Anti-C1S antibody and its uses

Abstract

Antibodies and antigen-binding fragments thereof that bind to C1s, inhibit the activity of C1s, and modulate the activity of at least one component in the classical pathway (CP) of complement activation; and methods of treating complement-mediated disorders using anti-C1s antibodies and fragments.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 342,544, filed May 16, 2022, and U.S. Provisional Application No. 63 / 221,131, filed Jul. 13, 2021, each of which is hereby incorporated by reference in its entirety.

[0003] Sequence Listing

[0004] This application contains a Sequence Listing that has been electronically submitted in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy, created on Jun. 28, 2022, is named SEQ_LIST_PIPL_1121-102PCT.txt and is 279,262 bytes in size. TECHNICAL FIELD

[0005] The present disclosure relates to antibodies and antigen-binding fragments that bind to C1s protein, and to the use of antibodies and antigen-binding fragments that bind to C1s protein for treating complement-mediated disorders. BACKGROUND OF THE DISCLOSURE

[0006] The complement system is a tightly regulated network of proteins that forms an important part of both the innate and adaptive immune systems. Through multiple activation pathways and a wide range of complement-mediated effector functions, the complement system plays a key role in inflammation, pathogen defense, and recovery after injury. Complement activation can have both beneficial and harmful effects. The defense responses mediated by complement activation can eliminate pathogens. Inappropriate, unwanted, excessive, or deficient complement activation is associated with disease conditions such as certain autoimmune diseases and inflammatory syndromes, reflecting the complex balancing act of complement between immune regulation and inflammatory tissue damage.

[0007] The complement system can be activated through three different pathways that converge at the effector stage, where C3 convertase activates complement C3 protein and triggers a cascade of downstream effects. Each pathway has different triggers, initiator molecules, and enzymatic cascades upstream of the production of C3 convertase: (1) The classical pathway (CP) can be triggered when an antibody–antigen complex binds to the C1 complex and C1 is activated; (2) The alternative pathway (AP) can be triggered by cell surface components that are foreign to the host; and (3) The mannose-binding lectin pathway (LP) can be triggered when a circulating lectin binds to mannose residues on the surface of a microorganism. After the upstream cascades of each pathway produce C3 convertase, subsequent cleavage of complement proteins C3 and C5: (1) generates anaphylatoxins C3a and C5a, which attract and activate effector immune cells to the site of antibody binding / complement activation; (2) mediates the deposition of C3b opsonins that mediate phagocytosis and lymphocyte activation; and (3) triggers the lytic pathway to form a membrane attack complex (MAC) that disrupts cell membranes and causes cell lysis.

[0008] The CP is not only crucial for antibody-mediated defense against foreign pathogens but is also known or suspected to be involved in triggering immune responses to the engagement of autoantibodies with autoantigens, i.e., autoimmune disorders. The CP is triggered by the activation of the C1 complex, which normally circulates in plasma as an inactive complex of its subcomponent proteins C1q, C1r, and C1s. After recognizing and binding to an antibody–antigen complex, the subcomponents of the C1 complex are sequentially activated in an enzymatic cascade, where the binding of C1q to the Fc regions of two antigen-bound antibodies activates the C1r subunit, which in turn activates C1s by cleaving the C1s zymogen into active C1s with two subcomponents (A chain and B chain) linked by an interchain disulfide bond. The activated C1s B chain is a serine protease that can then cleave serum C4 and C2 to form the C3 convertase C4b2a. Then, the formation of C3 convertase in the CP triggers the downstream effector processes common to all three complement pathways.

[0009] Complement component 1s (C1s) protein has a central role in the function of the classical pathway (CP), in which the zymogen of C1s in the inactive C1 complex must be cleaved into the active two-subunit (two-chain) C1s that can cleave and activate serum C4 in order for the downstream effector cascade to proceed. For example, in human patients, a deficiency in C1s protein can cause severe immune complex diseases because immune complex deposits cannot be efficiently cleared in the absence of functional C1, C2, or C4 and subsequent normal levels of C3b. Some autoimmune and / or inflammatory disorders are known or suspected to involve the spontaneous activation of the CP by complexes of autoantibodies (especially complement-fixing autoantibodies) that bind to autoantigens. Complement-mediated disorders are known to include paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), hereditary angioedema (HAE), age-related macular degeneration (AMD), and autoimmune hemolytic anemias such as cold agglutinin disease (CAD). The complement system, specifically the CP, is suspected of playing a role in certain autoimmune disorders such as immune thrombocytopenic purpura (ITP), an autoimmune hemorrhagic disorder characterized by isolated thrombocytopenia with a platelet count <150,000 / μL, which is suspected to be caused by the development of autoantibodies targeting auto-platelet antigens. The CP is also suspected of playing a role in neuromyelitis optica (NMO), an autoimmune disorder with inflammatory demyelinating lesions in the central nervous system (especially the spinal cord and optic nerves), which is thought to be caused by the binding of anti-aquaporin-4 (AQP4) autoantibodies to astrocytes. Therapeutic approaches for treating complement-mediated disorders include small molecule regulators such as protease inhibitors, although some known therapies (such as the C1 inhibitor C1 INH) are known to act beyond the regulation of the CP, including the regulation of the lectin pathway and the kinin, coagulation, and fibrinolytic systems. Therapeutic approaches using antibodies against components of the CP pathway are being explored. SUMMARY OF THE INVENTION

[0010] The present invention relates to novel antibodies and antigen-binding fragments that bind to human complement component 1s (C1s), and methods of making and using antibodies and antigen-binding fragments that bind to C1s.

[0011] The present disclosure provides anti-C1s antibodies, anti-C1s antibody fragments that bind to C1s, nucleic acids encoding the anti-C1s antibodies and anti-C1s antibody fragments, and methods for preparing and using the anti-C1s antibodies and anti-C1s antibody fragments. The term "anti-C1s antibody" in the present disclosure encompasses anti-C1s antibodies capable of binding to C1s and antigen-binding fragments thereof, specifically anti-C1s antibodies that specifically bind to C1s and antigen-binding fragments thereof. The anti-C1s antibodies disclosed herein are capable of binding to zymogen human C1s (native C1s). The anti-C1s antibodies disclosed herein are capable of binding to active human C1s. The anti-C1s antibodies disclosed herein are capable of inhibiting C1s. The anti-C1s antibodies disclosed herein are capable of binding to zymogen C1s and inhibiting C1s. The anti-C1s antibodies disclosed herein are capable of binding to active C1s and inhibiting C1s. The anti-C1s antibodies disclosed herein are recombinant anti-C1s antibodies or antigen-binding fragments thereof that are capable of specifically binding to human C1s and inhibiting C1s.

[0012] The present disclosure provides anti-C1s antibodies for in vivo, ex vivo or in vitro therapeutic and diagnostic uses. The anti-C1s antibodies disclosed herein can be used to treat conditions mediated by the function of C1s. The anti-C1s antibodies disclosed herein can be used to treat complement-mediated conditions, specifically conditions mediated by the classical pathway (CP) of complement activation. The anti-C1s antibodies disclosed herein are capable of binding to C1s and inhibiting C1s, which has an effect on the activity of the CP pathway, and the effect can be measured by the effect on the activity of at least one component of the CP downstream of C1s activation, where the component can be a molecule or biological process associated with the CP. The anti-C1s antibodies disclosed herein are capable of inhibiting the activity of the CP pathway. The anti-C1s antibodies disclosed herein can be used to treat conditions mediated by the function of the CP. The anti-C1s antibodies disclosed herein can be used to target C1s to selectively inhibit the CP in complement-mediated autoimmune and / or inflammatory conditions. The anti-C1s antibodies disclosed herein can be used to treat complement-mediated autoimmune and / or inflammatory conditions, including but not limited to immune thrombocytopenic purpura (ITP) or neuromyelitis optica (NMO).

[0013] In certain embodiments, the anti-C1s antibodies disclosed herein are capable of binding to zymogen (native) C1s and inhibiting the activation of C1s. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of binding to active C1s and inhibiting the activation of C1s. The anti-C1s antibodies disclosed herein are capable of inhibiting one or more effects of C1s activation on at least one downstream component. In certain embodiments, the downstream component is at least one of the cleavage of C2 and / or the cleavage of C4 and / or the formation of C3 convertase and / or the lysis of serum complement-induced antibody-sensitized cells, and the binding of the anti-C1s antibody to active C1s has the effect of inhibiting the cleavage of C2 and / or the cleavage of C4 and / or the formation of C3 convertase and / or the lysis of serum-induced antibody-sensitized cells.

[0014] In one aspect, provided are anti-C1s antibodies capable of binding to C1s and modulating at least one effect of CP activation triggered by an antibody-antigen complex. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of modulating at least one effect of CP activation triggered by an antibody-antigen complex in a dose-dependent manner. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of inhibiting at least one effect of CP activation triggered by an antibody-antigen complex in a dose-dependent manner. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of inhibiting the lysis of serum-induced antibody-sensitized cells in a dose-dependent manner.

[0015] In one aspect, provided are anti-C1s antibodies capable of having an effect of modulating the activity of at least one component involved in C4 cleavage and / or deposition triggered by an antibody-antigen complex. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of inhibiting the activity of at least one component involved in the deposition of C4 from serum triggered by IgM. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of inhibiting the deposition of human C4 from human serum triggered by IgM in a dose-dependent manner. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of modulating the activity of at least one component involved in the deposition of cynomolgus monkey C4 from cynomolgus monkey serum triggered by IgM in a dose-dependent manner. In certain embodiments, the anti-C1s antibodies disclosed herein are capable of inhibiting the deposition of cynomolgus monkey C4 from cynomolgus monkey serum triggered by IgM in a dose-dependent manner.

[0016] In another aspect, provided are anti-C1s antibodies that have a selective effect upstream of the generation and activity of common complement system effectors within the CP. In certain embodiments, the selective inhibition of the CP by the anti-C1s antibodies disclosed herein prevents C4 cleavage and downstream effects triggered by C4 cleavage. In certain embodiments, the anti-C1s antibodies disclosed herein prevent the formation of C3 convertase via the CP pathway.

[0017] In another aspect, anti-C1s antibodies that bind to pro-C1s and active C1s proteins are provided. In certain embodiments, the anti-C1s antibodies disclosed herein bind to pro-C1s and active C1s proteins with similar affinities. In certain embodiments, the anti-C1s antibodies disclosed herein specifically bind to pro-C1s and active C1s proteins with similar high affinities, wherein the KD value is in the low nanomolar to picomolar range.

[0018] In another aspect, anti-C1s antibodies are provided that exhibit cross-reactivity with at least one non-human C1s. In certain embodiments, the anti-C1s antibodies disclosed herein may be capable of binding to human C1s and rat C1s. In certain embodiments, the anti-C1s antibodies disclosed herein may be capable of binding to human C1s and cynomolgus monkey C1s. In certain embodiments, the anti-C1s antibodies disclosed herein may be capable of binding to human C1s, rat C1s, and cynomolgus monkey C1s. In certain embodiments, the anti-C1s antibodies disclosed herein do not exhibit detectable binding to mouse C1s.

[0019] In another aspect, anti-C1s antibodies are provided that bind to epitopes accessible on pro-C1s and active C1s in an internal region of C1s within a 151 amino acid region from residue Y272 to R422 (M151) in the N-terminal heavy chain (NHC), wherein the key residues include R316, K336, and the sub-region G390 to R422 of human C1s (SEQ ID NO:99).

[0020] The anti-C1s antibodies provided herein are recombinantly expressed and are recombinant antibodies. The anti-C1s antibodies provided herein can further be one or more of the following: humanized antibodies, chimeric antibodies, single-chain antibodies, Fab fragments, single-chain variable fragments (scFv), aptamers, single-domain antibodies (VHH, nanobodies, camelid antibodies) or any other C1s-binding fragment or variant. Specifically, the anti-C1s antibodies disclosed herein are humanized antibodies. In certain embodiments, the anti-C1s antibodies disclosed herein can include frameworks in which amino acids have been substituted into existing antibody frameworks, specifically to affect properties such as antigen-binding ability. In certain embodiments, the anti-C1s antibodies disclosed herein can include complementarity-determining regions (CDRs) from a source (parent) antibody that have been grafted (fused) into a framework from a different type (class) of antibody and / or from an organism different from the parent antibody (specifically a recipient human framework). In certain embodiments, the anti-C1s antibodies disclosed herein can include frameworks in which amino acids have been substituted, mutated or replaced in regions outside the CDRs to affect properties such as antigen binding or antibody structure, for example, in the variable region framework around the CDRs and / or in the constant region (specifically the Fc region). In certain embodiments, one or more of the CDRs have been substituted, mutated or replaced. In certain embodiments, the anti-C1s antibodies disclosed herein can be humanized anti-C1s antibody variants.

[0021] In certain embodiments, the anti-C1s antibodies disclosed herein include at least one polypeptide having an amino acid sequence as shown in Table 1 (“Variable Regions of Anti-C1s Antibodies”) or the Sequence Listing, or a sequence that is substantially the same as the amino acid sequence shown in Table 1 or the Sequence Listing (e.g., a sequence that is at least about 85%, 90%, 92%, 95%, 97%, or 98%, 99% identical). The anti-C1s antibodies disclosed herein can include at least one polypeptide having an amino acid sequence selected from the following, or a sequence that is substantially the same as a polypeptide having an amino acid sequence selected from the following (e.g., a sequence that is at least about 85%, 90%, 92%, 95%, 97%, or 98%, 99% identical): SEQ ID NO:1; SEQ ID NO:2; SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:6; SEQ ID NO:7; SEQ ID NO:8; SEQ ID NO:9; SEQ ID NO:11; SEQ ID NO:12; SEQ ID NO:13; SEQ ID NO:14; SEQ ID NO:16; SEQ ID NO:17; SEQ ID NO:18; SEQ ID NO:19; SEQ ID NO:21; SEQ ID NO:22; SEQ ID NO:23; SEQ ID NO:24; SEQ ID NO:26; SEQ ID NO:27; SEQ ID NO:28; SEQ ID NO:29; SEQ ID NO:31; SEQ ID NO:32; SEQ ID NO:33; SEQ ID NO:34; SEQ ID NO:36; SEQ ID NO:37; SEQ ID NO:38; SEQ ID NO:39; SEQ ID NO:41; SEQ ID NO:42; SEQ ID NO:43; SEQ ID NO:44; SEQ ID NO:46; SEQ ID NO:47; SEQ ID NO:48; SEQ ID NO:49; SEQ ID NO:51; SEQ ID NO:52; SEQ ID NO:53; SEQ ID NO:54; SEQ ID NO:56; SEQ ID NO:57; SEQ ID NO:58; SEQ ID NO:59; SEQ ID NO:61; SEQ ID NO:62; SEQ ID NO:63; SEQ ID NO:64; SEQ ID NO:66; SEQ ID NO:67; SEQ ID NO:68; SEQ ID NO:69; SEQ ID NO:71; SEQ ID NO:73; SEQ ID NO:75; SEQ ID NO:77;SEQ ID NO:79; SEQ ID NO:81; SEQ ID NO:83; SEQ ID NO:85; SEQ ID NO:87; SEQ ID NO:89; SEQ ID NO:91; SEQ ID NO:93; SEQ ID NO:95; SEQ ID NO:97; SEQ ID NO:129; SEQ ID NO:130; SEQ ID NO:131; SEQ ID NO:132; SEQ ID NO:133; SEQ ID NO:134; SEQ ID NO:135; SEQ ID NO:136; SEQ ID NO:137; SEQ ID NO:138; SEQ ID NO:139; or SEQ ID NO:140.;

[0022] In one embodiment, the anti-C1s antibodies disclosed herein include: a heavy chain variable region (VH) polypeptide having the amino acid sequence shown in SEQ ID NO:1 or a sequence substantially identical to SEQ ID NO:1; and a light chain variable region (VL) polypeptide having the amino acid sequence shown in SEQ ID NO:6 or a sequence substantially identical to SEQ ID NO:6. In one embodiment, the anti-C1s antibodies disclosed herein include: a heavy chain complementarity determining region 1 (HC CDR1) having the amino acid sequence shown in SEQ ID NO:2; a heavy chain complementarity determining region 2 (HC CDR2) having the amino acid sequence shown in SEQ ID NO:3; a heavy chain complementarity determining region 3 (HC CDR3) having the amino acid sequence shown in SEQ ID NO:4; a light chain complementarity determining region 1 (LC CDR1) having the amino acid sequence shown in SEQ ID NO:7; a light chain complementarity determining region 2 (LC CDR2) having the amino acid sequence shown in SEQ ID NO:8; and a light chain complementarity determining region 3 (LC CDR3) having the amino acid sequence shown in SEQ ID NO:9; or variants of the antibody including 1, 2, 3, 4, 5, or 6 amino acid substitutions in the CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified as 2-7 herein, which includes: an HC polypeptide having the amino acid sequence shown in SEQ ID NO:71; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO:73.

[0023] In one embodiment, the anti-C1s antibodies disclosed herein include: a VH polypeptide having the amino acid sequence shown in SEQ ID NO: 11 or a sequence substantially identical to SEQ ID NO: 11; and a VL polypeptide having the amino acid sequence shown in SEQ ID NO: 16 or a sequence substantially identical to SEQ ID NO: 16. In one embodiment, the anti-C1s antibodies disclosed herein include: an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 12; an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 13; an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 14; an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 17; an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 18; and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 19, or variants of the antibody comprising 1, 2, 3, 4, 5, or 6 amino acid substitutions in the CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified as 2-8 herein, which includes: an HC polypeptide having the amino acid sequence shown in SEQ ID NO: 75; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO: 77.

[0024] In one embodiment, the anti-C1s antibodies disclosed herein include: a VH polypeptide having the amino acid sequence shown in SEQ ID NO: 21 or a sequence substantially identical to SEQ ID NO: 21; and a VL polypeptide having the amino acid sequence shown in SEQ ID NO: 26 or a sequence substantially identical to SEQ ID NO: 26. In one embodiment, the anti-C1s antibodies disclosed herein include: an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 22; an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 23; an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 24; an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 27; an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 28; and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 29, or variants of the antibody comprising 1, 2, 3, 4, 5 or 6 amino acid substitutions in the CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified herein as hz2-7 (H1L2), which includes: an HC polypeptide having the amino acid sequence shown in SEQ ID NO: 79; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO: 81.

[0025] In one embodiment, the anti-C1s antibodies disclosed herein include: a VH polypeptide having the amino acid sequence shown in SEQ ID NO: 31 or a sequence substantially identical to SEQ ID NO: 31; and a VL polypeptide having the amino acid sequence shown in SEQ ID NO: 36 or a sequence substantially identical to SEQ ID NO: 36. In one embodiment, the anti-C1s antibodies disclosed herein include: an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 32; an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 33; an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 34; an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 37; an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 38; and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 39, or variants of the antibody that include 1, 2, 3, 4, 5, or 6 amino acid substitutions in the CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified herein as hz2-7 (H1L2 G131A), which includes: an HC polypeptide having the amino acid sequence shown in SEQ ID NO: 83; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO: 85.

[0026] In one embodiment, the anti-C1s antibodies disclosed herein include: an HC variable region polypeptide having the amino acid sequence shown in SEQ ID NO: 41 or a sequence substantially identical to SEQ ID NO: 41; and an LC variable region polypeptide having the amino acid sequence shown in SEQ ID NO: 46 or a sequence substantially identical to SEQ ID NO: 46. In one embodiment, the anti-C1s antibodies disclosed herein include: an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 42; an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 43; an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 44; an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 47; an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 48; and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 49, or variants of the antibody comprising 1, 2, 3, 4, 5, or 6 amino acid substitutions in the CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified herein as hz2-8 (H1L2), which includes: an HC polypeptide having the amino acid sequence shown in SEQ ID NO: 87; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO: 89.

[0027] In one embodiment, the anti-C1s antibodies disclosed herein include: a VH polypeptide having the amino acid sequence shown in SEQ ID NO: 51 or a sequence substantially identical to SEQ ID NO: 51; and a VL polypeptide having the amino acid sequence shown in SEQ ID NO: 56 or a sequence substantially identical to SEQ ID NO: 56. In one embodiment, the anti-C1s antibodies disclosed herein include: an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 52; an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 53; an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 54; an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 57; an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 58; and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 59, or variants of said antibody comprising 1, 2, 3, 4, 5 or 6 amino acid substitutions in said CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified herein as hz2-8 (H1L2 G80A / T82A), said antibody comprising: an HC polypeptide having the amino acid sequence shown in SEQ ID NO: 91; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO: 93.

[0028] In one embodiment, the anti-C1s antibodies disclosed herein include: a VH polypeptide having the amino acid sequence shown in SEQ ID NO: 61 or a sequence substantially identical to SEQ ID NO: 61; and a VL polypeptide having the amino acid sequence shown in SEQ ID NO: 66 or a sequence substantially identical to SEQ ID NO: 66. In one embodiment, the anti-C1s antibodies disclosed herein include: an HC CDR1 having the amino acid sequence shown in SEQ ID NO: 62; an HC CDR2 having the amino acid sequence shown in SEQ ID NO: 63; an HC CDR3 having the amino acid sequence shown in SEQ ID NO: 64; an LC CDR1 having the amino acid sequence shown in SEQ ID NO: 67; an LC CDR2 having the amino acid sequence shown in SEQ ID NO: 68; and an LC CDR3 having the amino acid sequence shown in SEQ ID NO: 69, or variants of said antibodies comprising 1, 2, 3, 4, 5 or 6 amino acid substitutions in said CDR regions. In a non-limiting embodiment, the anti-C1s antibody disclosed herein is the antibody identified herein as hz2-8 (H1L2 G80A), said antibody comprising: an HC polypeptide having the amino acid sequence shown in SEQ ID NO: 95; and an LC polypeptide having the amino acid sequence shown in SEQ ID NO: 97.

[0029] On the other hand, the CDR consensus sequences of the anti-C1s antibodies disclosed herein are provided. Table 1.b. discloses non-limiting exemplary embodiments of the CDR consensus sequences of anti-C1s antibodies. In one embodiment, an antibody is provided, wherein the VH comprises: HC CDR1, which has the amino acid sequence shown in SEQ ID NO: 129; HC CDR2, which has the amino acid sequence shown in SEQ ID NO: 130; and HC CDR3, which has the amino acid sequence shown in SEQ ID NO: 131, and the VL comprises: LC CDR1, which has the amino acid sequence shown in SEQ ID NO: 132; LC CDR2, which has the amino acid sequence shown in SEQ ID NO: 133; and LC CDR3, which has the amino acid sequence shown in SEQ ID NO: 134. In another embodiment, an antibody is provided, wherein the VH comprises: HC CDR1, which has the amino acid sequence shown in SEQ ID NO: 135; HC CDR2, which has the amino acid sequence shown in SEQ ID NO: 136; and HC CDR3, which has the amino acid sequence shown in SEQ ID NO: 137, and the VL comprises: LC CDR1, which has the amino acid sequence shown in SEQ ID NO: 138; LC CDR2, which has the amino acid sequence shown in SEQ ID NO: 139; and LC CDR3, which has the amino acid sequence shown in SEQ ID NO: 140.

[0030] On the other hand, anti-C1s antibodies (comprising variants and fragments as disclosed herein) are provided, which can be used for treating conditions mediated by CP activation, including but not limited to conditions mediated by CP activation triggered by antibody-antigen complexes, and specifically conditions associated with or characterized by CP activation by autoantibodies that bind to self-antigens. Methods and compositions for therapeutic use of the anti-C1s antibodies disclosed herein are provided, which therapeutic use includes but is not limited to treating conditions mediated by CP activation, specifically conditions characterized by CP activation by autoantibodies that bind to self-antigens. Methods and compositions for therapeutic use of the anti-C1s antibodies as disclosed herein are provided, which therapeutic use includes treating ITP and / or NMO. In certain embodiments, pharmaceutical compositions comprising the anti-C1s antibodies disclosed herein and suitable carriers and / or excipients are provided.

[0031] In another aspect, methods for treating complement-mediated disorders are provided, such methods comprising administering to a subject in need thereof an effective amount of an anti-C1s antibody disclosed herein. According to this aspect, methods for treating complement-mediated disorders comprise administering an effective amount of an anti-C1s antibody disclosed herein to have the effect of modulating or inhibiting one or more biological effects associated with CP activation triggered by an antibody-antigen complex. According to this aspect, an effective amount of the anti-C1s antibody is an amount of the antibody sufficient to cause a desired level of modulation or inhibition. In certain embodiments, the method comprises administering to a subject an effective amount of an anti-C1s antibody that results in modulating one or more biological effects associated with CP activation by autoantibodies binding to self-antigens, including but not limited to reducing platelet lysis triggered by anti-platelet antibodies and / or antibody-mediated platelet removal.

[0032] In another aspect, methods for treating diseases or disease states in which unwanted, abnormal, inappropriate, or excessive CP activation is involved are provided, such methods comprising administering to a subject in need thereof an effective amount of an anti-C1s antibody disclosed herein.

[0033] In another aspect, methods for diagnosing or screening for complement-mediated disorders in a subject are provided. Methods for diagnosing or screening for complement-mediated disorders can be practiced in vivo, ex vivo, or in vitro. In certain embodiments, the method comprises administering an anti-C1s antibody to a subject known or suspected of having a complement-mediated autoimmune disorder and measuring one or more biological effects or symptoms associated with the complement-mediated autoimmune disorder. In certain embodiments, the ex vivo or in vitro method comprises administering an anti-C1s antibody to a sample taken from a subject and measuring one or more biological effects or symptoms associated with the complement-mediated autoimmune disorder.

[0034] In another aspect, one or more isolated nucleic acid molecules are provided, the one or more isolated nucleic acid molecules encoding at least a portion of at least one anti-C1s antibody disclosed herein. In certain embodiments, the isolated nucleic acid molecule encoding at least a portion of at least one anti-C1s antibody among the anti-C1s antibodies disclosed herein comprises the nucleotide sequence shown in Table 2 herein or a sequence substantially identical to the nucleotide sequence shown in Table 2 (e.g., a sequence that is at least about 85%, 90%, 92%, 95%, 97% or 98%, 99% identical). In certain embodiments, the isolated nucleic acid molecule encoding at least one HC sequence among the heavy chain (HC) sequences of the anti-C1s antibodies disclosed herein may comprise a nucleotide sequence selected from at least one of the following: SEQ ID NO:5 or a sequence substantially identical to SEQ ID NO:5; SEQ ID NO:15 or a sequence substantially identical to SEQ ID NO:15; SEQ ID NO:25 or a sequence substantially identical to SEQ ID NO:25; SEQ ID NO:35 or a sequence substantially identical to SEQ ID NO:35; SEQ ID NO:45 or a sequence substantially identical to SEQ ID NO:45; SEQ ID NO:55 or a sequence substantially identical to SEQ ID NO:55; SEQ ID NO:65 or a sequence substantially identical to SEQ ID NO:65; SEQ ID NO:72 or a sequence substantially identical to SEQ ID NO:72; SEQ ID NO:76 or a sequence substantially identical to SEQ ID NO:76; SEQ ID NO:80 or a sequence substantially identical to SEQ ID NO:80; SEQ ID NO:84 or a sequence substantially identical to SEQ ID NO:84; SEQ ID NO:88 or a sequence substantially identical to SEQ ID NO:88; SEQ ID NO:92 or a sequence substantially identical to SEQ ID NO:92; or SEQ ID NO:96 or a sequence substantially identical to SEQ ID NO:96.

