has at least 80% homology with the wild-type SIRPγ shown in SEQ ID NO: 14.

[0016] In some implementations, the SIRPγ variant in this disclosure has a lower affinity for CD47 on the surface of erythrocytes.

[0017] In some implementations, the SIRPγ variants in this disclosure bind almost no to CD47 on the surface of red blood cells.

[0018] In some implementations, the SIRPγ variant in this disclosure has a higher affinity for CD47 expressed by tumor cells.

[0019] In some embodiments, the SIRPγ variant contains an amino acid mutation selected from one or more of K19E, Q52S, K53G, E54R, M72K, and N101D.

[0020] In some embodiments, the SIRPγ variant contains amino acid mutations of K19E, Q52S, K53G, E54R, M72K, and N101D.

[0021] In some implementations, the SIRPγ variant contains an N51A or N51M mutation.

[0022] In some embodiments, the SIRPγ variant contains L13V and / or M6I amino acid mutations.

[0023] In some embodiments, the SIRPγ variant contains one or more amino acid mutations selected from H56Q, N70E, R77K, S79Q, and M112V.

[0024] In some embodiments, the SIRPγ variant comprises an amino acid mutation selected from any of the following:

[0025] a) L13V, N51A, and S79Q;

[0026] b) L13V, N51A, and R77K;

[0027] c) L13V, N51A and M112V;

[0028] d) L13V, N51M, H56Q and N70E;

[0029] e)L13V, N51A and N70E;

[0030] f)M6I, N51M and R77K;

[0031] g)M6I, N51A and R77K;

[0032] h)M6I, N51A and N70E;

[0033] i) M6I, L13V, N51M and R77K;

[0034] j)M6I, L13V, N51A and R77K;

[0035] k)M6I, L13V, N51A and N70E;

[0036] l)N51A and R77K;

[0037] m)L13V, N51M and R77K;

[0038] n)L13V, N51M and N70E;

[0039] o)N51A;

[0040] p)L13V and N51M; and

[0041] r)N51M.

[0042] In some embodiments, the SIRPγ variant contains N51A and M112V amino acid mutations.

[0043] In some embodiments, the SIRPγ variant comprises: L14C, G115C, K19E, Q52S, K53G, E54R, M72K, and N101D amino acid mutations; and amino acid mutations selected from any of the following:

[0044] a) L13V, N51A, and S79Q;

[0045] b) L13V, N51A, and R77K;

[0046] c) L13V, N51A and M112V;

[0047] d) L13V, N51M, H56Q and N70E;

[0048] e)L13V, N51A and N70E;

[0049] f)M6I, N51M and R77K;

[0050] g)M6I, N51A and R77K;

[0051] h)M6I, N51A and N70E;

[0052] j)M6I, L13V, N51M and R77K;

[0053] k)M6I, L13V, N51A and R77K;

[0054] l)M6I, L13V, N51A and N70E;

[0055] m)N51A and R77K;

[0056] n)L13V, N51M and R77K;

[0057] o)L13V, N51M and N70E;

[0058] p)N51A;

[0059] q)L13V and N51M; and

[0060] r)N51M.

[0061] In some embodiments, the SIRPγ variant comprises amino acid mutations of Q8C, G107C, K19E, Q52S, K53G, E54R, M72K, and N101D; and amino acid mutations selected from any of the following:

[0062] a) L13V, N51A, and S79Q;

[0063] b) L13V, N51A, and R77K;

[0064] c) L13V, N51A and M112V;

[0065] d) L13V, N51M, H56Q and N70E;

[0066] e)L13V and N51M;

[0067] f)M6I, N51M and R77K;

[0068] g)M6I, N51A and R77K;

[0069] h)M6I, N51A and N70E;

[0070] j)M6I, L13V, N51M and R77K;

[0071] k)M6I, L13V, N51A and R77K;

[0072] l)M6I, L13V, N51A and N70E;

[0073] m)N51A and R77K;

[0074] n)L13V, N51M and N70E; and

[0075] o)N51A; and

[0076] p)N51A and M112V.

[0077] In some embodiments, the SIRPγ variant comprises amino acid mutations of L14C, G115C, K19E, Q52S, K53G, E54R, M72K, N101D, L13V, and N51M.

[0078] In some embodiments, the SIRPγ variant comprises amino acid mutations of L14C, G115C, M6I, K19E, Q52S, N51A, K53G, E54R, N70E, M72K, and N101D.

[0079] In some embodiments, the SIRPγ variants comprise amino acid mutations of Q8C, G107C, K19E, N51A, Q52S, K53G, E54R, M72K, N101D, and M112V.

[0080] In some implementations, the SIRPγ variants comprise amino acid mutations of Q8C, G107C, L13V, K19E, N51A, Q52S, K53G, E54R, M72K, R77K, and N101D.

[0081] In some embodiments, the aforementioned SIRPγ variant comprises the following amino acid sequence:

[0082] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0083] SEQ ID NO: 13 where:

[0084] X1 is selected from M or I; X2 is selected from Q or C; X3 is selected from L or V; X4 is selected from L or C; X5 is selected from K or E; X6 is selected from N, M, or A; X7 is selected from Q or S; X8 is selected from K or G; X9 is selected from E or R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 Selected from N or D; X16 Selected from G or C; X 17 Selected from M or V; X 18 Selected from G or C, and

[0085] i) When X4 is C, X 18 For C; or

[0086] ii) When X2 is C, X 16 The answer is C.

[0087] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0088] a) X5 is E;

[0089] b) X7 is S;

[0090] c) X8 is G;

[0091] d) X9 is R;

[0092] e)X 12 For K; or

[0093] f)X 15 The answer is D.

[0094] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein: X5 is E; X7 is S; X8 is G; X9 is R; X 12 Let K be the number of K; and X be the number of X. 15 The answer is D.

[0095] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, X6 is A or M.

[0096] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, X3 is V and / or X1 is I.

[0097] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0098] a)X 10 For Q;

[0099] b)X 11 E;

[0100] c)X 13 Let K be the value of K.

[0101] d)X 14 For Q; or

[0102] e)X 17 V.

[0103] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0104] a) X3 is V, X6 is A and X 14 For Q;

[0105] b) X3 is V, X6 is A and X 13 Let K be the value of K.

[0106] c) X3 is V, X6 is A and X 17 For V;

[0107] d) X3 is V, X6 is M, X 10 For Q and X 11 E;

[0108] e) X3 is V and X6 is M;

[0109] f) X1 is I, X6 is M and X 13 Let K be the value of K.

[0110] g) X1 is I, X6 is A and X 13 Let K be the value of K.

[0111] h) X1 is I, X6 is A and X 11 E;

[0112] i) X1 is I, X3 is V, X6 is M and X 13 Let K be the value of K.

[0113] j) X1 is I, X3 is V, X6 is A and X 13 Let K be the value of K.

[0114] k) X1 is I, X3 is V, X6 is A and X 11 E;

[0115] l)X6 represents A and X 13 Let K be the value of K.

[0116] m)X3 is V, X6 is M and X 13 Let K be the value of K.

[0117] n)X3 is V, X6 is M and X 11 E;

[0118] o)X3 is V, X6 is A and X 11 E;

[0119] p)X6 is A;

[0120] q) X3 is V and X6 is M; or

[0121] r)X6 is M.

[0122] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, X6 is A and X 17 V.

[0123] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0124] i) X4 is C, and X 18 The answer is C;

[0125] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0126] iii) Select any one of a) to k):

[0127] a) X3 is V and X6 is M;

[0128] b) X1 is I, X6 is M and X 13 Let K be the value of K.

[0129] c) X1 is I, X6 is A and X 13 Let K be the value of K.

[0130] d) X1 is I, X6 is A and X 11 E;

[0131] e) X1 is I, X3 is V, X6 is M and X 13 Let K be the value of K.

[0132] f) X1 is I, X3 is V, X6 is A and X 13 Let K be the value of K.

[0133] g) X1 is I, X3 is V, X6 is A and X 11 E;

[0134] h)X6 represents A and X 13 Let K be the value of K.

[0135] j) X3 is V, X6 is A and X 13 For K; or

[0136] k)X3 is V, X6 is M and X 11 The value is E.

[0137] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0138] i) X4 is C, and X 18 The answer is C;

[0139] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 D; and iii) are selected from any one of a) to f):

[0140] a) X3 is V, X6 is A and X 14 For Q;

[0141] b) X3 is V, X6 is A and X 17 For V;

[0142] c) X3 is V, X6 is M, X 10 For Q and X 11 E;

[0143] d) X3 is V, X6 is M and X 13 Let K be the value of K.

[0144] e) X3 is V, X6 is A and X 11 For E; or

[0145] f)X6 is A.

[0146] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein: i) X4 is C, and X 18 The answer is C;

[0147] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 D; and iii)X6 is M.

[0148] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein: i) X4 is C, and X 18 The answer is C;

[0149] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and iii) X3 is V and X6 is M

[0150] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein: i) X2 is C, and X 16 The answer is C;

[0151] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 D; and iii) are selected from any one of a) to o):

[0152] a) X3 is V, X6 is A and X 14 For Q;

[0153] b) X3 is V, X6 is A and X 13 Let K be the value of K.

[0154] c) X3 is V, X6 is A and X 17 For V;

[0155] d) X3 is V, X6 is M, X 10 For Q and X 11 E;

[0156] e) X3 is V, X6 is A and X 11 E;

[0157] f) X1 is I, X6 is M and X 13 Let K be the value of K.

[0158] g) X1 is I, X6 is A and X 13 Let K be the value of K.

[0159] h) X1 is I, X6 is A and X 11 E;

[0160] j) X1 is I, X3 is V, X6 is M and X 13 Let K be the value of K.

[0161] k) X1 is I, X3 is V, X6 is A and X 13 Let K be the value of K.

[0162] l) X1 is I, X3 is V, X6 is A and X 11 E;

[0163] m)X6 represents A and X 13 Let K be the value of K.

[0164] n)X6 is A; and;

[0165] o)X3 is V and X6 is M.

[0166] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein: i) X2 is C, and X 16 The answer is C;

[0167] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0168] iii) X6 represents A and X 17 V.

[0169] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0170] i) X4 is C, and X 18 The answer is C;

[0171] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0172] iii) X1 is I, X6 is A and X 11 For E; and

[0173] iv) X2 is selected from Q; X3 is selected from L or V; X6 is selected from N, M or A; X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; X 16 Selected from G; X 17 Selected from M or V.

[0174] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0175] i) X4 is C, and X 18 The answer is C;

[0176] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0177] iii) X3 is V, X6 is M; and

[0178] iv) X1 is selected from M or I; X2 is selected from Q; X4 is selected from L; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 16 Selected from G; X 17 Selected from M or V.

[0179] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0180] i) X4 is C, and X 18 The answer is C;

[0181] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0182] iii) X6 is A; X 17 For V; and

[0183] iv) X1 is selected from M or I; X3 is selected from L or V; X2 is selected from Q; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 16 Selected from G.

[0184] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0185] i) X2 is C, and X 16 The answer is C;

[0186] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0187] iii) X6 is A; X 17 For V; and

[0188] iv) X1 is selected from M or I; X3 is selected from L or V; X4 is selected from L; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 18 Selected from G.

[0189] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13 above, wherein:

[0190] i) X2 is C, and X 16 The answer is C;

[0191] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0192] iii) X3 is V; X6 is A; and X 13 For K

[0193] iv) X1 is selected from M or I; X4 is selected from L; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 17 For is selected from V or M; and X 18Selected from G.

[0194] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13, the SIRPγ variant has a higher affinity for CD47 compared to the wild type; preferably, the SIRPγ variant has an affinity of less than 2 × 10⁻⁶. -9 M's KD value binds to human CD47.

[0195] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13, the SIRPγ variant thereon has at least 85% sequence identity with the SIRPγ variant shown in any of the amino acid sequences in SEQ ID NO: 15-46 or SEQ ID NO: 105-106 (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity).

[0196] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13, the SIRPγ variant comprises any amino acid sequence selected from SEQ ID NO: 15-46.

[0197] In some embodiments of the SIRPγ variant shown in SEQ ID NO: 13, the SIRPγ variant comprises any amino acid sequence selected from SEQ ID NO: 105-106.

[0198] In some embodiments, this disclosure provides a fusion protein, wherein the fusion protein comprises any of the SIRPγ variants described above.

[0199] In some embodiments, the aforementioned fusion protein is a PD-1-SIRPγ fusion protein containing an anti-PD-1 antibody. In some embodiments, the PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein is a full-length antibody, selectable from any anti-PD-1 antibody, including but not limited to tislelizumab, camrelizumab, toripalimab, sintilimab, cemiplimab, pembrolizumab, nivolumab, prolgolimab, genolimzumab, dostarlimab, zimberelimab, AK-105, sasanlimab, MGD-013, HLX-10, spartalizumab, SCT-I10A, CS-1003, retifanlimab, or MEDI-0680, etc.

[0200] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein comprises:

[0201] The heavy chain variable region contains HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 1, 2, and 3, respectively; and

[0202] The light chain variable region includes LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 4, 5 and 6 respectively.

[0203] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein comprises:

[0204] The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 95, 96, and 97, respectively; and

[0205] The light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 98, 99 and 100, respectively.

[0206] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein comprises the heavy chain variable region of SEQ ID NO: 7 and the light chain variable region of SEQ ID NO: 8.

[0207] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein includes a heavy chain variable region as shown in SEQ ID NO: 101 and a light chain variable region as shown in SEQ ID NO: 102.

[0208] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein includes a constant region; preferably, it includes the heavy chain constant region of SEQ ID NO: 9 and the light chain constant region of SEQ ID NO: 10.

[0209] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein comprises:

[0210] Heavy chain, comprising the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 11; and

[0211] A light chain comprising the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 12.

[0212] In some embodiments, the anti-PD-1 antibody in the aforementioned PD-1-SIRPγ fusion protein comprises: a heavy chain containing the amino acid sequence of SEQ ID NO: 103, or an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 103; and

[0213] A light chain comprising the amino acid sequence of SEQ ID NO: 104, or an amino acid sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 104.

[0214] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains amino acid mutations of L14C, K19E, Q52S, K53G, E54R, M72K, N101D, and G115C.

[0215] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains amino acid mutations of Q8C, K19E, Q52S, K53G, E54R, M72K, N101D, and G107C.

[0216] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains an N51A or N51M amino acid mutation.

[0217] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein further comprises one or more amino acid mutations selected from M6I, L13V, N70E, and R77K.

[0218] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein further includes the M112V amino acid mutation.

[0219] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains an amino acid mutation selected from any of the following:

[0220] a) L13V and N51M;

[0221] b) M6I, N51M, and R77K;

[0222] c) M6I, N51A, and R77K;

[0223] d) M6I, N51A, and N70E;

[0224] e) N51A and R77K;

[0225] f) M6I, L13V, N51M, and R77K;

[0226] g)M6I, L13V, N51A and R77K;

[0227] h)M6I, L13V, N51A and N70E;

[0228] i)L13V, N51A and R77K;

[0229] j)L13V, N51M and R77K; and

[0230] k)L13V, N51M and N70E.

[0231] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains N51A and M112V amino acid mutations.

[0232] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises: L14C, K19E, Q52S, K53G, E54R, M72K, N101D, and G115C amino acid mutations; and amino acid mutations selected from any of the following:

[0233] a) L13V and N51M;

[0234] b) M6I, N51M, and R77K;

[0235] c) M6I, N51A, and R77K;

[0236] d) M6I, N51A, and N70E;

[0237] e) N51A and R77K;

[0238] f) M6I, L13V, N51M, and R77K;

[0239] g)M6I, L13V, N51A and R77K;

[0240] h)M6I, L13V, N51A and N70E;

[0241] i)L13V, N51A and R77K;

[0242] j)L13V, N51M and R77K; and

[0243] k)L13V, N51M and N70E.

[0244] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains amino acid mutations of L14C, G115C, K19E, Q52S, K53G, E54R, M72K, N101D, L13V, and N51M.

[0245] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein contains amino acid mutations of L14C, G115C, M6I, K19E, Q52S, N51A, K53G, E54R, N70E, M72K, and N101D.

[0246] In some embodiments, the SIRPγ variants in the aforementioned PD-1-SIRPγ fusion protein contain amino acid mutations of Q8C, G107C, K19E, N51A, Q52S, K53G, E54R, M72K, N101D, and M112V.

[0247] In some embodiments, the SIRPγ variant of the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0248] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0249] SEQ ID NO: 13 where:

[0250] X1 is selected from M or I; X2 is selected from Q or C; X3 is selected from L or V; X4 is selected from L or C; X5 is selected from K or E; X6 is selected from N, M, or A; X7 is selected from Q or S; X8 is selected from K or G; X9 is selected from E or R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 Selected from N or D; X16 selected from G or C; X 17 Selected from M or V; X 18 Selected from G or C, and

[0251] i) When X4 is C, X 18 For C; or

[0252] ii) When X2 is C, X 16 The answer is C.

[0253] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0254] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0255] SEQ ID NO: 13

[0256] in:

[0257] X1 is selected from M or I; X2 is Q; X3 is selected from L or V; X4 is C; X5 is selected from K or E; X6 is selected from N, M, or A; X7 is selected from Q or S; X8 is selected from K or G; X9 is selected from E or R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 Selected from N or D; X 16 For G; X 17 Selected from M or V; X 18 The answer is C.