[0035] In certain embodiments, an isolated nucleic acid molecule encoding at least one LC sequence in the light chain (LC) sequence of an anti-C1s antibody or an antigen-binding fragment thereof disclosed herein may comprise a nucleotide sequence selected from at least one of the following: SEQ ID NO:10 or a sequence substantially identical to SEQ ID NO:10; SEQ ID NO:20 or a sequence substantially identical to SEQ ID NO:20; or SEQ ID NO:30 or a sequence substantially identical to SEQ ID NO:30; SEQ ID NO:40 or a sequence substantially identical to SEQ ID NO:40; SEQ ID NO:50 or a sequence substantially identical to SEQ ID NO:50; SEQ ID NO:60 or a sequence substantially identical to SEQ ID NO:60; SEQ ID NO:70 or a sequence substantially identical to SEQ ID NO:70; SEQ ID NO:74 or a sequence substantially identical to SEQ ID NO:74; SEQ ID NO:78 or a sequence substantially identical to SEQ ID NO:78; SEQ ID NO:82 or a sequence substantially identical to SEQ ID NO:82; SEQ ID NO:86 or a sequence substantially identical to SEQ ID NO:86; SEQ ID NO:90 or a sequence substantially identical to SEQ ID NO:90; SEQ ID NO:94 or a sequence substantially identical to SEQ ID NO:94; or SEQ ID NO:98 or a sequence substantially identical to SEQ ID NO:98.

[0036] In another aspect, a vector is provided that comprises one or more nucleic acid molecules encoding at least one amino acid sequence of an anti-C1s antibody disclosed herein. In certain embodiments, a vector is provided that comprises one or more nucleic acid molecules encoding at least one sequence in the heavy chain (HC) or light chain (LC) sequence of an anti-C1s antibody disclosed herein.

[0037] In another aspect, at least one host cell is provided that contains a vector comprising one or more nucleic acid molecules encoding the amino acid sequence of an anti-C1s antibody disclosed herein. In certain embodiments, the host cell is transiently transfected with a vector comprising one or more nucleic acid molecules encoding the amino acid sequence of an anti-C1s antibody or an antigen-binding fragment thereof disclosed herein, wherein the host cell is capable of supporting vector expression and recombinant production of the anti-C1s antibody or an antigen-binding fragment thereof encoded by the vector. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 A schematic diagram showing the proposed role of CP and anti-C1s antibody, where Ag indicates antigen, IgG and IgM indicate antibody types that can bind to Ag, C1 indicates the C1 complex, where C1q, C1r, and C1s indicate the sub-component proteins of the C1 complex, anti-C1s indicates the anti-C1s antibody that binds to C1s, the labels C2a, C2b, C3, C3a, C3b, C4a, C4b, and C5 indicate the components of the CP cascade arranged in the schematic diagram of the steps in the cascade, and MAC indicates the membrane attack complex.

[0039] Figures 2A - 2D Results showing the ability of anti-C1s antibody to inhibit IgM-induced deposition of C4 from serum in a functional assay. Figure 2A Shows the effect of anti-C1s antibody 2-7, anti-C1s antibody 2-8, and the antibody from non-functional clone 3-30 on the deposition of IgM-induced human C4 from human serum (NHS) within the indicated antibody concentration range. Figure 2B Shows the effect of anti-C1s antibody 2-7 and anti-C1s antibody 2-8 on the deposition of IgM-induced cynomolgus monkey C4 from cynomolgus monkey serum (CMS) within the indicated antibody concentration range. In Figures 2A - 2B it, the symbols are used as follows: open circles represent the results using anti-C1s antibody 2-7; open triangles represent the results using anti-C1s antibody 2-8; open squares represent the results using anti-C1s antibody 3-30. Figure 2C Shows the effect of humanized anti-C1s antibody hz2-7 (H1L2), humanized anti-C1s antibody hz2-8 (H1L2), humanized anti-C1s antibody variant hz2-7 (H1L2 G131A), humanized anti-C1s antibody variant hz2-8 (H1L2 G80A), and humanized anti-C1s antibody variant hz2-8 (H1L2 G80A / T82A) on the deposition of IgM-induced human C4 from human serum (NHS) within the indicated antibody concentration range. Figure 2D Shows the effect of humanized anti-C1s antibody hz2-7 (H1L2), humanized anti-C1s antibody hz2-8 (H1L2), humanized anti-C1s antibody variant hz2-7 (H1L2 G131A), humanized anti-C1s antibody variant hz2-8 (H1L2 G80A), and humanized anti-C1s antibody variant hz2-8 (H1L2 G80A / T82A) on the deposition of IgM-induced cynomolgus monkey C4 from cynomolgus monkey serum (CMS) within the indicated antibody concentration range. In Figures 2C - 2DIn the figure, the symbols are used as follows: an open circle represents the result of using the anti-C1s antibody hz2-7 (H1L2); a solid black circle represents the result of using the anti-C1s antibody hz2-7 (H1L2G131A); an open triangle represents the result of using the anti-C1s antibody hz2-8 (H1L2); a solid black triangle represents the result of using the anti-C1s antibody hz2-8 (H1L2 G80A); and a triangle with a diagonal line represents the result of using the anti-C1s antibody hz2-8 (H1L2 G80A / T82A).

[0040] Figures 3A - 3V The measurement results of the binding affinity and cross-reactivity of the anti-C1s antibodies are shown. Figures 3A - 3D The measurement of the anti-C1s antibody affinity using the Bio-Layer Interferometry technology and the use of the AHC biosensor to measure the binding kinetics of the active human C1s protein by combining the antibody with the indicated concentration of active human C1s protein over the indicated time are shown, where the results are shown, Figure 3A showing the result of the binding of anti-C1s antibody 2-7 to active human C1s, Figure 3B showing the result of the binding of anti-C1s antibody 2-8 to active human C1s, Figure 3C showing the binding of the humanized anti-C1s antibody hz2-7 (H1L2) to active human C1s, and Figure 3D showing the binding of the humanized anti-C1s variant hz2-8 (H1L2 G80A / T82A) to active human C1s. Figure 3E is a summary table showing the KD, k on , k off and R 2 values calculated for each of the indicated combinations of antibody and target. Figures 3F - 3G The results of measuring the binding of the anti-C1s antibody to active and zymogen forms of C1s using ELISA are shown, where Figure 3F the results of the binding of the indicated concentrations of 2-7, 2-8, hz2-7 (H1L2) and hz2-8 (H1L2 G80A / T82A) and human IgG1 as a control to active human C1s are shown, and Figure 3G the results of the binding of the indicated concentrations of 2-7, 2-8, hz2-7 (H1L2) and hz2-8 (H1L2 G80A / T82A) and human IgG1 as a control to human C1s zymogen are shown. Figures 3H - 3Q The measurement of the anti-C1s antibody affinity using the Bio-Layer Interferometry technology and the determination of the anti-C1s antibody affinity from human (HuC1s, Figures 3H - 3K ), rat (RaC1s, Figures 3L - 3M) and the results of the binding kinetics of recombinant C1s derived from cynomolgus monkeys (CyC1s, Figures 3N - 3Q ), where Figure 3H shows the binding of 2-7 to human C1s (HuC1s), Figure 3I shows the binding of 2-8 to HuC1s, Figure 3J shows the binding of hz2-7 (H1L2) to HuC1s, Figure 3K shows the binding of hz2-8 (H1L2G80A / T82A) to HuC1s, Figure 3L shows the binding of 2-8 to RaC1s, Figure 3M shows the binding of hz2-8 (H1L2 G80A / T82A) to RaC1s, Figure 3N shows the binding of 2-7 to CyC1s, Figure 3O shows the binding of 2-8 to CyC1s, Figure 3P shows the binding of hz2-7 (H1L2) to CyC1s, and Figure 3Q shows the binding of hz2-8 (H1L2 G80A / T82A) to CyC1s. Figure 3R is a summary table showing the KD, k on , k off and R 2 values calculated for each of the indicated combinations of antibody and target. Figures 3S - 3V shows the results of anti-C1s antibody cross-reactivity measurements using ELISA, where the figures show the binding of the indicated concentrations of 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2 G80A / T82A), and IgG (control) to human C1s (HuC1s, Figure 3S ), mouse C1s (MoC1s, Figure 3T ), rat C1s (RatC1s, Figure 3U ), and cynomolgus monkey C1s (CyC1s, Figure 3V ). In Figures 3F - 3G and 3S-3V, the symbols are used as follows: open circles represent the results using anti-C1s antibody 2-7; open triangles represent the results using anti-C1s antibody 2-8; open squares represent the results using mouse IgG; filled black circles represent the results using anti-C1s antibody hz2-7 (H1L2); filled black triangles represent the results using anti-C1s antibody hz2-8 (H1L2 G80A / T82A); and filled black squares represent the results using human IgG1.

[0041] Figures 4A - 4D shows the results of measuring the binding of anti-C1s antibodies to full-length and different truncated forms of recombinant C1s using ELISA. Figure 4AIt shows that 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2) and IgG controls (mouse IgG, human IgG1) bind to full-length human C1s (HuC1s). Figure 4B It shows that 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2) and IgG controls (mouse IgG, human IgG1) bind to full-length mouse C1s (MoC1s). Figure 4C It shows that 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2) and IgG controls (mouse IgG, human IgG1) bind to truncated human C1s NHC (HuC1s(NHC)). Figure 4D It shows that 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2) and IgG controls (mouse IgG, human IgG1) bind to truncated human C1s M151 (HuC1s(M151)). In Figures 4A - 4D it, the symbols are used as follows: open circles represent the results using anti-C1s antibody 2-7; open triangles represent the results using anti-C1s antibody 2-8; open squares represent the results using mouse IgG; filled black circles represent the results using anti-C1s antibody hz2-7 (H1L2); filled black triangles represent the results using anti-C1s antibody hz2-8 (H1L2); and filled black squares represent the results using human IgG1.

[0042] Figures 5A - 5B It shows the binding to full-length and truncated forms (including deletion mutants) of C1s and their schematic comparison. Figure 5AImages showing the binding of the representative anti-C1s antibody 2-8 to full-length and different truncated forms of C1s as measured by Western blot. Under non-reducing (left blot) and reducing (right blot) conditions, 200 ng of full-length human C1s (HuC1s) in lane 1, 200 ng of full-length mouse C1s (MoC1s) in lane 2, 200 ng of truncated human C1s M151 (HuC1s(M151)) in lane 3, and 200 ng of truncated human C1s NHC (HuC1s(NHC)) in lane 4 were resolved on SDS-PAGE and subsequently transferred to a nitrocellulose membrane (blot). The blocked blot was incubated with antibody 2-8 and developed with an HRP-labeled secondary antibody. In the blot of the gel run without a reducing agent (left), lane 1 shows the binding of antibody 2-8 to full-length human C1s (HuC1s, upper band) and the autocleaved C1s heavy chain (lower band), lane 2 shows no binding of antibody 2-8 to full-length mouse C1s (MoC1s) (no detectable antibody binding), lane 3 shows the binding of antibody 2-8 to multiple isoforms of truncated human C1s M151 (HuC1s(M151)), and lane 4 shows the binding of antibody 2-8 to truncated human C1s NHC (HuC1s(NHC)). On the blot of the gel run with a reducing agent (right), lane 1 shows low-level binding of antibody 2-8 to the autocleaved C1s heavy chain from full-length human C1s (HuC1s), lane 2 shows no binding of antibody 2-8 to full-length mouse C1s (MoC1s) (no detectable binding), lane 3 shows the binding of antibody 2-8 to truncated human C1s M151 (HuC1s(M151)), and lane 4 shows the binding of antibody 2-8 to truncated human C1s NHC (HuC1s(NHC)). Figure 5B Schematic comparison and alignment of full-length human C1s, HuC1s (e.g., as exemplified in SEQ ID NO:99), human C1s deletion mutant 1 (M151) designated as HuC1s(M151) or M151 (e.g., as exemplified in SEQ ID NO:107), human deletion mutant 2 (NHC) designated as HuC1s(NHC) or NHC (e.g., as exemplified in SEQ ID NO:109), and human C1s deletion mutant 3 (NHCΔ33) designated as human C1s(NHCΔ33) or NHCΔ33 (e.g., as exemplified in SEQ ID NO:111) are shown. Figure 5B The positions of the point mutations introduced into HuC1s, M151, NHC, and NHCΔ33 in Example 6 are also shown.

[0043] Figures 6A - 6UShows the results of binding measurements of hz2-7 (H1L2), hz2-8 (H1L2 G80A), hz2-8 (H1L2 G80A / T82A), and human IgG1 (control) to NHCΔ33 and point mutants M1 - M20 of human C1s using ELISA, as follows: Figure 6A Shows binding to NHCΔ33; Figure 6B Shows binding to M1 (M277I); Figure 6C Shows binding to M2 (M277I and K331Q); Figure 6D Shows binding to M3 (M277I, P278S, and N329D); Figure 6E Shows binding to M4 (P280A and E282K); Figure 6F Shows binding to M5 (K336G); Figure 6G Shows binding to M6 (P280A, E282K, and K336G); Figure 6H Shows binding to M7 (D343Y and E351A); Figure 6I Shows binding to M8 (E351A); Figure 6J Shows binding to M9 (K336G, D343Y, and E351A); Figure 6K Shows binding to M10 (D283I and P285A); Figure 6L Shows binding to M11 (S349P); Figure 6M Shows binding to M12 (G381E and G382E); Figure 6N Shows binding to M13 (A292D); Figure 6O Shows binding to M14 (R316H); Figure 6P Shows binding to M15 (S360N); Figure 6Q Shows binding to M16 (R368H); Figure 6R Shows binding to M17 (N380H). Figure 6S Shows binding to M18 (V288T); Figure 6T Shows binding to M19 (Q303K); and Figure 6U Shows binding to M20 (A320S). In Figures 6A to 6U the following symbols are used as described: open circles represent the results using the anti-C1s antibody hz2-7 (H1L2); filled black triangles represent the results using the anti-C1s antibody hz2-8 (H1L2 G80A); triangles with a diagonal represent the results using the anti-C1s antibody hz2-8 (H1L2 G80A / T82A); and open squares represent the results using human IgG1 (HuIgG1).

[0044] Figures 7A - 7BResults showing the ability of anti-C1s antibodies to inhibit complement-mediated lysis of antibody-sensitized sheep red blood cells (EA cells) are shown. Results of anti-C1s antibodies hz2-7 (H1L2), hz2-8 (H1L2), hz2-8 (H1L2 G80A), and hz2-8 (H1L2 G80A / T82A) and isotype control human IgG1 on normal human serum (NHS, Figure 7A ) or cynomolgus monkey serum (CMS, Figure 7B )-induced EA cell lysis are shown. The symbols are used as follows: open circles represent results using anti-C1s antibody hz2-7 (H1L2); open triangles represent results using anti-C1s antibody hz2-8 (H1L2); solid black triangles represent results using anti-C1s antibody hz2-8 (H1L2 G80A); triangles with a diagonal line represent results using anti-C1s antibody hz2-8 (H1L2 G80A / T82A); and open squares represent results using human IgG1. DETAILED DESCRIPTION

[0045] The present invention relates to novel antibodies that bind to C1s and antigen-binding fragments thereof, and methods for preparing and using such novel antibodies and antigen-binding fragments thereof.

[0046] TERMS / DEFINITIONS

[0047] Unless otherwise defined, scientific and technical terms used in connection with the present invention shall have the meanings commonly understood by one of ordinary skill in the art. The use of singular terms ("a / an" or "the" or other use of a singular form of a term) includes plural referents, and the use of plural terms shall include the singular, unless the context clearly dictates otherwise. Thus, for example, reference to "an antibody" includes "one or more" antibodies or "multiple" such antibodies. All publications mentioned herein are hereby incorporated by reference in their entirety.

[0048] Generally, the terms and techniques of molecular biology, microbiology, cell and tissue culture, protein and nucleotide chemistry, and recombinant DNA technology that are known to those of ordinary skill in the art can be used in the antibodies, antigen-binding fragments, compositions, and methods disclosed herein. The techniques and procedures described herein are generally performed according to conventional methods well known in the art and as described in various general and more specific references, particularly Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.) and Ausubel et al. (1994) Current Protocols in Molecular Biology, Volumes I-III (John Wiley & Sons, N.Y.). Unless otherwise indicated herein, enzymatic reactions and purification techniques are performed according to the manufacturer's specifications or as commonly achieved in the art or as described herein. Techniques and methods for the preparation and formulation of pharmaceuticals and the treatment of subjects are described using conventional terms herein.

[0049] "Antibody" is used in the broadest sense to refer to a polypeptide or combination of polypeptides that recognize and bind an antigen through one or more immunoglobulin variable regions, where the immunoglobulin variable regions can be naturally occurring or non-naturally occurring, e.g., as a result of engineering, chimerization, humanization, optimization, CDR grafting, or affinity maturation.

[0050] "Antibodies" as disclosed herein can be full (complete, full-length) antibodies, single-chain antibodies or antigen-binding fragments having one or two chains, and can be naturally occurring and non-naturally occurring. Antibodies include at least sufficient complementarity determining regions (CDRs) interspersed with framework regions (FRs) for the antibody to recognize and bind an antigen. The anti-C1s antibodies disclosed herein can be, but are not limited to, at least one of the following: monoclonal antibodies, polyclonal antibodies, humanized antibodies, chimeric antibodies, single-chain antibodies, Fab fragments, single-chain variable fragments (scFvs), aptamers, single-domain antibodies (VHHs, nanobodies, camel antibodies), recombinant antibodies, modified antibodies having a peptide / other moiety linked to the antibody and / or additional amino acids added at the N-terminus or C-terminus, or other C1s-binding fragments or variants. A full antibody, full-length antibody, complete antibody, naturally occurring antibody or equivalent terms shall be understood to refer to a polypeptide, specifically a glycoprotein, which in its standard form comprises at least two heavy chains (HCs) and two light chains (LCs) interconnected by disulfide bonds. Each HC contains a heavy chain variable region (VH) and an HC constant region (CH), and each light chain contains a light chain variable region (VL) and an LC constant region (CL). The HC variable region VH and the LC variable region VL contain binding domains that interact with an antigen. The VH and VL regions can be further subdivided into CDR regions characterized by hypervariable regions interspersed with generally more conserved FR regions. Each VH and VL typically consists of three CDRs and four FRs arranged in the following order from the amino-terminus to the carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. It should be understood that the assignment of amino acids to each domain is made according to methods known in the art, specifically according to the definitions found in, inter alia, Sequences of Proteins of Immunological Interest, Kabat et al.; National Institutes of Health, Bethesda, Md.; 5th ed., NIH Publication No. 91-3242 (1991); Chothia et al., (1987) J Mol Biol 196:901-917 or Chothia et al., (1989) Nature 342:878-883. The constant region of an antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system and the classical complement system. Generally, an antibody includes at least heavy chain (HC) CDR1, CDR2, and CDR3 and light chain (LC) CDR1, CDR2, and CDR3 sequences, where any one of these sequences can be naturally occurring or non-naturally occurring.An antibody can include fewer CDR sequences as long as the antibody can recognize and bind to an antigen. Alternatively, a full antibody can be a single-chain antibody, such as a homodimer of only heavy chains where antigen binding is mediated by one variable domain.

[0051] The anti-C1s antibodies disclosed herein can be variants that include at least one altered CDR or framework sequence, where the CDR and / or framework sequences can be optimized by mutating the nucleic acid molecule encoding such framework sequence. The variants can be constructed from HC and LC portions independently derived from different sources. Techniques for generating variants include, but are not limited to, conservative amino acid substitutions, computer modeling, screening of candidate polypeptides individually or in combination, and codon optimization, and it should be understood that those skilled in the art are able to generate antibody variants as needed. The anti-C1s antibodies disclosed herein can be fragments. The antigen-binding function of an antibody can be carried out by fragments such as: Fab fragment; a monovalent fragment consisting of VL, VH, CL, and CH1 domains; F(ab)2 fragment; a bivalent fragment comprising two Fab fragments linked at the hinge region by a disulfide bond; Fd fragment consisting of VH and CH1 domains; single-chain variable fragment (scFv) consisting of VL and VH domains of a single arm of an antibody; single-domain antibody (dAb) fragment consisting of a VH domain; and isolated CDR (VHH, nanobody) or aptamer. The antigen-binding portion can be incorporated into single-domain antibodies, bispecific antibodies, trispecific antibodies, tetravalent antibodies, v-NARs, and bis-scFvs (see, for example, Hollinger and Hudson, 2005, Nature Biotechnology, 23(9):1126-1136). The antigen-binding portion of an antibody can be grafted onto a polypeptide-based scaffold to form a monobody (see, for example, U.S. Patent No. 6,703,199, which describes fibronectin polypeptide monobodies).

[0052] The term antibody encompasses a wide variety of polypeptide classes that can be biochemically distinguished. The "class" of an antibody can refer to the type of constant domain or constant region that its heavy chain possesses. Those skilled in the art will understand that there are five major classes of antibodies, namely, IgA, IgD, IgE, IgG, and IgM, and several of these antibodies can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2, each of which is well characterized and known to confer functional specialization. Modified versions of each of these classes and isotypes are readily distinguishable and are within the scope of this disclosure. All immunoglobulin classes are within the scope of this disclosure.

[0053] The term "chimeric" antibody refers to an antibody in which a portion of the heavy chain (HC) and / or light chain (LC) involved in forming the immunoreactive site is derived from a particular source or species, while the remaining portion of the HC and / or LC is derived from a different source or species. In certain embodiments, the target-binding region or site will be from a non-human source (e.g., mouse or non-human primate), and the constant region will be from a human source.

[0054] As used herein, the phrase "humanized antibody" refers to an antibody (immunoglobulin) or antibody variant that includes portions of an antibody (immunoglobulin), where the portions include amino acid sequences of different origins, with at least a portion including amino acid sequences of human origin. Humanized antibodies include portions derived from non-human antibodies, typically murine antibodies that bind to a target of interest, where the portion from the non-human antibody can be the entire variable region or a portion of the variable region from the "parent" non-human antibody, such as one or more CDRs from the "parent" non-human antibody. A humanized antibody can include one or more variable regions from a non-human antibody grafted (fused) onto a human-derived immunoglobulin framework, specifically a human variable region framework. A humanized antibody can include CDRs from the parent non-human antibody grafted (fused) into a framework that includes a portion of a variable region derived from a human immunoglobulin framework, specifically a recipient human framework or a human consensus framework. Techniques and principles for designing, preparing, and testing humanized antibodies are known (Jones PT, Dear PH, Foote J, Neuberger MS, Winter G. Replacing the complementarity-determining regions in a human antibody with those from a mouse. Nature 1986, 321(6069):522-5; Almagro JC, Fransson J. Humanization of antibodies. Front Biosci. 2008, 13:1619-33). It should be understood that changes can be made to one or more CDR sequences and / or the recipient framework at multiple positions in order to develop humanized antibodies with improved characteristics for a desired use, e.g., high affinity for a target, specificity for a selected epitope, avoidance of undesirable events such as isomerization or deamidation, low clearance rate, low toxicity, etc. The anti-C1s antibodies disclosed herein can be humanized variants.

[0055] "Binding" refers to the direct association between two molecules resulting from, for example, covalent, electrostatic, hydrophobic, and ionic and / or hydrogen bond interactions (including interactions such as salt bridges and water bridges). "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, the binding affinity as used herein refers to the intrinsic binding affinity reflecting the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). Affinity can be measured by conventional methods known in the art (including the methods described herein). The concentration calculated at approximately 50% of the maximum binding (calculated EC 50 ) can be used as an estimate of the affinity. The affinity of molecule X for its partner Y is typically represented by the dissociation constant (Kd or KD, representing k off / k on ) measured for the interaction. "Specific binding" or similar terms refer to high-affinity binding, specifically binding occurring with a measured affinity (Kd, KD) in at least the nanomolar range. The anti-C1s antibodies of the present disclosure specifically bind to the C1s protein.

[0056] "Substantially identical" as used to refer to a sequence that is substantially the same as an identified amino acid sequence or nucleotide sequence shall be understood to mean a sequence that is at least about 85%, 90%, 92%, 95%, 97%, or 98%, 99% identical to the amino acid sequence or nucleotide sequence, determined as the percentage of residues in the candidate amino acid or nucleic acid sequence that are identical to the residues in the identified reference sequence after aligning the two sequences and introducing gaps (if necessary) to achieve the maximum percentage of identity. Methods and computer programs for alignment are well known in the art, such as publicly available computer software / programs like Clustal Omega (from the European Bioinformatics Institute (EMBL-EBI) of the European Molecular Biology Laboratory, available at https: / / www.ebi.ac.uk / Tools / msa / clustalo / ); when using such software / programs, it is preferred to use the default parameters, for example, the default parameters for gap penalties and extension penalties.

[0057] "Subject" is a mammal, where mammals include but are not limited to primates (e.g., humans and non-human primates such as monkeys), domestic animals (e.g., cows, sheep, cats, dogs, pigs, llamas, and horses), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the subject is a human. The phrases "to a subject in need", "to a patient in need", "to a patient in need of treatment", or "to a subject in need of treatment" can include a subject who would benefit from administration of an anti-C1s antibody disclosed herein for treating a complement-mediated disorder. It should be understood that administration of an anti-C1s antibody to a "subject in need" can be interpreted to mean a subject known or suspected of having a complement-mediated disorder such as ITP or NMO based on indicators such as symptoms, family history, or genotype. It should be further understood that an anti-C1s antibody can be administered to a subject not known or not suspected of having a complement-mediated disorder, and the purposes can include but are not limited to prophylactic or preventive purposes, for screening, for diagnosis, for research purposes, or for achieving a result different from treating a disorder.

[0058] An "effective amount" of an anti-C1s antibody, e.g., in a pharmaceutical formulation, refers to an amount that is effective at a given dosage and for a necessary period of time to achieve a desired therapeutic or preventive result. It should be understood that an "effective amount" is intended to refer to the amount of an anti-C1s antibody or a pharmaceutical composition comprising an anti-C1s antibody that will elicit a biological response or a desired therapeutic effect in a cell, tissue, system, non-human animal subject, non-human mammalian subject, or human subject being measured. The terms "therapeutically effective amount", "pharmacologically effective amount", and "physiologically effective amount" are used interchangeably and refer to the amount of an anti-C1s antibody required to provide a threshold level of the active antibody in the bloodstream or target tissue. The exact amount will depend on a variety of factors, e.g., the particular anti-C1s antibody (active agent); the components and physical characteristics of the composition; the intended population of subjects / patients to be treated; considerations such as the disease state, age, sex, and weight of the subject, and the exact amount can be readily determined by one of ordinary skill in the art based on the information provided herein or otherwise available in the relevant literature. As used in this context, the terms "improve", "increase", or "decrease" indicate a value or parameter relative to a baseline measurement, such as the measurement in the same subject before initiation of the treatment described herein, or the measurement in a control individual (or control individuals) in the absence of the treatment described herein, or a value available in an information source such as a textbook, manual, or database.

[0059] The term "pharmaceutical composition" or "pharmaceutical formulation" refers to a preparation in a form that permits the biological activity of the active ingredient(s) contained therein, specifically the anti-C1s antibodies disclosed herein, to be effective. It should be understood that a pharmaceutical composition may contain more than one active ingredient, for example, more than one anti-C1s antibody or a combination of an anti-C1s antibody and another active ingredient acting on a different target, where the combination may be, but is not limited to, a combination of an anti-C1s antibody and another active ingredient having a desired effect on other complement pathways or other processes involved in inflammation, a combination of an anti-C1s antibody and a gene therapy agent, or a combination of an anti-C1s antibody and a protein targeting other targets (e.g., an Fc fusion protein). "Pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation that is non-toxic to a subject in addition to the active ingredient(s). It should be understood that a pharmaceutically acceptable carrier may be, but is not limited to, a buffer, an excipient, a stabilizer, an adjuvant, or a preservative.

[0060] As used herein, the term "treat" or "treating" or similar terms may refer to an outcome that is considered beneficial to a particular subject under a defined set of circumstances. Treating a complement-mediated disorder may non-exclusively refer to any one of alleviating, ameliorating, slowing down, interrupting, halting, mitigating, stopping, or reversing the progression or severity of existing symptoms, disorders, conditions, or diseases, and may further encompass preventing or delaying the onset of one or more symptoms of a complement-mediated disorder, and / or reducing the severity or frequency of one or more symptoms of a complement-mediated disorder. The term "treatment" or "method of treatment" or equivalent forms may encompass one or more uses of the anti-C1s antibodies disclosed herein, including but not limited to therapeutic use, prophylactic use, preventive use, diagnostic use, imaging use, and screening use.