[0258] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0259] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0260] SEQ ID NO: 13 where:

[0261] X1 is selected from M or I; X2 is Q; X3 is selected from L or V; X4 is C; X6 is selected from N, M, or A; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 16 For G; X 17 Selected from M or V; X 18 It is C; and:

[0262] a) X5 is E;

[0263] b) X7 is S;

[0264] c) X8 is G;

[0265] d) X9 is R;

[0266] e)X 12 For K; or

[0267] f)X 15 The answer is D.

[0268] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0269] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0270] SEQ ID NO: 13 where:

[0271] X1 is selected from M or I; X2 is Q; X3 is selected from L or V; X4 is C; X5 is E; X6 is selected from N, M, or A; X7 is S; X8 is G; X9 is R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 For D; X 16 For G; X 17 Selected from M or V; X 18 The answer is C.

[0272] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0273] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0274] SEQ ID NO: 13

[0275] in:

[0276] X1 is selected from M or I; X2 is Q; X3 is selected from L or V; X4 is C; X5 is E; X6 is selected from M or A; X7 is S; X8 is G; X9 is R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 For D; X 16 For G; X 17 Selected from M or V; X 18 The answer is C.

[0277] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0278] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0279] SEQ ID NO: 13 where:

[0280] i) X4 is C, and X 18 The answer is C;

[0281] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 10 H; X 12 K; X 15 For D; X 16 For G; and X 17 For M; and

[0282] iii) Select from any of the following:

[0283] a) X3 is V and X6 is M; and X1 is selected from M or I; X 11 Selected from N or E; X 13 Selected from R or K; and X 14 Selected from S or Q;

[0284] b) X1 is I, X6 is M and X 13 Let K be the number of elements; and X3 be selected from L or V, X 11 Selected from N or E; and X 14 Selected from S or Q;

[0285] c) X1 is I, X6 is A and X 13 Let K be the number of elements; and X3 be selected from L or V, X 11 Selected from N or E; and X 14 Selected from S or Q;

[0286] d) X1 is I, X6 is A and X 11 E; and X3 is selected from L or V, X 13 Selected from R or K and X 14 Selected from S or Q

[0287] e) X1 is I, X3 is V, X6 is M and X 13 Let K be the number of K; and X be the number of X.11 Selected from N or E; and X 14 Selected from S or Q;

[0288] f) X1 is I, X3 is V, X6 is A and X 13 Let K be the number of K; and X be the number of X. 11 Selected from N or E; and X 14 Selected from S or Q;

[0289] g) X1 is I, X3 is V, X6 is A and X 11 Let E be the value of X; and X be the value of X. 13 Selected from R or K; and X 14 Selected from S or Q;

[0290] h)X6 represents A and X 13 Let K be the number of elements in the array; and X1 be selected from M or I; X3 be selected from L or V, X... 11 Selected from N or E; and X 14 Selected from S or Q;

[0291] j) X3 is V, X6 is A and X 13 Let K be the number of elements; and X1 be selected from M or I; X 11 Selected from N or E; and X 14 Selected from S or Q; and

[0292] k)X3 is V, X6 is M and X 11 Let E be the value of X1; and X1 be selected from M or I; X 13 Selected from R or K; and X 14 Choose from S or Q.

[0293] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0294] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0295] SEQ ID NO: 13

[0296] in:

[0297] i) X4 is C, and X 18 The answer is C;

[0298] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 10 H; X 12 K; X 15 For D; X 16 For G; and X 17 For M; and

[0299] iii) X3 is V, X6 is M and X 13 Let K be a subset of M, and X1 be selected from M or I; X 11 Selected from N or E; and X 14 Choose from S or Q.

[0300] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0301] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0302] SEQ ID NO: 13 where:

[0303] i) X2 is C, and X 16 The answer is C;

[0304] ii) X4 is L, X5 is E; X7 is S; X8 is G; X9 is R; X 10 H; X 12 K; X 15 For D; and X 18 For G; and

[0305] iii) X1 is I or M, X3 is L or V, X6 is A, X 11 For N or E, and X 13 For R or K, X 14 Selected from S or Q, and X 17 V.

[0306] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein comprises the following amino acid sequence:

[0307] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0308] SEQ ID NO: 13

[0309] in:

[0310] i) X4 is C, and X 18 The answer is C;

[0311] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 10 H; X 12 K; X 15 For D; X 16 For G; X 14 Let S and X be the numbers. 17 For M; and

[0312] iii) Selected from any of the following mutations:

[0313] a) X3 is V and X6 is M; and X1 is selected from M or I; X 11 Selected from N or E; and X 13 Selected from R or K;

[0314] b) X1 is I, X6 is M and X 13 Let K be the number of elements; and X3 be selected from L or V, and X... 11 Selected from N or E;

[0315] c) X1 is I, X6 is A and X 13 Let K be the number of elements; and X3 be selected from L or V, and X... 11 Selected from N or E;

[0316] d) X1 is I, X6 is A and X 11 Let E be the value of X3; and X3 be selected from L or V, and X 13 Selected from R or K;

[0317] e) X1 is I, X3 is V, X6 is M and X 13 Let K be the number of K; and X be the number of X. 11 Selected from N or E;

[0318] f) X1 is I, X3 is V, X6 is A and X 13 Let K be the number of K; and X be the number of X. 11 Selected from N or E;

[0319] g) X1 is I, X3 is V, X6 is A and X 11 Let E be the value of X; and X be the value of X. 13 Selected from R or K;

[0320] h)X6 represents A and X 13 Let K be the number of elements in the array; and X1 be selected from M or I; X3 be selected from L or V, and X... 11 Selected from N or E;

[0321] j) X3 is V, X6 is A and X 13 Let K be the number of elements; and X1 be selected from M or I; and X 11 Selected from N or E; and

[0322] k)X3 is V, X6 is M and X 11 For E; and X1 is selected from M or I; and X 13 Selected from R or K.

[0323] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein, wherein the SIRPγ variant comprises the following amino acid sequence:

[0324] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0325] SEQ ID NO: 13

[0326] in:

[0327] i) X4 is C, and X 18 The answer is C;

[0328] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 10 H; X 12 K; X 15 For D; X 16 For G; and X 17 For M; and

[0329] iii) X3 is V, X6 is M and X 13 Let K be a subset of M, and X1 be selected from M or I; and X 11 Selected from N or E.

[0330] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein is directly or via a linker linked to the heavy or light chain of the anti-PD-1 antibody.

[0331] In some embodiments, the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein is linked to the heavy or light chain of the anti-PD-1 antibody via a linker, wherein the linker is a linker known in the art; preferably, the linker is selected from (G4S)xGn, where x is selected from 1 to 10 and n is selected from 0 to 10; more preferably, the linker comprises a sequence as shown in SEQ ID NO: 47 or 48.

[0332] In some embodiments, the N-terminus of the SIRPγ variant in the aforementioned PD-1-SIRPγ fusion protein is linked to the C-terminus of the anti-PD-1 antibody heavy chain.

[0333] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises:

[0334] A first peptide chain comprising any amino acid sequence selected from SEQ ID NO: 49-59; and / or

[0335] The second peptide chain contains the amino acid sequence of SEQ ID NO: 12.

[0336] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises:

[0337] A first peptide chain comprising an amino acid sequence selected from SEQ ID NO: 107 or 108; and / or

[0338] The second peptide chain contains the amino acid sequence of SEQ ID NO: 104.

[0339] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises two identical first peptide chains and second peptide chains, wherein:

[0340] A first peptide chain comprising any amino acid sequence selected from SEQ ID NO: 49-59; and / or

[0341] The second peptide chain contains the amino acid sequence of SEQ ID NO: 12.

[0342] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises two identical first peptide chains and second peptide chains, wherein:

[0343] The first peptide chain comprises an amino acid sequence selected from SEQ ID NO: 107 or 108; and / or

[0344] The second peptide chain contains the amino acid sequence of SEQ ID NO: 104;

[0345] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises two identical first peptide chains and second peptide chains, wherein:

[0346] The first peptide chain contains the amino acid sequence of SEQ ID NO: 49; and

[0347] The second peptide chain contains the amino acid sequence of SEQ ID NO: 12.

[0348] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises two identical first peptide chains and second peptide chains, wherein:

[0349] The first peptide chain contains the amino acid sequence of SEQ ID NO: 52; and the second peptide chain contains the amino acid sequence of SEQ ID NO: 12.

[0350] In some embodiments, the aforementioned PD-1-SIRPγ fusion protein comprises two identical first and second peptide chains, wherein:

[0351] The first peptide chain contains the amino acid sequence of SEQ ID NO: 108; and the second peptide chain contains the amino acid sequence of SEQ ID NO: 104.

[0352] In some embodiments, a fusion protein is provided comprising a human Fc domain monomer fused with the SIRPγ variant, which is a SIRPγ variant-Fc fusion protein.

[0353] In some embodiments, the Fc domain monomers in the aforementioned SIRPγ variant-Fc fusion protein are the Fc regions of human IgG1, IgG2, or IgG4.

[0354] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0355] i) L14C and G115C; or

[0356] ii) Q8C and G107C.

[0357] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein contains any amino acid mutation selected from K19E, Q52S, K53G, E54R, M72K, and N101D.

[0358] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein contains amino acid mutations of K19E, Q52S, K53G, E54R, M72K, and N101D.

[0359] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein contains an N51A or N51M amino acid mutation.

[0360] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein further comprises one or more amino acid mutations selected from M6I, L13V, H56Q, N70E, R77K, S79Q, and M112V.

[0361] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein contains an amino acid mutation selected from any of the following:

[0362] a) L13V, N51A, and S79Q;

[0363] b) L13V, N51A, and R77K;

[0364] c) L13V, N51A and M112V;

[0365] d) L13V, N51M, H56Q and N70E; e) L13V and N51M;

[0366] f)M6I, N51M and R77K;

[0367] g)M6I, N51A and R77K;

[0368] h)M6I, N51A and N70E;

[0369] i) M6I, L13V, N51M and R77K;

[0370] j)M6I, L13V, N51A and R77K;

[0371] k)M6I, L13V, N51A and N70E;

[0372] l)L13V, N51A and N70E; and

[0373] m)N51A.

[0374] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein contains an amino acid mutation selected from any of the following:

[0375] n)N51A and R77K; or

[0376] o)N51A and M112V.

[0377] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises an amino acid mutation selected from the following:

[0378] p)N51M.

[0379] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0380] i)L14C and G115C;

[0381] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0382] iii) Amino acid mutations selected from any of the following:

[0383] a) L13V, N51A, and S79Q;

[0384] b) L13V, N51A, and R77K;

[0385] c) L13V, N51A and M112V;

[0386] d) L13V, N51M, H56Q and N70E;

[0387] e)L13V, N51A and N70E;

[0388] f)M6I, N51M and R77K;

[0389] g)M6I, N51A and R77K;

[0390] h)M6I, N51A and N70E;

[0391] j)M6I, L13V, N51M and R77K;

[0392] k)M6I, L13V, N51A and R77K;

[0393] l)M6I, L13V, N51A and N70E;

[0394] m)N51A; and

[0395] n)L13V and N51M.

[0396] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0397] i)L14C and G115C;

[0398] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0399] iii) Amino acid mutations selected from any of the following:

[0400] o)N51A and R77K; and

[0401] p)N51A and M112V.

[0402] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0403] i)L14C and G115C;

[0404] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0405] iii)N51M.

[0406] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0407] i)L14C and G115C;

[0408] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0409] iii) Amino acid mutations selected from any of the following:

[0410] a) L13V, N51A, and S79Q;

[0411] b) L13V, N51A and M112V;

[0412] c) L13V, N51M, H56Q and N70E;

[0413] d) L13V, N51A, and N70E;

[0414] e)N51A; and

[0415] f)N51A and R77K.

[0416] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0417] i)Q8C and G107C;

[0418] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0419] iii) Amino acid mutations selected from any of the following:

[0420] a) L13V, N51A, and S79Q;

[0421] b) L13V, N51A, and R77K;

[0422] c) L13V, N51A and M112V;

[0423] d) L13V, N51M, H56Q and N70E;

[0424] e)L13V, N51A and N70E;

[0425] f)M6I, N51M and R77K;

[0426] g)M6I, N51A and R77K;

[0427] h)M6I, N51A and N70E;

[0428] j)M6I, L13V, N51M and R77K;

[0429] k)M6I, L13V, N51A and R77K;

[0430] l)M6I, L13V, N51A and N70E;

[0431] m)N51A; and

[0432] n)L13V and N51M.

[0433] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0434] i)Q8C and G107C;

[0435] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0436] iii) N51A and R77K.

[0437] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0438] i)L14C and G115C;

[0439] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0440] iii) L13V and N51M.

[0441] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0442] i)L14C and G115C;

[0443] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0444] iii) M6I, N51A and N70E.

[0445] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises:

[0446] i)Q8C and G107C;

[0447] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0448] iii) L13V, N51A and R77K.

[0449] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein, wherein the SIRPγ variant comprises:

[0450] i)Q8C and G107C;

[0451] ii) K19E, Q52S, K53G, E54R, M72K, and N101D; and

[0452] iii) N51A and M112V.

[0453] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0454] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0455] SEQ ID NO: 13

[0456] in:

[0457] X1 is selected from M or I; X2 is selected from Q or C; X3 is selected from L or V; X4 is selected from L or C; X5 is selected from K or E; X6 is selected from N, M, or A; X7 is selected from Q or S; X8 is selected from K or G; X9 is selected from E or R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 Selected from N or D; X 16 Selected from G or C; X 17 Selected from M or V; X 18 Selected from G or C, and

[0458] i) When X4 is C, X 18 For C; or

[0459] ii) When X2 is C, X 16 The answer is C.

[0460] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0461] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX16 PGTEX 17 ALX 18 AKPS;

[0462] SEQ ID NO: 13

[0463] in:

[0464] X1 is selected from M or I; X3 is selected from L or V; X5 is selected from K or E; X6 is selected from N, M or A; X7 is selected from Q or S; X8 is selected from K or G; X9 is selected from E or R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 Selected from N or D; X 17 Selected from M or V; and:

[0465] i) X4 is C, and X 18 For C; or

[0466] ii) X2 is C, and X 16 The answer is C.

[0467] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0468] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0469] SEQ ID NO: 13

[0470] in:

[0471] X1 is selected from M or I; X3 is selected from L or V; X5 is E; X6 is selected from N, M or A; X7 is S; X8 is G; X9 is R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 K; X 13Selected from R or K; X 14 Selected from S or Q; X 15 For D; X 17 Selected from M or V; and:

[0472] i) X4 is C, and X 18 For C; or

[0473] ii) X2 is C, and X 16 The answer is C.

[0474] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0475] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0476] SEQ ID NO: 13

[0477] in:

[0478] X1 is selected from M or I; X3 is selected from L or V; X5 is E; X6 is selected from M or A; X7 is S; X8 is G; X9 is R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 For D; X 17 Selected from M or V; and:

[0479] i) X4 is C, and X 18 For C; or

[0480] ii) X2 is C, and X 16 The answer is C.

[0481] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein, wherein the SIRPγ variant comprises the following amino acid sequence:

[0482] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0483] SEQ ID NO: 13

[0484] in:

[0485] i) X4 is C, and X 18 The answer is C;

[0486] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 12 K; X 15 For D; and X 16 G; and iii) are selected from any one of the following a) to n):

[0487] a) X3 is V, X6 is A, X 14 Let Q be the number of elements in the array; and X1 be selected from M or I; X3 be selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; and X 17 Selected from M or V;

[0488] b) X3 is V, X6 is A, X 13 Let K be the number of elements; and X1 be selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0489] c) X3 is V, X6 is A, X 17 Let V be the variable; and X1 be selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K and X 14 Selected from S or Q;

[0490] d) X3 is V, X6 is M, X 10 For Q and X 11For E; and X1 is selected from M or I; X3 is selected from L or V; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0491] e) X3 is V and X6 is M; and X1 is selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0492] f) X1 is I, X6 is M and X 13 Let K be the number of elements in the array; and X3 be selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V; and

[0493] g) X1 is I, X6 is A and X 13 Let K be the number of elements in the array; and X3 be selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0494] h) X1 is I, X6 is A and X 11 E; and X3 is selected from L or V; X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0495] j) X1 is I, X3 is V, X6 is M and X 13 Let K be the number of K; and X be the number of X. 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0496] k) X1 is I, X3 is V, X6 is A and X 13 Let K be the number of K; and X be the number of X. 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V; and

[0497] l) X1 is I, X3 is V, X6 is A and X 11 Let E be the value of X; and X be the value of X. 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0498] m)X3 is V, X6 is A and X 11 E; X1 is selected from M or I; X3 is selected from L or V; X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V; and

[0499] n)X6 is A; and X1 is selected from M or I; X3 is selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V.

[0500] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0501] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0502] SEQ ID NO: 13

[0503] in:

[0504] i) X4 is C, and X 18 The answer is C;

[0505] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 15 For D; and X 16 For G; and

[0506] iii) X6 is selected from M, and X1 is selected from M or I; X3 is selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V.

[0507] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0508] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0509] SEQ ID NO: 13

[0510] in:

[0511] i) X4 is C, and X 18 The answer is C;

[0512] ii) X2 is Q; X5 is E; X7 is S; X8 is G; X9 is R; X 12 K; X 15 For D; and X 16 G; and iii) are selected from o) to r):

[0513] o)X6 represents A and X 13 Let K be the number of K, and X be the number of K. 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0514] p)X3 is V, X6 is M and X 13 Let K be the number of K, and X be the number of K. 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17Selected from M or V;

[0515] q) X3 is V, X6 is M and X 11 Let E be the integer, and X be the integer. 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V; and

[0516] r)X6 represents A and X 17 Let V be X, and X be X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0517] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0518] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0519] SEQ ID NO: 13

[0520] in:

[0521] i) X2 is C, and X 16 The answer is C;

[0522] ii) X2 is Q; X4 is L; X5 is E; X7 is S; X8 is G; X9 is R; X 12 K; X 15 For D; X 16 For G and X 18 For G; and

[0523] iii) Select any one of a) to n):

[0524] a) X3 is V, X6 is A, X 14 Let Q be the number of elements in the array; and X1 be selected from M or I; X3 be selected from L or V; X 10Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; and X 17 Selected from M or V;

[0525] b) X3 is V, X6 is A, X 13 Let K be the number of elements; and X1 be selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0526] c) X3 is V, X6 is A, X 17 Let V be the variable; and X1 be selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K and X 14 Selected from S or Q;

[0527] d) X3 is V, X6 is M, X 10 For Q and X 11 For E; and X1 is selected from M or I; X3 is selected from L or V; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0528] e) X3 is V, X6 is A and X 11 E; X1 is selected from M or I; X3 is selected from L or V; X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0529] f) X1 is I, X6 is M and X 13 Let K be the number of elements in the array; and X3 be selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0530] g) X1 is I, X6 is A and X 13 Let K be the number of elements in the array; and X3 be selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0531] h) X1 is I, X6 is A and X 11 E; and X3 is selected from L or V; X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0532] j) X1 is I, X3 is V, X6 is M and X 13 Let K be the number of K; and X be the number of X. 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0533] k) X1 is I, X3 is V, X6 is A and X 13 Let K be the number of K; and X be the number of X. 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0534] l) X1 is I, X3 is V, X6 is A and X 11 For E and X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V;

[0535] m)X6 is A; and X1 is selected from M or I; X3 is selected from L or V; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V; and

[0536] n) X3 is V and X6 is M; and X1 is selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; and X 17 Selected from M or V.

[0537] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0538] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0539] SEQ ID NO: 13

[0540] in:

[0541] i) X2 is C, and X 16 The answer is C;

[0542] ii) X2 is Q; X4 is L; X5 is E; X7 is S; X8 is G; X9 is R; X 12 K; X 15 For D; X 16 For G and X 18 For G; and

[0543] iii) Selected from either o) or p):

[0544] o)X6 is A, X 13 Let K be the number of elements; and X1 be selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; and X 17 Selected from M or V; or

[0545] p)X6 represents A and X 17 Let V be the variable; and X1 be selected from M or I; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K and X 14 Selected from S or Q;

[0546] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0547] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0548] SEQ ID NO: 13

[0549] i) X4 is C, and X 18 The answer is C;

[0550] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0551] iii) X1 is I, X6 is A and X 11 For E; and

[0552] iv) X2 is selected from Q; X3 is selected from L or V; X6 is selected from N, M or A; X 10 Selected from H or Q; X 13 Selected from R or K; X 14 Selected from S or Q; X 16 Selected from G; X 17 Selected from M or V.

[0553] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0554] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0555] SEQ ID NO: 13

[0556] i) X4 is C, and X 18 The answer is C;

[0557] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0558] iii) X3 is V, X6 is M; and

[0559] iv) X1 is selected from M or I; X2 is selected from Q; X4 is selected from L or C; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 16 Selected from G; X 17 Selected from M or V.

[0560] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0561] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0562] SEQ ID NO: 13

[0563] i) X2 is C, and X 16 The answer is C;

[0564] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0565] iii) X6 is A; X 17 For V; and

[0566] iv) X1 is selected from M or I; X3 is selected from L or V; X4 is selected from L or C; X 10 Selected from H or Q; X 11 Selected from N or E; X 13 Selected from R or K; X 14 Selected from S or Q; X 18Selected from G.

[0567] In some embodiments, the SIRPγ variant in the aforementioned SIRPγ variant-Fc fusion protein comprises the following amino acid sequence:

[0568] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0569] SEQ ID NO: 13

[0570] i) X2 is C, and X 16 The answer is C;

[0571] ii) X5 is E; X7 is S; X8 is G; X9 is R; X 12 For K; and X 15 For D; and

[0572] iii) X3 is V; X6 is A and X 13 Let K be the value of K.

[0573] iv) X1 is selected from M or I; X3 is selected from L or V; X4 is selected from L or C; X 10 Selected from H or Q; X 11 Selected from N or E; X 14 Selected from S or Q; X 17 Selected from M or V; and X 18 Selected from G.

[0574] In some embodiments, the Fc domain monomer in the aforementioned SIRPγ variant-Fc fusion protein comprises an Fc variant, wherein the Fc variant contains at least one amino acid mutation.

[0575] In some embodiments, the Fc variant described in the aforementioned SIRPγ variant-Fc fusion protein exhibits eliminated or reduced binding to the Fcγ receptor compared to the wild-type form of the human IgG Fc region. In some embodiments, the Fc variant exhibits eliminated or reduced binding to CD16a, CD32a, CD32b, CD32c, and CD64 Fcγ receptors compared to the wild-type form of the human IgG Fc region. In some embodiments, the Fc variant exhibits eliminated or reduced binding to C1q compared to the wild-type form of human IgG Fc. In some embodiments, the Fc variant exhibits eliminated or reduced binding to the Fcγ receptor compared to the wild-type human IgG4 Fc region.

[0576] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein, wherein the Fc domain monomer comprises one or more amino acid mutations of L234A, L235A, G237A, and N297A.

[0577] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein, wherein the Fc domain monomer contains amino acid mutations of L234A, L235A, G237A, and N297A.

[0578] In some embodiments, two Fc domain monomers may form an Fc domain dimer, wherein each Fc domain monomer is independently selected from (i) a human IgG1 Fc region containing mutations L234A, L235A, G237A, and N297A; (ii) a human IgG2 Fc region containing mutations A330S, P331S, and N297A; (iii) a human IgG4 Fc region containing mutations S228P, E233P, F234V, L235A, and N297A; or (iv) a human IgG4 Fc region containing mutations F234A and L235A; the amino acid mutation sites of the above-mentioned Fc domains are numbered using the EU numbering rule.

[0579] In some embodiments, the two Fc domain monomers are identical (i.e., homodimers). In some embodiments, the two Fc domain monomers are different (i.e., heterodimers). In some embodiments, at least one of the Fc domain monomers in the Fc domain dimer is a human IgG1 Fc region containing mutations of L234A, L235A, G237A, and N297A. In some embodiments, at least one of the Fc domain monomers in the Fc domain dimer is a human IgG2 Fc region containing mutations of A330S, P331S, and N297A. In some embodiments, at least one of the Fc domain monomers in the Fc domain dimer is a human IgG4 Fc region containing mutations of S228P, E233P, F234V, L235A, and N297A. In some embodiments, at least one of the Fc domain monomers in the Fc domain dimer is a human IgG4 Fc region containing F234A and L235A.

[0580] In some embodiments, the Fc domain monomer in the aforementioned SIRPγ variant-Fc fusion protein contains the amino acid sequence of SEQ ID NO: 60 or 61.

[0581] In some embodiments, the Fc domain monomer in the aforementioned SIRPγ variant-Fc fusion protein contains the amino acid sequence of SEQ ID NO: 109.

[0582] In some embodiments, the SIRPγ variant described in the aforementioned SIRPγ variant-Fc fusion protein is linked to the N-terminus or C-terminus of the Fc domain monomer via a peptide bond or linker.

[0583] In some embodiments, a linker (e.g., a spacer region) is inserted between the SIRPγ variant and the Fc domain monomer. In some embodiments, the disclosed high-affinity SIRPγ variant is fused to an Fc domain monomer that cannot form a dimer. In some embodiments, the disclosed SIRPγ variant is fused to an Fc domain monomer capable of forming a dimer (e.g., a heterodimer) with another Fc domain monomer. In some embodiments, the disclosed SIRPγ variant is fused to an Fc domain monomer, and this fusion protein forms a homodimer. In some embodiments, the disclosed SIRPγ variant is fused to a first Fc domain monomer, and a different protein or peptide (e.g., an antibody variable region) is fused to a second Fc domain monomer. In some embodiments, the SIRPγ variant is linked to a first Fc domain monomer, and a therapeutic protein (e.g., a cytokine, interleukin, antigen, steroid, anti-inflammatory agent, or immunomodulator) is linked to a second Fc domain monomer. In some embodiments, the first and second Fc domain monomers form a heterodimer.

[0584] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein comprises an Fc domain dimer, wherein the C-terminus of the SIRPγ variant is linked to the N-terminus of the Fc domain monomer via a linker; preferably, the linker is Gm(G4S)xGn, where x, m, and n are selected from integers from 0 to 10, and x, m, and n are not simultaneously 0.

[0585] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein comprises an Fc domain dimer, wherein the C-terminus of the SIRPγ variant is linked to the N-terminus of the Fc domain monomer via a peptide bond.

[0586] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein comprises an Fc domain dimer, wherein the N-terminus of the SIRPγ variant is linked to the C-terminus of the Fc domain monomer via a peptide bond.

[0587] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein comprises an Fc domain dimer, wherein the N-terminus of the SIRPγ variant is linked to the C-terminus of the Fc domain monomer via a linker.

[0588] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein comprises any amino acid sequence selected from SEQ ID NO: 62-90.

[0589] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein comprises any amino acid sequence selected from SEQ ID NO: 110-113.

[0590] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein is a homodimer containing two identical amino acid sequences selected from any of SEQ ID NO: 62-90.

[0591] In some embodiments, the aforementioned SIRPγ variant-Fc fusion protein is a homodimer containing two identical amino acid sequences selected from any of SEQ ID NO: 110-113.

[0592] In some embodiments, this disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of any of the preceding SIRPγ variants, or any of the preceding fusion proteins, and one or more pharmaceutically acceptable carriers, diluents, buffers, or excipients.

[0593] In some embodiments, the therapeutically effective amount is 0.1-3000 mg of any of the preceding SIRPγ variants or any of the preceding fusion proteins in a unit dose of the composition.

[0594] In some embodiments, this disclosure provides a nucleic acid molecule that encodes a polypeptide or a fusion protein as described in any of the foregoing embodiments.

[0595] In some implementations, this disclosure provides an expression vector comprising the aforementioned nucleic acid molecules.

[0596] In some embodiments, this disclosure provides a host cell comprising the aforementioned expression vector. In some embodiments, the host cell may be selected from prokaryotic cells and eukaryotic cells, preferably eukaryotic cells, and more preferably mammalian cells. In some embodiments, the host cell is a non-human mammalian cell. In some embodiments, the host cell does not include human embryonic cells, and the mammalian cells include, but are not limited to, CHO, 293, and NSO.

[0597] In some embodiments, this disclosure provides a method for treating an individual suffering from a disease or condition, the method comprising administering to the individual a therapeutically effective amount of any of the preceding SIRPγ variants, or any of the preceding fusion proteins, or the preceding pharmaceutical compositions, or the preceding nucleic acid molecules, or the preceding carriers.

[0598] In some implementations, the aforementioned method also includes administering a therapeutically effective amount of a CD38 antagonist to an individual suffering from a disease or condition.

[0599] In some embodiments, the aforementioned method further includes administering a therapeutically effective amount of a CD38 antagonist to an individual suffering from a disease or condition, wherein the CD38 antagonist is an anti-CD38 antibody.

[0600] In some embodiments, the anti-CD38 antibody described in the foregoing method comprises:

[0601] The heavy chain variable region includes HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 119, 120 and 121 respectively; and the light chain variable region includes LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 122, 123 and 124 respectively.

[0602] In some embodiments, the anti-CD38 antibody described in the foregoing method comprises a heavy chain variable region as shown in SEQ ID NO: 117 and a light chain variable region as shown in SEQ ID NO: 118.

[0603] In some embodiments, the anti-CD38 antibody described in the foregoing method comprises a heavy chain as shown in SEQ ID NO: 115 and a light chain as shown in SEQ ID NO: 116.

[0604] In some embodiments, this disclosure provides the use of any of the foregoing SIRPγ variants, or any of the foregoing fusion proteins, or the foregoing pharmaceutical compositions, or the foregoing nucleic acid molecules, or the foregoing expression vectors in the preparation of medicaments for treating diseases or conditions.

[0605] In some embodiments, this disclosure provides the use of any of the foregoing SIRPγ variants, or any of the foregoing fusion proteins, or the foregoing pharmaceutical compositions, or the foregoing nucleic acid molecules, or the foregoing expression vectors in combination with a CD38 antagonist (such as an anti-CD38 antibody) in the preparation of a medicament for treating a disease or condition.

[0606] In some embodiments, this disclosure provides a SIRPγ variant of any of the foregoing, or a fusion protein of any of the foregoing, or a pharmaceutical composition of the foregoing, or a nucleic acid molecule of the foregoing, or an expression vector of the foregoing, for use as a medicament for treating a disease or condition.

[0607] In some implementations, the aforementioned disease or condition is cancer, an autoimmune disease, or an inflammatory disease.

[0608] In some implementations, the aforementioned diseases or conditions, wherein the cancer is selected from: solid tumors, hematologic cancers, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, multiple myeloma, bladder cancer, pancreatic cancer, cervical cancer, endometrial cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, bronchial cancer, liver cancer, ovarian cancer, colorectal cancer, gastric cancer, gallbladder cancer, gastrointestinal stromal tumor cancer, thyroid cancer, head and neck cancer, oropharyngeal cancer, esophageal cancer, melanoma, non-melanoma skin cancer, Merkel cell carcinoma, virus-induced cancer, neuroblastoma, breast cancer, and prostate cancer. The list includes renal cell carcinoma, renal pelvis carcinoma, leukemia, lymphoma, sarcoma, glioma, and brain tumors; wherein the autoimmune disease or inflammatory disease is selected from multiple sclerosis, rheumatoid arthritis, spondyloarthritis, systemic lupus erythematosus, antibody-mediated inflammation or autoimmune diseases, graft-versus-host disease, sepsis, diabetes, psoriasis, atherosclerosis, Sjögren's syndrome, progressive systemic sclerosis, scleroderma, acute coronary syndrome, ischemia-reperfusion injury, Crohn's disease, endometriosis, glomerulonephritis, myasthenia gravis, idiopathic pulmonary fibrosis, asthma, acute respiratory distress syndrome (ARDS), vasculitis, and inflammatory autoimmune myositis. In some embodiments, the aforementioned diseases or conditions are associated with CD47 and / or PD-1.

[0609] The exemplary anti-PD-1 antibody sequences, preparation methods, and related properties disclosed herein are all described in PCT application No. PCT / CN2020 / 074098, the entire contents of which are incorporated herein by reference.

[0610] For example, the anti-PD-1 antibody is Hu23-11, whose sequence is as follows:

[0611] Table 1. CDR of anti-PD-1 antibodies

[0612]

[0613]

[0614] Hu23-11 Heavy Chain Variable Region:

[0615] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSS

[0616] SEQ ID NO: 7

[0617] >Variable region of Hu23-11 light chain:

[0618] DIVMTQTPLSLPVTPGEPASISCRSSQSLVHSTGNTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGGGTKVEIK

[0619] SEQ ID NO: 8

[0620] >Constant region of Hu23-11 heavy chain:

[0621] ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP P CPAPE AA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0622] SEQ ID NO: 9

[0623] >Constant region of Hu23-11 light chain:

[0624] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0625] SEQ ID NO: 10

[0626] >Hu23-11 heavy antibody chain:

[0627] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0628] SEQ ID NO: 11

[0629] >Hu23-11 light chain

[0630] DIVMTQTPLSLPVTPGEPASISCRSSQSLVHSTGNTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0631] SEQ ID NO: 12

[0632] >Hu33-5 heavy chain variable region:

[0633] EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVATISGGGVDTYYQDNVQGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPYGHGYFDVWGQGTTVTVSS

[0634] SEQ ID NO: 101

[0635] >Hu33-5 light chain variable region:

[0636] DIQMTQSPSSLSASVGDRVTITCRASQDINNFLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGGGTKVEIK

[0637] SEQ ID NO: 102

[0638] >Hu33-5 heavy chain:

[0639] EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVATISGGGVDTYYQDNVQGRFT ISRDNAKNSLYLQMNSLRAEDTAVYYCASPYGHGYFDVWGQGTTVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0640] SEQ ID NO: 103

[0641] >Hu33-5 light chain:

[0642] DIQMTQSPSSLSASVGDRVTITCRASQDINNFLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGSGT DYTLTISSLQPEDFATYYCQQGNTLPWTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0643] SEQ ID NO: 104 BRIEF DESCRIPTION OF THE DRAWINGS

[0644] Figure 1The experimental results show the binding of the SIRPγ fusion protein to human erythrocytes;

[0645] Figures 2A - 2F Experimental results showing the binding of different SIRPγ fusion proteins to Karpass 299 tumor cells are presented separately;

[0646] Figure 3 The results of the pharmacodynamic experiments of PD-1-SIRPγ fusion protein in a mouse Karpas299 model reconstructed from human PBMCs are shown.