[0061] As used herein, a "vector" is a nucleic acid molecule capable of propagating a nucleic acid to which a vector sequence is linked in a host cell into which the vector is introduced. A vector capable of directing the expression of a nucleic acid operably linked thereto is referred to herein as an "expression vector".

[0062] Anti-C1S antibody

[0063] Antibodies and antigen-binding fragments are provided that are capable of binding to C1s and modulating the activity of C1s, thereby having the effect of modulating the activity of at least one component involved in at least one complement-mediated disorder, specifically at least one disorder associated with CP activation by autoantibodies that bind to self-antigens. Antibodies and antigen-binding fragments are provided that are capable of binding to C1s and modulating the activity of C1s, thereby having the effect of modulating the activity of at least one component of CP involved in ITP and / or NMO. Anti-C1s antibodies that are capable of binding to C1s and modulating the activity of C1s can be used in methods and compositions for treating complement-mediated disorders associated with CP activation by autoantibodies that bind to self-antigens, specifically ITP.

[0064] Once an antibody or fragment specific for C1s is provided, the desired biological activity of modulating the activity of C1s or having an effect on modulating at least one component involved in a complement-mediated disorder can be tested by several methods known to those skilled in the art.

[0065] It should be understood that as used herein, "modulate" or similar terms may refer to one or more effects that may occur when the anti-C1s antibodies disclosed herein bind to their target molecule C1s. "Modulate" and its equivalents may refer to different modes of action and effects depending on the component being considered, i.e., modulate may refer to neutralize, reverse, inhibit, block, reduce, antagonize, agonize, amplify, enhance, or otherwise alter the complement system, specifically the activity of at least one component involved in the CP. It should be understood that as used herein, "inhibit C1s" or "inhibition of C1s" or similar terms refer to inhibiting at least one activity or function of C1s involved in the complement cascade, such that inhibiting C1s has downstream effects, such as inhibiting the activation of enzymes involved in the complement cascade, the association or dissociation of molecules involved in the complement cascade, the generation of downstream effectors that require C1s activation, or the degree of lysis of serum complement-induced antibody-sensitized cells. It should be understood that as used herein, "C1s inhibition" or similar terms do not require demonstration of the specific activity or function of C1s that is inhibited when the C1s antibody of the present invention binds to the C1s target molecule, since measurements showing inhibition of processes or effectors known to be downstream of C1s activation can be used as surrogate measurements or indicators of C1s activity or function, and thus as an indicator of inhibition of C1s activity or function. Thus, a C1s antibody capable of binding and inhibiting C1s is an antibody capable of, inter alia, inhibiting the activity of the classical pathway (CP) of complement activation, inhibiting downstream effects of C1s activation, inhibiting IgM-induced deposition of C4 from serum, inhibiting antibody-antigen complex-mediated C4 cleavage, inhibiting antibody-antigen complex-mediated deposition of C4b, inhibiting the formation of C3 convertase, inhibiting downstream effects of CP activation, etc.

[0066] It should be understood that the term "component" can refer not only to the target molecule C1s, but also to molecules, processes or pathways involved in complement-mediated effects downstream of C1s activation, specifically downstream molecules, processes or pathways involved in CP-mediated effects downstream of C1s activation. It should be understood that by targeting C1s with an anti-C1s antibody, CP can be selectively targeted, such that CP-mediated effects can be specifically or selectively inhibited. This selective targeting effect of CP is also understood to keep the alternative and lectin pathways intact against infection. It should be understood that the anti-C1s antibody as disclosed herein can be used to therapeutically target CP activation triggered by antibody-antigen complexes and to modulate the activity of CP according to CP activation triggered by antibody-antigen complexes. The anti-C1s antibody as disclosed herein can be used to therapeutically target at least one component involved in complement-mediated disorders, specifically at least one component involved in disorders mediated by CP, specifically at least one component involved in CP-mediated autoimmune disorders, specifically at least one component involved in ITP and / or NMO. Without wishing to be bound by a particular mechanism of action, targeting C1s with the anti-C1s antibody as disclosed herein should be understood to specifically inhibit CP-mediated cell lysis, which is the basis of autoimmune pathologies that result in effects such as the destruction of self platelets, red blood cells or astrocytes. In certain embodiments, the anti-C1s antibody as disclosed herein can be used to inhibit the CP pathway in a dose-dependent manner. In certain embodiments, the anti-C1s antibody as disclosed herein can be used to inhibit the effects of CP activation triggered by antibody-antigen complexes in a dose-dependent manner.

[0067] In certain embodiments, the anti-C1s antibody as disclosed herein can be used to therapeutically target CP activation triggered by antibody-antigen complexes and to modulate the activity of CP downstream of CP activation triggered by antibody-antigen complexes. Specifically, the anti-C1s antibody as disclosed herein can be used to therapeutically target CP activation triggered by antibody-antigen complexes and to inhibit the activity of CP downstream of CP activation triggered by antibody-antigen complexes.

[0068] Therapeutically targeting at least one component involved in disorders mediated by CP with the anti-C1s antibody as disclosed herein should be understood to allow precise modulation of the activity of CP and downstream effects of CP activity. Specifically, therapeutically targeting at least one component involved in disorders mediated by CP with the anti-C1s antibody as disclosed herein should be understood to allow precise inhibition of the activity of CP and downstream effects of CP activity.

[0069] The anti-C1s antibodies disclosed herein allow for the development of treatments that can be customized for each subject (e.g., dose, frequency of administration), where the treatment can be easily continued and interrupted and combined with other therapies. In certain strategic embodiments, the anti-C1s antibodies disclosed herein can be combined with other therapies that can address multiple therapeutic targets and / or address the deficiencies or undesirable effects of one of the therapies in a combination therapy.

[0070] Exemplary embodiments of anti-C1s antibodies and their uses

[0071] Non-limiting exemplary embodiments of the anti-C1s antibodies of the present invention are currently disclosed, specifically in the examples, tables, and figures.

[0072] Antibodies capable of specifically binding to C1s and inhibiting C1s function.

[0073] A functional cascade can be used to identify and characterize the anti-C1s antibodies of the present invention, where the first step in the cascade involves screening clones attacked by C1s to obtain antibodies capable of binding to C1s, followed by sequencing of the homologous variable regions, recombinant expression of chimeric antibodies with VH and VL from murine clones ligated to human constant regions, purification of the recombinant antibodies, and screening of the antibodies using functional assays.

[0074] As demonstrated by the exemplary embodiments disclosed in Example 2, recombinant chimeric antibodies capable of modulating C1s function were obtained, as measured by an assay for the deposition of IgM-induced C4 from serum. Exemplary recombinant chimeric anti-C1s antibodies capable of inhibiting C1s function were obtained, as measured by the ability to inhibit the deposition of IgM-induced human C4 from human serum and the ability to inhibit the deposition of IgM-induced cynomolgus monkey C4 from cynomolgus monkey serum. Exemplary recombinant chimeric anti-C1s antibodies capable of inhibiting C1s function in a dose-dependent manner were obtained.

[0075] Humanized variants

[0076] When administered to human subjects, humanized antibodies that incorporate CDRs from non-human sources into a human antibody framework are expected to be non-immunogenic. As demonstrated by the exemplary embodiments disclosed in Example 2, humanized anti-C1s antibody variants were successfully generated, tested, optimized, and selected. Humanization of the 2-7 and 2-8 antibodies yielded humanized anti-C1s antibody variants, with the antibodies identified as hz2-7 (H1L2) and hz2-8 (H1L2) having the highest in vitro activity. After initial design and testing, variants showing desired antigen binding affinity and cross-reactivity were selected for further evaluation and development, including but not limited to modifying some of the parental CDR sequences to avoid potential unwanted events such as aspartic acid isomerization and asparagine deamidation and selected modifications of the framework sequences.

[0077] Anti-C1s antibodies with high affinity for biologically relevant targets

[0078] Exemplary anti-C1s antibodies show high affinity for biologically relevant targets. Anti-C1s antibodies show high affinity binding to both the zymogen (native) C1s protein and the active C1s protein. As demonstrated by the exemplary embodiments of affinity measurements performed using a variety of methods disclosed in Examples 2 and 3 and shown in Figures 2 and 3, antibodies 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2), hz2-8 (H1L2 G80A), and hz2-8 (H1L2 G80A / T82A) exhibit favorable affinity characteristics.

[0079] Anti-C1s antibodies with cross-reactivity to non-human targets

[0080] It is desirable that therapeutically useful antibodies or antibody fragments have sufficient cross-reactivity with non-human targets (non-human homologs) from sources associated with additional studies such as preclinical efficacy studies, animal models of disease, toxicology studies, etc., such that the antibody or antibody fragment should recognize, for example, rat homologs and / or primate homologs, such as homologs from cynomolgus monkeys. As demonstrated by exemplary embodiments in the functional screening assays disclosed in Example 2 and shown in Figure 2, antibodies 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2), hz2-8 (H1L2 G80A), and hz2-8 (H1L2G80A / T82A) inhibit IgM-induced deposition of human C4 from human serum and IgM-induced deposition of cynomolgus monkey C4 from cynomolgus monkey serum, thereby providing antibodies that are functionally cross-reactive with cynomolgus monkey C1s in the cynomolgus monkey system against human C1s production. As demonstrated by exemplary embodiments of binding affinity measurements disclosed in Example 3 and shown in Figure 3, antibodies 2-7, 2-8, hz2-7 (H1L2), hz2-8 (H1L2), and hz2-8 (H1L2G80A / T82A) exhibit detectable binding and high affinity for human C1s and cynomolgus monkey C1s, while antibodies 2-8 and hz2-8 (H1L2 G80A / T82A) also exhibit detectable binding and fairly high affinity for rat C1s.

[0081] Anti-C1s antibodies capable of selectively inhibiting the classical pathway (CP)

[0082] For CP-mediated disorders, such as those triggered by complement-fixing autoantibodies that bind to antigens, it is desirable to selectively inhibit the CP at a step upstream of the generation of common complement system effectors, specifically the C3 convertase. Selectively inhibiting the CP should be understood as inhibiting the action of CP activation triggered by the antibody-antigen complex while leaving the alternative and lectin complement pathways intact to combat infection. For CP-mediated disorders, selectively inhibiting the CP at a step upstream of the generation of the C3 convertase should be understood as selectively inhibiting the unwanted action of CP activation associated with the disorder while leaving the alternative and lectin complement pathways substantially intact to combat infection. The anti-C1s antibodies of the present invention have a selective action within the CP by inhibiting C1s (i.e., inhibiting at least one C1s activity or function), thereby acting upstream of the formation of common complement system effectors.

[0083] As demonstrated by the exemplary embodiments disclosed in Example 2, the anti-C1s antibodies disclosed herein are capable of binding to C1s and inhibiting C1s functions involved in IgM-induced C4 deposition from human serum and cynomolgus monkey serum, wherein it should be understood that inhibiting C4 deposition in a test assay indicates at least one of the following: C4 cleavage did not occur; C3 convertase formation was inhibited (prevented); and / or no downstream effects mediated by the common complement pathway effector C3 were produced by CP activation. Specifically, the anti-C1s antibodies disclosed herein are capable of specifically binding to C1s and inhibiting at least one activity or function of C1s such that IgM-induced C4 deposition from human serum and cynomolgus monkey serum is inhibited, C4 cleavage is inhibited, C3 convertase formation is inhibited (prevented), and CP activation is inhibited.

[0084] The exemplary embodiments disclosed in Example 7 demonstrate that the anti-C1s antibodies disclosed herein are capable of modulating C1s functions involved in the complement classical pathway (CP). Sheep red blood cells coated with rabbit anti-sheep red blood cell antiserum (hemolysin), also known as antibody-sensitized sheep red blood cells (“EA” or “EA cells”), have traditionally been used to assay the activity of the complement classical pathway (CP) in serum samples. Commercially available EA cells (Complement Technology, #B202) and a known assay system were used to study complement-mediated EA lysis in response to exposure to serum. The humanized anti-C1s antibodies hz2-7 (H1L2), hz2-8 (H1L2), hz2-8 (H1L2 G80A), and hz2-8 (H1L2G80A / T82A) inhibited complement-mediated EA lysis in a dose-dependent manner (Example 7, Figure 7A and 7B ), wherein the antibody hz2-8 (H1L2) was the most effective inhibitor, the antibodies hz2-8 (H1L2 G80A) and hz2-8 (H1L2G80A / T82A) were also highly effective inhibitors, and the antibody hz2-7 (H1L2) was less effective and showed species dependence.

[0085] The anti-C1s antibodies disclosed herein showed different inhibition levels under different conditions, e.g., using different antibodies under the same assay conditions, or using the same antibody under different assay conditions, such as using sera from different species. Thus, the anti-C1s antibodies disclosed herein provide tools and avenues for studying the roles of specific epitopes and binding interactions in the CP pathway.

[0086] Composition

[0087] Compositions are provided that include the anti-C1s antibodies of the present invention and a pharmaceutically acceptable carrier or excipient suitable for the intended use of each composition. Such carriers include, but are not limited to: saline, buffers, glucose, water, glycerol, ethanol, excipients, stabilizers, preservatives, or combinations thereof. It should be understood that the pharmaceutical formulation should match the mode of administration.

[0088] The anti-C1s antibodies disclosed herein can be administered by any suitable means, including but not limited to injection or parenteral administration. Parenteral administration can include intramuscular, intravenous, intraarterial, intraperitoneal, subcutaneous, intraspinal (including epidural or intrathecal), intraocular, intracerebral, intraventricular, intracardiac, intradermal / intracutaneous, intraarticular, intralymphatic, or intraosseous administration. The anti-C1s antibodies disclosed herein can be formulated into compositions for introduction into the circulatory system by parenteral administration, specifically by intravenous or intraarterial administration. The anti-C1s antibodies disclosed herein can be administered using a device, or as a depot, or in the form of a sustained release formulation (e.g., a semipermeable matrix of a solid hydrophobic polymer containing the antibody, or microcapsules) to allow slow and / or measured and / or local delivery. The anti-C1s antibodies disclosed herein can be formulated and administered using colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in a coarse emulsion.

[0089] Methods

[0090] Methods are provided for treating complement-mediated disorders using an effective amount of the anti-C1s antibodies disclosed herein. Without wishing to be bound by a particular mechanism of action, methods are provided for precisely targeting and inhibiting C1s using the anti-C1s antibodies disclosed herein, thereby allowing precise targeting of C1s to selectively modulate the activity of the CP. Methods for precisely targeting and inhibiting C1s can be methods of using the route of administration to target the location or system involved in the disorder, methods of using the timing of administration to target critical periods, and combinations of these methods. Methods are provided for treating complement-mediated disorders in which the CP is activated by complement-fixing autoantibodies that bind to self-antigens, and in which such activation of the CP results in harmful effects on tissues carrying the self-antigen. Methods are provided for treating disorders associated with unwanted activation of the CP, wherein such methods use an effective amount of the anti-C1s antibodies disclosed herein to selectively modulate the activity of CP processes such as C4 cleavage and deposition, thereby resulting in unwanted attack and destruction of self-tissues. Specifically, methods are provided for treating autoimmune conditions by inhibiting the downstream effects of CP activation by complexes of complement-fixing autoantibodies that bind to self-antigens on platelets.

[0091] Methods and compositions are provided for precisely targeting and inhibiting C1s by using an administration route to target a location or system involved in a disorder. Without being bound by a particular mechanism of action, methods and compositions are provided for treating disorders characterized by undesirable, abnormal, inappropriate, or excessive CP activation in the circulatory system that can be targeted by introducing an anti-C1s antibody into the circulatory system, for example, by intravenous or intra-arterial administration. Without being bound by a particular mechanism of action, methods and compositions are provided for treating disorders characterized by undesirable, abnormal, inappropriate, or excessive CP activation in the central nervous system (CNS) that can be targeted by introducing an anti-C1s antibody into one or more CNS tissues or structures by intraspinal (including epidural or intrathecal) administration, intraocular administration, intracerebral administration, or intraventricular administration. Without being bound by a particular mechanism of action, methods and compositions are provided for treating disorders characterized by undesirable, abnormal, inappropriate, or excessive CP activation in one or more joints that can be targeted by introducing an anti-C1s antibody into one or more joints by intra-articular administration.

[0092] Methods are provided for treating immune thrombocytopenic purpura (ITP) by precisely targeting C1s using an effective amount of an anti-C1s antibody disclosed herein. Compositions of the anti-C1s antibody disclosed herein can be formulated for introduction into the circulatory system, for example, by intravenous and / or intra-arterial administration, and introduced by a method that allows targeting and inhibition of C1s in the circulatory system. Without being bound by a particular mechanism of action, the methods using the anti-C1s antibody disclosed herein can inhibit antibody-mediated platelet removal by inhibiting CP. Without being bound by a particular mechanism of action, the methods using the anti-C1s antibody disclosed herein can inhibit the deposition of C4 on platelets, which can lead to undesirable platelet destruction. When the CP pathway has been activated by complement-fixing autoantibodies bound to autoantigens on platelets, the methods provided herein allow precise targeting of C1s, where inhibition of C1s activity interrupts the CP pathway and prevents downstream effects, including but not limited to inhibition of IgM-induced deposition of C4 from serum; inhibition of antibody-antigen complex-mediated C4 cleavage; inhibition of antibody-antigen complex-mediated deposition of C4b on platelets; platelet destruction; platelet elimination; thrombocytopenia; and bleeding disorders.

[0093] Provided are methods of treating neuromyelitis optica (NMO) by precisely targeting C1s using an effective amount of an anti-C1s antibody disclosed herein. Compositions of the anti-C1s antibody disclosed herein can be formulated for targeted introduction into the CNS, specifically for intraspinal (e.g., epidural or intrathecal) or intracerebral / intraventricular introduction, and are introduced by a method that allows targeting and inhibition of C1s in the CNS where complement-dependent cytotoxicity (CDC) may occur. Without being bound by a particular mechanism of action, methods using the anti-C1s antibody disclosed herein can inhibit complement-dependent cytotoxicity (CDC) triggered by autoantibodies binding to autoantigens, including but not limited to inhibiting antibody-antigen complex-mediated C4 cleavage; inhibiting antibody-antigen complex-mediated deposition of C4b on astrocytes; inhibiting downstream effects of antibody-antigen complex-mediated CP activation and inhibiting downstream inflammation leading to, for example, oligodendrocyte and neuron damage, as well as inflammatory demyelinating lesions in the CNS, specifically the spinal cord and optic nerve.

[0094] The methods and compositions provided herein allow for precise timing of treatment. Without being bound by a particular mechanism of action, the anti-C1s antibody disclosed herein can be administered during the acute phase of the disorder to inhibit CP-mediated processes during the acute phase of the disorder. Without being bound by a particular mechanism of action, selectively inhibiting CP-mediated processes during the acute phase of the disorder using an anti-C1s antibody can interrupt acute events and / or prevent progression of the disorder, and then allow CP to resume normal activity after the anti-C1s antibody has dissociated from the C1s target.

[0095] The methods and compositions provided herein allow for precise targeting by timing and location of treatment. Without being bound by a particular mechanism of action, the timing of administration of the anti-C1s antibody disclosed herein can occur before the acute phase of the disorder (e.g., in response to indicators of a possible acute phase onset) or during the acute phase to selectively inhibit CP-mediated processes during the acute phase without inhibiting the function of other complement pathways, and the route of administration can be selected to target specific tissues, systems, or structures involved in the disorder. Without being bound by a particular mechanism of action, inhibiting CP-mediated processes before or during the acute phase of the disorder using an anti-C1s antibody can prevent or interrupt acute events without inhibiting the normal function of other complement pathways, and then allow CP to resume normal activity after the anti-C1s antibody has dissociated from the C1s target.

[0096] The methods and compositions provided herein allow for control of the amount of therapeutic agent present at one or more time points during treatment. The anti-C1s antibodies disclosed herein can inhibit C1s activity and selectively inhibit CP pathway activity in a dose-dependent manner. Without being bound by a particular mechanism of action, a dose (“effective amount”) sufficient to improve at least one biological effect or symptom of the disorder can depend on factors specific to the subject in need thereof such that treatment can include determining the dose of anti-C1s antibody required to constitute an effective amount for the subject to be treated and administering to the subject an effective amount of anti-C1s antibody, wherein the level of the therapeutic agent (anti-C1s antibody) is expected to decline after treatment due to dissociation from the C1s target and clearance from the blood.

[0097] Methods for treating complement-mediated disorders as provided herein include administering to a subject in need thereof an effective amount of an anti-C1s antibody disclosed herein, wherein administering the effective amount of anti-C1s antibody improves at least one biological effect or symptom associated with the disorder. Methods for treating complement-mediated disorders associated with autoantibody-triggered CP activation are provided, wherein administering to a subject in need thereof an effective amount of an anti-C1s antibody disclosed herein inhibits the CP pathway and prevents downstream effects of unwanted CP activation.

[0098] The following examples are provided to illustrate and not limit the claimed invention.

[0099] Examples

[0100] Example 1: Antibody Production and Identification of Antibodies That Bind to C1S

[0101] The generation of novel monoclonal antibodies against C1s was performed by GenScript USA, Inc. (Piscataway, NJ) under contract using in vivo rodent immunization and single B cell cloning technology. The active form of the C1s protein (CompTech A104 C1s enzyme, CompTech) was used as an antigen to immunize BALB / cJ and SJL mice (The Jackson Laboratories). Sufficient plasma titers as determined by enzyme-linked immunosorbent assay (ELISA) were obtained, which triggered downstream antibody recovery and screening activities. Secondary lymphoid organs were harvested for CD138 plasma cell enrichment and then loaded on On a 14K chip (Berkeley Lights, Inc., Emeryville, CA), single B cells secreting antibodies that specifically bind to C1s antigen were screened. Positive single B cell clones were selected and exported for subsequent variable domain sequencing and recombinant antibody expression. Full-length recombinant chimeric antibodies were obtained by expression in ExpiCHO cells using pcDNA3.4-VH-CH (where VH is from the positive B cell clone (mouse) and CH is IgG1 (human)) and pcDNA3.4-VL-CL (where VL is from the positive B cell clone (mouse) and CL is κ (human)). The functional activities of the recombinantly expressed and purified antibodies were further screened in an in vitro cell-free system. From the obtained positive C1s-binding B cell clones, 159 clones were successfully exported for antibody sequencing, and 39 clones were sequenced with a pair of homologous heavy and light chains. All 39 anti-C1s clones were recombinantly expressed and purified, and functional screening was performed.

[0102] Example 2. Functional Screening of Anti-C1S Antibodies

[0103] The anti-C1s antibodies from all 39 anti-C1s clones were recombinantly expressed and purified, and then subjected to a functional screening assay to identify anti-C1s antibodies capable of inhibiting IgM-induced C4 deposition. Briefly, under conditions suitable for IgM-induced C4 deposition from serum, the anti-C1s antibodies were incubated with serum in IgM-coated plates, and then the amount of deposited C4 was detected. According to the functional model, this functional assay serves as a surrogate assay for C1s activity, where C4 must be cleaved by active C1s before C4b deposition (specifically, C4b deposition on target cells such as platelets), such that measuring the effect of the anti-C1s antibody on IgM-induced C4 deposition from serum is used to report the effect of the anti-C1s antibody on C1s activity. This functional assay serves as a surrogate assay for C1s activity (C1s function, C1s bioactivity) in antibody-antigen complex-mediated CP activation and the downstream effects of this CP activation (see Figure 1 ). Anti-C1s antibodies capable of inhibiting IgM-induced C4 deposition were selected for further study.

[0104] Functional screening was performed by first coating 300 ng / well of IgM from human serum (Sigma-Aldrich I8260) in 96-well plates overnight. The next morning, the plates were blocked with gelatin blocking buffer (1% in PBS, Alfa Aesar J62755) for 1 hour before adding various concentrations of anti-C1s antibodies diluted in gelatin blocking buffer to each well. After briefly equilibrating the anti-C1s antibodies by gently shaking the plates, NHS (normal human serum, CompTech NHS) was added to each well to a final concentration of 1.25% (v / v). The plates were mixed well and incubated at 37 °C for 1 hour to allow IgM-induced C4 deposition. Then, the plates were washed thoroughly before incubating for 1 hour with goat anti-human C4 (CompTech A205) diluted in gelatin blocking buffer. After incubation, the plates were washed again and incubated for 1 hour with rabbit anti-goat IgG (H+L) secondary antibody HRP-conjugated (Invitrogen 81-1620) diluted in gelatin blocking buffer. Finally, the plates were washed with PBS and developed with ELISA liquid substrate (Sigma-Aldrich T4444), and the reaction was stopped by adding an equal volume of ELISA liquid substrate of 1 M H2SO4. The bound antibody was measured by absorbance at OD 450nm to determine the amount of deposited C4. Figure 2A Results are shown for recombinant antibodies from clone 2-7 (designated anti-C1s antibody 2-7) and from clone 2-8 (designated anti-C1s antibody 2-8) of the top candidates, as well as for an antibody from one of the non-functional clones (3-30) that binds to C1s but does not measurably inhibit C1s function in the functional assay. The half-maximal inhibitor concentration (IC50) was determined for each antibody, which shows the ability to inhibit IgM-induced C4 deposition, where the IC 50 values for anti-C1s antibody 2-7 and anti-C1s antibody 2-8 were between 0.3 μg / ml and 0.7 μg / ml.

[0105] The functional screening assay was repeated using normal cynomolgus monkey serum (CMS, CompTech NCYS) as described above. Figure 2B Results are shown that anti-C1s antibodies 2-7 and 2-8 also effectively inhibit IgM-induced deposition of cynomolgus monkey C4, where the IC 50 of anti-C1s antibody 2-7 was approximately 0.3 μg / ml, and the IC 50 of anti-C1s antibody 2-8 was approximately 0.5 μg / ml.

[0106] Generation and screening of humanized anti-C1s antibody variants

[0107] Humanization of the parental antibody was carried out by transplanting CDRs onto the human antibody framework. First, homology modeling of the 3D structure of the parental antibody was performed to establish a structural model of the parental antibody. The amino acid sequence of the variable fragment framework was identified based on overall sequence identity, matching VH-VL interface positions, standard positions of similarly classified CDRs (Kabat numbering), and removal of potential N-glycosylation sites. Humanized antibodies were designed by generating multiple hybrid sequences that fused selected portions of the parental antibody sequence with the human framework sequence. The isotypes selected to form the humanized antibodies were IgG1 for the heavy chain and IgG1κ for the light chain. Using the 3D model, these humanized sequences were systematically analyzed by eye and computer modeling to isolate the sequences most likely to retain antigen binding. The goal was to maximize the amount of human sequence in the final humanized antibody while maintaining the original antibody specificity. Then, a humanized variant pairing the humanized VH and VL was expressed in ExpiCHO cells using the pcDNA3.4-VL-CL-IRES-VH-CH vector, where IRES is the "internal ribosome entry site" that allows for co-expression of the light and heavy chains, and the humanized variant was designed and constructed. Then, the expressed recombinant humanized variant was purified for affinity analysis.

[0108] As part of the affinity analysis, in one round of designing, generating, and testing variants, three VH variants with the VH-CDR of the parental antibody were generated at corresponding positions in three different human IgG1-derived frameworks, and three VL (VK) variants with the VL-CDR of the parental antibody were generated at corresponding positions in three different human IgG1κ-derived frameworks. A total of nine (9) humanized variants representing each combination of VH and VL (VK) variants were prepared according to a 3VH x 3VK matrix, and the antigen-binding properties (k on 、k off 、KD) of the humanized variants were evaluated.

[0109] Variants showing desired antigen-binding affinity based on a cutoff value were selected for further evaluation and development. In some cases, the parental CDR sequences were modified to avoid potential undesired events such as aspartic acid isomerization or asparagine deamidation.

[0110] The humanized anti-C1s antibodies hz2-7 (H1L2) and hz2-8 (H1L2) were selected from all the tested humanized variants because they had excellent ability to inhibit IgM-induced C4 deposition from NHS and CMS (see Figure 2C and 2D ). Comparison of the IC 50 values showed that the antibody hz2-7 (H1L2) inhibited IgM-induced C4 from NHS ( Figure 2D) and CMS( Figure 2C ) does not deposit as effectively as its parental antibody 2-7( Figure 2A (NHS), Figure 2B (CMS)), but the antibody hz2-8 (H1L2) inhibits IgM-induced C4 deposition from NHS( Figure 2C ) and CMS( Figure 2D ) more effectively than its parental antibody 2-8( Figure 2A (NHS), Figure 2B (CMS)).

[0111] Figure 2C The results of the inhibition of IgM-induced C4 deposition from NHS by the humanized antibodies hz2-7 (H1L2) and hz2-8 (H1L2) and the variants hz2-7 (H1L2G131A), hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A) are shown. For C1s-binding antibodies capable of inhibiting IgM-induced C4 deposition from NHS, the following IC 50 values( Figure 2C ) were calculated: the IC 50 of hz2-7 (H1L2) was 1.1 μg / ml; the IC 50 of hz2-8 (H1L2) was 0.2 μg / ml; the IC 50 of hz2-8 (H1L2G80A) was 0.5 μg / ml; and the IC 50 of hz2-8 (H1L2 G80A / T82A) was 0.4 μg / ml. As Figure 2C shown, the variant hz2-7 (H1L2 G131A) became functionally inactivated after introducing the mutation G131A located within CDR3, and thus the IC 50 value of the functionally inactivated variant hz2-7 (H1L2G131A) was not calculated. Both variants hz2-8 (H1L2G80A) and hz2-8 (H1L2 G80A / T82A) were functionally active, but both were less effective than hz2-8 (H1L2)( Figure 2C ).

[0112] Figure 2D The results of the inhibition of IgM-induced C4 deposition from CMS by the humanized antibodies hz2-7 (H1L2) and hz2-8 (H1L2) and the variants hz2-7 (H1L2G131A), hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A) are shown. Figure 2C and Figure 2D The comparison shows that when using CMS( Figure 2D ) instead of NHS( Figure 2C) When this was done, results of the same pattern were obtained. For C1s-binding antibodies capable of inhibiting IgM-induced C4 deposition from CMS, the following IC 50 values ( Figure 2D ) were calculated: The IC 50 of hz2-7 (H1L2) was 0.9 μg / ml; the IC 50 of hz2-8 (H1L2) was 0.2 μg / ml; the IC 50 of hz2-8 (H1L2 G80A) was 0.5 μg / ml; and the IC 50 of hz2-8 (H1L2 G80A / T82A) was 0.4 μg / ml. The IC 50 value of the functionally inactivated variant hz2-7 (H1L2 G131A) was not calculated.

[0113] The antibody hz2-8 (H1L2G80A / T82A), in which both the aspartic acid isomerization and asparagine deamidation sites had been removed, was selected for further development.

[0114] Table 1.a. below shows the SEQ ID NOs assigned to the heavy chain (HC) and light chain (LC) of each full-length antibody used in the examples, and presents the VH amino acid sequence and SEQ ID NO: of each antibody, as well as the VL amino acid sequence and SEQ ID NO:, with the CDRs indicated by underlining.

[0115]

[0116]

[0117]

[0118] Example 3. Binding Affinity of Anti-C1S Antibodies

[0119] Anti-C1s Antibody Affinity and Binding Kinetics: Measurements Using Biolayer Interferometry

[0120] Using the RED96e system (Sartorius AG), biolayer interferometry technology was used for anti-C1s antibody affinity measurement and binding kinetics determination. For the first baseline, a prehydrated anti-human IgG Fc capture (AHC) biosensor was first equilibrated in 1x KB (kinetic buffer, 1x PBS pH 7.4 + 0.02% Tween-20 + 0.1% BSA) for 120 seconds, and then with a 10 mg / ml anti-C1s antibody (2-7, Figure 3A ; 2-8, Figure 3B ; hz2-7 (H1L2), Figure 3C; hz2-8 (H1L2 G80A / T82A), Figure 3D ) was loaded onto the AHC biosensor for 240 seconds. Then, a second baseline signal was established for 120 seconds before associating with various concentrations of active human C1s (CompTech A104) for 240 to 360 seconds. Finally, the analyte was dissociated in 1x KB for 360 seconds. Anti-C1s antibodies 2-7 and 2-8 were also analyzed against human pro-C1s (CompTech A103) (data not shown). Data analysis was performed using HT software (Sartorius AG) for data analysis. KD, k on , k off and R 2 values were calculated for each antibody and target combination, as Figure 3E summarized, showing high-affinity binding, where the KD values were in the low nanomolar to picomolar range. Figure 3E Further shown was that anti-C1s antibodies 2-7 and 2-8 gave exactly the same calculated values for pro-C1s and active C1s proteins, where the K D values were between 0.14 nM and 1.2 nM.

[0121] Anti-C1s antibody binding affinity: Measurement using ELISA

[0122] ELISA was performed by first coating a 96-well plate overnight with 200 ng / well of human active C1s (CompTech A104, Figure 3F ) or human pro-C1s (CompTech A103, Figure 3G ) protein. The next morning, the plate was blocked for 1 hour with SuperBlock T20 blocking buffer (Thermo Fisher Scientific Inc., catalog number 37536) with gentle shaking. Then, various concentrations of anti-C1s antibodies diluted in SuperBlock T20 blocking buffer were added to the plate and incubated for 1 hour with gentle shaking. Human IgG1 (BioXCell BP0297) was used as a negative control. The plate was washed before incubating for 1 hour with an HRP-conjugated secondary antibody diluted in SuperBlock T20 blocking buffer (goat anti-human IgG antibody, HRP conjugate (Millipore AP309P)) with gentle shaking. Finally, the plate was washed and developed with an ELISA liquid substrate (Sigma-Aldrich), and the reaction was stopped by adding an equal volume of 1M H2SO4 to the ELISA liquid substrate. Bound antibody was measured by absorbance at OD 450nm .

[0123] Figure 3F and Figure 3G together demonstrate that top candidates 2-7 and 2-8, and their humanized variants hz2-7 (H1L2) and hz2-8 (H1L2 G80A / T82A), respectively show strong binding to human active C1s (CompTech A104)( Figure 3F ) and human pro-C1s (CompTech A103)( Figure 3G ), with half-maximal effective concentration (EC 50 ) values ranging between 8 ng / ml and 13 ng / ml. The EC 50 assays for each antibody and target combination were as follows: for antibody 2-7, the EC 50 for human active C1s was 12 ng / ml and the EC 50 for human pro-C1s was 9 ng / ml; for antibody 2-8, the EC 50 for human active C1s was 12 ng / ml and the EC 50 for human pro-C1s was 10 ng / ml; for variant hz2-7 (H1L2), the EC 50 for human active C1s was 13 ng / ml and the EC 50 for human pro-C1s was 12 ng / ml; and for variant hz2-8 (H1L2 G80A / T82A), the EC 50 for human active C1s was 10 ng / ml and the EC 50 for human pro-C1s was 8 ng / ml. Control assays using the same concentration of human IgG1 showed that these patterns do not reflect non-specific binding( Figures 3F - 3G ).

[0124] Anti-C1s antibody cross-reactivity: Measurements using biolayer interferometry

[0125] For the first baseline, the pre-hydrated anti-human IgG Fc capture (AHC) biosensor was first equilibrated in 1x KB (kinetic buffer, 1x PBS pH 7.4 + 0.02% Tween-20 + 0.1% BSA) for 120 seconds, followed by incubation with 10 mg / ml anti-C1s antibody (2-7, Figure 3H and 3N ; 2-8, Figure 3I 、 3L and 3O; hz2-7 (H1L2), Figure 3J and 3P ; hz2-8 (H1L2 G80A / T82A), Figure 3K 、 3MAnd 3Q) were loaded onto the AHC biosensor for 240 seconds. Then, a second baseline signal was established for 120 seconds before associating with various concentrations of human C1s (produced in-house (SEQ ID NO:99), Figures 3H - 3K as shown in), rat C1s (produced in-house (SEQ ID NO:105), Figures 3L - 3M as shown in), or cynomolgus monkey C1s (produced in-house (SEQ ID NO:103), Figures 3N - 3Q as shown in) for 240 to 360 seconds. Finally, the analyte was dissociated in 1x KB for 360 seconds. Data analysis was performed using Data Analysis HT software (Sartorius). KD, k on , k off and R 2 values were calculated for each antibody and target combination as summarized in Figure 3R . Top candidates 2-7 and 2-8 and their humanized variants hz2-7 (H1L2) and hz2-8 (H1L2G80A / T82A) each showed strong binding to human C1s and cynomolgus monkey C1s, but did not show binding to mouse C1s ( Figure 3R ), where K D was between 0.012 nM and 3.5 nM. Anti-C1s antibody 2-7 and its humanized variant hz2-7 (H1L2) did not show binding to rat C1s, while 2-8 and its humanized variant hz2-8 (H1L2 G80A / T82A) showed binding to rat C1s, where K D values were between 16 nM and 17 nM ( Figure 3R ). The binding affinity of 2-8 and hz2-8 (H1L2 G80A / T82A) for rat C1s was weaker than that for human C1s and cynomolgus monkey C1s ( Figure 3R ).

[0126] Anti-C1s antibody cross-reactivity: Measurement using ELISA

[0127] ELISA was performed by first coating 96-well plates with 200 ng / well of human C1s (identified as full-length human C1s, HuC1s, wt human C1s) (produced in-house (SEQ ID NO:99), Figure 3S ), mouse C1s (produced in-house (SEQ ID NO:101), Figure 3T ), rat C1s (produced in-house (SEQ ID NO:105), Figure 3U ), or cynomolgus monkey C1s (produced in-house (SEQ ID NO:103), Figure 3V)Performed by overnight protein coating of a 96-well plate. The next morning, the plate was blocked with SuperBlock T20 blocking buffer (Thermo Scientific 37536) for 1 hour with gentle shaking. Then, various concentrations of anti-C1s antibodies diluted in SuperBlock T20 blocking buffer were added to the plate and incubated for 1 hour with gentle shaking. Mouse IgG (Sigma Aldrich I5381) and human IgG1 (BioXCell BP0297) were used as negative controls. The plate was washed before incubation for 1 hour with HRP-conjugated secondary antibodies (goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody HRP (Invitrogen G-21040) or goat anti-human IgG antibody, HRP conjugate (Millipore AP309P)) diluted in SuperBlock T20 blocking buffer with gentle shaking. Finally, the plate was washed and developed with ELISA liquid substrate (Sigma Aldrich), and the reaction was stopped by adding an equal volume of 1M H2SO4 to the ELISA liquid substrate. Binding antibodies were measured by absorbance at OD 450nm The top candidates 2-7 and 2-8 and their humanized variants hz2-7 (H1L2) and hz2-8 (H1L2G80A / T82A) each showed strong binding to human C1s( Figure 3S ) and cynomolgus monkey C1s( Figure 3V ), but no binding to mouse C1s( Figure 3T ), where EC 50 was between 4 ng / ml and 15 ng / ml. Anti-C1s 2-7 and its humanized variant hz2-7 (H1L2) did not show binding to rat C1s, while 2-8 and its humanized variant hz2-8 (H1L2 G80A / T82A) showed weaker binding to rat C1s( Figure 3U ), where EC 50 was between 63 ng / ml and 105 ng / ml.

[0128] Example 4. Binding of anti-C1S antibodies to full-length and truncated C1S measured by ELISA

[0129] Binding of anti-C1s antibodies to full-length human C1s protein, full-length mouse C1s protein, the NHC fragment of human C1s protein( Figure 5B for comparison) and the middle fragment of human C1s located within NHC (identified as "M151" or "huC1sM151"), which encompasses 151 amino acids at residues 272 to 422 of human C1s, upstream of the autocleavage site R422-I423( Figure 5BFor comparison). Constructs for expressing full-length human C1s (HuC1s or "wt human C1s") (SEQ ID NO: 99), full-length mouse C1s (moC1s) (SEQ ID NO: 101), human C1s deletion mutant 1 (huC1sM151) (SEQ ID NO: 107), and human C1s deletion mutant 2 (huC1sNHC) corresponding to the NHC of C1s (SEQ ID NO: 109) were developed, with a C-terminal 6xHis tag on each expressed protein. All expression and purification of full-length and truncated C1s constructs (SEQ ID NO: 100, 102, 108, 110) and the resulting proteins (SEQ ID NO: 99, 101, 107, 109) were carried out in-house.

[0130] ELISA was performed by first coating a 96-well plate with 200 ng / well of full-length or truncated C1s protein overnight. The next morning, with gentle shaking, the plate was blocked with SuperBlock T20 blocking buffer (Thermo Fisher Scientific 37536) for 1 hour. Then, various concentrations of anti-C1s antibodies diluted in SuperBlock T20 blocking buffer were added to the plate and incubated for 1 hour with gentle shaking. Mouse IgG (Sigma-Aldrich I5381) and human IgG1 (BioXCell BP0297) were used as negative controls. The plate was washed before incubating for 1 hour with an HRP-conjugated secondary antibody (goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody HRP (Invitrogen G-21040) or goat anti-human IgG antibody, HRP conjugate (Millipore AP309P)) diluted in SuperBlock T20 blocking buffer with gentle shaking. Finally, the plate was washed and developed with ELISA liquid HRP substrate (Sigma-Aldrich), and the reaction was stopped by adding 1 M H2SO4 in the same volume as the ELISA liquid substrate. The bound antibody was measured by absorbance at OD 450nm . The results are shown in Figure 4. Top candidates 2-7 and 2-8 and their humanized variants hz2-7 (H1L2) and hz2-8 (H1L2) each showed strong binding to full-length human C1s ( Figure 4A ), but no binding to full-length mouse C1s ( Figure 4B ). All anti-C1s antibodies showed higher EC 50 ( Figure 4C -D) for binding to the two truncated human C1s (NHC and M151) compared to full-length C1s, indicating that human C1s (M151) retains the major binding sites for anti-C1s and that the flanking regions can contribute to maintaining the correct (or preferred) conformation.

[0131] Example 5. Binding of anti-C1S antibody to full-length and truncated C1S demonstrated by Western blot

[0132] Western blot analysis was performed by first resolving 200 ng / lane of full-length or truncated C1s (human C1s, mouse C1s, NHC of human C1s, and M151 of human C1s) on SDS-PAGE with or without a reducing agent (Invitrogen B0004) as follows: in lane 1, 200 ng of full-length human C1s (HuC1s; SEQ ID NO:99); in lane 2, 200 ng of full-length mouse C1s (MoC1s; SEQ ID NO:101); in lane 3, 200 ng of truncated human C1s M151 (HuC1s(M151); SEQ ID NO:107); and in lane 4, 200 ng of truncated human C1s NHC (HuC1s(NHC); SEQ ID NO:109). The proteins were then transferred to a nitrocellulose membrane and blocked overnight with 5% blotto (ChemCruz sc-2325) diluted in TBST (Teknova T1688). The next morning, with gentle shaking, the blocked membrane was first incubated with 1 μg / ml of anti-C1s antibody diluted in 5% blotto-TBST for 1 hour. Then, with gentle shaking, the membrane was washed and incubated with an HRP-conjugated secondary antibody diluted in 5% blotto-TBST (goat anti-human IgG antibody, HRP conjugate (Millipore AP309P)) for 1 hour. Finally, the membrane was washed thoroughly and developed using SuperSignal West Dura Extended Duration Substrate (Thermo Fisher Scientific 34075) and imaged under Azure Biosystems. Representative results are shown in Figure 5A which presents the results for anti-C1s antibody 2-8,

[0133] Where the blot on the left is from a gel run under non-reducing conditions, and the blot on the right is from a gel run under reducing conditions. On the blot of the gel run without a reducing agent (left), lane 1 shows antibody 2-8 binding to two (2) major bands of full-length human C1s (HuC1s), with the top band corresponding to full-length HuC1s and the bottom band corresponding to the auto-cleaved heavy chain, lane 2 shows no detectable antibody 2-8 binding to full-length mouse C1s (MoC1s), lane 3 shows antibody 2-8 binding to multiple bands of truncated human C1s M151 (HuC1s(M151)), and lane 4 shows antibody 2-8 binding to several high molecular weight bands of truncated human C1s NHC (HuC1s(NHC)). In the blot of the gel run with a reducing agent (right), lane 1 shows low level binding of antibody 2-8 to the band corresponding to the autocleaved heavy chain of full-length human C1s (HuC1s), lane 2 shows no detectable antibody 2-8 to full-length mouse C1s (MoC1s), lane 3 shows low level binding of antibody 2-8 to a monomer of truncated human C1s M151 (HuC1s(M151)), and lane 4 shows low level binding of antibody 2-8 to a monomer of truncated human C1s NHC (HuC1s(NHC)).

[0134] The results using anti-C1s antibodies 2-7, hz2-7(H1L2) and hz2-8(H1L2) showed that Figure 5A The results for antibody 2-8 showed the same binding pattern as shown in (data not shown). These results show that the binding sites of anti-C1s antibody 2-7, anti-C1s antibody 2-8, anti-C1s antibody hz2-7 (H1L2) and anti-C1s antibody hz2-8 (H1L2) are located in the human C1sM151 region on NHC.

[0135] Example 6. Epitope mapping of anti-C1S antibodies using truncated C1S and C1S with point mutations

[0136] As indicated in Examples 4-5, the epitope of the anti-C1s antibodies disclosed herein is located within human C1s (M151) (SEQ ID NO: 107). Human C1s (M151) is the last 151 amino acids upstream of the auto-cleavage site (R422-I423) located within the human C1s NHC. To further define the epitope within human C1s (M151), an additional truncated C1s mutant was designed. This mutant was designated human C1s deletion mutant 3 (NHCΔ33) (SEQ ID NO: 111) and lacks the G390-R422 segment of the human C1s NHC ( Figure 5BThe last 33 amino acids corresponding to the (used for comparison). C1s(NHCΔ33) (SEQ ID NO:111) was cloned and expressed with a C-terminal 6xHis tag, and its expression and purification were carried out internally. Figure 5B Shows a schematic comparison and alignment of the full-length human C1s, HuC1s (SEQ ID NO:99), human C1s deletion mutant 1 (M151), HuC1s (M151) (SEQ ID NO:107), human deletion mutant 2 (NHC), HuC1s (NHC) (SEQ ID NO:109), and human C1s deletion mutant 3 (NHCΔ33), human C1s (NHCΔ33) (SEQ ID NO:111) used in these experiments.

[0137] As described in Example 4, the binding of various anti-C1s antibodies to human C1s (NHCΔ33) (SEQ ID NO:111) was carried out by ELISA, and the results are shown in Figure 6A . Hz2-7 (H1L2) showed little binding to human C1s (NHCΔ33), while hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A) showed strong binding to C1s (NHCΔ33), where the EC 50 values were 8 ng / ml and 15 ng / ml, respectively. As expected, human IgG1 showed no binding. This different binding profile of the humanized 2-7 variant to the humanized 2-8 variant strongly indicates that antibodies 2-7 and 2-8 bind to different epitopes within human C1s (M151), as demonstrated in Example 4. The results indicate that the epitope of hz2-7 (H1L2) is within the 33 amino acids upstream of the cleavage site (G390-R422), and the epitopes of hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A) are within the remaining part (Y272-A389) of M151.

[0138] To identify the epitopes of hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A), various point mutations were introduced into human C1s either individually or in double / triple combinations, and 20 human C1s mutants named M1-M20 (SEQ ID NO:112-128 and 141-143) were obtained, as shown in Table 2.

[0139] For all point mutations, the residue at a certain position of human C1s was replaced with the corresponding residue of murine C1s. Among the 22 mutated residues, 6 residues were identical between human C1s and rat C1s, 10 residues were identical between rat C1s and murine C1s, and the remaining 6 residues were unique residues at the corresponding positions of human C1s, rat C1s, and murine C1s. Human C1s M1-M20 (SEQ ID NO: 112-128 and 141-143) was cloned with a C-terminal 6xHis tag and expressed and purified internally.

[0140] Table 2: Point mutations of human C1s and the corresponding residues of rat and murine C1s

[0141]

[0142] As described in Example 4, the binding of anti-C1s antibodies to mutant M1-M20 of human C1s was measured by ELISA, and the results are shown in Figures 6B - 6U . All anti-C1s humanized variants, including hz2-7 (H1L2), hz2-8 (H1L2 G80A), and hz2-8 (H1L2G80A / T82A), maintained strong binding properties to most (16 out of 20) C1s mutants, including M1 ( Figure 6B ), M2 ( Figure 6C ), M3 ( Figure 6D ), M4 ( Figure 6E ), M7 ( Figure 6H ), M8 ( Figure 6I ), M10 ( Figure 6K ), M11 ( Figure 6L ), M12 ( Figure 6M ), M13 ( Figure 6N ), M15 ( Figure 6P ), M16 ( Figure 6Q ), M17 ( Figure 6R ), M18 ( Figure 6S ), M19 ( Figure 6T ), and M20 ( Figure 6U)。Isotype control human IgG1 did not show binding to any of the mutants. These results strongly suggest that 20 of the 22 point mutations (i.e., M277I, P278S, P280A, E282K, D283I, P285A, V288T, A292D, Q303K, A320S, N329D, K331Q, D343Y, S349P, E351A, S360N, R368H, N380H, G381E, and G382E) have no effect on the binding of hz2-7 (H1L2), hz2-8 (H1L2 G80A), and hz2-8 (H1L2 G80A / T82A) to C1s. Thus, these 20 residues are not critical binding sites for any of the anti-C1s antibodies examined.

[0143] As Figure 6A shown, the binding site of hz2-7 (H1L2) is within the last 33 amino acids (G390 - R422) of the C1s NHC. M1 - M20 of human C1s is not expected to affect the binding of hz2-7 (H1L2) as none of the point mutations are within this region. Surprisingly, M14 of human C1s (SEQ ID NO:125) containing only the single point mutation R316H completely abolished the binding of hz2-7 (H1L2), while having no effect on the binding of hz2-8 (H1L2 G80A) or hz2-8 (H1L2 G80A / T82A) ( Figure 6O ). Human IgG1 did not show binding to M14 as expected ( Figure 6O ). These results strongly suggest that in addition to G390 - R422, R316 is another key residue involved in the binding of hz2-7 (H1L2), further confirming the previous indication that the binding sites of hz2-7 (H1L2) are different from those of hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A).

[0144] In addition, as Figure 6J shown, mutant M9 (SEQ ID NO:120) encompassing the point mutations K336G, D343Y, and E351A completely abolished the binding of hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A), while hz2-7 (H1L2) retained binding with high potency. As previously demonstrated, D343Y and E351A are not important for the binding of any of the anti-C1s antibodies examined ( Figures 6H - 6I)。Therefore, K336 is (or may be) a key residue for the binding of hz2-8 (H1L2G80A) and hz2-8 (H1L2G80A / T82A). In addition to M9, the K336G point mutation also exists in M5 and M6 of human C1s, either alone (M5) or in combination with other non-essential residues (M6). As Figures 6F - 6G shown in

[0145] , the K336G single mutation results in a significant reduction in the binding of hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A), but has no effect on hz2-7 (H1L2), which confirms that K336 is a key residue for the binding of hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A) to C1s and constitutes a unique epitope of the 2-8 antibody.

[0146] Example 7. Inhibition of Complement-Mediated Lysis of Antibody-Sensitized Sheep Red Blood Cells by Anti-C1S Antibodies

[0147] Sheep red blood cells coated with rabbit anti-sheep red blood cell antiserum (hemolysin), also known as antibody-sensitized sheep red blood cells (EA), have traditionally been used to assay the activity of the classical pathway of complement in serum samples, where these assays allow the determination of the CH50 titer of serum. Using commercially available EA (Complement Technology, #B202), the inhibition of anti-C1s antibodies on complement-mediated lysis of EA was evaluated. Briefly, humanized anti-C1s variants including hz2-7 (H1L2), hz2-8 (H1L2), hz2-8 (H1L2G80A), and hz2-8 (H1L2 G80A / T82A) were serially diluted in GVB++ buffer (Complement Technology, catalog #B100, manufacturer's ingredient list: 0.1% gelatin, 5 mM barbital, 145 mM NaCl, 0.025% NaN3, 0.15 mM calcium chloride, 0.5 mM magnesium chloride, pH 7.3). Then, the diluted anti-C1s antibodies were mixed with normal human serum (NHS, Figure 7A ) or cynomolgus monkey serum (CMS, Figure 7B ) to achieve a final serum concentration of 1%. Before adding EA cells, the antibody and serum mixture was equilibrated at room temperature. For each reaction, approximately 0.5 - 1x10 8EA cells. To determine the 0% lysis background control value, EA cells were mixed only with buffer ("serum-free control"). To determine the 100% lysis control value, GVB++ buffer was replaced with water. EA cells were gently mixed with antibodies and serum, and the mixture was then incubated at 37 °C for 1 hour with periodic mixing. After incubation, the remaining EA cells were pelleted by spinning, and the absorbance of the supernatant was measured by measuring OD 540nm To calculate the percentage of lysis that occurred under specific incubation conditions, the value of the 0% lysis background was first subtracted from each absorbance reading, and the result was then normalized relative to the 100% lysis control (subtracting the 0% lysis background) to determine the percentage of lysis that occurred.

[0148] Results of human complement inhibition by anti-C1s antibodies are shown in Figure 7A . Antibody hz2-8 (H1L2) was the most effective inhibitor of complement-mediated EA lysis, with an IC 50 of 0.2 μg / ml. Antibodies hz2-8 (H1L2 G80A) and hz2-8 (H1L2 G80A / T82A), which are sequence-optimized variants of hz2-8 (H1L2), also effectively inhibited human complement-mediated EA lysis, with similar IC 50 values between 0.4 μg / ml and 0.5 μg / ml. hz2-7 (H1L2) was the weakest inhibitor in this case, with an IC 50 of 4.1 μg / ml, and maximum inhibition reached only about 50% at the highest concentration tested (4 μg / ml). The results of cynomolgus monkey complement inhibition shown in Figure 7B were similar to the human serum results, where hz2-8 (H1L2) was the most effective inhibitor of cynomolgus monkey complement-mediated EA lysis, with an IC 50 of 0.1 μg / ml, and its variants hz2-8 (H1L2 G80A) and hz2-8 (H1L2G80A / T82A) also maintained high inhibitory potency, with similar IC 50 values between 0.3 μg / ml and 0.4 μg / ml. Hz2-7 (H1L2) was more effective at inhibiting cynomolgus monkey complement than human complement, with an IC 50 of 0.8 μg / ml, and the degree of inhibition reached maximum ( Figure 7B ). These results strongly support the role of various anti-C1s antibodies in inhibiting the classical complement pathway, which is consistent with the findings from Example 2.