[0647] Figure 4A A schematic diagram of the structure of the PD-1-SIRPγ fusion protein is shown. Figure 4B This diagram shows the structure of SIRPγ connected to the C-terminus of Fc. Figure 4C The diagram shows the structure of SIRPγ connected to the N-terminus of Fc.

[0648] Figure 5 The results of the fusion protein disclosed herein in a hemagglutination assay are shown.

[0649] Figure 6A The effect of fusion protein 4656 on stimulating PBMC release of IFNγ was demonstrated; Figure 6B The effects of fusion proteins 4658 and 4646 on stimulating PBMC release of IFNγ are shown.

[0650] Figure 7 This study demonstrates the therapeutic effect of the combination of the fusion protein and the anti-CD38 antibody on MOLP-8 cell xenografts in mice.

[0651] Figure 8 This study demonstrates the therapeutic effects of different fusion proteins on MDA-MB-231 cell xenografts in mice. Detailed Implementation

[0652] the term

[0653] The terminology used herein is for descriptive purposes only and is not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0654] The singular forms “a,” “an,” and “the” used in the specification and claims include plural references unless the context clearly indicates otherwise.

[0655] Unless the context clearly requires otherwise, throughout the specification and claims, the words “comprising,” “having,” “including,” etc., should be understood as having an inclusive meaning, rather than an exclusive or exhaustive meaning; that is, the meaning of “including but not limited to.”

[0656] The three-letter and single-letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968).

[0657] The term “SIRPγ” refers to any SIRPγ polypeptide or fragment thereof capable of binding to CD47. The amino acid sequence of the wild-type SIRPγ peptide is shown in SEQ ID NO: 14.

[0658] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to polymers of amino acid residues. These terms also apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as to both naturally occurring and non-naturally occurring amino acid polymers. Unless otherwise stated, a particular polypeptide sequence also implicitly encompasses variants with conserved modifications.

[0659] The "SIRPγ variant" comprises amino acid substitutions, deletions, or insertions (or combinations thereof) compared to the wild-type SIRPγ peptide, and the SIRPγ variant exhibits enhanced activity in binding to cell surface CD47 relative to the wild-type SIRPγ peptide. In some embodiments, the fusion protein formed by the SIRPγ variant is more stable and less prone to breakage. In some embodiments, the amino acid mutation occurs at one or more amino acid residues selected from M6, Q8, L13, L14, K19, N51, Q52, K53, E54, H56, N70, M72, R77, S79, N101, G107, M112, and G115. In some embodiments, the “SIRPγ variant” comprises at least three amino acid mutations selected from M6I, Q8C, L13V, L14C, K19E, N51M or N51A, Q52S, K53G, E54R, H56Q, N70E, M72K, R77K, S79Q, N101D, G107C, M112V, or G115C. In some embodiments, the aforementioned SIRPγ variant, relative to the wild-type SIRPγ peptide shown in SEQ ID NO: 14, comprises the following amino acid mutations:

[0660] i)L14C and G115C;

[0661] ii) Q8C and G107C; or

[0662] iii) Q8C and G115C.

[0663] The term "high-affinity SIRPγ variant" refers to a peptide containing a SIRPγ mutant that binds to CD47 with a higher affinity than wild-type SIRPγ.

[0664] In some specific implementations, the SIRPγ variant comprises a sequence as shown in SEQ ID NO: 13:

[0665] EEELQX1IX2PEKLX3X4VTVGX5TATLHCTVTSLLPVGPVLWFRGVGPGRELIYX6X7X8X9GX 10 FPRVTTVSDLTKRX 11 NX 12 DFSIX 13 IX 14 SITPADVGTYYCVKFRKGSPEX 15 VEFKSX 16 PGTEX 17 ALX 18 AKPS;

[0666] SEQ ID NO: 13

[0667] in:

[0668] X1 is selected from M or I; X2 is selected from Q or C; X3 is selected from L or V; X4 is selected from L or C; X5 is selected from K or E; X6 is selected from N, M, or A; X7 is selected from Q or S; X8 is selected from K or G; X9 is selected from E or R; X 10 Selected from H or Q; X 11 Selected from N or E; X 12 Selected from M or K; X 13 Selected from R or K; X 14 Selected from S or Q; X 15 Selected from N or D; X16 selected from G or C; X 17 Selected from M or V; X 18 Selected from G or C, and

[0669] i) When X4 is C, X 18 The answer is C;

[0670] ii) When X2 is C, X 16 For C; or

[0671] iii) When X2 is C, X 18 The answer is C.

[0672] Exemplary amino acid substitution sites included in the SIRPγ variants of this disclosure are shown in Table 2:

[0673]

[0674] The term "amino acid mutation" encompasses amino acid substitution, deletion, insertion, and modification. Any combination of substitution, deletion, insertion, and modification can be performed to achieve the final construct, provided the final construct possesses the desired properties. Amino acid sequence deletions and insertions include amino and / or carboxyl terminus deletions and amino acid insertions. A specific amino acid mutation is an amino acid substitution. In one embodiment, an amino acid mutation is a non-conservative amino acid substitution, i.e., replacing one amino acid with another amino acid that has a different structure and / or chemical properties. Amino acid substitutions include substitutions with non-naturally occurring amino acids or with derivatives of 20 naturally occurring amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5-hydroxylysine). Amino acid mutations can be generated using genetic or chemical methods known in the art. Genetic methods can include site-directed mutagenesis, PCR, gene synthesis, etc. Methods other than genetic engineering that alter amino acid side chain groups, such as chemical modification, are also expected to be available. Various names may be used herein to refer to the same amino acid mutation.

[0675] "Conservative" amino acid substitution refers to the interchangeability of residues with similar side chains. For example, a group of amino acids with aliphatic side chains includes glycine, alanine, valine, leucine, and isoleucine; a group of amino acids with aliphatic-hydroxyl side chains includes serine and threonine; a group of amino acids with amide-containing side chains includes asparagine and glutamine; a group of amino acids with aromatic side chains includes phenylalanine, tyrosine, and tryptophan; a group of amino acids with basic side chains includes lysine, arginine, and histidine; and a group of amino acids with sulfur-containing side chains includes cysteine ​​and methionine. Preferred conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine.

[0676] The term "fusion protein" generally refers to a protein obtained by fusing two or more proteins or peptides. Genes or nucleic acid molecules encoding the two or more proteins or peptides may be linked together to form a fusion gene or fused nucleic acid molecule that encodes the fusion protein. Translation of the fusion gene produces a single peptide that has the properties of at least one, or even each, of the two or more proteins or peptides prior to fusion. The terms fusion protein and recombinant fusion protein are used herein with the same meaning. Fusion proteins described herein generally contain at least two domains (A and B) and optionally include a third component, a linker between the two domains. The generation of recombinant fusion proteins is known in the art and generally involves removing the stop codon from a cDNA sequence encoding a first protein or peptide, and then attaching a cDNA sequence of a second protein in a frame-compliant manner via ligation or overlap extension PCR. This DNA sequence is then expressed by a cell as a single protein. This protein may be engineered to include the complete sequences of the two original proteins or peptides, or only a portion thereof. The fusion protein in this disclosure refers to a fusion protein comprising the SIRPγ variant of this disclosure. In some embodiments, the fusion protein is a PD-1-SIRPγ variant fusion protein, which is a tetrapeptide structure comprising an anti-PD-1 antibody and a SIRPγ variant, wherein the N-terminus of the SIRPγ variant is linked to the C-terminus of the anti-PD-1 antibody heavy chain via a linker. In other embodiments, the fusion protein is a SIRPγ-Fc fusion protein, wherein the C-terminus of the SIRPγ variant is linked to the N-terminus of Fc via a peptide bond or a linker.

[0677] The term "link" in which the SIRPγ variant is attached to the polypeptide chain of the anti-PD-1 antibody refers to an effective connection between the polypeptides, including, for example, a connection via peptide bonds or a linker. This connection does not result in the loss of function of either the SIRPγ peptide or the anti-PD-1 antibody.

[0678] "Linker" or "connector" refers to a linker polypeptide sequence used to connect protein domains or different proteins or different polypeptides. It usually has a certain degree of flexibility, and the use of linkers will not cause the original function of the protein domain to be lost.

[0679] The term “and / or”, such as “X and / or Y”, should be understood to mean “X and Y” or “X or Y” and should be used to provide clear support for both meanings or either meaning.

[0680] The terms “programmed cell death 1,” “programmed cell death 1,” “protein PD-1,” “PD-1,” “PDCD1,” and “hPD-1” are used interchangeably and include variants, isotypes, species homologs of human PD-1, and analogs that share at least one epitope with PD-1. The complete PD-1 sequence is available in GenBank accession number U64863.

[0681] The term “programmed death-ligand-1 (PD-L1)” is one of the two cell surface glycoprotein ligands of PD-1 (the other being PD-L2), which downregulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term “PD-L1” includes human PD-L1 (hPD-L1), variants, isotypes, and interspecies homologs of hPD-L1, as well as five analogs that share at least one common epitope with hPD-L1. The complete hPD-L1 sequence is available in GenBank accession number Q9NZQ7.

[0682] The terms “CD47”, “integrin-associated protein (IAP)”, “ovarian cancer antigen OA3”, and “Rh-associated antigen” are synonymous and used interchangeably to refer to human CD47 including the CD47 shown in NP_001768.1, as well as any natural polymorphisms of human CD47, such as single nucleotide polymorphisms (SNPs) or splice variants.

[0683] The term “antibody” as used in this article is used in the broadest sense and covers a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, and antibody fragments (or antigen-binding fragments, or antigen-binding portions), as long as they exhibit the desired antigen-binding activity.

[0684] "Natural antibodies" refer to naturally occurring immunoglobulin molecules with different structures. For example, natural IgG antibodies are heterotetraglycoproteins of approximately 150,000 Daltons, composed of two identical light chains and two identical heavy chains linked by disulfide bonds. From the N to the C-terminus, each heavy chain has a variable region (VH), also called a variable heavy domain or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from the N to the C-terminus, each light chain has a variable region (VL), also called a variable light domain or light chain variable domain, followed by a constant light (CL) domain.

[0685] The terms “full-length antibody,” “intact antibody,” and “all antibody” are used interchangeably in this document to refer to antibodies that have a structure substantially similar to that of natural antibodies or that have a heavy chain containing an Fc region as defined herein.

[0686] The term "variable region" or "variable domain" refers to the domain in the antibody heavy or light chain involved in antibody-antigen binding. VH and VL each contain four conserved frame regions (FRs) and three complementarity-determining regions (CDRs). The terms "complementarity-determining region" and "CDR" refer to the regions within the variable domain that primarily facilitate antigen binding; "frame" or "FR" refers to the variable domain residues other than the CDR residues. VH contains three CDR regions: HCDR1, HCDR2, and HCDR3; VL contains three CDR regions: LCDR1, LCDR2, and LCDR3. Each VH and VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A single VH or VL may be sufficient to confer antigen-binding specificity. Furthermore, antibodies binding to a specific antigen can be isolated by screening libraries of complementary VLs or VHs from the VH or VL of the antibody binding that antigen.

[0687] The amino acid sequence boundaries of CDRs can be determined using various well-known schemes, such as the "Kabat" numbering rule (see Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD), the "Chothia" numbering rule (see Al-Lazikani et al., (1997) JMB 273: 927-948), and the ImMunoGenTics (IMGT) numbering rule (Lefranc MP, Immunologist, 7, 132-136 (1999); Lefranc, MP et al., Dev. Comp. Immunol., 27, 55-77 (2003), etc.). The relationships between numbering systems, including, for example, the Kabat numbering and the unique IMGT numbering system, are well known to those skilled in the art and are shown in Table 3 below.

[0688] Table 3. Relationships between CDR numbering systems

[0689] Kabat + Chothia IMGT Kabat Chothia HCDR1 26-35 26-35 31-35 26-32 HCDR2 50-65 51-57 50-65 52-56 HCDR3 95-102 93-102 95-102 95-102 LCDR1 24-34 27-32 24-34 26-32 LCDR2 50-56 50-52 50-56 50-52 LCDR3 89-97 89-97 89-97 91-96

[0690] Unless otherwise stated, the variable regions and CDR sequences in this disclosure embodiment are subject to the "Kabat" numbering rule.

[0691] The "class" of an antibody refers to the type of constant region possessed by its heavy chain. Based on the amino acid sequence of its constant region, antibody light chains include two types: kappa (κ) and lambda (λ). According to the different amino acid compositions and sequences of the constant region of the antibody heavy chain, antibodies can be divided into five classes, or antibody isotypes: IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε chains, respectively. Within the same class of Ig, differences in the amino acid composition of its hinge region and the number and position of disulfide bonds in its heavy chain can further lead to different subclasses; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. Each of the five classes of Ig can have either a κ chain or a λ chain. The “conventional variants” of the human antibody heavy chain constant region and human antibody light chain constant region described in this disclosure refer to prior art variants of the heavy chain constant region or light chain constant region derived from humans that do not alter the structure and function of the antibody variable region. Exemplary variants include IgG1, IgG2, IgG3, and IgG4 heavy chain constant region variants that involve site-specific modifications and amino acid substitutions in the heavy chain constant region. In some embodiments, the substitutions are YTE mutations, L234A and / or L235A mutations, S228P mutations, and / or mutations that acquire a knock-in-hole structure. These mutations have been shown to impart novel properties to the antibody without altering the function of the antibody variable region. In some embodiments, the antibody is an IgG1 isotype with P329, P234, and P235 mutations in the hinge region to reduce effector function. In some embodiments, the antibody is an IgG2 isotype. In some embodiments, the antibody is an IgG4 isotype with an S228P mutation in the hinge region to improve the stability of the IgG4 antibody.

[0692] The term "antibody fragment" refers to a molecule that is distinct from the intact antibody but contains a portion of the intact antibody that binds to the antigen to which the intact antibody is bound. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, single-domain antibodies, biantibodies, linear antibodies, single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

[0693] The terms "Fc domain," "Fc region," or "fragment crystallizable region" are used to define the C-terminal region of an antibody heavy chain, including the native sequence Fc region and variant Fc regions. In some embodiments, the Fc region of the human IgG heavy chain is defined as an amino acid residue extending from the Cys226 position or from Pro230 to its carboxyl terminus. The boundary of the Fc region of the antibody heavy chain can also vary, for example, by omitting the C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) or by omitting the C-terminal glycine and lysine of the Fc region (residues 446 and 447 according to the EU numbering system). Therefore, in some embodiments, the composition of a complete fusion protein may include a population of fusion proteins with all K447 residues removed and / or G446+K447 residues removed. In some embodiments, the composition of a complete fusion protein may include a population of fusion proteins without the removal of K447 residues and / or G446+K447 residues. In some embodiments, the composition of the complete fusion protein comprises a population of fusion proteins containing and without K447 residues and / or G446+K447 residues. Suitable native sequence Fc regions for the fusion proteins described herein include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4. Unless otherwise specified herein, the amino acid residues in the Fc region or constant region are numbered according to the EU numbering system, also known as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0694] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a specific source or species, while the remaining portion of the heavy and / or light chain is derived from a different source or species.

[0695] The term "humanized" antibody refers to an antibody that retains the reactivity of a non-human antibody while exhibiting lower immunogenicity in humans. This can be achieved, for example, by retaining the non-human CDR region and replacing the rest of the antibody with its human counterpart (i.e., the framework portion of the constant region and the variable region).

[0696] The term "human antibody" refers to an antibody possessing an amino acid sequence that corresponds to the amino acid sequence of antibodies generated by humans or human cells, or to the amino acid sequence of antibodies derived from non-human sources using sequences encoded by human antibody sets or other human antibodies. The definition of human antibody explicitly excludes humanized antibodies containing non-human antigen-binding residues.

[0697] The term "KD" refers to the dissociation constant, which is derived from the ratio of kd to ka (i.e., kd / ka) and expressed as a molar concentration (M). The KD value of an antibody can be determined using methods well-established in the art. Methods for determining antibody KD include measuring surface plasmon resonance using a biosensing system, such as a system, or measuring affinity in solution by solution equilibrium titration (SET).

[0698] The term "effector function" refers to biological activities that can be attributed to the antibody's Fc region (either the native Fc region or the Fc region of an amino acid sequence variant) and vary across antibody isotypes. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0699] The term "antigen" refers to a molecule or molecular moiety that can be bound by a selective binder, such as an antigen-binding protein (including, for example, an antibody), and is further capable of being used in animals to produce antibodies that can bind to that antigen. An antigen may have one or more epitopes that can interact with different antigen-binding proteins (e.g., antibodies).

[0700] The term "affinity" or "binding affinity" refers to the strength of the binding interaction between two molecules. Generally, binding affinity refers to the sum of the strengths of non-covalent interactions between a molecule and its binding partner, such as high-affinity SIRP-γ peptide variants and CD47. Unless otherwise indicated, binding affinity refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of the binding pair. The binding affinity between two molecules is typically described by the dissociation constant (KD) or association constant (KA).

[0701] The term "less than" refers to a numerically smaller KD value relative to the stated KD value, along with an increased binding affinity. As used herein, the term "greater than" refers to a numerically larger KD value relative to the stated KD value, along with a decreased binding affinity.

[0702] The terms "anti-PD-1 antibody" and "PD-1-binding antibody" refer to antibodies capable of binding to PD-1 with sufficient affinity, making them usable as diagnostic and / or therapeutic agents targeting PD-1. In one embodiment, the binding extent of an anti-PD-1 antibody to an unrelated, non-PD-1 protein is less than about 10% of the antibody's binding to PD-1, for example, through... Measured by surface plasmon resonance assay. In some embodiments, the antibody binding to PD-1 has the following dissociation constant (KD): < about 1 μM, < about 100 nM, < about 10 nM, < about 1 nM, < about 0.1 nM, < about 0.01 nM, or < about 0.001 nM (e.g., 10⁻⁸ M or less, e.g., 10⁻⁸ M to 10⁻¹² M, e.g., 10⁻⁹ M to 10⁻¹⁰ M). In some embodiments, the anti-PD-1 antibody binds to conserved PD-1 epitopes from PD-1 of different species.

[0703] "About" means within an acceptable range of error for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. In the context of a particular measurement, result, or implementation, unless otherwise expressly stated in the embodiments or elsewhere in the specification, "about" means within one standard deviation, or up to 5%, according to convention in the art.

[0704] The terms “antibody-dependent cell cytotoxicity,” “antibody-dependent cell-mediated cytotoxicity,” or “ADCC” refer to mechanisms that induce cell death that rely on the interaction between antibody-coated target cells and lytic effector cells (such as natural killer (NK) cells, monocytes, macrophages, and neutrophils) via Fcγ receptors (FcγR) expressed on the effector cells. For example, NK cells express FcγRIIIa, while monocytes express FcγRI, FcγRII, and FcγRIIIa. The ADCC activity of the antibodies described herein can be assessed in vitro using cells expressing the antigen as target cells and NK cells as effector cells. Cell lysis is detected based on the release of markers (e.g., radioactive substrates, fluorescent dyes, or native intracellular proteins) from lysed cells.

[0705] The term “antibody-dependent phagocytosis” (“ADCP”) refers to the mechanism by which antibody-coated target cells are eliminated through internalization by phagocytes (such as macrophages or dendritic cells).

[0706] The term "complement-dependent cytotoxicity" or "CDC" refers to a mechanism that induces cell death in which the Fc effector domain of a target-binding antibody binds to and activates the complement component C1q. C1q then activates the complement cascade, leading to target cell death. Activation of complement can also result in the deposition of complement components on the surface of target cells, which promote CDC by binding to complement receptors (e.g., CR3) on leukocytes.

[0707] The term "nucleic acid" is used interchangeably with the term "polynucleotide" herein and refers to deoxyribonucleotides or ribonucleotides and their polymers in single-stranded or double-stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, or non-naturally occurring, have similar binding properties to a reference nucleic acid, and are metabolized in a manner similar to that of a reference nucleotide. Examples of such analogs include, but are not limited to, thiophosphates, aminophosphates, methylphosphonates, chiral methylphosphonates, 2-O-methylribonucleotides, and peptide-nucleic acids (PNAs). "Isolated" nucleic acids refer to nucleic acid molecules that have been separated from components of their native environment. Isolated nucleic acids include nucleic acid molecules found in cells that typically contain such molecules but are present outside the chromosome or at a chromosomal location different from their native chromosomal location.

[0708] Unless otherwise stated, a particular nucleic acid sequence also implicitly encompasses variants of its conserved modifications (e.g., degenerate codon substitutions) and complementary sequences, as well as explicitly stated sequences. Specifically, as detailed below, degenerate codon substitutions can be obtained by generating sequences in which the third position of one or more selected (or all) codons is substituted with a mixture of bases and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., J. Biol. Chem. 260:2605-2608, 1985; and Rossolini et al., Mol. Cell. Probes 8:91-98, 1994).

[0709] The term "sequence identity" refers to the degree (percentage) to which the amino acids / nucleic acids of two sequences are identical at equivalent positions when two sequences are optimally aligned, with gaps introduced where necessary to obtain the maximum percentage of sequence identity, without considering any conserved substitutions as part of sequence identity. To determine the percentage of sequence identity, alignment can be performed in a variety of ways within the scope of the art, such as using publicly available computer software like BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR) software. Those skilled in the art can determine the parameters suitable for measuring alignment, including any algorithms required to achieve maximum alignment across the full length of the sequences being compared.

[0710] The terms "conservatively modified variant" or "conservative substitution" refer to the replacement of an amino acid in a protein with another amino acid having similar characteristics (e.g., charge, side chain size, hydrophilicity / hydrophobicity, ossicular structure, and rigidity), such that such a change can generally be made without altering the protein's biological activity. Those skilled in the art will appreciate that, generally, a single amino acid substitution in a non-essential region of a polypeptide does not substantially alter its biological activity (see, for example, Watson et al., (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Ed.)). When applied to nucleic acid sequences, the term "conservatively modified variant" refers to nucleic acids that encode the same or substantially the same amino acid sequence, or, where the nucleic acid does not encode an amino acid sequence, substantially the same sequence. Due to the degeneracy of the genetic code, any given protein can be encoded by multiple functionally identical nucleic acids. For example, the codons GCA, GCC, GCG, and GCU all encode the amino acid alanine. Therefore, at each position where the codon designates alanine, that codon can be changed to any of the corresponding codons without altering the encoded polypeptide. Such nucleic acid variations are “silent variations,” and they are a type of conserved modification variation. Each nucleic acid sequence encoding a polypeptide described herein also represents every possible silent variation of the nucleic acid. Those skilled in the art will recognize that each codon in the nucleic acid (except for AUG—usually the only codon for methionine; and TGG—usually the only codon for tryptophan) can be modified to produce a functionally identical molecule. Therefore, each silent variation of the nucleic acid encoding the polypeptide is implicitly contained in each of the said sequences.

[0711] The term "vector" refers to a polynucleotide molecule capable of transporting another polynucleotide linked to it. One type of vector is a "plasmid," which is a circular double-stranded DNA loop in which an additional DNA segment can be attached. Another type of vector is a viral vector, such as adeno-associated virus vectors (AAV or AAV2), in which an additional DNA segment can be attached to the viral genome. Some vectors are capable of autonomous replication in the host cells to which they are introduced (e.g., bacterial vectors with bacterial origins of replication and attachable mammalian vectors). Other vectors (e.g., non-attached mammalian vectors) can integrate into the host cell's genome after introduction into the host cell, thereby replicating along with the host genome.

[0712] The term "expression vector" or "expression construct" refers to a nucleic acid sequence suitable for transformation of host cells and containing one or more heterologous coding regions that direct and / or control (together with the host cell) the expression of the vector. Expression constructs may include, but are not limited to, sequences that affect or control transcription, translation, and, in the presence of introns, influence RNA splicing of coding regions that are operatively linked to them.

[0713] As used herein, “operably linked” means that the components to which the term applies are in a relationship that allows them to perform their inherent functions under suitable conditions. For example, a control sequence in a vector that is “operably linked” to a protein-coding sequence is linked to it, thereby achieving expression of the protein-coding sequence under conditions compatible with the transcriptional activity of that control sequence.

[0714] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include “transformers” and “transformed cells,” which include primary transformed cells and their derived progeny, regardless of passage number. Progeny may not be identical to parental cells in their nucleic acid contents and may contain mutations. This includes mutant progeny with the same function or biological activity as those screened or selected in the initially transformed cells. Host cells include prokaryotic and eukaryotic host cells, with eukaryotic host cells including, but not limited to, mammalian cells, insect cell lines, plant cells, and fungal cells. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cattle, horse, and hamster cells, including but not limited to Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, young hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, and HEK-293 cells.Fungal cells include yeast and filamentous fungal cells, including, for example, *Pichia pastoris*, *Pichia finlandica*, *Pichia trehalophila*, *Pichia koclamae*, *Pichiamembranaefaciens*, *Pichia minuta* (Ogataea minuta, *Pichia lindneri*), *Pichia xiaopuntiae*, *Pichia thermotolerans*, *Pichia salictaria*, *Pichia guercuum*, *Pichia pijperi*, *Pichia stiptis*, *Pichia methanolica*, *Pichia* genus, *Saccharomyces cerevisiae*, *Saccharomyces* genus, and *Hansenula*. The fungi include *Candida polymorpha*, *Kluyveromyces lactis*, *Candida albicans*, *Aspergillus nidulans*, *Aspergillus niger*, *Aspergillus oryzae*, *Trichoderma reesei*, *Chrysosporium lucknowense*, *Fusarium sp.*, *Fusarium gramineum*, *Fusarium venenatum*, *Physcomitrella patens*, and *Neurospora crassa*. Pichia, any Saccharomyces, Hansenula polymorpha, any Kluyveromyces, Candida albicans, any Aspergillus, Trichoderma reesei, Chrysosporium lucknowense, any Fusarium, Yarrowia lipolytica, and Neurospora crassa.

[0715] As used in this application, the terms “cell,” “cell line,” and “cell culture” are used interchangeably, and all such names include progeny. Therefore, the terms “transformer” and “transformed cell” include primary subject 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 progeny will have identical DNA contents. This includes mutant progeny that have the same function or biological activity as the original transformed cells from which they were selected.

[0716] "Optional" or "optionally" means that the event or circumstances described below may, but do not have to, occur, including the circumstances in which the event or circumstances may or may not occur.

[0717] The term "pharmaceutical composition" means a mixture containing one or more antibodies or antigen-binding fragments described herein, along with other chemical components, such as physiological / pharmaceutical carriers and excipients.

[0718] The term "pharmaceutically acceptable carrier" refers to a component in a pharmaceutical formulation that is distinct from the active ingredient and non-toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0719] The term "packaging insert" is used to refer to the instruction leaflet typically included in the commercial packaging of a therapeutic product, which contains information about indications, usage, dosage, administration, combination therapy, contraindications, and / or warnings about the use of such a therapeutic product.

[0720] The terms “subject” or “individual” include both humans and non-human animals. Non-human animals include all vertebrates (e.g., mammals and non-mammals) such as non-human primates (e.g., cynomolgus monkeys), sheep, dogs, cattle, chickens, amphibians, and reptiles. Unless otherwise specified, the terms “patient” or “subject” are used interchangeably herein. As used herein, the term “cynomolgus monkey” refers to the cynomolgus monkey (Macaca fascicularis). In some embodiments, the individual or subject is a human.

[0721] "Administration" or "giving," when applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, refers to the contact between an exogenous drug, therapeutic agent, diagnostic agent, or composition and the animal, human, subject, cell, tissue, organ, or biological fluid.

[0722] The term "sample" refers to a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present within the subject's body. Exemplary samples include biological fluids such as blood, serum and serous fluid, plasma, lymph, urine, saliva, cystic fluid, tears, excretions, sputum, mucosal secretions of secretory tissues and organs, vaginal secretions, ascites, pleura, pericardium, peritoneum, fluids in the abdominal cavity and other body cavities, fluids collected by bronchoalveolar lavage fluid, synovial fluid, liquid solutions in contact with the subject or biological sources, such as cell and organ culture media (including cell or organ conditioned media), lavage fluids, tissue biopsy samples, fine-needle aspiration, surgically removed tissue, organ cultures, or cell cultures.

[0723] "Treatment" (and its grammatical variations) refers to a clinical intervention that attempts to alter the natural processes of the individual being treated, and may be implemented for prevention or during a clinicopathological process. The desired effects of treatment include, but are not limited to, preventing the onset or recurrence of disease, alleviating symptoms, reducing / decreasing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, improving or alleviating the disease state, and resolving or improving prognosis. In some implementations, the antibodies disclosed herein are used to delay the onset of disease or slow its progression.

[0724] An "effective dose" is generally an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate such symptoms and / or underlying causes, prevent the occurrence of symptoms and / or underlying causes, and / or improve or mitigate damage caused by or associated with a disease state (e.g., lung disease). In some embodiments, an effective dose is a therapeutically effective dose or a preventatively effective dose. A "therapeuticly effective dose" is an amount sufficient to treat a disease state or symptom, particularly a state or symptom associated with that disease state, or otherwise prevent, inhibit, delay, or reverse the progression of the disease state or any other undesirable symptom associated with that disease. A "preventatively effective dose" is an amount that, when administered to a subject, will have a predetermined preventative effect, such as preventing or delaying the onset (or recurrence) of the disease state, or reducing the likelihood of the onset (or recurrence) of the disease state or related symptoms. A complete therapeutic or preventative effect may not occur after the administration of a single dose, but may occur after the administration of a series of doses. Therefore, a therapeutically or preventatively effective dose may be administered in a single or multiple-dose manner. "Therapeutic effective dose" and "preventive effective dose" can vary depending on factors such as an individual's disease state, age, sex, and weight, as well as the ability of the treatment agent or combination of treatment agents to elicit the desired response in the individual. Exemplary indicators of an effective treatment agent or combination of treatment agents include, for example, improved health status in the patient.

[0725] The term “diseases associated with CD47 and / or PD-1 activity” refers to any disease or disorder caused by and / or associated with CD47 and / or PD-1 activity, such as any disease or disorder caused by and / or associated with increased or decreased CD47 and / or PD-1 activity.

[0726] The terms “cancer” and “cancerous” refer to or describe a physiological disorder typical of mammals characterized by unregulated cell growth. This definition includes both benign and malignant cancers. “Early-stage cancer” or “early-stage tumor” refers to non-invasive or metastatic cancer, or cancer classified as stage 0, I, or II. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma (including medulloblastoma and retinoblastoma), sarcoma (including liposarcoma and synovial cell sarcoma), neuroendocrine tumors (including carcinoid tumors, gastrinomas, and islet cell carcinomas), mesothelioma, Schwann cell tumors (including acoustic neuroma), meningioma, adenocarcinoma, melanoma, and leukemia or lymphoid malignancies. More specific examples of this type of cancer include, but are not limited to, squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma and squamous cell carcinoma of the lung, peritoneal cancer, hepatocellular carcinoma, gastric cancer (including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer (or hepatic carcinoma), bladder cancer, hepatoma, breast cancer (including metastatic breast cancer), colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer (or renal cancer), prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, testicular cancer, esophageal cancer, bile duct tumors, and head and neck cancer and multiple myeloma.

[0727] "Autoimmune disease" or "inflammatory disease" refers to any disease, condition, or syndrome in which an excessive or unregulated inflammatory response results in excessive inflammatory symptoms, damage to host tissues, or loss of tissue function. In some embodiments disclosed herein, the autoimmune disease or inflammatory disease mentioned herein is selected from multiple sclerosis, rheumatoid arthritis, spondyloarthritis, systemic lupus erythematosus, antibody-mediated inflammation or autoimmune diseases, graft-versus-host disease, sepsis, diabetes, psoriasis, atherosclerosis, Sjögren's syndrome, progressive systemic sclerosis, scleroderma, acute coronary syndrome, ischemia-reperfusion, Crohn's disease, endometriosis, glomerulonephritis, myasthenia gravis, idiopathic pulmonary fibrosis, asthma, acute respiratory distress syndrome (ARDS), vasculitis, and inflammatory autoimmune myositis.

[0728] The term "antagonist" is used in the broadest sense and includes any molecule that partially or completely blocks, inhibits, or neutralizes the biological activity of the natural CD38 polypeptide. Suitable antagonist molecules include, in particular, antagonist antibodies or antibody fragments (e.g., antigen-binding fragments), fragments or amino acid sequence variants of natural polypeptides, peptides, antisense oligonucleotides, small organic molecules, etc.

[0729] Example

[0730] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the disclosure. Experimental methods in the embodiments of this disclosure that do not specify specific conditions are generally performed under conventional conditions, such as those described in Cold Spring Harbor's Antibody Technology Manual or Molecular Cloning Manual; or under conditions recommended by the raw material or commercial manufacturer. Reagents that do not specify their source are commercially available, conventional reagents.

[0731] Example 1. Screening and preparation of SIRPγ mutants

[0732] To obtain a high-affinity CD47 receptor, the CD47 receptor (SIRPγ) was maturated using yeast display and phage display technologies. Based on SIRPγ, an affinity maturation library targeting CD47 binding was designed and prepared, and high-affinity SIRPγ mutants were screened from it. Simultaneously, based on the crystal structure of SIRPγ, intrachain disulfide bonds were designed using computer simulation (Molecular Operating Environment software).