[0149] Although the present invention has been described with reference to certain specific embodiments thereof, those skilled in the art will understand that changes and modifications can be made to the specifically described embodiments without departing from the principles of the present invention, the principles of which are intended to be limited only by the scope of the appended claims. Sequence Listing <110> Mabwell Therapeutics, Inc. <120> Anti-C1S Antibody and Its Use <130> 1121-102PCT <140> <141> <150> 63 / 342,544 <151> 2022-05-16 <150> 63 / 221,131 <151> 2021-07-13 <160> 143 <170> PatentIn version 3.5 <210> 1 <211> 141 <212> PRT <213> Mus musculus <220> <223> Protein sequence of antibody 2-7 VH <400> 1 Met Tyr Leu Gly Leu Asn Cys Val Phe Ile Val Phe Leu Leu Lys Gly 1 5 10 15 Val Gln Ser Glu Val Lys Leu Glu Glu Ser Gly Gly Gly Leu Val Pro 20 25 30 Pro Gly Gly Ser Met Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe 35 40 45 Ser Asn Tyr Phe Met Asn Trp Val Arg Gln Ser Pro Glu Lys Gly Leu 50 55 60 Glu Trp Val Ala Glu Ile Arg Leu Lys Phe Thr Asn Tyr Ala Thr His 65 70 75 80 Tyr Ala Glu Ser Val Glu Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser 85 90 95 Lys Ser Ser Val Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr 100 105 110 Gly Ile Tyr Tyr Cys Thr Arg Asp Tyr Gly Ser Arg Asn Gly Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser 130 135 140 <210> 2 <211> 10 <212> PRT <213> Mus musculus <220> <223> 2-7 HC CDR-1 <400> 2 Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn 1 5 10 <210> 3 <211> 19 <212> PRT <213> Mus musculus <220> <223> 2-7 HC CDR-2 <400> 3 Glu Ile Arg Leu Lys Phe Thr Asn Tyr Ala Thr His Tyr Ala Glu Ser 1 5 10 15 Val Glu Gly <210> 4 <211> 11 <212> PRT <213> Mus musculus <220> <223> 2-7 HC CDR-3 <400> 4 Asp Tyr Gly Ser Arg Asn Gly Tyr Phe Asp Tyr 1 5 10 <210> 5 <211> 423 <212> DNA <213> Mus musculus <220> <223> Nucleotide sequence of antibody 2-7 VH <400> 5 atgtacttgg gactgaactg tgtattcata gtttttctct taaaaggtgt ccagagtgaa 60 gtgaagcttg aggagtctgg aggaggcttg gtgccacctg gaggatccat gaaactctcc 120 tgtgttgcct ctggattcac tttcagtaac tactttatga actgggtccg ccagtctcca 180 gagaaggggc ttgagtgggt tgctgaaatt agattgaaat ttactaatta tgcaacacat 240 tatgcggagt ctgtggaagg gaggttcacc atctcaagag atgattccaa aagtagtgtc 300 tacctccaaa tgaacaactt aagagctgaa gacactggca tttattactg taccagggac 360 tacggtagta ggaatgggta ctttgactac tggggccaag gcaccactct cacagtctcc 420 tca 423 <210> 6 <211> 132 <212> PRT <213> House mouse (Mus musculus) <220> <223> Protein sequence of antibody 2-7 VL <400> 6 Met Arg Cys Leu Ala Glu Phe Leu Gly Leu Leu Val Leu Trp Ile Pro 1 5 10 15 Gly Ala Ile Gly Asp Ile Val Met Thr Gln Ala Ala Pro Ser Val Pro 20 25 30 Val Thr Pro Gly Glu Ser Val Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr Trp Phe Leu Gln Arg 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe 85 90 95 Thr Leu Arg Val Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 115 120 125 Leu Glu Ile Lys 130 <210> 7 <211> 16 <212> PRT <213> Mus musculus <220> <223> 2-7 LC CDR-1 <400> 7 Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr 1 5 10 15 <210> 8 <211> 7 <212> PRT <213> Mus musculus <220> <223> 2-7 LC CDR-2 <400> 8 Arg Met Ser Asn Leu Ala Ser 1 5 <210> 9 <211> 9 <212> PRT <213> Mus musculus <220> <223> 2-7 LC CDR-3 <400> 9 Met Gln His Leu Glu Tyr Pro Tyr Thr 1 5 <210> 10 <211> 396 <212> DNA <213> Mus musculus <220> <223> Nucleotide sequence of antibody 2-7 VL <400> 10 atgaggtgcc tagctgagtt cctggggctg cttgtgctct ggatccctgg agccattggg 60 gatattgtga tgactcaggc tgcaccctct gtacctgtca ctcctggaga gtcagtatcc 120 atctcctgca ggtctagtaa gagtctcctg catagtaacg gcaacacttt cttgtattgg 180 ttcctacaga ggccaggcca gtctcctcag ctcctgatat atcggatgtc caaccttgcc 240 tcaggagtcc cagacaggtt cagtggcagt gggtcaggaa ctgctttcac actgagagtc 300 agtagagtgg aggctgagga tgtgggtatt tattactgta tgcaacatct agaatatccg 360 tacacgttcg gaggggggac caagctggaa ataaaa 396 <210> 11 <211> 135 <212> PRT <213> Mus musculus <220> <223> Protein sequence of antibody 2 - 8 VH <400> 11 Met Lys Cys Ser Trp Val Ile Phe Phe Leu Met Ala Val Val Thr Gly 1 5 10 15 Val Asn Ser Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg 20 25 30 Pro Gly Ala Leu Val Lys Leu Ser Cys Lys Ala Ser Gly Phe Asn Ile 35 40 45 Lys Glu Tyr Tyr Met His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu 50 55 60 Glu Trp Ile Gly Trp Ile Asp Pro Glu Asn Gly Asp Thr Ile Tyr Asp 65 70 75 80 Pro Lys Phe Gln Gly Lys Ala Ser Ile Thr Ala Asp Thr Ser Ser Asn 85 90 95 Thr Ala Tyr Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala Tyr Trp Gly Gln Gly 115 120 125 Thr Leu Val Thr Val Ser Ala 130 135 <210> 12 <211> 10 <212> PRT <213> Mus musculus <220> <223> 2-8 HC CDR-1 <400> 12 Gly Phe Asn Ile Lys Glu Tyr Tyr Met His 1 5 10 <210> 13 <211> 17 <212> PRT <213> Mus musculus <220> <223> 2-8 HC CDR-2 <400> 13 Trp Ile Asp Pro Glu Asn Gly Asp Thr Ile Tyr Asp Pro Lys Phe Gln 1 5 10 15 Gly <210> 14 <211> 7 <212> PRT <213> Mus musculus <220> <223> 2-8 HC CDR-3 <400> 14 Ser Arg Leu Phe Phe Ala Tyr 1 5 <210> 15 <211> 405 <212> DNA <213> Mus musculus <220> <223> Nucleotide sequence of antibody 2-8 VH <400> 15 atgaaatgca gctgggtcat cttcttcctg atggcagtgg ttacaggggt caattcagag 60 gttcagctgc agcagtctgg ggctgaactt gtgaggccag gggccttagt caagttgtcc 120 tgcaaagctt ctggcttcaa cattaaagag tattatatgc attgggtgaa gcagaggcct 180 gaacagggcc tggagtggat tggatggatt gatcctgaga atggtgatac tatatatgac 240 ccgaagttcc agggcaaggc cagtataaca gcagacacat cctccaacac agcctacctg 300 caactcagca gcctgacatc tgaggacact gccgtctatt actgtgctag atcgagacta 360 ttctttgctt actggggcca agggactctg gtcactgtct ctgca 405 <210> 16 <211> 127 <212> PRT <213> Mus musculus <220> <223> Protein sequence of antibody 2-8 VL <400> 16 Met Glu Ser Gln Ile Gln Val Leu Val Phe Val Phe Leu Trp Leu Ser 1 5 10 15 Gly Val Asp Gly Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser 20 25 30 Thr Ser Val Gly Asp Arg Val Ser Phe Thr Cys Lys Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Val Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp 65 70 75 80 Arg Phe Thr Gly Thr Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser 85 90 95 Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 115 120 125 <210> 17 <211> 11 <212> PRT <213> Mus musculus <220> <223> 2-8 LC CDR-1 <400> 17 Lys Ala Ser Gln Asp Val Arg Thr Ala Val Asp 1 5 10 <210> 18 <211> 7 <212> PRT <213> Mus musculus <220> <223> 2-8 LC CDR-2 <400> 18 Ser Ala Ser Tyr Arg Tyr Thr 1 5 <210> 19 <211> 9 <212> PRT <213> Mus musculus <220> <223> 2-8 LC CDR-3 <400> 19 Gln Gln Gln Tyr Thr Thr Pro Tyr Thr 1 5 <210> 20 <211> 381 <212> DNA <213> Mus musculus <220> <223> Nucleotide sequence of antibody 2-8 VL <400> 20 atggagtcac agattcaggt ccttgtattc gtgtttctct ggttgtctgg tgttgacgga 60 gacattgtaa tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 120 ttcacctgca aggccagtca ggatgtgcgc actgctgtag actggtatca acagaaacca 180 ggacaatctc ctaaactact gatttactcg gcatcctacc ggtacactgg agtccctgat 240 cgcttcactg gcactggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 300 gaagacctgg cagtttactt ctgtcagcaa caatatacta ctccgtacac gttcggaggg 360 gggaccaagc tggaaataaa a 381 <210> 21 <211> 146 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of antibody hz2-7 (H1L2) VH <400> 21 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Leu Glu Ser Gly 20 25 30 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 35 40 45 Ser Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn Trp Val Arg Gln Ala 50 55 60 Pro Gly Lys Gly Leu Glu Trp Val Ser Glu Ile Arg Leu Lys Phe Thr 65 70 75 80 Asn Tyr Ala Thr His Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile 85 90 95 Ser Arg Asp Asp Ser Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu 100 105 110 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Asp Tyr Gly Ser 115 120 125 Arg Asn Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 130 135 140 Ser Ser 145 <210> 22 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2) HC CDR-1 <400> 22 Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn 1 5 10 <210> 23 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic peptide <220> <223> hz2-7(H1L2) HC CDR-2 <400> 23 Glu Ile Arg Leu Lys Phe Thr Asn Tyr Ala Thr His Tyr Ala Asp Ser 1 5 10 15 Val Lys Gly <210> 24 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic peptide <220> <223> hz2-7(H1L2) HC CDR-3 <400> 24 Asp Tyr Gly Ser Arg Asn Gly Tyr Phe Asp Tyr 1 5 10 <210> 25 <211> 438 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-7 (H1L2) VH <400> 25 atggacccca agggcagcct gagctggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgagctacg gcgaggttca gctgctcgag agcggaggtg gtctggtgca gcctggcggt 120 agcctgaggc tgagctgcgc cgcaagcggc tttaccttca gcaactactt tatgaattgg 180 gtgaggcagg caccgggcaa gggcctggag tgggtaagcg agatcaggct gaagttcacc 240 aattacgcta cccattacgc cgacagcgtg aagggccgct tcaccatcag cagggacgac 300 tctaagagca ccctgtacct gcagatgaac tcactgaggg ccgaggacac cgccgtgtac 360 tactgcacta gggactacgg cagcaggaac ggctacttcg actactgggg ccagggcact 420 ctggtcaccg tgagcagc 438 <210> 26 <211> 132 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of antibody hz2-7 (H1L2) VL <400> 26 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro 20 25 30 Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr Trp Phe Leu Gln Lys 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95 Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 115 120 125 Leu Glu Ile Lys 130 <210> 27 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2) LC CDR-1 <400> 27 Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr 1 5 10 15 <210> 28 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2) LC CDR-2 <400> 28 Arg Met Ser Asn Leu Ala Ser 1 5 <210> 29 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2) LC CDR-3 <400> 29 Met Gln His Leu Glu Tyr Pro Tyr Thr 1 5 <210> 30 <211> 396 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-7 (H1L2) VL <400> 30 atggagaccg acaccctgct gctctgggtg ctgctgctct gggtgcccgg ctccaccgga 60 gacatcgtga tgacccagac acccctgagc ctgcccgtga cccccggtga gcccgccagc 120 atcagctgca ggtcaagcaa gagcctcctg cacagcaacg gcaacacctt tctgtactgg 180 ttcctgcaga agccgggaca gagcccccag ctgctgatct acaggatgag caacctggcc 240 tctggcgtgc ccgacaggtt cagcggcagt ggcagcggaa ccgacttcac cctgaagatt 300 agcagggtgg aggccgagga cgtgggcgtg tactactgta tgcagcacct ggagtacccc 360 tacaccttcg gcggagggac caaactggaa atcaag 396 <210> 31 <211> 146 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of antibody hz2-7 (H1L2 G131A) VH <400> 31 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Leu Glu Ser Gly 20 25 30 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 35 40 45 Ser Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn Trp Val Arg Gln Ala 50 55 60 Pro Gly Lys Gly Leu Glu Trp Val Ser Glu Ile Arg Leu Lys Phe Thr 65 70 75 80 Asn Tyr Ala Thr His Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile 85 90 95 Ser Arg Asp Asp Ser Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu 100 105 110 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Asp Tyr Gly Ser 115 120 125 Arg Asn Ala Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 130 135 140 Ser Ser 145 <210> 32 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2 G131A) HC CDR-1 <400> 32 Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn 1 5 10 <210> 33 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2 G131A) HC CDR-2 <400> 33 Glu Ile Arg Leu Lys Phe Thr Asn Tyr Ala Thr His Tyr Ala Asp Ser 1 5 10 15 Val Lys Gly <210> 34 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2 G131A) HC CDR-3 <400> 34 Asp Tyr Gly Ser Arg Asn Ala Tyr Phe Asp Tyr 1 5 10 <210> 35 <211> 438 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-7 (H1L2 G131A) VH <400> 35 atggacccca agggcagcct gagctggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgagctacg gcgaggttca gctgctcgag agcggaggtg gtctggtgca gcctggcggt 120 agcctgaggc tgagctgcgc cgcaagcggc tttaccttca gcaactactt tatgaattgg 180 gtgaggcagg caccgggcaa gggcctggag tgggtaagcg agatcaggct gaagttcacc 240 aattacgcta cccattacgc cgacagcgtg aagggccgct tcaccatcag cagggacgac 300 tctaagagca ccctgtacct gcagatgaac tcactgaggg ccgaggacac cgccgtgtac 360 tactgcacta gggactacgg cagcaggaac gcctacttcg actactgggg ccagggcact 420 ctggtcaccg tgagcagc 438 <210> 36 <211> 132 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of antibody hz2-7 (H1L2 G131A) VL <400> 36 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro 20 25 30 Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr Trp Phe Leu Gln Lys 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95 Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 115 120 125 Leu Glu Ile Lys 130 <210> 37 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2 G131A) LC CDR-1 <400> 37 Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr 1 5 10 15 <210> 38 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2 G131A) LC CDR-2 <400> 38 Arg Met Ser Asn Leu Ala Ser 1 5 <210> 39 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-7(H1L2 G131A) LC CDR-3 <400> 39 Met Gln His Leu Glu Tyr Pro Tyr Thr 1 5 <210> 40 <211> 396 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-7 (H1L2 G131A) VL <400> 40 atggagaccg acaccctgct gctctgggtg ctgctgctct gggtgcccgg ctccaccgga 60 gacatcgtga tgacccagac acccctgagc ctgcccgtga cccccggtga gcccgccagc 120 atcagctgca ggtcaagcaa gagcctcctg cacagcaacg gcaacacctt tctgtactgg 180 ttcctgcaga agccgggaca gagcccccag ctgctgatct acaggatgag caacctggcc 240 tctggcgtgc ccgacaggtt cagcggcagt ggcagcggaa ccgacttcac cctgaagatt 300 agcagggtgg aggccgagga cgtgggcgtg tactactgta tgcagcacct ggagtacccc 360 tacaccttcg gcggagggac caaactggaa atcaag 396 <210> 41 <211> 140 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of antibody hz2-8 (H1L2) VH <400> 41 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Gln Val Gln Leu Val Gln Ser Gly 20 25 30 Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 35 40 45 Ser Gly Phe Asn Ile Lys Glu Tyr Tyr Met His Trp Val Arg Gln Ala 50 55 60 Pro Glu Gln Gly Leu Glu Trp Met Gly Trp Ile Asp Pro Glu Asn Gly 65 70 75 80 Asp Thr Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala 85 90 95 Asp Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser 100 105 110 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 130 135 140 <210> 42 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8(H1L2) HC CDR-1 <400> 42 Gly Phe Asn Ile Lys Glu Tyr Tyr Met His 1 5 10 <210> 43 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8(H1L2) HC CDR-2 <400> 43 Trp Ile Asp Pro Glu Asn Gly Asp Thr Ile Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 44 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8(H1L2) HC CDR-3 <400> 44 Ser Arg Leu Phe Phe Ala Tyr 1 5 <210> 45 <211> 420 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-8 (H1L2) VH <400> 45 atggacccca agggcagcct gagctggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgagctacg gccaggtgca gctggtgcag agtggggccg aggtgaagaa gccgggagcc 120 agcgtgaagg tgagctgcaa ggccagcggc ttcaacatca aggagtacta catgcactgg 180 gtgaggcaag cgccggaaca gggcctggag tggatgggct ggatcgaccc cgagaacggc 240 gataccatct acgcccagaa gttccagggc agggtgacca tcaccgctga caccagcacc 300 aacaccgcct acatggagct gagcagcctg agaagcgagg acacagccgt gtactactgc 360 gccaggtcac gcttgttctt cgcctactgg ggccaaggca ccctggtgac cgtgagctct 420 <210> 46 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of antibody hz2-8 (H1L2) VL <400> 46 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Lys Ser Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 115 120 125 <210> 47 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8(H1L2) LC CDR-1 <400> 47 Arg Ala Ser Gln Asp Val Arg Thr Ala Leu Asp 1 5 10 <210> 48 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8(H1L2) LC CDR-2 <400> 48 Ser Ala Ser Tyr Arg Tyr Ser 1 5 <210> 49 <211> 9 <212> PRT <213> Synthetic sequence <220> <223> Description of synthetic sequence: synthetic peptide <220> <223> hz2-8(H1L2) LC CDR-3 <400> 49 Gln Gln Gln Tyr Thr Thr Pro Tyr Thr 1 5 <210> 50 <211> 381 <212> DNA <213> Synthetic sequence <220> <223> Description of synthetic sequence: synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-8 (H1L2) VL <400> 50 atggagaccg acaccctgct gctctgggtg ctgctgctct gggtgcccgg ctccaccgga 60 gacatccaga tgacccaaag cccaaagagc ctgagcgcca gcgtgggcga cagggtgacg 120 atcacctgca gagccagcca ggacgtgagg accgccctgg attggtatca gcagaagccc 180 ggtcagagcc ccaagctcct gatctacagc gcaagctaca ggtacagcgg cgtgcccagc 240 aggttcagcg gctccggcag cggcaccgac ttcactctta ccatcagctc actgcagccc 300 gaggacttcg ccacctacta ctgccagcag cagtacacca ccccctacac cttcggccag 360 ggcaccaagt tggagatcaa g 381 <210> 51 <211> 140 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of antibody hz2-8 (H1L2 G80A / T82A) <400> 51 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Gln Val Gln Leu Val Gln Ser Gly 20 25 30 Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 35 40 45 Ser Gly Phe Asn Ile Lys Glu Tyr Tyr Met His Trp Val Arg Gln Ala 50 55 60 Pro Glu Gln Gly Leu Glu Trp Met Gly Trp Ile Asp Pro Glu Asn Ala 65 70 75 80 Asp Ala Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala 85 90 95 Asp Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser 100 105 110 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 130 135 140 <210> 52 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A / T82A) HC CDR-1 <400> 52 Gly Phe Asn Ile Lys Glu Tyr Tyr Met His 1 5 10 <210> 53 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A / T82A) HC CDR-2 <400> 53 Trp Ile Asp Pro Glu Asn Ala Asp Ala Ile Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 54 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A / T82A) HC CDR-3 <400> 54 Ser Arg Leu Phe Phe Ala Tyr 1 5 <210> 55 <211> 420 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-8 (H1L2 G80A / T82A) VH <400> 55 atggacccta agggctctct gtcttggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgtcttatg gccaggtgca gctggtgcag tctggcgccg aagtgaaaaa gcctggcgcc 120 tctgtgaagg tgtcctgcaa ggcctctggc ttcaacatca aagaatatta catgcactgg 180 gtccgacagg cccctgagca aggattggaa tggatgggct ggatcgaccc cgagaacgcc 240 gatgccatct acgcccagaa attccagggc agagtgacca tcaccgccga cacctctacc 300 aacaccgcct acatggaact gtccagcctg agatctgagg acaccgccgt gtactactgc 360 gcccggtcca gactgttctt cgcctattgg ggccagggca ccctggtcac agtttcttcc 420 <210> 56 <211> 127 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of antibody hz2-8 (H1L2 G80A / T82A) VL <400> 56 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Lys Ser Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 115 120 125 <210> 57 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A / T82A) LC CDR-1 <400> 57 Arg Ala Ser Gln Asp Val Arg Thr Ala Leu Asp 1 5 10 <210> 58 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A / T82A) LC CDR-2 <400> 58 Ser Ala Ser Tyr Arg Tyr Ser 1 5 <210> 59 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A / T82A) LC CDR-3 <400> 59 Gln Gln Gln Tyr Thr Thr Pro Tyr Thr 1 5 <210> 60 <211> 381 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Nucleotide sequence of the VL of antibody hz2-8 (H1L2 G80A / T82A) <400> 60 atggaaaccg acacactgct gctgtgggtg ctgctcttgt gggtgccagg atctaccggc 60 gacatccaga tgacccagtc tccaaagtct ctgtccgcct ccgtgggcga cagagtgacc 120 atcacctgta gagcctctca ggacgtgcgg accgctctgg attggtatca gcagaagcct 180 ggccagtctc ctaagctgct gatctactcc gcctcctacc ggtactctgg cgtgccctcc 240 agattctccg gctctggctc tggcaccgac tttaccctga caatctccag cctgcagcct 300 gaggacttcg ccacctacta ctgccagcaa cagtacacca caccttacac cttcggccag 360 ggcaccaagc tggaaatcaa g 381 <210> 61 <211> 140 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of the VH of antibody hz2-8 (H1L2 G80A) <400> 61 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Gln Val Gln Leu Val Gln Ser Gly 20 25 30 Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 35 40 45 Ser Gly Phe Asn Ile Lys Glu Tyr Tyr Met His Trp Val Arg Gln Ala 50 55 60 Pro Glu Gln Gly Leu Glu Trp Met Gly Trp Ile Asp Pro Glu Asn Ala 65 70 75 80 Asp Thr Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala 85 90 95 Asp Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser 100 105 110 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 130 135 140 <210> 62 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A) HC CDR-1 <400> 62 Gly Phe Asn Ile Lys Glu Tyr Tyr Met His 1 5 10 <210> 63 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A) HC CDR-2 <400> 63 Trp Ile Asp Pro Glu Asn Ala Asp Thr Ile Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 64 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A) HC CDR-3 <400> 64 Ser Arg Leu Phe Phe Ala Tyr 1 5 <210> 65 <211> 420 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-8 (H1L2 G80A) VH <400> 65 atggacccta agggctctct gtcttggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgtcttatg gccaggtgca gctggtgcag tctggcgccg aagtgaaaaa gcctggcgcc 120 tctgtgaagg tgtcctgcaa ggcctctggc ttcaacatca aagaatatta catgcactgg 180 gtccgacagg cccctgagca aggattggaa tggatgggct ggatcgaccc cgagaacgcc 240 gataccatct acgcccagaa attccagggc agagtgacca tcaccgccga cacctctacc 300 aacaccgcct acatggaact gtccagcctg agatctgagg acaccgccgt gtactactgc 360 gcccggtcca gactgttctt cgcctattgg ggccagggca ccctggtcac agtttcttcc 420 <210> 66 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of antibody hz2-8 (H1L2 G80A) VL <400> 66 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Lys Ser Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 115 120 125 <210> 67 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A) LC CDR-1 <400> 67 Arg Ala Ser Gln Asp Val Arg Thr Ala Leu Asp 1 5 10 <210> 68 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic peptide <220> <223> hz2-8 (H1L2 G80A) LC CDR-2 <400> 68 Ser Ala Ser Tyr Arg Tyr Ser 1 5 <210> 69 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic peptide <220> <223> hz2-8 (H1L2 G80A) LC CDR-3 <400> 69 Gln Gln Gln Tyr Thr Thr Pro Tyr Thr 1 5 <210> 70 <211> 381 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: synthetic polynucleotide <220> <223> Nucleotide sequence of antibody hz2-8 (H1L2 G80A) VL <400> 70 atggaaaccg acacactgct gctgtgggtg ctgctcttgt gggtgccagg atctaccggc 60 gacatccaga tgacccagtc tccaaagtct ctgtccgcct ccgtgggcga cagagtgacc 120 atcacctgta gagcctctca ggacgtgcgg accgctctgg attggtatca gcagaagcct 180 ggccagtctc ctaagctgct gatctactcc gcctcctacc ggtactctgg cgtgccctcc 240 agattctccg gctctggctc tggcaccgac tttaccctga caatctccag cctgcagcct 300 gaggacttcg ccacctacta ctgccagcaa cagtacacca caccttacac cttcggccag 360 ggcaccaagc tggaaatcaa g 381 <210> 71 <211> 471 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Full-length antibody 2-7 HC protein sequence <400> 71 Met Tyr Leu Gly Leu Asn Cys Val Phe Ile Val Phe Leu Leu Lys Gly 1 5 10 15 Val Gln Ser Glu Val Lys Leu Glu Glu Ser Gly Gly Gly Leu Val Pro 20 25 30 Pro Gly Gly Ser Met Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe 35 40 45 Ser Asn Tyr Phe Met Asn Trp Val Arg Gln Ser Pro Glu Lys Gly Leu 50 55 60 Glu Trp Val Ala Glu Ile Arg Leu Lys Phe Thr Asn Tyr Ala Thr His 65 70 75 80 Tyr Ala Glu Ser Val Glu Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser 85 90 95 Lys Ser Ser Val Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr 100 105 110 Gly Ile Tyr Tyr Cys Thr Arg Asp Tyr Gly Ser Arg Asn Gly Tyr Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 72 <211> 1413 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody 2-7 HC nucleotide sequence <400> 72 atgtacttgg gactgaactg tgtattcata gtttttctct taaaaggtgt ccagagtgaa 60 gtgaagcttg aggagtctgg aggaggcttg gtgccacctg gaggatccat gaaactctcc 120 tgtgttgcct ctggattcac tttcagtaac tactttatga actgggtccg ccagtctcca 180 gagaaggggc ttgagtgggt tgctgaaatt agattgaaat ttactaatta tgcaacacat 240 tatgcggagt ctgtggaagg gaggttcacc atctcaagag atgattccaa aagtagtgtc 300 tacctccaaa tgaacaactt aagagctgaa gacactggca tttattactg taccagggac 360 tacggtagta ggaatgggta ctttgactac tggggccaag gcaccactct cacagtctcc 420 tcagccagca ccaagggccc ttccgtgttt ccactggccc cctcctctaa atccacatct 480 ggcggcaccg ccgccctggg ctgtctggtg aaggactact tcccagagcc tgtgacagtg 540 tcctggaact ctggcgccct gacatccggc gtgcacacat ttccagccgt gctgcagagc 600 tccggcctgt acagcctgtc tagcgtggtg acagtgccct cctctagcct gggcacacag 660 acctatatct gcaacgtgaa tcacaagcca agcaatacca aggtggacaa gaaggtggag 720 cccaagtcct gtgataagac acacacctgc cccccttgtc ctgctcccga gctgctgggc 780 ggccctagcg tgttcctgtt tccacccaag cctaaggaca ccctgatgat ctcccggaca 840 cccgaggtga cctgcgtggt ggtggacgtg tctcacgagg atcctgaggt gaagttcaac 900 tggtatgtgg atggcgtgga ggtgcacaat gccaagacca agcccagaga ggagcagtac 960 aactctacat atagggtggt gagcgtgctg accgtgctgc accaggactg gctgaacggc 1020 aaggagtata agtgcaaggt gtccaataag gccctgcccg cccccatcga gaagacaatc 1080 agcaaggcca agggccagcc tcgggagcca caggtgtaca ccctgcctcc atccagagac 1140 gagctgacaa agaaccaggt gtctctgaca tgtctggtga agggcttcta tcctagcgat 1200 atcgccgtgg agtgggagtc caatggccag ccagagaaca attacaagac cacaccccct 1260 gtgctggact ccgatggctc cttctttctg tattccaagc tgaccgtgga taagtctcgg 1320 tggcagcagg gcaacgtgtt cagctgttcc gtgatgcacg aagccctgca taatcactat 1380 actcagaaat ccctgtccct gtcacctgga aag 1413 <210> 73 <211> 238 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody 2-7 LC protein sequence <400> 73 Met Arg Cys Leu Ala Glu Phe Leu Gly Leu Leu Val Leu Trp Ile Pro 1 5 10 15 Gly Ala Ile Gly Asp Ile Val Met Thr Gln Ala Ala Pro Ser Val Pro 20 25 30 Val Thr Pro Gly Glu Ser Val Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr Trp Phe Leu Gln Arg 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe 85 90 95 Thr Leu Arg Val Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 115 120 125 Leu Glu Ile Lys Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro 130 135 140 Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu 145 150 155 160 Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn 165 170 175 Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser 180 185 190 Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala 195 200 205 Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly 210 215 220 Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 74 <211> 714 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody 2-7 LC nucleotide sequence <400> 74 atgaggtgcc tagctgagtt cctggggctg cttgtgctct ggatccctgg agccattggg 60 gatattgtga tgactcaggc tgcaccctct gtacctgtca ctcctggaga gtcagtatcc 120 atctcctgca ggtctagtaa gagtctcctg catagtaacg gcaacacttt cttgtattgg 180 ttcctacaga ggccaggcca gtctcctcag ctcctgatat atcggatgtc caaccttgcc 240 tcaggagtcc cagacaggtt cagtggcagt gggtcaggaa ctgctttcac actgagagtc 300 agtagagtgg aggctgagga tgtgggtatt tattactgta tgcaacatct agaatatccg 360 tacacgttcg gaggggggac caagctggaa ataaaaacag tggccgcccc aagcgtgttc 420 atctttcccc cttccgacga gcagctgaag tctggcaccg ccagcgtggt gtgcctgctg 480 aacaacttct accctcggga ggccaaggtc cagtggaagg tggataacgc cctgcagtct 540 ggcaatagcc aggagtccgt gaccgagcag gactctaagg atagcacata ttccctgtct 600 agcaccctga cactgagcaa ggccgattac gagaagcaca aggtgtatgc ctgtgaagtc 660 acccatcagg ggctgtcatc acccgtcact aagtcattca atcgcggaga atgc 714 <210> 75 <211> 465 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody 2-8 HC protein sequence <400> 75 Met Lys Cys Ser Trp Val Ile Phe Phe Leu Met Ala Val Val Thr Gly 1 5 10 15 Val Asn Ser Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg 20 25 30 Pro Gly Ala Leu Val Lys Leu Ser Cys Lys Ala Ser Gly Phe Asn Ile 35 40 45 Lys Glu Tyr Tyr Met His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu 50 55 60 Glu Trp Ile Gly Trp Ile Asp Pro Glu Asn Gly Asp Thr Ile Tyr Asp 65 70 75 80 Pro Lys Phe Gln Gly Lys Ala Ser Ile Thr Ala Asp Thr Ser Ser Asn 85 90 95 Thr Ala Tyr Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala Tyr Trp Gly Gln Gly 115 120 125 Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 130 135 140 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 145 150 155 160 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 165 170 175 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 180 185 190 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 195 200 205 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 210 215 220 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 225 230 235 240 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 245 250 255 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 260 265 270 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 275 280 285 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 290 295 300 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 305 310 315 320 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 325 330 335 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 340 345 350 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 355 360 365 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 370 375 380 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 385 390 395 400 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 405 410 415 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 420 425 430 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 435 440 445 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 450 455 460 Lys 465 <210> 76 <211> 1395 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody 2-8 HC nucleotide sequence <400> 76 atgaaatgca gctgggtcat cttcttcctg atggcagtgg ttacaggggt caattcagag 60 gttcagctgc agcagtctgg ggctgaactt gtgaggccag gggccttagt caagttgtcc 120 tgcaaagctt ctggcttcaa cattaaagag tattatatgc attgggtgaa gcagaggcct 180 gaacagggcc tggagtggat tggatggatt gatcctgaga atggtgatac tatatatgac 240 gaacagggcc tggagtggat tggatggatt gatcctgaga atggtgatac tatatatgac 240 ccgaagttcc agggcaaggc cagtataaca gcagacacat cctccaacac agcctacctg 300 ccgaagttcc agggcaaggc cagtataaca gcagacacat cctccaacac agcctacctg 300 caactcagca gcctgacatc tgaggacact gccgtctatt actgtgctag atcgagacta 360 caactcagca gcctgacatc tgaggacact gccgtctatt actgtgctag atcgagacta 360 ttctttgctt actggggcca agggactctg gtcactgtct ctgcagccag caccaagggc 420 ttctttgctt actggggcca agggactctg gtcactgtct ctgcagccag caccaagggc 420 ccttccgtgt ttccactggc cccctcctct aaatccacat ctggcggcac cgccgccctg 480 ccttccgtgt ttccactggc cccctcctct aaatccacat ctggcggcac cgccgccctg 480 ggctgtctgg tgaaggacta cttcccagag cctgtgacag tgtcctggaa ctctggcgcc 540 ggctgtctgg tgaaggacta cttcccagag cctgtgacag tgtcctggaa ctctggcgcc 540 ctgacatccg gcgtgcacac atttccagcc gtgctgcaga gctccggcct gtacagcctg 600 ctgacatccg