[0733] >SIRPγ peptide wild type

[0734] EEELQMIQPEKLLLVTVGKTATLHCTVTSLLPVGPVLWFRGVGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRISSITPADVGTYYCVKFRKGSPENVEFKSGPGTEMALGAKPS

[0735] SEQ ID NO: 14

[0736] After screening, the following SIRPγ variants were finally obtained:

[0737] S-46:

[0738] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0739] SEQ ID NO:15

[0740] >S-56:

[0741] EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0742] SEQ ID NO:16

[0743] >S-57:

[0744] EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0745] SEQ ID NO:17

[0746] >S-58:

[0747] EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0748] SEQ ID NO:18

[0749] >S-59:

[0750] EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0751] SEQ ID NO:19

[0752] >S-60:

[0753] EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0754] SEQ ID NO:20

[0755] >S-61:

[0756] EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0757] SEQ ID NO:21

[0758] >S-62:

[0759] EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0760] SEQ ID NO:22

[0761] >S-63:

[0762] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0763] SEQ ID NO:23

[0764] >S-64:

[0765] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0766] SEQ ID NO:24

[0767] >S-65:

[0768] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0769] SEQ ID NO:25

[0770] >S-66:

[0771] EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0772] SEQ ID NO:26

[0773] >S-8Q:

[0774] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRIQSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0775] SEQ ID NO:27

[0776] >S-10Q:

[0777] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0778] SEQ ID NO:28

[0779] >S-12Q:

[0780] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEVALGAKPS

[0781] SEQ ID NO:29

[0782] >S-14Q:

[0783] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGQFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0784] SEQ ID NO:30

[0785] >S-15Q:

[0786] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0787] SEQ ID NO:31

[0788] >S-20Q

[0789] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0790] SEQ ID NO:32

[0791] >S-21Q

[0792] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0793] SEQ ID NO:33

[0794] >S-8L

[0795] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRIQSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0796] SEQ ID NO:34

[0797] >S-12L

[0798] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEVALCAKPS

[0799] SEQ ID NO:35

[0800] >S-14L

[0801] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGQFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0802] SEQ ID NO:36

[0803] >S-15L

[0804] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0805] SEQ ID NO:37

[0806] >S-20L

[0807] EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0808] SEQ ID NO:38

[0809] >S-30

[0810] EEELQIICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0811] SEQ ID NO:39

[0812] >S-31

[0813] EEELQIICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0814] SEQ ID NO:40

[0815] >S-32

[0816] EEELQIICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0817] SEQ ID NO:41

[0818] >S-33

[0819] EEELQIICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0820] SEQ ID NO:42

[0821] >S-34

[0822] EEELQIICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0823] SEQ ID NO:43

[0824] >S-35

[0825] EEELQIICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0826] SEQ ID NO:44

[0827] >S-36

[0828] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0829] SEQ ID NO:45

[0830] >S-37

[0831] EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0832] SEQ ID NO: 46

[0833] Further mutations of S-10Q and S-12Q yielded the following mutants:

[0834] >S-10Qm:

[0835] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS

[0836] SEQ ID NO: 105

[0837] >S-12Qm

[0838] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEVALGAKPS

[0839] SEQ ID NO: 106

[0840] Example 2. Construction and expression of PD-1-SIRPγ fusion protein

[0841] SIRPγ mutants are linked to anti-PD-1 antibodies directly or via linkers, wherein the SIRPγ mutant can be linked to the C-terminus or N-terminus of the heavy or light chain of the anti-PD-1 antibody to construct different forms of fusion proteins. The linker can be (G4S)nGx, where x and n are independently selected from integers from 1 to 10.

[0842] An example connector is as follows:

[0843] Table 4. Connecting Subsequences

[0844]

[0845] For example, the SIRPγ mutant is linked to the C-terminus of the heavy chain of an anti-PD-1 antibody via a linker. Then, the resulting heavy chain of the anti-PD-1 antibody containing the SIRPγ mutant is recombined with the light chain of the anti-PD-1 antibody to obtain a tetrapeptide fusion protein containing two identical polypeptides (structure shown in [link to structure]). Figure 4A ).

[0846] Based on the following sequences, heavy chain gene fragments containing the SIRPγ mutant fusion protein and light chain gene fragments containing the anti-PD-1 antibody were synthesized. The heavy chain gene and light chain gene were cloned into eukaryotic expression vectors, respectively, to form heavy chain plasmids and light chain plasmids. These expression vectors were then introduced into eukaryotes to express the antibody fusion protein. The cell culture supernatant was purified using a MabSelect Sure affinity column to obtain the expression product.

[0847] The exemplary amino acid sequence of the light chain of the anti-PD-1 antibody fusion protein in this disclosure is as follows:

[0848] (1) The relevant sequence of the fusion protein obtained by fusing with Hu-23-11 antibody:

[0849] >Antibody fusion protein light chain amino acid sequence (fusion protein second chain)

[0850] DIVMTQTPLSLPVTPGEPASISCRSSQSLVHSTGNTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGGGTKV EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0851] SEQ ID NO: 12

[0852] The exemplary amino acid sequence of the anti-PD-1 antibody fusion protein heavy chain in this disclosure is as follows:

[0853] >4646 Heavy chain amino acid sequence (first chain of the fusion protein)

[0854] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGAGGGGSGGGGSGGGGSGGGGSG EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFP RVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0855] SEQ ID NO: 49 [[ID=⑦]]

[0856] >4656 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0857] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0858] SEQ ID NO: 50

[0859] >4657 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0860] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0861] SEQ ID NO: 51

[0862] >4658 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0863] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVT TVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0864] SEQ ID NO: 52

[0865] >4659 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0866] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0867] SEQ ID NO: 53

[0868] >4660 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0869] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0870] SEQ ID NO: 54

[0871] >4661 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0872] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0873] SEQ ID NO: 55

[0874] >4662 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0875] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVT TVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0876] SEQ ID NO: 56

[0877] >4663 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0878] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0879] SEQ ID NO: 57

[0880] >4664 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0881] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVT TVSDLTKRNNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0882] SEQ ID NO: 58

[0883] >4665 Heavy chain amino acid sequence (the first chain of the fusion protein)

[0884] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYEMHWVRQAPGQGLEWMGLIDPETGGTVYNQKFKDRVTITADKSTSTAYMELSSLRSEDTAVYYCARERFSYYGSTSDWYFDVWGQ GTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQ VYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVT TVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0885] SEQ ID NO: 59

[0886] (2) Relevant sequence of the fusion protein obtained by fusing with Hu-33-5 antibody:

[0887] >Fusion protein second chain (light chain):

[0888] DIQMTQSPSSLSASVGDRVTITCRASQDINNFLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGSGT DYTLTISSLQPEDFATYYCQQGNTLPWTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0889] SEQ ID NO: 104

[0890] >5048 First strand sequence (second strand of fusion protein):

[0891] EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVATISGGGVDTYYQDNVQGRFT ISRDNAKNSLYLQMNSLRAEDTAVYYCASPYGHGYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTK RENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0892] SEQ ID NO: 107

[0893] >5047 First strand sequence:

[0894] EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVATISGGGVDTYYQDNVQGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPYGHGYFDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGG EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTK RNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEVALGAKPS

[0895] SEQ ID NO: 108

[0896] Example 3. Construction and expression of SIRPγ-Fc fusion protein

[0897] A SIRPγ mutant is linked to an Fc to construct a SIRPγ-Fc fusion protein. In some embodiments, the Fc includes an Fc variant containing at least one amino acid mutation.

[0898] An example Fc domain monomer sequence is as follows:

[0899] Fc-1:

[0900] ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

[0901] SEQ ID NO: 60

[0902] Fc-2:

[0903] DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0904] SEQ ID NO: 61

[0905] Fc-3:

[0906] ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSI EKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

[0907] SEQ ID NO: 109

[0908] For example, the SIRPγ-Fc fusion protein is a dipeptide structure containing two identical polypeptide chains, wherein the N-terminus of the SIRPγ mutant is fused directly or via a linker to the C-terminus of the Fc, or the C-terminus of the SIRPγ mutant is fused directly to the N-terminus of the Fc, to obtain the SIRPγ-Fc fusion protein (structure shown in [link to structure]). Figure 4B and Figure 4C ).

[0909] Different gene fragments were synthesized based on the following sequences, and the genes were cloned into eukaryotic expression vectors. These expression vectors were then introduced into eukaryotes to express the SIRPγ-Fc fusion protein. The cell culture supernatant was purified using a MabSelect Sure affinity column to obtain the expression product.

[0910] The exemplary amino acid sequence of the SIRPγ-Fc fusion protein disclosed herein is as follows:

[0911] >4666

[0912] ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGGSGGGG EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVT TVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[0913] SEQ ID NO: 62

[0914] >8QCGC

[0915] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRIQSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0916] SEQ ID NO:63

[0917] >10QCGC

[0918] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0919] SEQ ID NO:64

[0920] >12QCGC

[0921] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEVALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0922] SEQ ID NO:65

[0923] >14QCGC

[0924] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGQFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0925] SEQ ID NO:66

[0926] >15QCGC

[0927] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0928] SEQ ID NO:67

[0929] >20QCGC

[0930] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0931] SEQ ID NO:68

[0932] >21QCGC

[0933] EEELQMICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0934] SEQ ID NO:69

[0935] >8LCGC

[0936] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRIQSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0937] SEQ ID NO:70

[0938] >10LCGC

[0939] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0940] SEQ ID NO:71

[0941] >12LCGC

[0942] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEVALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0943] SEQ ID NO:72

[0944] >14LCGC

[0945] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGQFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0946] SEQ ID NO:73

[0947] >15LCGC

[0948] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0949] SEQ ID NO:74

[0950] >20LCGC

[0951] EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0952] SEQ ID NO:75

[0953] >21LCGC

[0954] EEELQMIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0955] SEQ ID NO:76

[0956] >56-Fc

[0957] EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0958] SEQ ID NO:77

[0959] >57-Fc

[0960] EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0961] SEQ ID NO:78

[0962] >58-Fc

[0963] EEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0964] SEQ ID NO:79

[0965] >60-Fc

[0966] EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0967] SEQ ID NO:80

[0968] >61-Fc

[0969] EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0970] SEQ ID NO:81

[0971] >62-Fc

[0972] EEELQIIQPEKLVCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0973] SEQ ID NO:82

[0974] >30-Fc

[0975] EEELQIICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0976] SEQ ID NO:83

[0977] >31-Fc

[0978] EEELQIICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0979] SEQ ID NO:84

[0980] >32-Fc

[0981] EEELQIICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0982] SEQ ID NO:85

[0983] >33-Fc

[0984] EEELQIICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0985] SEQ ID NO:86

[0986] >34-Fc

[0987] EEELQIICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0988] SEQ ID NO:87

[0989] >35-Fc

[0990] EEELQIICPEKLVLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR ENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0991] SEQ ID NO:88

[0992] >36-Fc

[0993] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0994] SEQ ID NO:89

[0995] >37-Fc

[0996] EEELQMIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYMSGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0997] SEQ ID NO:90

[0998] >4854(10QCGCm)

[0999] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIKISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEMALGAKPS DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1000] SEQ ID NO:110

[1001] >4855(12QCGCm)

[1002] EEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKR NNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEVALGAKPSDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1003] SEQ ID NO:111

[1004] >4924

[1005] ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGGEEELQIIQPEKLLCVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRENKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSGPGTEMALCAKPS

[1006] SEQ ID NO:112

[1007] >4845

[1008] ESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSGGGGSGGGGSGGGGEEELQMICPEKLLLVTVGETATLHCTVTSLLPVGPVLWFRGVGPGRELIYASGRGHFPRVTTVSDLTKRNNKDFSIRISSITPADVGTYYCVKFRKGSPEDVEFKSCPGTEVALGAKPS

[1009] SEQ ID NO: 113

[1010] The following proteins were also prepared and purified using conventional methods as control proteins:

[1011] HX009 is a bispecific antibody against CD47 and PD-1, prepared according to WO2019109357, and its sequence is as follows:

[1012] >HX009 heavy chain amino acid sequence

[1013] EVQLVQSGGGLVQPGGSLKLSCAASGFTFSSYGMSWVRQAPGKGLDWVATISGGGRDTYYPDSVKGRFTISRDNSKNNLYLQMNSLRAEDTALYYCARQKGEAWFAYWGQGTLVTVSAASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSGGGGSERGETGPEEELQVIQPDKSVSVAAGESAILHCTVTSLIPVGPIQWFRGAGPARELIYNQKEGHFPRVTTVSESTKRENMDFSISISNITPADAGTYYCVKFRKGSPDTEFKSGAGTELSVRAKPS

[1014] SEQ ID NO: 91

[1015] > Amino acid sequence of the light chain of HX009

[1016] DIVLTQSPASLAVSPGQRATITCRASESVDNYGISFMNWFQQKPGQPPKLLIYAASNKGTGVPARFSGSGSGTDFTLNINPMEEEDTAMYFCQQSKEVPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1017] SEQ ID NO: 92

[1018] ALX148 is a SIRPα-Fc fusion protein, prepared according to US10259859, and its sequence is as follows;

[1019] >ALX148

[1020] EEELQVIQPDKSVLVAAGETATLRCTITSLFPVGPIQWFRGAGPGRELIYNQREGPFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKPSDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1021] SEQ ID NO: 93

[1022] >SIRPγWT-Fc

[1023] EEELQMIQPEKLLLVTVGKTATLHCTVTSLLPVGPVLWFRGVGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRISSITPADVGTYYCVKFRKGSPENVEFKSGPGTEMALGAKPSEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1024] SEQ ID NO: 94

[1025] >CD47-Fc

[1026] QLLFNKTKSVEFTFCNDTVVIPCFVTNMEAQNTTEVYVKWKFKGRDIYTFDGALNKSTVPTDFSSAKIEVSQLLKGDASLKMDKSDAVSHTGNYTCEVTELTREGETIIELKYRVVSWFSPNEEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[1027] SEQ ID NO: 114

[1028] This disclosure also uses an unrelated anti-HIV antibody, C25, as a negative control.

[1029] The anti-CD38 antibody used in the test case is the hu11E antibody disclosed in patent WO2020052546, and its sequence is as follows:

[1030] >hu11E antibody heavy chain:

[1031] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGMHWVRQAPGKGLEWVAFISSGSSSIYYADTVKGRFTISRDNAKSSLYLQMTSLRAEDTAVYYCARNYVSSYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIAKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1032] SEQ ID NO: 115

[1033] >hu11E antibody light chain:

[1034] DIVMTQSPDSLAVSLGERATINCRASENVDNYGISFMHWYQQKPGQPPKLLIYRASNLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSNKDPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1035] SEQ ID NO: 116

[1036] >hu11E antibody heavy chain variable region:

[1037] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGMHWVRQAPGKGLEWVAFISSGSSSIYYADTVKGRFTISRDNAKSSLYLQMTSLRAEDTAVYYCARNYVSSYGYFDYWGQGTTVTVSS

[1038] SEQ ID NO: 117

[1039] >hu11E antibody light chain variable region:

[1040] DIVMTQSPDSLAVSLGERATINCRASENVDNYGISFMHWYQQKPGQPPKLLIYRASNLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSNKDPLTFGGGTKVEIK

[1041] SEQ ID NO: 118

[1042] CDR region of hu11E antibody:

[1043] >HCDR1:DYGMH (SEQ ID NO: 119)

[1044] >HCDR2:FISSGSSSIYYADTVKG(SEQ ID NO: 120)

[1045] >HCDR3:NYVSSYGYFDY(SEQ ID NO: 121)

[1046] >LCDR1:RASENVDNYGISFMH (SEQ ID NO: 122)

[1047] >LCDR2:RASNLES(SEQ ID NO: 123)

[1048] >LCDR3:QQSNKDPLT (SEQ ID NO: 124).

[1049] Test case

[1050] Test Example 1. BIAcore Affinity Assay for PD-1-SIRPγ Fusion Protein

[1051] IgG was captured using a Protein A biosensor chip (Cat.#29127556, GE) through affinity binding. Different antigens (hCD47: Cat.#12283-H08H, Lot.#LC12OC2201, SB; hPD-1: Cat.#10377-H08H, Lot.#LC13AU2111, SB) were allowed to flow over the chip surface. A Biacore T200 instrument was used to detect the reaction signals between the fusion protein and different antigens in real time, obtaining binding and dissociation curves. After dissociation in each experimental cycle, the biosensor chip was washed and regenerated using 10 mM Glycine-HCl pH 1.5 buffer. The experimental buffer system was 1×HBS-EP buffer solution (Cat#BR-1001-88, GE). After the experiment, the data were fitted using a (1:1) Langmuir model with GE Biacore T200 Evaluation version 3.0 software to obtain affinity values, as shown in Table 5.

[1052] The results showed that the SIRPγ variant fusion protein in this application had a significantly higher affinity for human CD47 than the wild-type SIRPγ peptide (the affinity of wild-type SIRPγ for CD47 was about 23 μM). The affinity of the PD-1-SIRPγ fusion protein for human PD-1 was comparable to that of the naked PD-1 anti-hu23-11 and higher than that of the bispecific antibody HX009 against CD47 and PD-1. The affinity of the SIRPγ-Fc fusion protein for human CD47 was significantly higher than that of the wild-type SIRPγ peptide.

[1053] Table 5. Affinity of fusion proteins

[1054]

[1055]

[1056] Test Example 2. Experiment on the binding of fusion protein to human red blood cells

[1057] Fresh, healthy human blood was mixed with an equal volume of PBS and centrifuged at 300g for 5 minutes to obtain cell clusters. Red blood cells were then washed 3-5 times with PBS. The cells were resuspended in FACS buffer (PBS + 5% BSA) and the cell density was adjusted to 2 × 10⁻⁶ cells / mL. 6Cells were seeded at a rate of 100 μL / well in 96-well round-bottom plates (3795#, corning), followed by the addition of different concentrations of antibody or fusion protein, with C25 as a negative control. Cells were incubated at 4°C for 1 hour. After washing twice with FACS buffer (PBS + 2% FBS), secondary antibody (Alexa488 goat anti-human IgG antibody: Invitrogen, CAT#A11013) was added, and the cells were incubated on ice in the dark for 30 minutes. Finally, cells were washed twice with FACS buffer and resuspended. The plates were read from FACS Cantoll. Results are shown in [Figure number missing]. Figure 1 .

[1058] FACS results showed that the control CD47 antibody hu5F9 (prepared according to US9017675) and the SIRPα-Fc fusion protein ALX148 both had strong binding ability to native CD47 on the surface of human erythrocytes, while the PD-1-SIRPγ fusion protein in this application had almost no binding ability to native CD47 on the surface of human erythrocytes, or only a weak binding ability. SIRPγ-Fc's binding to native CD47 on the surface of erythrocytes was weaker than that of the control antibody and the fusion protein, suggesting the safety advantages of the aforementioned SIRPγ-Fc and PD-1-SIRPγ fusion proteins.

[1059] Test Example 3. Fusion Protein Binding to Tumor Cells Experiment

[1060] Karpas 299 cells (Shanghai Zeye Biotechnology Co., Ltd.) were cultured in DMEM F12 medium (containing 10% fetal bovine serum) at a density of 1×10⁻⁶. 6 Karpas 299 cells / mL were blocked with 5% BSA, and the sample was added to a concentration of 10 μg / mL. After washing twice, Alexa Fluor 488-goat anti-human (H+L) antibody (Invitrogen, CAT#A11013) was added, and after washing twice, the fluorescence signal value was read by flow cytometry. Results are shown below. Figures 2A-2F .

[1061] FACS results showed that the PD-1-SIRPγ fusion protein and SIRPγ-Fc fusion protein disclosed herein have a strong binding ability to native CD47 on the surface of Karpas 299 cells, which is superior to the control bispecific antibody HX009.

[1062] Test Example 4. Efficacy experiment of SIRPγ fusion protein in a human PBMC-reconstructed mouse Karpas299 model.

[1063] This test case uses the Karpas299 mouse model (Beijing Biocytogen Gene Biotechnology Co., Ltd.) reconstituted with human PBMCs to evaluate the antitumor efficacy of PD-1-SIRPγ fusion protein in mice.

[1064] Karpas299 cells (1×10⁻⁶) 5 Cells / mouse / 100μL, containing 50μLMatrigel) were subcutaneously inoculated into the right rib area of ​​female NDG mice. Simultaneously, PBMCs from two donors were mixed at a 1:1 ratio and injected at 5×10⁻⁶ cells / mouse. 6 Cells / 100μL / mouse were injected intraperitoneally into mice. When the tumor volume in the tumor-bearing mice reached 160mm²... 3 Mice were randomly divided into groups of 7-8 mice each. The day of grouping was defined as Day 0 of the experiment. Starting on Day 0, the test samples were administered intraperitoneally twice a week for a total of four injections over two weeks. Tumor volume and animal weight were monitored and recorded twice weekly. All data were plotted and statistically analyzed using Excel and GraphPadPrism 5 software.

[1065] The formula for calculating tumor volume (V) is: V = 1 / 2 × a × b 2 Where a and b represent length and width, respectively.

[1066] The relative tumor proliferation rate T / C (%) = (T-T0) / (C-C0) × 100, where T and C are the tumor volumes of the treatment group and the control group at the end of the experiment; T0 and C0 are the tumor volumes at the beginning of the experiment.

[1067] Tumor inhibition rate TGI (%) = 1 - T / C (%).

[1068] Results (see) Figure 3 As shown in Table 6, after 11 days of administration, the tumor inhibition rate of PD-1 antibody hu23-11-8.5mpk was only 10.24%. The tumor inhibition rates of high-dose (4.4mpk) and low-dose (0.4mpk) Fc-SIRPγ fusion protein 4666 were 77.05% (p<0.001) and 33.04% (p<0.01), respectively. In contrast, the tumor inhibition rate of bispecific antibody HX009-10mpk was 60.00% (p<0.001).

[1069] The tumor inhibition rates of high-dose 10mpk and low-dose 1mpk of PD-1-SIRPγ fusion protein 4646 were 92.14% (p<0.001) and 41.59% (p<0.01), respectively, which were significantly better than those of high-dose PD-1 antibody hu23-11 and bispecific antibody HX009 at the same dose.

[1070] Table 6. Tumor inhibition rates of different proteins

[1071] protein dose Tumor inhibition rate % (Day 11) hu23-11 8.5mpk 10.24 4666 4.4mpk 77.05 4666 0.4mpk 33.04 HX009 10mpk 60 4646 10mpk 92.14 4646 1mpk 41.59

[1072] Test Example 5. Erythrocyte Agglutination Assay of Fusion Protein

[1073] Fresh, healthy human blood was diluted 100-fold with PBS (B320#, Shanghai Yuanpei Biotechnology Co., Ltd.). The diluted whole blood was spread into 96-well round-bottom plates (3795#, corning), 30 μL / well. Then, antibodies or bifunctional fusion proteins of varying concentrations were added in equal volumes. After mixing, the plates were incubated at 37°C for 4–6 hours. Erythrocyte sedimentation was observed using a high-content microscope. No hemagglutination was indicated by clear red dots, while hemagglutination was indicated by diffuse patterns.

[1074] Each sample was diluted 1:3 from the first column (0.25 mg / mL) to the 10th column. The 11th column contains PBS blank wells without antibody. Results are shown below. Figure 5 .

[1075] The results showed that, under the same conditions, neither the PD-1-SIRPγ fusion protein nor the SIRPγ-Fc fusion protein induced erythrocyte agglutination at the tested concentrations. This suggests the safety advantages of the PD-1-SIRPγ fusion protein and SIRPγ-Fc fusion protein disclosed herein.

[1076] Test Example 6. PD-1-SIRPγ bifunctional fusion protein stimulation of PBMC-T lymphocytes to secrete IFNγ experiment.

[1077] To investigate the effect of the PD-1-SIRPγ bifunctional fusion protein on the function of human primary T lymphocytes, human peripheral blood mononuclear cells (PBMCs) were collected and purified. After in vitro stimulation with tuberculin (TB) for 5 days, the secretion level of the cytokine IFNγ was measured. The experimental procedure is briefly described below:

[1078] PBMCs were obtained from fresh blood using Ficoll-Hypaque (17-5442-02, GE) density gradient centrifugation (Stem Cell Technologies) and cultured in RPMI 1640 (SH30809.01, GE) medium supplemented with 10% (v / v) FBS (10099-141, Gibco) at 37°C and 5% CO2.

[1079] Freshly isolated and purified PBMCs were adjusted to a density of 2 × 10⁶ cells / mL using RPMI 1640 medium. 6Cells were cultured at 20 mL of 20 mL cell suspension with 40 μL of tuberculin (97-8800, Synbiotics) added, and incubated at 37°C in a 5% CO2 incubator for 5 days. On day 5, the cultured cells were collected, centrifuged, and resuspended in fresh RPMI 1640 medium, adjusting the density to 1.1 × 10⁻⁶ cells / mL. 6 Cells were seeded at a density of 90 μL / mL into 96-well cell culture plates. Serially diluted antibody or fusion protein samples were added simultaneously, diluted with PBS (B320, Shanghai Yuanpei Biotechnology Co., Ltd.), 10 μL per well. The cell culture plates were incubated at 37°C in a 5% CO2 incubator for 3 days. After incubation, the cell culture plates were removed, and the cell culture supernatant was collected by centrifugation (4000 rpm, 10 min). The IFN-γ level was detected using an ELISA kit (Human IFN-γ Detection Kit: EHC102g.96, Xinbosheng). Specific procedures were followed according to the reagent instructions. Results are shown in Tables 7-8 and 8. Figure 6A and Figure 6B .

[1080] Table 7. Results of fusion protein activating PBMC secretion of IFNγ

[1081] sample EC50(nM) Emax% 4656 0.156 156.517 hu23-11 5.431 117.468 HX009 0.136 100

[1082] Table 8. Results of fusion protein activating PBMC secretion of IFNγ

[1083] sample EC50(nM) Emax% 4646 0.154 134.811 4658 0.277 147.607 HX009 0.145 100

[1084] The results showed that all bifunctional fusion proteins could activate the secretion of IFN-γ, and were superior to the control bispecific antibody HX009.

[1085] Test Example 7. The therapeutic effect of SIRPγ-Fc fusion protein on human multiple myeloma cell line MOLP-8 xenografts.

[1086] 5 × 10 MOLP-8 cells 6 Cells / mouse / 200μL (containing 50% Matrigel) were subcutaneously injected into the right rib area of ​​98 Balb / c nude mice. When the tumor volume of the tumor-bearing mice reached 160mm², the cells were injected into the tumor. 3 Mice were randomly divided into 5 groups of 7 mice each (the analyte concentrations were maintained at equimolar levels). The day of group assignment was defined as Day 0 of the experiment. Initially, mice received intraperitoneal injections of the respective fusion proteins or antibodies twice weekly. From Day 8 onwards, the frequency of administration of 4855 and 4845 was increased to three times weekly, while administration of the CD38 antibody hu11E was discontinued. The administration period was 17 days (Day 17). Tumor volume and animal weight were monitored and recorded twice weekly. When the tumor volume exceeded 1500 mmHg... 3 The experimental endpoint was to euthanize the tumor-bearing animals when most tumors ulcerated or the animals lost 20% of their body weight.

[1087] The CD38 antibody used in the experiment was the hu11E antibody disclosed in patent WO2020052546.

[1088] All data were plotted and statistically analyzed using Excel and GraphPad Prism 5 software.

[1089] The formula for calculating tumor volume (V) is: V = 1 / 2 × a × b 2 Where a and b represent length and width, respectively.

[1090] The relative tumor proliferation rate T / C (%) = (T-T0) / (C-C0) × 100, where T and C are the tumor volumes of the treatment group and the control group at the end of the experiment; T0 and C0 are the tumor volumes at the beginning of the experiment.

[1091] Tumor inhibition rate TGI (%) = 1 - T / C (%).

[1092] At the end of the experiment, tumor-bearing mice were euthanized, tumors were removed, and tumor weight was measured. The results showed that the weight of the removed tumor generally correlated with the trend of tumor volume. Tumor-bearing mice tolerated all specific antibodies and their monoclonal antibodies well; their weight fluctuated only slightly throughout the administration process, and no significant drug-induced weight loss or other symptoms occurred. The results are shown in Table 9 and... Figure 7 .

[1093] Table 9. Efficacy of SIRPγ-Fc fusion protein against subcutaneous xenografts in MOLP-8 mice

[1094]

[1095] Note: Day 0: First administration time; Seven days after administration, the dosing frequency for 4855 and 4845 was increased to three times a week, and CD38 antibody administration was discontinued; *p<0.05 vs Vehicle by student T test.

[1096] Experimental results showed that in the MOLP-8 mouse subcutaneous xenograft model of human multiple myeloma cells, CD38-specific antibody hu11E, SIRPγ-Fc assay molecule 4855, and 4845, used alone, all had a certain inhibitory effect on the growth of MOLP-8 mouse subcutaneous xenografts, with tumor inhibition rates of 84.86%, 33.89%, and 32.58%, respectively. The combination of CD38 antibody hu11E and 4855 resulted in a tumor inhibition rate >100%, which was significantly different from the CD38 antibody alone group (p<0.05).

[1097] Test Example 8. The therapeutic effect of PD-1-SIRPγ bifunctional fusion protein on human breast cancer cell MDA-MB-231 xenografts.

[1098] MDA-MB-231 cells (ATCC) 3×10 6 200 μL / mouse (containing 50% Matrigel) of cells were subcutaneously injected into the right rib area of ​​NOD / SCID mice. The tumor volume of the tumor-bearing mice reached an average of 178 mmHg. 3 Mice were randomly divided into 9 groups: PBS, 4646 (10 mpk), 5048 (10 mpk), 5047 (10 mpk), 4658 (10 mpk, 3 mpk), 4924 (4.4 mpk), HX009 (10 mpk), and hu23-11 (8.6 mpk), with 7 mice in each group. The day of grouping was defined as Day 0 of the experiment. On Day 0, PBMCs from two volunteers who had been stimulated with CD3 antibody for 3 days were mixed at a 1:1 ratio and inoculated with 5 × 10⁻⁶ ppm. 5 100 μL of cells per mouse tumor tissue was injected. The remaining PBMCs were discontinued from stimulation and cultured for another week. After one week, the cells were injected at 5 × 10⁻⁶ cells per mouse tumor tissue. 6 Cells / 100μL / mouse were injected intraperitoneally into tumor-bearing mice, considered as round 1 injection. Two rounds of PBMC injections were administered until the end of the experiment. Starting from Day 0, the tested fusion proteins or antibodies were injected intraperitoneally twice weekly. Tumor volume and animal weight were monitored and recorded twice weekly. When the tumor volume exceeded 1000 mmHg... 3 The experimental endpoint was to euthanize the tumor-bearing animals when most tumors ulcerated or the animals lost 20% of their body weight.

[1099] Data was recorded using Excel 2007 statistical software. Mean values ​​were calculated as averages (avg); standard deviation (SD) values ​​were calculated as standard deviations (STDEV); and summative efficiency (SEM) values ​​were calculated as STDEV / SQRT. Graphpad Prism 8 was used for chart processing and statistical analysis. P-values ​​for differences between groups were calculated using analysis of variance.

[1100] Tumor volume: V = 1 / 2 × L 长 ×L 短 2

[1101] Relative tumor proliferation rate: T / C (%) = [(T-T0) / (C-C0)] × 100%

[1102] Tumor inhibition rate: TGI (%) = 1 - T / C (%).

[1103] Where T0 and T are the tumor volumes in the treatment group at the beginning and end of the experiment, respectively. C0 and C are the tumor volumes in the solvent control group (vehicle) at the beginning and end of the experiment, respectively.

[1104] In the experiment, the test substance was administered intraperitoneally twice a week for a total of four weeks. Tumor volume data on day 32 after group administration were plotted and statistically analyzed. The results are shown in Table 10 and Figure 8 .

[1105] Table 10. Efficacy of fusion protein against MDA-MB-231 xenografts

[1106]

[1107] The results showed that, at the same dose of 10 mpk, 4646, 5048, 5047, and 4658 had significantly better inhibitory effects on MDA-MB-231 xenografts than the control bifunctional molecule HX009. The tumor-inhibiting effect of 4658 was dose-dependent.

[1108] Test Example 9. Degree of aggregation of fusion proteins

[1109] Aggregation degree was monitored by SEC-HPLC. A Waters e2695 chromatograph was used with a Waters Xbridge BEH 200A SEC column. The mobile phase was PBS (adjusted to pH 6.8 with dilute hydrochloric acid). 50 μg of protein was injected, and isocratic elution was performed at a flow rate of 0.5 mL / min. The results are shown in Table 11.