gcgtgcacac atttccagcc gtgctgcaga gctccggcct gtacagcctg 600 tctagcgtgg tgacagtgcc ctcctctagc ctgggcacac agacctatat ctgcaacgtg 660 tctagcgtgg tgacagtgcc ctcctctagc ctgggcacac agacctatat ctgcaacgtg 660 aatcacaagc caagcaatac caaggtggac aagaaggtgg agcccaagtc ctgtgataag 720 aatcacaagc caagcaatac caaggtggac aagaaggtgg agcccaagtc ctgtgataag 720 acacacacct gccccccttg tcctgctccc gagctgctgg gcggccctag cgtgttcctg 780 acacacacct gccccccttg tcctgctccc gagctgctgg gcggccctag cgtgttcctg 780 tttccaccca agcctaagga caccctgatg atctcccgga cacccgaggt gacctgcgtg 840 tttccaccca agcctaagga caccctgatg atctcccgga cacccgaggt gacctgcgtg 840 gtggtggacg tgtctcacga ggatcctgag gtgaagttca actggtatgt ggatggcgtg 900 gtggtggacg tgtctcacga ggatcctgag gtgaagttca actggtatgt ggatggcgtg 900 gaggtgcaca atgccaagac caagcccaga gaggagcagt acaactctac atatagggtg 960 gtgagcgtgc tgaccgtgct gcaccaggac tggctgaacg gcaaggagta taagtgcaag 1020 gtgtccaata aggccctgcc cgcccccatc gagaagacaa tcagcaaggc caagggccag 1080 cctcgggagc cacaggtgta caccctgcct ccatccagag acgagctgac aaagaaccag 1140 gtgtctctga catgtctggt gaagggcttc tatcctagcg atatcgccgt ggagtgggag 1200 tccaatggcc agccagagaa caattacaag accacacccc ctgtgctgga ctccgatggc 1260 tccttctttc tgtattccaa gctgaccgtg gataagtctc ggtggcagca gggcaacgtg 1320 ttcagctgtt ccgtgatgca cgaagccctg cataatcact atactcagaa atccctgtcc 1380 ctgtcacctg gaaag 1395 <210> 77 <211> 233 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody 2-8 LC protein sequence <400> 77 Met Glu Ser Gln Ile Gln Val Leu Val Phe Val Phe Leu Trp Leu Ser 1 5 10 15 Gly Val Asp Gly Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser 20 25 30 Thr Ser Val Gly Asp Arg Val Ser Phe Thr Cys Lys Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Val Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp 65 70 75 80 Arg Phe Thr Gly Thr Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser 85 90 95 Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Thr 115 120 125 Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu 130 135 140 Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro 145 150 155 160 Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly 165 170 175 Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr 180 185 190 Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His 195 200 205 Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val 210 215 220 Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 78 <211> 699 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody 2-8 LC nucleotide sequence <400> 78 atggagtcac agattcaggt ccttgtattc gtgtttctct ggttgtctgg tgttgacgga 60 gacattgtaa tgacccagtc tcacaaattc atgtccacat cagtaggaga cagggtcagc 120 ttcacctgca aggccagtca ggatgtgcgc actgctgtag actggtatca acagaaacca 180 ggacaatctc ctaaactact gatttactcg gcatcctacc ggtacactgg agtccctgat 240 cgcttcactg gcactggatc tgggacggat ttcactttca ccatcagcag tgtgcaggct 300 gaagacctgg cagtttactt ctgtcagcaa caatatacta ctccgtacac gttcggaggg 360 gggaccaagc tggaaataaa aacagtggcc gccccaagcg tgttcatctt tcccccttcc 420 gacgagcagc tgaagtctgg caccgccagc gtggtgtgcc tgctgaacaa cttctaccct 480 cgggaggcca aggtccagtg gaaggtggat aacgccctgc agtctggcaa tagccaggag 540 tccgtgaccg agcaggactc taaggatagc acatattccc tgtctagcac cctgacactg 600 agcaaggccg attacgagaa gcacaaggtg tatgcctgtg aagtcaccca tcaggggctg 660 tcatcacccg tcactaagtc attcaatcgc ggagaatgc 699 <210> 79 <211> 475 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-7 (H1L2) HC protein sequence <400> 79 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Leu Glu Ser Gly 20 25 30 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 35 40 45 Ser Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn Trp Val Arg Gln Ala 50 55 60 Pro Gly Lys Gly Leu Glu Trp Val Ser Glu Ile Arg Leu Lys Phe Thr 65 70 75 80 Asn Tyr Ala Thr His Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile 85 90 95 Ser Arg Asp Asp Ser Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu 100 105 110 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Asp Tyr Gly Ser 115 120 125 Arg Asn Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 130 135 140 Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser 145 150 155 160 Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys 165 170 175 Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu 180 185 190 Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu 195 200 205 Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr 210 215 220 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val 225 230 235 240 Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro 245 250 255 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 260 265 270 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 275 280 285 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 290 295 300 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 305 310 315 320 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 325 330 335 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 340 345 350 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 355 360 365 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 370 375 380 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 385 390 395 400 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 405 410 415 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 420 425 430 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 435 440 445 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 450 455 460 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 465 470 475 <210> 80 <211> 1428 <212> DNA <213> Synthetic sequence <220> <223> Description of synthetic sequence: synthetic polynucleotide <220> <223> Full-length antibody hz2-7 (H1L2) HC nucleotide sequence <400> 80 atggacccca agggcagcct gagctggaga atcctgctgt tcctgagcct ggccttcgag 60 Met-Asp-Pro-Lys-Gly-Ser-Pro-Glu-Leu-Glu-Asn-Pro-Cys-Val-Pro-Glu-Pro-Ala-Phe-Glu 60 ctgagctacg gcgaggttca gctgctcgag agcggaggtg gtctggtgca gcctggcggt 120 Leu-Ser-Tyr-Gly-Glu-Val-Gln-Leu-Leu-Glu-Ser-Gly-Gly-Val-Leu-Val-Gln-Leu-Ala-Gly 120 agcctgaggc tgagctgcgc cgcaagcggc tttaccttca gcaactactt tatgaattgg 180 Ser-Leu-Glu-Ala-Glu-Leu-Arg-Arg-Lys-Arg-Phe-Thr-Phe-Gln-Thr-Tyr-Phe-Met-Asn-Trp 180 gtgaggcagg caccgggcaa gggcctggag tgggtaagcg agatcaggct gaagttcacc 240 Val-Glu-Gln-Ala-Thr-Gly-Lys-Gly-Leu-Glu-Trp-Gly-Lys-Ala-Glu-Ile-Arg-Ala-Glu-Phe-Thr 240 aattacgcta cccattacgc cgacagcgtg aagggccgct tcaccatcag cagggacgac 300 Asn-Tyr-Ala-Thr-His-Tyr-Ala-Asp-Ser-Val-Lys-Gly-Pro-Leu-Thr-Ile-Ser-Arg-Asp-Asp 300 tctaagagca ccctgtacct gcagatgaac tcactgaggg ccgaggacac cgccgtgtac 360 Ser-Lys-Ser-Thr-Leu-Tyr-Leu-Gln-Met-Asn-Leu-Thr-Glu-Gly-Pro-Glu-Asp-Thr-Ala-Val-Tyr 360 tactgcacta gggactacgg cagcaggaac ggctacttcg actactgggg ccagggcact 420 Tyr-Cys-Thr-Arg-Asp-Tyr-Gly-Gln-Gln-Glu-Arg-Leu-Phe-Asp-Tyr-Trp-Gly-Pro-Gly-Thr 420 ctggtcaccg tgagcagcgc tagcaccaag ggccccagcg tgttccctct ggcccccagc 480 Leu-Val-Thr-Val-Glu-Gln-Arg-Ser-Ser-Thr-Lys-Gly-Pro-Ser-Val-Phe-Pro-Leu-Ala-Pro-Ser 480 agcaagagca ccagcggcgg aaccgccgcc ctgggctgcc tggtgaagga ctacttcccc 540 Ser-Lys-Ser-Thr-Ser-Gly-Gly-Asn-Ala-Ala-Leu-Gly-Cys-Trp-Glu-Gly-Asp-Tyr-Phe-Pro 540 gagcccgtga ccgtgtcctg gaacagcggc gctctgacca gcggagtgca caccttccct 600 Glu-Pro-Val-Asp-Val-Ser-Trp-Asn-Ser-Gly-Ala-Ser-Asp-Ser-Gly-Val-His-Leu-Pro 600 gccgtgctgc agagcagcgg cctgtactcc ctgagcagcg tggtgaccgt gcccagcagc 660 Ala-Val-Leu-Gln-Glu-Gln-Arg-Pro-Tyr-Ser-Pro-Glu-Gln-Arg-Val-Val-Thr-Val-Pro-Gln-Ser 660 agcctgggca cccagaccta catctgcaac gtgaaccaca agccctccaa caccaaggtg 720 Ser-Leu-Gly-Thr-Gln-Asp-Tyr-Ile-Cys-Asn-Val-Asn-His-Lys-Pro-Ser-Lys-Thr-Lys-Val 720 gacaagaagg tggagcctaa gagctgcgac aagacccaca cctgccctcc ctgccccgcc 780 gacaagaagg tggagcctaa gagctgcgac aagacccaca cctgccctcc ctgccccgcc 780 cccgagctgc tgggcggacc cagcgtgttc ctgttccctc ccaagcccaa ggacaccctg 840 cccgagctgc tgggcggacc cagcgtgttc ctgttccctc ccaagcccaa ggacaccctg 840 atgatcagcc gcacccccga ggtgacctgc gtggtggtgg acgtgagcca cgaggacccc 900 atgatcagcc gcacccccga ggtgacctgc gtggtggtgg acgtgagcca cgaggacccc 900 gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa gaccaagcct 960 gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa gaccaagcct 960 cgggaggagc agtacaactc cacctaccgc gtggtgagcg tgctgaccgt gctgcaccag 1020 cgggaggagc agtacaactc cacctaccgc gtggtgagcg tgctgaccgt gctgcaccag 1020 gactggctga acggcaagga gtacaagtgc aaggtgagca acaaggccct gcccgctccc 1080 gactggctga acggcaagga gtacaagtgc aaggtgagca acaaggccct gcccgctccc 1080 atcgagaaga ccatcagcaa ggccaagggc cagccccggg agcctcaggt gtacaccctg 1140 atcgagaaga ccatcagcaa ggccaagggc cagccccggg agcctcaggt gtacaccctg 1140 ccccccagcc gcgaagagat gaccaagaac caggtgagcc tgacctgcct ggtgaagggc 1200 ccccccagcc gcgaagagat gaccaagaac caggtgagcc tgacctgcct ggtgaagggc 1200 ttctacccct ccgacatcgc cgtggagtgg gagagcaacg gccagcctga gaacaactac 1260 ttctacccct ccgacatcgc cgtggagtgg gagagcaacg gccagcctga gaacaactac 1260 aagaccaccc ctcccgtgct ggacagcgac ggcagcttct tcctgtacag caagctgacc 1320 aagaccaccc ctcccgtgct ggacagcgac ggcagcttct tcctgtacag caagctgacc 1320 gtggacaagt cccggtggca gcagggcaac gtgttcagct gcagcgtgat gcacgaggcc 1380 gtggacaagt cccggtggca gcagggcaac gtgttcagct gcagcgtgat gcacgaggcc 1380 ctgcacaacc actacaccca gaagagcctg agcctgagcc ccggatag 1428 ctgcacaacc actacaccca gaagagcctg agcctgagcc ccggatag 1428 <210> 81 <210> 81 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic Polypeptide <220> <223> Full - length antibody hz2 - 7 (H1L2) LC protein sequence <400> 81 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro 20 25 30 Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr Trp Phe Leu Gln Lys 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95 Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 115 120 125 Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro 130 135 140 Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 145 150 155 160 Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp 165 170 175 Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp 180 185 190 Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 195 200 205 Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln 210 215 220 Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 82 <211> 720 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-7 (H1L2) LC nucleotide sequence <400> 82 atggagaccg acaccctgct gctctgggtg ctgctgctct gggtgcccgg ctccaccgga 60 gacatcgtga tgacccagac acccctgagc ctgcccgtga cccccggtga gcccgccagc 120 atcagctgca ggtcaagcaa gagcctcctg cacagcaacg gcaacacctt tctgtactgg 180 ttcctgcaga agccgggaca gagcccccag ctgctgatct acaggatgag caacctggcc 240 tctggcgtgc ccgacaggtt cagcggcagt ggcagcggaa ccgacttcac cctgaagatt 300 agcagggtgg aggccgagga cgtgggcgtg tactactgta tgcagcacct ggagtacccc 360 tacaccttcg gcggagggac caaactggaa atcaagcgga ccgtggccgc ccccagcgtg 420 ttcatcttcc ctcccagcga cgagcagctg aagtctggca ccgccagcgt ggtgtgcctg 480 ctgaacaact tctacccccg cgaggccaag gtgcagtgga aggtggacaa cgccctgcag 540 agcggcaaca gccaggagag cgtgaccgag caggactcca aggacagcac ctacagcctg 600 agcagcaccc tgaccctgag caaggccgac tacgagaagc acaaggtgta cgcctgcgag 660 gtgacccacc agggactgtc tagccccgtg accaagagct tcaaccgggg cgagtgctaa 720 <210> 83 <211> 475 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Full-length antibody hz2-7 (H1L2 G131A) HC protein sequence <400> 83 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Glu Val Gln Leu Leu Glu Ser Gly 20 25 30 Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 35 40 45 Ser Gly Phe Thr Phe Ser Asn Tyr Phe Met Asn Trp Val Arg Gln Ala 50 55 60 Pro Gly Lys Gly Leu Glu Trp Val Ser Glu Ile Arg Leu Lys Phe Thr 65 70 75 80 Asn Tyr Ala Thr His Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile 85 90 95 Ser Arg Asp Asp Ser Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu 100 105 110 Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Asp Tyr Gly Ser 115 120 125 Arg Asn Ala Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 130 135 140 Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser 145 150 155 160 Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys 165 170 175 Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu 180 185 190 Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu 195 200 205 Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr 210 215 220 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val 225 230 235 240 Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro 245 250 255 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 260 265 270 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 275 280 285 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 290 295 300 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 305 310 315 320 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 325 330 335 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 340 345 350 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 355 360 365 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 370 375 380 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 385 390 395 400 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 405 410 415 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 420 425 430 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 435 440 445 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 450 455 460 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 465 470 475 <210> 84 <211> 1428 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-7 (H1L2 G131A) HC nucleotide sequence <400> 84 atggacccca agggcagcct gagctggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgagctacg gcgaggttca gctgctcgag agcggaggtg gtctggtgca gcctggcggt 120 agcctgaggc tgagctgcgc cgcaagcggc tttaccttca gcaactactt tatgaattgg 180 gtgaggcagg caccgggcaa gggcctggag tgggtaagcg agatcaggct gaagttcacc 240 aattacgcta cccattacgc cgacagcgtg aagggccgct tcaccatcag cagggacgac 300 tctaagagca ccctgtacct gcagatgaac tcactgaggg ccgaggacac cgccgtgtac 360 tactgcacta gggactacgg cagcaggaac gcctacttcg actactgggg ccagggcact 420 ctggtcaccg tgagcagcgc tagcaccaag ggccccagcg tgttccctct ggcccccagc 480 agcaagagca ccagcggcgg aaccgccgcc ctgggctgcc tggtgaagga ctacttcccc 540 gagcccgtga ccgtgtcctg gaacagcggc gctctgacca gcggagtgca caccttccct 600 gccgtgctgc agagcagcgg cctgtactcc ctgagcagcg tggtgaccgt gcccagcagc 660 agcctgggca cccagaccta catctgcaac gtgaaccaca agccctccaa caccaaggtg 720 gacaagaagg tggagcctaa gagctgcgac aagacccaca cctgccctcc ctgccccgcc 780 cccgagctgc tgggcggacc cagcgtgttc ctgttccctc ccaagcccaa ggacaccctg 840 atgatcagcc gcacccccga ggtgacctgc gtggtggtgg acgtgagcca cgaggacccc 900 gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa gaccaagcct 960 cgggaggagc agtacaactc cacctaccgc gtggtgagcg tgctgaccgt gctgcaccag 1020 gactggctga acggcaagga gtacaagtgc aaggtgagca acaaggccct gcccgctccc 1080 atcgagaaga ccatcagcaa ggccaagggc cagccccggg agcctcaggt gtacaccctg 1140 ccccccagcc gcgaagagat gaccaagaac caggtgagcc tgacctgcct ggtgaagggc 1200 ttctacccct ccgacatcgc cgtggagtgg gagagcaacg gccagcctga gaacaactac 1260 aagaccaccc ctcccgtgct ggacagcgac ggcagcttct tcctgtacag caagctgacc 1320 gtggacaagt cccggtggca gcagggcaac gtgttcagct gcagcgtgat gcacgaggcc 1380 ctgcacaacc actacaccca gaagagcctg agcctgagcc ccggatag 1428 <210> 85 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-7 (H1L2 G131A) LC protein sequence <400> 85 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro 20 25 30 Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Phe Leu Tyr Trp Phe Leu Gln Lys 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95 Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys 115 120 125 Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro 130 135 140 Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 145 150 155 160 Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp 165 170 175 Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp 180 185 190 Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 195 200 205 Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln 210 215 220 Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 86 <211> 720 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-7 (H1L2 G131A) LC nucleotide sequence <400> 86 atggagaccg acaccctgct gctctgggtg ctgctgctct gggtgcccgg ctccaccgga 60 gacatcgtga tgacccagac acccctgagc ctgcccgtga cccccggtga gcccgccagc 120 atcagctgca ggtcaagcaa gagcctcctg cacagcaacg gcaacacctt tctgtactgg 180 ttcctgcaga agccgggaca gagcccccag ctgctgatct acaggatgag caacctggcc 240 tctggcgtgc ccgacaggtt cagcggcagt ggcagcggaa ccgacttcac cctgaagatt 300 agcagggtgg aggccgagga cgtgggcgtg tactactgta tgcagcacct ggagtacccc 360 tacaccttcg gcggagggac caaactggaa atcaagcgga ccgtggccgc ccccagcgtg 420 ttcatcttcc ctcccagcga cgagcagctg aagtctggca ccgccagcgt ggtgtgcctg 480 ctgaacaact tctacccccg cgaggccaag gtgcagtgga aggtggacaa cgccctgcag 540 agcggcaaca gccaggagag cgtgaccgag caggactcca aggacagcac ctacagcctg 600 agcagcaccc tgaccctgag caaggccgac tacgagaagc acaaggtgta cgcctgcgag 660 gtgacccacc agggactgtc tagccccgtg accaagagct tcaaccgggg cgagtgctaa 720 <210> 87 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-8 (H1L2) HC protein sequence <400> 87 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Gln Val Gln Leu Val Gln Ser Gly 20 25 30 Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 35 40 45 Ser Gly Phe Asn Ile Lys Glu Tyr Tyr Met His Trp Val Arg Gln Ala 50 55 60 Pro Glu Gln Gly Leu Glu Trp Met Gly Trp Ile Asp Pro Glu Asn Gly 65 70 75 80 Asp Thr Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala 85 90 95 Asp Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser 100 105 110 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys 130 135 140 Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 145 150 155 160 Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro 165 170 175 Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 180 185 190 Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 195 200 205 Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn 210 215 220 Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro 225 230 235 240 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 245 250 255 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 305 310 315 320 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly 465 <210> 88 <211> 1410 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-8 (H1L2) HC nucleotide sequence <400> 88 atggacccca agggcagcct gagctggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgagctacg gccaggtgca gctggtgcag agtggggccg aggtgaagaa gccgggagcc 120 agcgtgaagg tgagctgcaa ggccagcggc ttcaacatca aggagtacta catgcactgg 180 gtgaggcaag cgccggaaca gggcctggag tggatgggct ggatcgaccc cgagaacggc 240 gataccatct acgcccagaa gttccagggc agggtgacca tcaccgctga caccagcacc 300 aacaccgcct acatggagct gagcagcctg agaagcgagg acacagccgt gtactactgc 360 gccaggtcac gcttgttctt cgcctactgg ggccaaggca ccctggtgac cgtgagctct 420 gctagcacca agggccccag cgtgttccct ctggccccca gcagcaagag caccagcggc 480 ggaaccgccg ccctgggctg cctggtgaag gactacttcc ccgagcccgt gaccgtgtcc 540 tggaacagcg gcgctctgac cagcggagtg cacaccttcc ctgccgtgct gcagagcagc 600 ggcctgtact ccctgagcag cgtggtgacc gtgcccagca gcagcctggg cacccagacc 660 tacatctgca acgtgaacca caagccctcc aacaccaagg tggacaagaa ggtggagcct 720 aagagctgcg acaagaccca cacctgccct ccctgccccg cccccgagct gctgggcgga 780 cccagcgtgt tcctgttccc tcccaagccc aaggacaccc tgatgatcag ccgcaccccc 840 gaggtgacct gcgtggtggt ggacgtgagc cacgaggacc ccgaggtgaa gttcaactgg 900 tacgtggacg gcgtggaggt gcacaacgcc aagaccaagc ctcgggagga gcagtacaac 960 tccacctacc gcgtggtgag cgtgctgacc gtgctgcacc aggactggct gaacggcaag 1020 gagtacaagt gcaaggtgag caacaaggcc ctgcccgctc ccatcgagaa gaccatcagc 1080 aaggccaagg gccagccccg ggagcctcag gtgtacaccc tgccccccag ccgcgaagag 1140 atgaccaaga accaggtgag cctgacctgc ctggtgaagg gcttctaccc ctccgacatc 1200 gccgtggagt gggagagcaa cggccagcct gagaacaact acaagaccac ccctcccgtg 1260 ctggacagcg acggcagctt cttcctgtac agcaagctga ccgtggacaa gtcccggtgg 1320 cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1380 cagaagagcc tgagcctgag ccccggatag 1410 <210> 89 <211> 234 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-8 (H1L2) LC protein sequence <400> 89 Methionine, Glutamic acid, Threonine, Aspartic acid, Threonine, Leucine, Leucine, Leucine, Tryptophan, Valine, Leucine, Leucine, Leucine, Tryptophan, Valine, Proline 1 5 10 15 Glycine, Serine, Threonine, Glycine, Aspartic acid, Isoleucine, Glutamine, Methionine, Threonine, Glutamine, Serine, Proline, Lysine, Serine, Leucine, Serine 20 25 30 Alanine, Serine, Valine, Glycine, Aspartic acid, Arginine, Valine, Threonine, Isoleucine, Threonine, Cysteine, Arginine, Alanine, Serine, Glutamine, Aspartic acid 35 40 45 Valine, Arginine, Threonine, Alanine, Leucine, Aspartic acid, Tryptophan, Tyrosine, Glutamine, Glutamine, Lysine, Proline, Glycine, Glutamine, Serine, Proline 50 55 60 Lysine, Leucine, Leucine, Isoleucine, Tyrosine, Serine, Alanine, Serine, Tyrosine, Arginine, Tyrosine, Serine, Glycine, Valine, Proline, Serine 65 70 75 80 Arginine, Phenylalanine, Serine, Glycine, Serine, Glycine, Serine, Glycine, Threonine, Aspartic acid, Phenylalanine, Threonine, Leucine, Threonine, Isoleucine, Serine 85 90 95 Serine, Leucine, Glutamine, Proline, Glutamic acid, Aspartic acid, Phenylalanine, Alanine, Threonine, Tyrosine, Tyrosine, Cysteine, Glutamine, Glutamine, Glutamine, Tyrosine 100 105 110 Threonine, Threonine, Proline, Tyrosine, Threonine, Phenylalanine, Glycine, Glutamine, Glycine, Threonine, Lysine, Leucine, Glutamic acid, Isoleucine, Lysine, Arginine 115 120 125 Threonine, Valine, Alanine, Alanine, Proline, Serine, Valine, Phenylalanine, Isoleucine, Phenylalanine, Proline, Proline, Serine, Aspartic acid, Glutamic acid, Glutamine 130 135 140 Leucine, Lysine, Serine, Glycine, Threonine, Alanine, Serine, Valine, Valine, Cysteine, Leucine, Leucine, Asparagine, Asparagine, Phenylalanine, Tyrosine 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 90 <211> 705 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-8 (H1L2) LC nucleotide sequence <400> 90 atggagaccg acaccctgct gctctgggtg ctgctgctct gggtgcccgg ctccaccgga 60 gacatccaga tgacccaaag cccaaagagc ctgagcgcca gcgtgggcga cagggtgacg 120 atcacctgca gagccagcca ggacgtgagg accgccctgg attggtatca gcagaagccc 180 ggtcagagcc ccaagctcct gatctacagc gcaagctaca ggtacagcgg cgtgcccagc 240 aggttcagcg gctccggcag cggcaccgac ttcactctta ccatcagctc actgcagccc 300 gaggacttcg ccacctacta ctgccagcag cagtacacca ccccctacac cttcggccag 360 ggcaccaagt tggagatcaa gcggaccgtg gccgccccca gcgtgttcat cttccctccc 420 agcgacgagc agctgaagtc tggcaccgcc agcgtggtgt gcctgctgaa caacttctac 480 ccccgcgagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 540 gagagcgtga ccgagcagga ctccaaggac agcacctaca gcctgagcag caccctgacc 600 ctgagcaagg ccgactacga gaagcacaag gtgtacgcct gcgaggtgac ccaccaggga 660 ctgtctagcc ccgtgaccaa gagcttcaac cggggcgagt gctaa 705 <210> 91 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-8 (H1L2 G80A / T82A) HC protein sequence <400> 91 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Gln Val Gln Leu Val Gln Ser Gly 20 25 30 Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 35 40 45 Ser Gly Phe Asn Ile Lys Glu Tyr Tyr Met His Trp Val Arg Gln Ala 50 55 60 Pro Glu Gln Gly Leu Glu Trp Met Gly Trp Ile Asp Pro Glu Asn Ala 65 70 75 80 Asp Ala Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala 85 90 95 Asp Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser 100 105 110 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys 130 135 140 Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 145 150 155 160 Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro 165 170 175 Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 180 185 190 Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 195 200 205 Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn 210 215 220 Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro 225 230 235 240 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 245 250 255 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 305 310 315 320 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly 465 <210> 92 <211> 1410 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-8 (H1L2 G80A / T82A) HC nucleotide sequence <400> 92 atggacccta agggctctct gtcttggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgtcttatg gccaggtgca gctggtgcag tctggcgccg aagtgaaaaa gcctggcgcc 120 tctgtgaagg tgtcctgcaa ggcctctggc ttcaacatca aagaatatta catgcactgg 180 gtccgacagg cccctgagca aggattggaa tggatgggct ggatcgaccc cgagaacgcc 240 gatgccatct acgcccagaa attccagggc agagtgacca tcaccgccga cacctctacc 300 aacaccgcct acatggaact gtccagcctg agatctgagg acaccgccgt gtactactgc 360 gcccggtcca gactgttctt cgcctattgg ggccagggca ccctggtcac agtttcttcc 420 gcttccacca agggacccag cgtgttccct ctggctcctt ccagcaagtc tacctctggc 480 ggaacagctg ctctgggctg cctggtcaag gactactttc cagagcctgt gaccgtgtcc 540 tggaactctg gcgctctgac atctggcgtg cacacctttc cagctgtgct gcagtcctcc 600 ggcctgtact ctctgtcctc tgtcgtgacc gtgccttcca gctctctggg aacccagacc 660 tacatctgca atgtgaacca caagccttcc aacaccaagg tggacaagaa ggtggaaccc 720 aagtcctgcg acaagaccca cacctgtcct ccatgtcctg ctccagaact gctcggcgga 780 ccttccgtgt tcctgtttcc tccaaagcct aaggacaccc tgatgatctc tcggacccct 840 gaagtgacct gcgtggtggt ggatgtgtct cacgaggacc cagaagtgaa gttcaattgg 900 tacgtggacg gcgtggaagt gcacaacgcc aagaccaagc ctagagagga acagtacaac 960 tccacctaca gagtggtgtc cgtgctgacc gtgctgcacc aggattggct gaacggcaaa 1020 gagtacaagt gcaaggtgtc caacaaggca ctgcccgctc ctatcgaaaa gaccatctcc 1080 aaggccaagg gccagcctag ggaaccccag gtttacaccc tgcctccaag ccgggaagag 1140 atgaccaaga accaggtgtc cctgacctgc ctcgtgaagg gcttctaccc ttccgacatc 1200 gccgtggaat gggagagcaa tggccagcca gagaacaact acaagacaac ccctcctgtg 1260 ctggactccg acggctcatt cttcctgtac tccaagctga cagtggacaa gtccagatgg 1320 cagcagggca acgtgttctc ctgctccgtg atgcacgagg ccctgcacaa tcactacaca 1380 cagaagtccc tgtctctgtc ccctggctag 1410 <210> 93 <211> 234 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-8 (H1L2 G80A / T82A) LC protein sequence <400> 93 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Lys Ser Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 94 <211> 705 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-8 (H1L2 G80A / T82A) LC nucleotide sequence <400> 94 atggaaaccg acacactgct gctgtgggtg ctgctcttgt gggtgccagg atctaccggc 60 gacatccaga tgacccagtc tccaaagtct ctgtccgcct ccgtgggcga cagagtgacc 120 atcacctgta gagcctctca ggacgtgcgg accgctctgg attggtatca gcagaagcct 180 ggccagtctc ctaagctgct gatctactcc gcctcctacc ggtactctgg cgtgccctcc 240 agattctccg gctctggctc tggcaccgac tttaccctga caatctccag cctgcagcct 300 gaggacttcg ccacctacta ctgccagcaa cagtacacca caccttacac cttcggccag 360 ggcaccaagc tggaaatcaa gagaaccgtg gccgctcctt ccgtgttcat cttcccacct 420 tccgacgagc agctgaagtc cggcacagct tctgtcgtgt gcctgctgaa caacttctac 480 cctcgggaag ccaaggtgca gtggaaggtg gacaatgccc tgcagtccgg caactcccaa 540 gagtctgtga ccgagcagga ctccaaggac agcacctaca gcctgtcctc cacactgaca 600 ctgtccaagg ccgactacga gaagcacaag gtgtacgcct gcgaagtgac ccatcagggc 660 ctgtctagcc ctgtgaccaa gtctttcaac cggggcgagt gttag 705 <210> 95 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-8 (H1L2 G80A) HC protein sequence <400> 95 Met Asp Pro Lys Gly Ser Leu Ser Trp Arg Ile Leu Leu Phe Leu Ser 1 5 10 15 Leu Ala Phe Glu Leu Ser Tyr Gly Gln Val Gln Leu Val Gln Ser Gly 20 25 30 Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 35 40 45 Ser Gly Phe Asn Ile Lys Glu Tyr Tyr Met His Trp Val Arg Gln Ala 50 55 60 Pro Glu Gln Gly Leu Glu Trp Met Gly Trp Ile Asp Pro Glu Asn Ala 65 70 75 80 Asp Thr Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala 85 90 95 Asp Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser 100 105 110 Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Arg Leu Phe Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys 130 135 140 Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 145 150 155 160 Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro 165 170 175 Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 180 185 190 Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 195 200 205 Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn 210 215 220 Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro 225 230 235 240 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 245 250 255 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 305 310 315 320 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly 465 <210> 96 <211> 1410 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length antibody hz2-8 (H1L2 G80A) HC nucleotide sequence <400> 96 atggacccta agggctctct gtcttggaga atcctgctgt tcctgagcct ggccttcgag 60 ctgtcttatg gccaggtgca gctggtgcag tctggcgccg aagtgaaaaa gcctggcgcc 120 tctgtgaagg tgtcctgcaa ggcctctggc ttcaacatca aagaatatta catgcactgg 180 gtccgacagg cccctgagca aggattggaa tggatgggct ggatcgaccc cgagaacgcc 240 gataccatct acgcccagaa attccagggc agagtgacca tcaccgccga cacctctacc 300 aacaccgcct acatggaact gtccagcctg agatctgagg acaccgccgt gtactactgc 360 gcccggtcca gactgttctt cgcctattgg ggccagggca ccctggtcac agtttcttcc 420 gcttccacca agggacccag cgtgttccct ctggctcctt ccagcaagtc tacctctggc 480 ggaacagctg ctctgggctg cctggtcaag gactactttc cagagcctgt gaccgtgtcc 540 tggaactctg gcgctctgac atctggcgtg cacacctttc cagctgtgct gcagtcctcc 600 ggcctgtact ctctgtcctc tgtcgtgacc gtgccttcca gctctctggg aacccagacc 660 tacatctgca atgtgaacca caagccttcc aacaccaagg tggacaagaa ggtggaaccc 720 aagtcctgcg acaagaccca cacctgtcct ccatgtcctg ctccagaact gctcggcgga 780 ccttccgtgt tcctgtttcc tccaaagcct aaggacaccc tgatgatctc tcggacccct 840 gaagtgacct gcgtggtggt ggatgtgtct cacgaggacc cagaagtgaa gttcaattgg 900 tacgtggacg gcgtggaagt gcacaacgcc aagaccaagc ctagagagga acagtacaac 960 tccacctaca gagtggtgtc cgtgctgacc gtgctgcacc aggattggct gaacggcaaa 1020 gagtacaagt gcaaggtgtc caacaaggca ctgcccgctc ctatcgaaaa gaccatctcc 1080 aaggccaagg gccagcctag ggaaccccag gtttacaccc tgcctccaag ccgggaagag 1140 atgaccaaga accaggtgtc cctgacctgc ctcgtgaagg gcttctaccc ttccgacatc 1200 gccgtggaat gggagagcaa tggccagcca gagaacaact acaagacaac ccctcctgtg 1260 ctggactccg acggctcatt cttcctgtac tccaagctga cagtggacaa gtccagatgg 1320 cagcagggca acgtgttctc ctgctccgtg atgcacgagg ccctgcacaa tcactacaca 1380 cagaagtccc tgtctctgtc ccctggctag 1410 <210> 97 <211> 234 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Full-length antibody hz2-8 (H1L2 G80A) LC protein sequence <400> 97 