[1110] Table 11. Purity of Fusion Proteins

[1111] Fusion protein SEC monomers (%) 8QCGC 96.48 10QCGC 97.68 4854 99.06 12QCGC 97 4855 98.69 14QCGC 96.78 15QCGC 98.58 20QCGC 96.98

[1112] Upon examination, the SEC purity of the fused molecules disclosed herein is all above 95%. sequence list <110> Jiangsu Hengrui Medicine Co., Ltd. Shanghai Hengrui Medicine Co., Ltd. <120> SIRPγ variants and their fusion proteins <130> 721102CPCT <150> CN202010922561.2 <151> 2020-09-04 <150> CN202110985121.6 <151> 2021-08-25 <160> 124 <170> SIPOSequenceListing 1.0 <210> 1 <211> 5 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 HCDR1 <400> 1 Asp Tyr Glu Met His 1 5 <210> 2 <211> 17 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 HCDR2 <400> 2 Leu Ile Asp Pro Glu Thr Gly Gly Thr Val Tyr Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 3 <211> 16 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 HCDR3 <400> 3 Glu Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe Asp Val 1 5 10 15 <210> 4 <211> 16 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 LCDR1 <400> 4 Arg Ser Ser Gln Ser Leu Val His Ser Thr Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 5 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 LCDR2 <400> 5 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 6 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 LCDR3 <400> 6 Phe Gln Gly Ser His Val Pro Tyr Thr 1 5 <210> 7 <211> 125 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 Heavy Chain Variable Region <400> 7 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 8 <211> 112 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 Light Chain Variable Region <400> 8 Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser 20 25 30 Thr Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 9 <211> 326 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 heavy chain constant region <400> 9 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser 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 Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro 100 105 110 Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115 120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp 145 150 155 160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 165 170 175 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 Leu Ser Leu Ser Leu Gly 325 <210> 10 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Hu23-11 Light Chain Constant Region <400> 10 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> 11 <211> 451 <212> PRT <213> Artificial Sequence <220> <SI21> CHAIN <223> Hu23-11 heavy chain <400> 11 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly 450 <210> 12 <211> 219 <212> PRT [[ID=​​​​​​​​​Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser 20 25 30 Thr Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 115 120 125 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 130 135 140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 145 150 155 160 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185 190 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200 205 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 13 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> SIRPγ variant general formula <220> <221> UNSURE <222> (6)..(6) <223> Xaa is selected from Met or Ile. <220> <221> UNSURE <222> (8)..(8) <223> Xaa is selected from Gln or Cys. <220> <221> UNSURE <222> (13)..(13) <223> Xaa is selected from Leu or Val. <220> <221> UNSURE <222> (14)..(14) <223> Xaa is selected from Leu or Cys. <220> <221> UNSURE <222> (19)..(19) <223> Xaa is selected from Lys or Glu. <220> <221> UNSURE <222> (51)..(51) <223> Xaa is selected from Asn, Ala, or Met. <220> <221> UNSURE <222> (52)..(52) <223> Xaa is selected from Gln or Ser. <220> <221> UNSURE <222> (53)..(53) <223> Xaa is selected from Lys or Gly. <220> <221> UNSURE <222> (54)..(54) <223> Xaa is selected from Glu or Arg. <220> <221> UNSURE <222> (56)..(56) <223> Xaa is selected from His or Gln. <220> <221> UNSURE <222> (70)..(70) <223> Xaa is selected from Asn or Glu. <220> <221> UNSURE <222> (72)..(72) <223> Xaa is selected from Met or Lys. <220> <221> UNSURE <222> (77)..(77) <223> Xaa is selected from Arg or Lys. <220> <221> UNSURE <222> (79)..(79) <223> Xaa is selected from Ser or Gln. <220> <221> UNSURE <222> (101)..(101) <223> Xaa is selected from Asn or Asp. <220> <221> UNSURE <222> (107)..(107) <223> Xaa is selected from Gly or Cys. <220> <221> UNSURE <222> (112)..(112) <223> Xaa is selected from Met or Val. <220> <221> UNSURE <222> (115)..(115) <223> Xaa is selected from Gly or Cys. <400> 13 Glu Glu Glu Leu Gln Xaa Ile Xaa Pro Glu Lys Leu Xaa Xaa Val Thr 1 5 10 15 Val Gly Xaa Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Xaa Xaa Xaa Xaa Gly Xaa Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Xaa Asn Xaa Asp Phe Ser Ile Xaa Ile Xaa Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Xaa Val Glu Phe Lys Ser Xaa Pro Gly Thr Glu Xaa 100 105 110 Ala Leu Xaa Ala Lys Pro Ser 115 <210> 14 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> SIRPγ peptide wild type <400> 14 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Lys Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Asn Gln Lys Glu Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Met Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asn Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 15 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-46 <400> 15 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 16 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-56 <400> 16 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 17 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-57 <400> 17 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 18 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-58 <400> 18 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 19 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-59 <400> 19 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 20 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑60 <400> 20 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 21 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑61 <400> 21 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 22 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑62 <400> 22 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 23 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑63 <400> 23 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 24 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-64 <400> 24 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 25 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑65 <400> 25 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 26 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑66 <400> 26 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 27 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-8Q <400> 27 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Gln Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 28 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-10Q <400> 28 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 29 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-12Q <400> 29 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Val 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 30 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-14Q <400> 30 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly Gln Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 31 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-15Q <400> 31 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 [[ID=……]]Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 32 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-20Q <400> 32 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 33 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-21Q <400> 33 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 34 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-8L <400> 34 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Gln Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 35 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-12L <400> 35 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Val 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 36 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-14L <400> 36 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly Gln Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 37 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-15L<00​​​​​​​​​​​​​​​​​​​​​​Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 38 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-20L <400> 38 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 39 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-30 <400> 39 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 40 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-31 <400> 40 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 41 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-32 <400> 41 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 42 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-33 <400> 42 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 43 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-34 <400> 43 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 44 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S‑35 <400> 44 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 45 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-36 <400> 45 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser 115 <210> 46 <211> 119 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> S-37 <400> 46 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser 115 <210> 47 <211> 21 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Linker 1 <400> 47 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1 5 10 15 I Gly Gly Gly Ser Gly 20 U <210> 48 <211> 19 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> Linker 2 <400> 48 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1 5 10 15 Gly Gly Gly <210> 49 <211> 592 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4646 Heavy Chain Amino Acid Sequence <400> 49 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Gly Gly Gly Gly Ser Gly Glu Glu Glu Leu Gln Met Ile 465 470 475 480 Gln Pro Glu Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu 485 490 495 His Cys Thr Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe 500 505 510 Arg Gly Val Gly Pro Gly Arg Glu Leu Ile Tyr Met Ser Gly Arg Gly 515 520 525 His Phe Pro Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn 530 535 540 Lys Asp Phe Ser Ile Arg Ile Ser Ser Ile Thr Pro Ala Asp Val Gly 545 550 555 560 Thr Tyr Tyr Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu 565 570 575 Phe Lys Ser Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 590 <210> 50 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4656 heavy chain amino acid sequence <400> 50 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu 465 470 475 480 Lys Leu Leu Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Met Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 51 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> Amino acid sequence of the 4657 heavy chain <400> 51 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu 465 470 475 480 Lys Leu Leu Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Ala Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 52 <211> 589 <212> PRT <213> Artificial Sequence [[ID=四十二]]<220>[[ID=四十三]] [[ID=四十四]]<221> CHAIN[[ID=四十五]] [[ID=四十六]]<223> 4658 Heavy Chain Amino Acid Sequence[[ID=四十七]] <400> 52 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu 465 470 475 480 Lys Leu Leu Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Ala Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe 530 535 540 Ser Ile Arg Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 53 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4659 Heavy Chain Amino Acid Sequence <400> 53 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Met Ile Gln Pro Glu 465 470 475 480 Lys Leu Leu Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Ala Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 54 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4660 Heavy Chain Amino Acid Sequence <400> 54 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu 465 470 475 480 Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Met Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 55 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4661 Heavy chain amino acid sequence <400> 55 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu 465 470 475 480 Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Ala Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu 465 470 475 480 Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Ala Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe 530 535 540 Ser Ile Arg Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 57 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4663 Heavy Chain Amino Acid Sequence <400> 57 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Met Ile Gln Pro Glu 465 470 475 480 Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Ala Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 58 ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Met Ile Gln Pro Glu 465 470 475 480 Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Met Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe 530 535 540 Ser Ile Lys Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 555 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 59 <211> 589 <212> PRT <213> Artificial Sequence <220> <221> CHAIN <223> 4665 Heavy Chain Amino Acid Sequence <400> 59 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 Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Asp Pro Glu Thr Gly Gly Thr Val 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 Arg Phe Ser Tyr Tyr Gly Ser Thr Ser Asp Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Glu Glu Glu Leu Gln Met Ile Gln Pro Glu 465 470 475 480 Lys Leu Val Cys Val Thr Val Gly Glu Thr Ala Thr Leu His Cys Thr 485 490 495 Val Thr Ser Leu Leu Pro Val Gly Pro Val Leu Trp Phe Arg Gly Val 500 505 510 Gly Pro Gly Arg Glu Leu Ile Tyr Met Ser Gly Arg Gly His Phe Pro 515 520 525 Arg Val Thr Thr Val Ser Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe 530 535 540 Ser Ile Arg Ile Ser Ser Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr 545 550 550 560 Cys Val Lys Phe Arg Lys Gly Ser Pro Glu Asp Val Glu Phe Lys Ser 565 570 575 Gly Pro Gly Thr Glu Met Ala Leu Cys Ala Lys Pro Ser 580 585 <210> 60 <211> 229 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <22​​​​​​​​​​​​​​​​​​​​​​​​​​ Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 130 135 140 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 145 150 155 160 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu 180 185 190 Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 195 200 205 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 Leu Ser Leu Gly Lys 225 <210> 61 <211> 227 <212> PRT <213> Artificial Sequence <220> <221> DOMAIN <223> Fc‑2 <400> 61 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly 1 5 10 15 Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 20 25 30 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 35 40 45 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 50 55 60 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr 65 70 75 80 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 85 90 95 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 100 105 110 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 115 120 125 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 130 135 140 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 145 150 155 160 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 165 170 175 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 180 185 190 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 195 200 205 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 210 215 220 Pro Gly Lys 225 <210> 62 <211> 358 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 4666 <400> 62 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala 1 5 10 15 Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 65 70 75 80 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 85 90 95 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 130 135 140 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 145 150 155 160 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu 180 185 190 Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 195 200 205 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 Leu Ser Leu Gly Lys Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu 225 230 235 240 Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Cys Val Thr Val 245 250 255 Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro Val 260 265 270 Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu Ile 275 280 285 Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser Asp 290 295 300 Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser Ile 305 310 315 320 Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys Gly 325 330 335 Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met Ala 340 345 350 Leu Cys Ala Lys Pro Ser 355 <210> 63 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 8QCGC <400> 63 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Gln Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 64 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 10QCGC <400> 64 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 65 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 12QCGC <400> 65 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Val 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 66 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 14QCGC <400> 66 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly Gln Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 67 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 15QCGC <400> 67 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 68 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 20QCGC <400> 68 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 69 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 21QCGC <400> 69 Glu Glu Glu Leu Gln Met Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 70 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 8LCGC <400> 70 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Gln Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 71 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 10LCGC <400> 71 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 72 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 12LCGC <400> 72 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Val 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 73 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 14LCGC <400> 73 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly Gln Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 74 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 15LCGC <400> 74 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 75 <211> 346[[ID=3**]] <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 20LCGC <400> 75 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 76 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 21LCGC <400> 76 Glu Glu Glu Leu Gln Met Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 77 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 56‑Fc <400> 77 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 78 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 57-Fc <400> 78 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345<(0006316)>[[ID=(17)]]<210> 79 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 58-Fc <400> 79 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Leu Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 It should be noted that in the translation, for the tags like -8 which are required to be preserved exactly as-is, the ones in the original text seem to have some inconsistent notations in the input (e.g., <(000631)> and <(0006316)> which might be incorrect in the original). I've translated them as accurately as possible while keeping the original form as much as possible. If there are specific rules or corrections regarding these tags, it may affect the translation result.Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 80 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 60‑Fc <400> 80 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 81 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 61‑Fc <400> 81 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 82 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 62-Fc <400> 82 Glu Glu Glu Leu Gln Ile Ile Gln Pro Glu Lys Leu Val Cys Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Gly Pro Gly Thr Glu Met 100 105 110 Ala Leu Cys Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 83 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 30‑Fc <400> 83 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 84 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 31‑Fc <400> 84 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 85 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 32‑Fc <400> 85 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Leu Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 86 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 33‑Fc <400> 86 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Met Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 87 <211> 346 <212> PRT <213> Artificial Sequence <220> <221> PEPTIDE <223> 34-Fc <400> 87 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Asn Asn Lys Asp Phe Ser Ile Lys Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 225 230 235 240 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 245 250 255 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 260 265 270 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 275 280 285 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 290 295 300 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 305 310 315 320 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 325 330 335 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 88 <211> 346 <212> PRT <213> Artificial Sequence <400> 88 Glu Glu Glu Leu Gln Ile Ile Cys Pro Glu Lys Leu Val Leu Val Thr 1 5 10 15 Val Gly Glu Thr Ala Thr Leu His Cys Thr Val Thr Ser Leu Leu Pro 20 25 30 Val Gly Pro Val Leu Trp Phe Arg Gly Val Gly Pro Gly Arg Glu Leu 35 40 45 Ile Tyr Ala Ser Gly Arg Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Asp Leu Thr Lys Arg Glu Asn Lys Asp Phe Ser Ile Arg Ile Ser Ser 65 70 75 80 Ile Thr Pro Ala Asp Val Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Glu Asp Val Glu Phe Lys Ser Cys Pro Gly Thr Glu Met 100 105 110 Ala Leu Gly Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys 115 120 125 Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro 130 135 140 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 145 150 155 160 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 165 170 175 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 180 185 190 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 195 200 205 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 210 215 220 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gl...

Claims

1. A SIRPy variant, the amino acid sequence of which is set forth in SEQ ID NO: 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 106.

2. The SIRPg variant of claim 1, wherein the SIRPg variant binds to human CD47 with a KD of less than 2 x 10 -9 M.

3. A fusion protein, which consists of a SIRPy variant according to claim 1, an anti-PD-1 antibody, and a linker, the N-terminus of the SIRPy variant is connected to the C-terminus of the heavy chain of the anti-PD-1 antibody via the linker, wherein: 1) the anti-PD-1 antibody comprises: a heavy chain, the amino acid sequence of which is set forth in SEQ ID NO: 11; and a light chain, the amino acid sequence of which is set forth in SEQ ID NO: 12; and the amino acid sequence of the SIRPy variant is set forth in SEQ ID NO: 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25; or 2) the anti-PD-1 antibody comprises: a heavy chain, the amino acid sequence of which is set forth in SEQ ID NO: 103; and a light chain, the amino acid sequence of which is set forth in SEQ ID NO: 104; and the amino acid sequence of the SIRPy variant is set forth in SEQ ID NO: 18 or 106.

4. The fusion protein according to claim 3, wherein the fusion protein consists of two identical first peptide chains and two identical second peptide chains, wherein: the first peptide chain has the amino acid sequence set forth in SEQ ID NO: 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, or 59; and the second peptide chain has the amino acid sequence set forth in SEQ ID NO: 12; or the first peptide chain has the amino acid sequence set forth in SEQ ID NO: 107 or 108; and the second peptide chain has the amino acid sequence set forth in SEQ ID NO:

104.

5. The fusion protein according to claim 4, wherein the fusion protein consists of two identical first peptide chains and two identical second peptide chains, wherein: the first peptide chain has the amino acid sequence set forth in SEQ ID NO: 49; and the second peptide chain has the amino acid sequence set forth in SEQ ID NO: 12; or the first peptide chain has the amino acid sequence set forth in SEQ ID NO: 52; and the second peptide chain has the amino acid sequence set forth in SEQ ID NO: 12; or the first peptide chain has the amino acid sequence set forth in SEQ ID NO: 108; and the second peptide chain has the amino acid sequence set forth in SEQ ID NO:

104.

6. A fusion protein, wherein: the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the SIRPy variant is connected to the N-terminus of the Fc domain monomer via a peptide bond; or the fusion protein consists of a SIRPy variant according to claim 1, and a human Fc domain monomer, the C-terminus of the The fusion protein consists of a SIRPy variant according to claim 1, a human Fc domain monomer and a linker, the N-terminus of the SIRPy variant is connected to the C-terminus of the Fc domain monomer via the linker; wherein the amino acid sequence of the Fc domain monomer is set forth in SEQ ID NO: 60, 61 or 109; and the amino acid sequence of the SIRPy variant is set forth in SEQ ID NO: 15, 23, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 106.

7. The fusion protein according to claim 6, wherein the fusion protein is a homodimer consisting of two identical Fc domain monomers selected from the group consisting of the amino acid sequences set forth in SEQ ID NO: 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 83, 84, 85, 86, 87, 88, 111 or 113.

8. A pharmaceutical composition comprising a therapeutically effective amount of the fusion protein according to any one of claims 3 to 7, and one or more pharmaceutically acceptable carriers, buffers or excipients.

9. A nucleic acid molecule encoding the SIRPy variant according to claim 1 or 2, or the fusion protein according to any one of claims 3 to 7.

10. An expression vector comprising the nucleic acid molecule of claim 9.

11. A host cell comprising the expression vector of claim 10, wherein the host cell is selected from the group consisting of CHO, 293 and NSO cells.

12. Use of the fusion protein according to any one of claims 3 to 5 in the manufacture of a medicament for the treatment of a disease or disorder, wherein the disease or disorder is selected from the group consisting of multiple myeloma, colon and rectum cancer and lymphoma.

13. Use of the fusion protein according to claim 6 or 7 in the manufacture of a medicament for the treatment of a disease or disorder, wherein the disease or disorder is selected from the group consisting of multiple myeloma, colon and rectum cancer, breast cancer and lymphoma.

14. Use of the fusion protein according to any one of claims 3 to 5 in combination with an anti-CD38 antibody in the manufacture of a medicament for the treatment of a disease or disorder, wherein the disease or disorder is selected from the group consisting of multiple myeloma, colon and rectum cancer and lymphoma; wherein the anti-CD38 antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3, and a light chain variable region comprising LCDR1, LCDR2 and LCDR3, wherein: the amino acid sequences of HCDR1, HCDR2 and HCDR3 of the heavy chain variable region are set forth in SEQ ID NOs: 119, 120 and 121, respectively; and the amino acid sequences of LCDR1, LCDR2 and LCDR3 of the light chain variable region are set forth in SEQ ID NOs: 122, 123 and 124, respectively.

15. The use of claim 14, wherein the amino acid sequence of the variable region of the heavy chain of the anti-CD38 antibody is set forth in SEQ ID NO: 117, and the amino acid sequence of the variable region of the light chain is set forth in SEQ ID NO:

118.

16. The use of claim 15, wherein the anti-CD38 antibody comprises a heavy chain and a light chain, wherein the amino acid sequence of the heavy chain is set forth in SEQ ID NO: 115, and the amino acid sequence of the light chain is set forth in SEQ ID NO: 116.

Images