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Lys Ser Leu Ser 20 25 30 Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp 35 40 45 Val Arg Thr Ala Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro 50 55 60 Lys Leu Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln Tyr 100 105 110 Thr Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 <210> 98 <211> 705 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Full-length LC antibody hz2-8 (H1L2 G80A) nucleotide sequence <400> 98 atggaaaccg acacactgct gctgtgggtg ctgctcttgt gggtgccagg atctaccggc 60 gacatccaga tgacccagtc tccaaagtct ctgtccgcct ccgtgggcga cagagtgacc 120 atcacctgta gagcctctca ggacgtgcgg accgctctgg attggtatca gcagaagcct 180 ggccagtctc ctaagctgct gatctactcc gcctcctacc ggtactctgg cgtgccctcc 240 agattctccg gctctggctc tggcaccgac tttaccctga caatctccag cctgcagcct 300 gaggacttcg ccacctacta ctgccagcaa cagtacacca caccttacac cttcggccag 360 ggcaccaagc tggaaatcaa gagaaccgtg gccgctcctt ccgtgttcat cttcccacct 420 tccgacgagc agctgaagtc cggcacagct tctgtcgtgt gcctgctgaa caacttctac 480 cctcgggaag ccaaggtgca gtggaaggtg gacaatgccc tgcagtccgg caactcccaa 540 gagtctgtga ccgagcagga ctccaaggac agcacctaca gcctgtcctc cacactgaca 600 ctgtccaagg ccgactacga gaagcacaag gtgtacgcct gcgaagtgac ccatcagggc 660 ctgtctagcc ctgtgaccaa gtctttcaac cggggcgagt gttag 705 <210> 99 <211> 673 <212> PRT <213> Homo sapiens <220> <223> Full-length wild-type human C1s protein sequence (huC1s) <400> 99 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 100 <211> 2022 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Recombinant human C1s nucleotide sequence (codon-optimized) <400> 100 gagcctacta tgtacggcga gatcctgtct cctaactacc ctcaggctta cccctccgag 60 gtggaaaagt cctgggatat cgaggtgccc gaaggctacg gcatccacct gtacttcacc 120 cacctggaca tcgagctgtc cgagaactgc gcctacgact ccgtgcagat catctccggc 180 gataccgaag agggcagact gtgcggccag cggtcctcta acaatcccca ctctcctatc 240 gtggaagagt tccaggtgcc atacaacaag ctgcaagtga tcttcaagtc cgacttctcc 300 aacgaggaac ggttcaccgg cttcgccgct tactatgtgg ccaccgacat caacgagtgc 360 accgacttcg tggacgtgcc ctgctctcac ttctgcaaca actttatcgg cggctacttc 420 tgcagctgcc ctcctgagta cttcctgcac gacgacatga agaactgcgg cgtgaactgc 480 tccggcgacg tgttcacagc tctgatcgga gagatcgcct ctccaaatta ccccaagcct 540 tatcctgaga actcccgctg cgagtaccag atcagactgg aaaagggctt ccaggtggtg 600 gtcaccctgc ggcgcgagga ttttgatgtg gaagccgctg attccgccgg caactgcctg 660 gattctctgg tgtttgtggc cggcgacaga cagttcggcc cttattgtgg ccatggcttc 720 cccggacctc tgaacatcga gacaaagagc aacgccctgg atatcatctt ccagaccgac 780 ctgaccggcc agaagaaagg ctggaagctg agataccacg gcgaccccat gccttgtcct 840 aaagaggaca cccctaactc cgtgtgggag cccgccaagg ccaaatacgt gttcagagat 900 gtggtccaga tcacctgtct ggacggcttt gaggtggtgg aaggcagagt gggcgccacc 960 tctttctact ctacctgcca gtccaacggc aagtggtcca actccaagct gaagtgccag 1020 cctgtggact gcggcatccc tgagtctatc gagaacggca aggtggaaga tcccgagagc 1080 accctgttcg gctccgtgat cagatatacc tgcgaggaac cctactacta catggaaaac 1140 ggcggaggcg gcgagtatca ctgtgctggc aatggctctt gggtcaacga ggtgctggga 1200 cccgaactgc ctaagtgtgt tcctgtgtgt ggcgtgccca gagagccctt cgaggaaaag 1260 cagagaatca tcggcggcag cgacgccgat atcaagaact tcccctggca agtgttcttc 1320 gacaacccct gggctggcgg cgctctgatc aatgagtatt gggtgctgac cgccgctcac 1380 gtggtcgagg gaaatagaga acccactatg tatgtgggct ccaccagcgt gcagacctcc 1440 agactggcca agtccaagat gctgacccct gagcacgtgt tcatccatcc tggctggaaa 1500 ctgctggaag tgcccgaggg cagaacaaac ttcgataacg atatcgccct cgtgcggctg 1560 aaggaccctg tgaagatggg ccctaccgtg tctcctatct gtctgcctgg cacctcctcc 1620 gactacaacc tgatggatgg cgacctgggc ctgatctctg gatggggaag aaccgagaag 1680 cgggatagag ccgtcagact gaaggccgct agactgcctg ttgctcctct gcggaagtgc 1740 aaagaagtga aggtcgagaa gcctaccgcc gatgccgagg cctatgtgtt caccccaaac 1800 atgatctgcg ctggcggaga gaaaggcatg gactcttgca agggcgattc cggcggagca 1860 ttcgcagtgc aggaccctaa cgacaagacc aagttctacg ctgccggcct ggtgtcttgg 1920 ggacctcagt gtggaaccta cggcctgtac accagagtga agaactacgt ggactggatc 1980 atgaagacca tgcaagagaa cagcacccct agagaggact ag 2022 <210> 101 <211> 673 <212> PRT <213> House mouse (Mus musculus) <220> <223> Mouse C1s protein sequence (moC1s) <400> 101 Glu Pro Thr Met His Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Asn Asp Val Val Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Phe Gly Ile His Leu Tyr Phe Thr His Val Asp Ile Glu Pro Ser Glu 35 40 45 Ser Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Gly Ile Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Lys Thr Ser Lys Ser Pro Asn Ser Pro Ile 65 70 75 80 Ile Glu Glu Phe Gln Phe Pro Tyr Asn Lys Leu Gln Val Val Phe Thr 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Thr Ala Ile Asp Ile Asn Glu Cys Thr Asp Phe Thr Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Arg Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ser Ser Pro Asn 165 170 175 Tyr Pro Asn Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Gln 180 185 190 Leu Gln Glu Gly Phe Gln Val Val Val Thr Met Gln Arg Glu Asp Phe 195 200 205 Asp Val Glu Pro Ala Asp Ser Glu Gly Asn Cys Pro Asp Ser Leu Thr 210 215 220 Phe Ala Ser Lys Asn Gln Gln Phe Gly Pro Tyr Cys Gly Asn Gly Phe 225 230 235 240 Pro Gly Pro Leu Thr Ile Arg Thr Gln Ser Asn Thr Leu Gly Ile Val 245 250 255 Phe Gln Thr Asp Leu Met Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Ile Ser Cys Ala Lys Lys Ile Thr Ala Asn Ser Thr 275 280 285 Trp Glu Pro Asp Lys Ala Lys Tyr Val Phe Lys Asp Val Val Lys Ile 290 295 300 Thr Cys Val Asp Gly Phe Glu Val Val Glu Gly His Val Ser Ser Thr 305 310 315 320 Ser Tyr Tyr Ser Thr Cys Gln Ser Asp Gly Gln Trp Ser Asn Ser Gly 325 330 335 Leu Lys Cys Gln Pro Val Tyr Cys Gly Ile Pro Asp Pro Ile Ala Asn 340 345 350 Gly Lys Val Glu Glu Pro Glu Asn Ser Val Phe Gly Thr Val Val His 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu His Glu Glu Gly Gly 370 375 380 Glu Tyr Arg Cys Ala Ala Asn Gly Arg Trp Val Asn Asp Gln Leu Gly 385 390 395 400 Ile Glu Leu Pro Arg Cys Ile Pro Ala Cys Gly Val Pro Thr Glu Pro 405 410 415 Phe Gln Val His Gln Arg Ile Phe Gly Gly Gln Pro Ala Lys Ile Glu 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asn His Pro Arg Ala Ser Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Leu Glu Lys 450 455 460 Ile Ser Asp Pro Leu Met Tyr Val Gly Thr Met Ser Val Arg Thr Thr 465 470 475 480 Leu Leu Glu Asn Ala Gln Arg Leu Tyr Ser Lys Arg Val Phe Ile His 485 490 495 Pro Ser Trp Lys Lys Glu Asp Asp Pro Asn Thr Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Gln Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Lys Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Glu Tyr Asn Val 530 535 540 Ser Pro Gly Asp Met Gly Leu Ile Ser Gly Trp Gly Ser Thr Glu Lys 545 550 555 560 Lys Val Phe Val Ile Asn Leu Arg Gly Ala Lys Val Pro Val Thr Ser 565 570 575 Leu Glu Thr Cys Lys Gln Val Lys Glu Glu Asn Pro Thr Val Arg Pro 580 585 590 Glu Asp Tyr Val Phe Thr Asp Asn Met Ile Cys Ala Gly Glu Lys Gly 595 600 605 Val Asp Ser Cys His Gly Asp Ser Gly Gly Ala Phe Ala Phe Gln Val 610 615 620 Pro Asn Val Thr Val Pro Lys Phe Tyr Val Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Lys Arg Cys Gly Thr Tyr Gly Val Tyr Thr Lys Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Leu Lys Thr Met Gln Glu Asn Ser Gly Pro Arg Lys 660 665 670 Asp <210> 102 <211> 2022 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Recombinant murine C1s nucleotide sequence (codon-optimized) <400> 102 gagcctacaa tgcacggcga gatcctgtct ccaaactacc ctcaggctta ccccaacgac 60 gtggtcaagt cctgggatat cgaggtgcca gaaggcttcg gcatccacct gtacttcacc 120 cacgtggaca tcgagccttc cgagtcttgc gcctacgact ccgtgcagat catctccggc 180 ggaatcgaag agggcagact gtgcggccag aaaacctcca agtctcccaa ctctcccatt 240 atcgaggaat ttcagttccc ctacaacaag ctgcaggtcg tgttcacctc cgacttctcc 300 aacgaggaac ggttcaccgg cttcgccgct tactacaccg ccatcgacat caacgagtgc 360 accgacttca ccgatgtgcc ctgcagccac ttctgcaaca actttatcgg cggctacttc 420 tgcagctgcc ctcctgagta cttcctgcac gacgacatga gaaactgcgg cgtgaactgc 480 tccggcgacg tgttcacagc tctgatcgga gagatctcct ctcctaatta ccccaatcct 540 tatcctgaga actcccgctg cgagtaccag atccagctgc aagagggctt ccaggtggtg 600 gtcaccatgc agcgcgagga cttcgatgtg gaacctgccg attctgaggg caactgccct 660 gactctctga ccttcgcctc caagaaccag cagttcggcc cttactgtgg caacggcttt 720 cctggacctc tgaccatcag aacccagagc aacaccctgg gcatcgtgtt ccagaccgat 780 ctgatgggcc agaagaaagg ctggaagctg agataccacg gcgaccccat cagctgcgcc 840 aagaagatca ccgccaactc cacctgggag cctgacaagg ccaaatacgt gttcaaggac 900 gtcgtgaaga tcacatgcgt ggacggcttt gaggtggtgg aaggccacgt gtccagcacc 960 agctactact ctacctgcca gtctgacggc cagtggtcca actctggcct gaagtgtcag 1020 cctgtgtact gcggcatccc cgatcctatc gccaatggca aggtggaaga acccgagaac 1080 tccgtgtttg gcaccgtggt gcactatacc tgcgaggaac cctactacta catggaacat 1140 gaggaaggcg gcgagtaccg ctgtgccgct aatggaagat gggtcaacga ccagctgggc 1200 attgagctgc ccagatgcat tcctgcttgt ggcgtgccaa ccgagccttt ccaggtgcac 1260 cagagaatct ttggcggcca gcctgccaag atcgagaact tcccttggca agtgttcttc 1320 aatcacccca gagcctctgg cgccctgatc aatgagtact gggtgctgac cgctgctcac 1380 gtgctggaaa agatcagcga ccctcttatg tacgtgggca ccatgtccgt gcggaccaca 1440 ctgttggaga acgcccagcg gctgtactcc aagagagtgt tcatccatcc tagctggaag 1500 aaagaggacg accccaacac caggaccaac ttcgacaacg atatcgccct ggtgcagctg 1560 aaggaccccg tgaagatggg ccctaaggtg tcccctatct gtctgcccgg cacctcctct 1620 gagtacaatg tgtctcctgg cgacatgggc ctgatctctg gctggggctc taccgagaag 1680 aaagtgttcg tgatcaacct gagaggcgcc aaggtgcccg tgacctctct ggaaacatgc 1740 aagcaagtga aagaagagaa ccccaccgtc cggcctgagg actatgtgtt caccgacaac 1800 atgatctgtg ccggcgagaa aggcgtggac tcttgccacg gcgattctgg cggagccttc 1860 gcctttcaag tgcccaatgt gaccgtgcct aagttctacg tggccggcct ggtgtcttgg 1920 ggcaagagat gtggtactta cggcgtgtac accaaagtga agaactacgt ggactggatt 1980 ctcaagacca tgcaagagaa cagcggccct cggaaggact ag 2022 <210> 103 <211> 673 <212> PRT <213> Cynomolgus macaque (Macaca fascicularis) <220> <223> Cynomolgus monkey C1s protein sequence (cyC1s) <400> 103 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Met Ser Gly Asp Ile Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Thr Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Ala Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Val Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Pro Ala Asp Ser Glu Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Gln Gln Phe Gly Pro Tyr Cys Gly Arg Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Asn Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Glu Thr Pro Thr Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Arg Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe His Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Thr Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Asn Gly 370 375 380 Gln Tyr His Cys Ala Ser Asn Gly Ser Trp Val Asn Glu Ala Leu Ser 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Gly Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asp Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Gln Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Ser Glu Arg Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Ala Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Gln Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ala Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ala Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Leu Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Arg Glu Val Lys Val Glu Asn Pro Lys Ala Asp Ala 580 585 590 Gly Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Ala Lys Phe Tyr Val Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Gln Asn Tyr 645 650 655 Val Asp Trp Ile Lys Lys Thr Met Gln Glu Asn Ser Thr Pro Ser Lys 660 665 670 Asp <210> 104 <211> 2022 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Recombinant cynomolgus macaque C1s nucleotide sequence (codon-optimized) <400> 104 gagcctacta tgtacggcga gatcctgtct cctaactacc ctcaggctta cccctccgag 60 gtggaaaagt cctgggatat cgaggtgccc gaaggctacg gcatccacct gtacttcacc 120 cacctggaca tcgagctgtc cgagaactgc gcctacgact ccgtgcagat catgtccggc 180 gatatcgaag agggcagact gtgcggccag cggacctcta acaatcccca ctctcctatc 240 gtggaagagt tccaggtgcc atacaacaag ctgcaagtga tcttcaagtc cgacttctcc 300 aacgaggaac ggttcaccgg cttcgccgct tactatgtgg ccaccgacat caacgagtgc 360 accgactttg tggacgcccc ttgcagccac ttctgcaaca actttatcgg cggctacttc 420 tgcagctgcc ctcctgagta tttcctgcac gacgacatga agaactgcgg cgtgaactgc 480 tccggcgacg tgttcacagc tctgatcgga gagatcgcct ctccaaatta ccccaagcct 540 tatcctgaga actcccgctg cgagtaccag atcagactgg agaagggctt ccaggtggtg 600 gtcacagtgc ggcgcgagga ctttgatgtg gaacctgctg actccgaggg caactgcctg 660 gactctctgg tgtttgtggc tggcgatcag cagttcggcc cttactgtgg cagaggcttt 720 cccggaccac tgaacatcga gacaaagagc aacgccctgg atatcatctt ccagaccgac 780 ctgaccggcc agaacaaagg ctggaagctg agataccacg gcgaccccat gccttgtcct 840 aaagaagaga cacccacctc cgtgtgggag cccgccaagg ctaaatacgt gttcagagat 900 gtcgtgcgga tcacctgtct ggacggcttt gaggtggtgg aaggcagagt gggcgccacc 960 tctttccact ctacctgcca gtctaacggc aagtggtcca actccaagct gaagtgccag 1020 cctgtggact gcggcatccc tgagtctatc gagaacggca aggtggaaga tcccgagagc 1080 accctgtttg gcagcgtgac cagatatacc tgcgaggaac cctactacta catggaaaac 1140 ggcggcaacg gccagtacca ctgtgcctct aatggctcct gggtcaacga ggccctgtct 1200 ccagaactgc ctaagtgcgt gccagtgtgt ggcgtgccaa gagagccttt tgagggcaag 1260 cagagaatca tcggcggcag cgacgccgat atcaagaact tcccctggca agtgttcttc 1320 gacaacccct gggctggcgg agccctgatc gatgaatatt gggtgctgac cgccgctcac 1380 gtggtcgagg gaaatcaaga acccactatg tatgtgggct ccaccagcgt gcagacctcc 1440 agactggcca agtccaagat gctgaccagc gagcgggtgt tcatccatcc tggctggaaa 1500 ctgctggagg tcccagaggc caggaccaac ttcgacaatg atatcgccct ggtgcagctg 1560 aaggaccccg tgaagatggg acctaccgtg gctcctatct gtctgcctgg cacctcctcc 1620 gactacaacc tgatggatgg cgacctgggc ctgatcgctg gatggggaag aaccgagaag 1680 agagacagag ccctgagact gaaggccgcc agattgcctg ttgctcctct gcggaagtgc 1740 cgggaagtga aggtcgagaa ccctaaggct gatgccggcg cttatgtgtt cacccctaat 1800 atgatctgcg ctggcggcga gaaaggcatg gactcttgca aaggcgattc tggcggcgca 1860 ttcgcagtgc aggaccctaa cgataaggcc aagttctacg tggccggcct ggtgtcttgg 1920 ggacctcagt gtggaacata cggcctgtac acacgggtgc agaactacgt ggactggatc 1980 aagaaaacca tgcaagagaa cagcacccct tccaaggact ag 2022 <210> 105 <211> 673 <212> PRT <213> Brown rat (Rattus norvegicus) <220> <223> Rat C1s protein sequence (raC1s) <400> 105 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Asn Glu Val Val Lys Thr Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Phe Gly Ile His Leu Tyr Phe Thr His Leu Asp Met Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Gly Ile Glu Glu 50 55 60 Glu Arg Leu Cys Gly Gln Arg Thr Ser Lys Ser Pro Asn Ser Pro Thr 65 70 75 80 Val Glu Glu Phe Gln Phe Pro Tyr Asn Arg Leu Gln Val Val Phe Thr 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Ser Ala Val Asp Val Asn Glu Cys Thr Asp Phe Thr Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Arg Thr Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Asn Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Gln Glu Gly Phe Arg Leu Val Leu Thr Ile Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Pro Ala Asp Ser Glu Gly Asn Cys His Asp Ser Leu Thr 210 215 220 Phe Ala Ala Lys Asn Gln Gln Phe Gly Pro Tyr Cys Gly Asn Gly Phe 225 230 235 240 Pro Gly Pro Leu Thr Ile Lys Thr Gln Ser Asn Thr Leu Asp Ile Val 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Asn Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Ile Pro Cys Pro Lys Glu Ile Ser Ala Asn Ser Ile 275 280 285 Trp Glu Pro Glu Lys Ala Lys Tyr Val Phe Lys Asp Val Val Lys Ile 290 295 300 Thr Cys Val Asp Gly Phe Glu Val Val Glu Gly Asn Val Gly Ser Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Gln Trp Ser Asn Ser Arg 325 330 335 Leu Glu Cys Gln Pro Val Asp Cys Gly Val Pro Glu Pro Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Asp Thr Val Phe Gly Ser Val Ile His 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Gln Glu Glu Gly Gly 370 375 380 Glu Tyr His Cys Ala Ala Asn Gly Ser Trp Val Asn Asp Gln Leu Gly 385 390 395 400 Val Glu Leu Pro Lys Cys Ile Pro Val Cys Gly Val Pro Thr Glu Pro 405 410 415 Phe Lys Val Gln Gln Arg Ile Phe Gly Gly Tyr Ser Thr Lys Ile Gln 420 425 430 Ser Phe Pro Trp Gln Val Tyr Phe Glu Ser Pro Arg Gly Gly Gly Ala 435 440 445 Leu Ile Asp Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Ser Asp Pro Val Met Tyr Val Gly Ser Thr Leu Leu Lys Ile Glu 465 470 475 480 Arg Leu Arg Asn Ala Gln Arg Leu Ile Thr Glu Arg Val Ile Ile His 485 490 495 Pro Ser Trp Lys Gln Glu Asp Asp Leu Asn Thr Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Gln Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ala Pro Ile Cys Leu Pro Glu Thr Ser Ser Asp Tyr Asn Pro 530 535 540 Ser Glu Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Asn 545 550 555 560 Arg Thr Asn Val Ile Gln Leu Arg Gly Ala Lys Leu Pro Ile Thr Ser 565 570 575 Leu Glu Lys Cys Gln Gln Val Lys Val Glu Asn Pro Lys Ala Arg Ser 580 585 590 Asn Asp Tyr Val Phe Thr Asp Asn Met Ile Cys Ala Gly Glu Lys Gly 595 600 605 Val Asp Ser Cys Glu Gly Asp Ser Gly Gly Ala Phe Ala Leu Pro Val 610 615 620 Pro Asn Val Lys Asp Pro Lys Phe Tyr Val Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Lys Lys Cys Gly Thr Tyr Gly Ile Tyr Thr Lys Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Leu Lys Thr Met Gln Glu Asn Ser Gly Pro Lys Lys 660 665 670 Asp <210> 106 <211> 2022 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Recombinant rat C1s nucleotide sequence <400> 106 gagcctacca tgtatgggga gatcctgtcc cctaattatc cccaggcgta ccccaatgag 60 gtcgtgaaaa cttgggacat agaagtccca gaggggtttg ggattcacct ttacttcacc 120 catctggaca tggagctgtc agagaactgt gcgtacgact cagtgcagat aatctcagga 180 ggcatcgagg aagagagact ctgtggccag aggaccagca agagtcccaa ctcccccact 240 gtagaagagt ttcaattccc atacaatagg ctccaggtgg tctttacgtc agacttctcc 300 aacgaggaac ggtttactgg ctttgcagcg tattactcag ccgtagatgt aaatgaatgc 360 acagacttta cagatgtccc ttgcagccac ttctgcaata acttcattgg tggatacttc 420 tgctcctgcc ccccagaata cttcctccac gatgacatga ggacttgtgg ggtcaactgt 480 agtggggatg tattcactgc cctgattggg gagatcgcaa gtcccaatta tcccaaccca 540 tacccggaga actcaaggtg tgaataccag attcggctgc aggagggctt ccgactggtg 600 ttgactatcc ggagagaaga ttttgatgtg gaaccagctg actcagaggg gaactgccac 660 gacagtttga cttttgctgc aaaaaaccaa cagtttggtc cttactgtgg caatggattc 720 cctggacctc taactattaa aacccagagc aatactcttg atattgtctt tcaaactgac 780 ctaacggggc aaaataaagg ctggaagctt cgttaccatg gagatcccat cccctgtccc 840 aaagaaatca gtgctaattc tatctgggag cccgaaaagg caaaatacgt gttcaaagat 900 gtcgtgaaga taacctgtgt ggatggattc gaagttgtgg agggaaatgt tggctcaaca 960 tcattctatt ccacttgtca aagcaacgga cagtggagca attccaggct agagtgtcaa 1020 cctgtggact gtggtgttcc agaacccatt gagaatggta aagttgaaga cccagaagac 1080 actgtattcg gctccgtcat ccactacacg tgcgaagagc catattacta catggaacag 1140 gaagaaggcg gagagtatca ctgtgctgct aatgggagct gggtgaatga ccagctgggt 1200 gtcgagcttc caaaatgtat tccagtctgt ggagtaccca ccgagccctt taaagtacag 1260 cagaggatat ttggaggata ctctacaaag attcaaagtt ttccttggca ggtctacttt 1320 gagtcccccc gaggtggcgg ggctcttatc gatgagtact gggtgctgac ggccgctcac 1380 gttgtggagg gaaactctga cccagtgatg tatgtcgggt ccacacttct gaaaatagag 1440 cggttgagaa atgcccagag gctcatcact gaacgtgtga ttattcatcc cagctggaaa 1500 caagaggacg acctgaatac acggacaaat tttgacaatg acattgccct ggtgcagctc 1560 aaagaccctg tgaaaatggg acccactgtt gcccccatct gcctgccaga aacctcctca 1620 gactacaacc cctcagaggg tgacctgggg ctgatctctg ggtggggccg aacagagaat 1680 agaaccaatg ttattcaact cagaggggcg aagttaccca taacatcttt agaaaagtgc 1740 cagcaggtga aagtggaaaa cccgaaagcg aggtcaaacg actatgtttt cactgacaac 1800 atgatctgtg ctggggaaaa gggtgtggac agctgtgaag gtgacagcgg aggggctttt 1860 gctctgccgg tccccaatgt caaggacccc aaattctatg tggctggcct ggtgtcctgg 1920 gggaaaaagt gtgggaccta tgggatctac acaaaggtaa agaactacgt ggactggatc 1980 ctgaaaacta tgcaggagaa tagtgggccc aagaaggact ag 2022 <210> 107 <211> 147 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s deletion mutant 1 (huC1sM151) <400> 107 Tyr His Gly Asp Pro Lys Glu Asp Thr Pro Asn Ser Val Trp Glu Pro 1 5 10 15 Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile Thr Cys Leu 20 25 30 Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr Ser Phe Tyr 35 40 45 Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys Leu Lys Cys 50 55 60 Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn Gly Lys Val 65 70 75 80 Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg Tyr Thr Cys 85 90 95 Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly Glu Tyr His 100 105 110 Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly Pro Glu Leu 115 120 125 Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro Phe Glu Glu 130 135 140 Lys Gln Arg 145 <210> 108 <211> 456 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polynucleotide <220> <223> Nucleotide sequence of recombinant human C1s deletion mutant 1 (huC1sM151) <400> 108 taccacggcg accccatgcc ttgtcctaaa gaggacaccc ctaactccgt gtgggagccc 60 gccaaggcca aatacgtgtt cagagatgtg gtccagatca cctgtctgga cggctttgag 120 gtggtggaag gcagagtggg cgccacctct ttctactcta cctgccagtc caacggcaag 180 tggtccaact ccaagctgaa gtgccagcct gtggactgcg gcatccctga gtctatcgag 240 aacggcaagg tggaagatcc cgagagcacc ctgttcggct ccgtgatcag atatacctgc 300 gaggaaccct actactacat ggaaaacggc ggaggcggcg agtatcactg tgctggcaat 360 ggctcttggg tcaacgaggt gctgggaccc gaactgccta agtgtgttcc tgtgtgtggc 420 gtgcccagag agcccttcga ggaaaagcag agatag 456 <210> 109 <211> 428 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Human C1s deletion mutant 2 (huC1sNHC) protein sequence <400> 109 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg His His His His His His 420 425 <210> 110 <211> 1269 <212> DNA <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polynucleotide <220> <223> Nucleotide sequence of recombinant human C1s deletion mutant 2 (huC1sNHC) <400> 110 gagcctacta tgtacggcga gatcctgtct cctaactacc ctcaggctta cccctccgag 60 gtggaaaagt cctgggatat cgaggtgccc gaaggctacg gcatccacct gtacttcacc 120 cacctggaca tcgagctgtc cgagaactgc gcctacgact ccgtgcagat catctccggc 180 gataccgaag agggcagact gtgcggccag cggtcctcta acaatcccca ctctcctatc 240 gtggaagagt tccaggtgcc atacaacaag ctgcaagtga tcttcaagtc cgacttctcc 300 aacgaggaac ggttcaccgg cttcgccgct tactatgtgg ccaccgacat caacgagtgc 360 accgacttcg tggacgtgcc ctgctctcac ttctgcaaca actttatcgg cggctacttc 420 tgcagctgcc ctcctgagta cttcctgcac gacgacatga agaactgcgg cgtgaactgc 480 tccggcgacg tgttcacagc tctgatcgga gagatcgcct ctccaaatta ccccaagcct 540 tatcctgaga actcccgctg cgagtaccag atcagactgg aaaagggctt ccaggtggtg 600 gtcaccctgc ggcgcgagga ttttgatgtg gaagccgctg attccgccgg caactgcctg 660 gattctctgg tgtttgtggc cggcgacaga cagttcggcc cttattgtgg ccatggcttc 720 cccggacctc tgaacatcga gacaaagagc aacgccctgg atatcatctt ccagaccgac 780 ctgaccggcc agaagaaagg ctggaagctg agataccacg gcgaccccat gccttgtcct 840 aaagaggaca cccctaactc cgtgtgggag cccgccaagg ccaaatacgt gttcagagat 900 gtggtccaga tcacctgtct ggacggcttt gaggtggtgg aaggcagagt gggcgccacc 960 tctttctact ctacctgcca gtccaacggc aagtggtcca actccaagct gaagtgccag 1020 cctgtggact gcggcatccc tgagtctatc gagaacggca aggtggaaga tcccgagagc 1080 accctgttcg gctccgtgat cagatatacc tgcgaggaac cctactacta catggaaaac 1140 ggcggaggcg gcgagtatca ctgtgctggc aatggctctt gggtcaacga ggtgctggga 1200 cccgaactgc ctaagtgtgt tcctgtgtgt ggcgtgccca gagagccctt cgaggaaaag 1260 cagagatag 1269 <210> 111 <211> 389 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Human C1s deletion mutant 3 (NHCD33) protein sequence <400> 111 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala 385 <210> 112 <211> 673 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M1 (M277I)-His <400> 112 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Ile Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 113 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M2 (M277I, K331Q)-His <400> 113 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Ile Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Gln Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 114 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M3 (M277I, P278, N329D)-His <400> 114 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Ile Ser Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asp Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 115 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M4 (P280A, E282K)-His <400> 115 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Ala Lys Lys Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 116 <211> 673 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M5 (K336G)-His <400> 116 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Gly 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 117 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M6 (P280A, E282K, K336G)-His <400> 117 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Ala Lys Lys Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Gly 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 118 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of human C1s M7 (D343Y, E351A)-His <400> 118 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Tyr Cys Gly Ile Pro Glu Ser Ile Ala Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 119 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: synthetic polypeptide <220> <223> Protein sequence of human C1s M8 (E351A)-His <400> 119 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Ala Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 120 <211> 673 <212> PRT <213> Artificial sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M9 (K336G, D343Y, E351A)-His <400> 120 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Gly 325 330 335 Leu Lys Cys Gln Pro Val Tyr Cys Gly Ile Pro Glu Ser Ile Ala Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 121 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M10 (D283I, P285A)-His <400> 121 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Ile Thr Ala Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 122 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M11 (S349P)-His <400> 122 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Pro Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 123 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M12 (G381E, G382E)-His <400> 123 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Glu Glu Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 124 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M13 (A292D)-His <400> 124 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Asp Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 125 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M14 (R316H)-His <400> 125 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly His Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 126 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M15 (S360N)-His <400> 126 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Asn Thr Leu Phe Gly Ser Val Ile Arg 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 620 Asp Pro Asn Asp Lys Thr Lys Phe Tyr Ala Ala Gly Leu Val Ser Trp 625 630 635 640 Gly Pro Gln Cys Gly Thr Tyr Gly Leu Tyr Thr Arg Val Lys Asn Tyr 645 650 655 Val Asp Trp Ile Met Lys Thr Met Gln Glu Asn Ser Thr Pro Arg Glu 660 665 670 Asp <210> 127 <211> 673 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequence: Synthetic polypeptide <220> <223> Protein sequence of human C1s M16 (R368H)-His <400> 127 Glu Pro Thr Met Tyr Gly Glu Ile Leu Ser Pro Asn Tyr Pro Gln Ala 1 5 10 15 Tyr Pro Ser Glu Val Glu Lys Ser Trp Asp Ile Glu Val Pro Glu Gly 20 25 30 Tyr Gly Ile His Leu Tyr Phe Thr His Leu Asp Ile Glu Leu Ser Glu 35 40 45 Asn Cys Ala Tyr Asp Ser Val Gln Ile Ile Ser Gly Asp Thr Glu Glu 50 55 60 Gly Arg Leu Cys Gly Gln Arg Ser Ser Asn Asn Pro His Ser Pro Ile 65 70 75 80 Val Glu Glu Phe Gln Val Pro Tyr Asn Lys Leu Gln Val Ile Phe Lys 85 90 95 Ser Asp Phe Ser Asn Glu Glu Arg Phe Thr Gly Phe Ala Ala Tyr Tyr 100 105 110 Val Ala Thr Asp Ile Asn Glu Cys Thr Asp Phe Val Asp Val Pro Cys 115 120 125 Ser His Phe Cys Asn Asn Phe Ile Gly Gly Tyr Phe Cys Ser Cys Pro 130 135 140 Pro Glu Tyr Phe Leu His Asp Asp Met Lys Asn Cys Gly Val Asn Cys 145 150 155 160 Ser Gly Asp Val Phe Thr Ala Leu Ile Gly Glu Ile Ala Ser Pro Asn 165 170 175 Tyr Pro Lys Pro Tyr Pro Glu Asn Ser Arg Cys Glu Tyr Gln Ile Arg 180 185 190 Leu Glu Lys Gly Phe Gln Val Val Val Thr Leu Arg Arg Glu Asp Phe 195 200 205 Asp Val Glu Ala Ala Asp Ser Ala Gly Asn Cys Leu Asp Ser Leu Val 210 215 220 Phe Val Ala Gly Asp Arg Gln Phe Gly Pro Tyr Cys Gly His Gly Phe 225 230 235 240 Pro Gly Pro Leu Asn Ile Glu Thr Lys Ser Asn Ala Leu Asp Ile Ile 245 250 255 Phe Gln Thr Asp Leu Thr Gly Gln Lys Lys Gly Trp Lys Leu Arg Tyr 260 265 270 His Gly Asp Pro Met Pro Cys Pro Lys Glu Asp Thr Pro Asn Ser Val 275 280 285 Trp Glu Pro Ala Lys Ala Lys Tyr Val Phe Arg Asp Val Val Gln Ile 290 295 300 Thr Cys Leu Asp Gly Phe Glu Val Val Glu Gly Arg Val Gly Ala Thr 305 310 315 320 Ser Phe Tyr Ser Thr Cys Gln Ser Asn Gly Lys Trp Ser Asn Ser Lys 325 330 335 Leu Lys Cys Gln Pro Val Asp Cys Gly Ile Pro Glu Ser Ile Glu Asn 340 345 350 Gly Lys Val Glu Asp Pro Glu Ser Thr Leu Phe Gly Ser Val Ile His 355 360 365 Tyr Thr Cys Glu Glu Pro Tyr Tyr Tyr Met Glu Asn Gly Gly Gly Gly 370 375 380 Glu Tyr His Cys Ala Gly Asn Gly Ser Trp Val Asn Glu Val Leu Gly 385 390 395 400 Pro Glu Leu Pro Lys Cys Val Pro Val Cys Gly Val Pro Arg Glu Pro 405 410 415 Phe Glu Glu Lys Gln Arg Ile Ile Gly Gly Ser Asp Ala Asp Ile Lys 420 425 430 Asn Phe Pro Trp Gln Val Phe Phe Asp Asn Pro Trp Ala Gly Gly Ala 435 440 445 Leu Ile Asn Glu Tyr Trp Val Leu Thr Ala Ala His Val Val Glu Gly 450 455 460 Asn Arg Glu Pro Thr Met Tyr Val Gly Ser Thr Ser Val Gln Thr Ser 465 470 475 480 Arg Leu Ala Lys Ser Lys Met Leu Thr Pro Glu His Val Phe Ile His 485 490 495 Pro Gly Trp Lys Leu Leu Glu Val Pro Glu Gly Arg Thr Asn Phe Asp 500 505 510 Asn Asp Ile Ala Leu Val Arg Leu Lys Asp Pro Val Lys Met Gly Pro 515 520 525 Thr Val Ser Pro Ile Cys Leu Pro Gly Thr Ser Ser Asp Tyr Asn Leu 530 535 540 Met Asp Gly Asp Leu Gly Leu Ile Ser Gly Trp Gly Arg Thr Glu Lys 545 550 555 560 Arg Asp Arg Ala Val Arg Leu Lys Ala Ala Arg Leu Pro Val Ala Pro 565 570 575 Leu Arg Lys Cys Lys Glu Val Lys Val Glu Lys Pro Thr Ala Asp Ala 580 585 590 Glu Ala Tyr Val Phe Thr Pro Asn Met Ile Cys Ala Gly Gly Glu Lys 595 600 605 Gly Met Asp Ser Cys Lys Gly Asp Ser Gly Gly Ala Phe Ala Val Gln 610 615 62...

Claims

1. A recombinant anti-C1s antibody or antigen-binding fragment thereof that is capable of specifically binding to human C1s and inhibiting C1s, wherein the recombinant anti-C1s antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), the VH comprises three heavy chain complementarity determining regions (HC CDRs), and the VL comprises three light chain complementarity determining regions (LC CDRs), wherein: (a) the VH comprises: HC CDR1 consisting of the amino acid sequence shown in SEQ ID NO:129, HC CDR2 consisting of the amino acid sequence shown in SEQ ID NO:3, and HC CDR3 consisting of the amino acid sequence shown in SEQ ID NO:4, and the VL comprises: LC CDR1 consisting of the amino acid sequence shown in SEQ ID NO:132, LC CDR2 consisting of the amino acid sequence shown in SEQ ID NO:133, and LC CDR3 consisting of the amino acid sequence shown in SEQ ID NO:134, or (b) the VH comprises: HC CDR1 consisting of the amino acid sequence shown in SEQ ID NO:22, HC CDR2 consisting of the amino acid sequence shown in SEQ ID NO:129, and HC CDR3 consisting of the amino acid sequence shown in SEQ ID NO:4, and the VL comprises: LC CDR1 consisting of the amino acid sequence shown in SEQ ID NO:132, LC CDR2 consisting of the amino acid sequence shown in SEQ ID NO:133, and LC CDR3 consisting of the amino acid sequence shown in SEQ ID NO:

134.

2. The anti-C1s antibody according to claim 1, wherein the antibody is capable of binding to residues K336, R316 of human C1s (SEQ ID NO:99) or a region encompassing A390 - R422.

3. The anti-C1s antibody according to claim 1, wherein the anti-C1s antibody is capable of inhibiting the activity of the classical pathway (CP) of complement activation.

4. The anti-C1s antibody according to claim 3, wherein the anti-C1s antibody is capable of performing at least one of the following: inhibiting downstream effects of C1s activation, inhibiting IgM-induced deposition of C4 from serum, inhibiting antibody-antigen complex-mediated C4 cleavage, inhibiting antibody-antigen complex-mediated C4b deposition, inhibiting formation of C3 convertase, and inhibiting downstream effects of CP activation.

5. The anti-C1s antibody according to claim 1, wherein the antibody exhibits cross-reactivity with at least one non-human C1s.

6. The anti-C1s antibody according to claim 5, wherein the non-human C1s is cynomolgus monkey C1s.

7. The anti-C1s antibody according to claim 6, wherein the anti-C1s antibody exhibits cross-reactivity with rat C1s.

8. The anti-C1s antibody according to claim 1, wherein the antibody is at least one of a chimeric antibody and its antigen-binding fragment.

9. The anti-C1s antibody according to claim 1, wherein the antibody is at least one of a humanized antibody and an antigen-binding fragment thereof.

10. A recombinant anti-C1s antibody or an antigen-binding fragment thereof, which is capable of specifically binding to human C1s and inhibiting C1s, wherein the anti-C1s antibody is one of the following: a. An antibody, the antibody comprising: a heavy chain (HC) polypeptide, which consists of the amino acid sequence of SEQ ID NO:71; and a light chain (LC) polypeptide, which consists of the amino acid sequence of SEQ ID NO:73; b. An antibody, which comprises: an HC polypeptide, which consists of the amino acid sequence of SEQ ID NO:79; and an LC polypeptide, which consists of the amino acid sequence of SEQ ID NO:81; or c. An antibody, which comprises: an HC polypeptide, which consists of the amino acid sequence of SEQ ID NO:83; and an LC polypeptide, which consists of the amino acid sequence of SEQ ID NO:

85.

11. A recombinant anti-C1s antibody or an antigen-binding fragment thereof, which is capable of specifically binding to human C1s and inhibiting C1s, wherein the anti-C1s antibody is one of the following: a. An antibody, the antibody comprising: a heavy chain variable region (VH), which consists of the amino acid sequence of SEQ ID NO:1; and a light chain variable region (VL), which consists of the amino acid sequence of SEQ ID NO:6; b. An antibody, which comprises: a VH, which consists of the amino acid sequence of SEQ ID NO:21; and a VL, which consists of the amino acid sequence of SEQ ID NO:26; or c. An antibody, which comprises: a VH, which consists of the amino acid sequence of SEQ ID NO:31; and a VL, which consists of the amino acid sequence of SEQ ID NO:

36.

12. The anti-C1s antibody according to claim 1, wherein the antibody comprises HC CDR1 of SEQ ID NO:22, HC CDR2 of SEQ ID NO:23, HC CDR3 of SEQ ID NO:24, LC CDR1 of SEQ ID NO:27, LC CDR2 of SEQ ID NO:28, and LC CDR3 of SEQ ID NO:

29.

13. The anti-C1s antibody according to claim 1, wherein the antibody comprises HC CDR1 of SEQ ID NO:52, HC CDR2 of SEQ ID NO:53, HC CDR3 of SEQ ID NO:54, LC CDR1 of SEQ ID NO:57, LC CDR2 of SEQ ID NO:58, and LC CDR3 of SEQ ID NO:

59.

14. Use of an anti-C1s antibody according to claim 1 in the preparation of a medicament for treating a complement-mediated disorder in a subject in need thereof, wherein the medicament inhibits the activity of the CP, wherein the complement-mediated disorder is an autoimmune disorder, and the autoimmune disorder is selected from one of immune thrombocytopenic purpura (ITP) and neuromyelitis optica (NMO).

15. The use according to claim 14, wherein the medicament inhibits at least one of C1s activation, antibody-antigen complex-mediated C4 cleavage, and formation of common complement pathway effectors.

16. The use according to claim 14, wherein the medicament is prepared for administration during the acute phase of the complement-mediated disorder.

17. A pharmaceutical composition comprising the anti-C1s antibody according to claim 1 and a pharmaceutically acceptable carrier and / or excipient.

18. An isolated nucleic acid molecule encoding at least a portion of the anti-C1s antibody according to claim 1, wherein the nucleic acid comprises at least one nucleotide sequence selected from the following: a. a heavy chain (HC) nucleotide sequence selected from: a nucleotide sequence consisting of the sequence of SEQ ID NO:5, a nucleotide sequence consisting of the sequence of SEQ ID NO:15, a nucleotide sequence consisting of the sequence of SEQ ID NO:25, a nucleotide sequence consisting of the sequence of SEQ ID NO:35, a nucleotide sequence consisting of the sequence of SEQ ID NO:45, a nucleotide sequence consisting of the sequence of SEQ ID NO:55, and a nucleotide sequence consisting of the sequence of SEQ ID NO:65, and b. a light chain (LC) nucleotide sequence selected from: a nucleotide sequence consisting of the sequence of SEQ ID NO:10, a nucleotide sequence consisting of the sequence of SEQ ID NO:20, a nucleotide sequence consisting of the sequence of SEQ ID NO:30, a nucleotide sequence consisting of the sequence of SEQ ID NO:40, a nucleotide sequence consisting of the sequence of SEQ ID NO:50, a nucleotide sequence consisting of the sequence of SEQ ID NO:60, and a nucleotide sequence consisting of the sequence of SEQ ID NO:

70.

19. A vector comprising the nucleic acid molecule according to claim 18.

20. A host cell comprising the vector according to claim 19.

Images