Soluble complement receptor type 1 variant and uses thereof

Abstract

The present application provides methods of inhibiting complement activity in a subject, the methods comprising administering to the subject a soluble complement receptor type 1 (sCR1) variant.

Description

[0001] Relevant application data

[0002] This application claims priority to Australian Patent Application No. 2018901703, filed on May 16, 2018, entitled “Soluble complement receptortype I variants and uses thereof”. The entire contents of that Australian patent application are incorporated herein by reference.

[0003] sequence list

[0004] This application is filed together with the sequence list in electronic form. The entire contents of the sequence list are incorporated herein by reference. Technical Field

[0005] This disclosure relates to type 1 soluble complement receptor variants and their uses. Background Technology

[0006] The complement system is part of the innate immune system and includes many cell surface soluble proteins that play an important role in eliminating foreign microorganisms while protecting the host from complement-related damage.

[0007] The complement system comprises soluble components C1-C9. It is activated when its major components are fragmented and these fragments, alone or in combination with other proteins, activate other complement proteins, leading to a proteolytic cascade. Activation of the complement system results in increased vascular permeability, chemotaxis of phagocytes, activation of inflammatory cells, opsonization of foreign granules, direct cell killing, and tissue damage.

[0008] All three pathways of the complement system ultimately lead to the formation of the membrane attack complex (MAC) (comprising complement components C5b, C6, C7, C8, and C9) and the release of anaphylactic toxins C3a and C5a. However, each pathway is triggered differently. The classical pathway is triggered in response to the antigen-antibody complex and involves the activation of complement component C4 by complement component C1s, leading to the successive activation of complement components C2, C3, and C5 and the formation of MAC. The lectin pathway involves the activation of mannan-binding lectin serine protease 1 (MASP1) and MASP2 via the binding of mannose-binding lectin (MBL) to the corresponding carbohydrate on the pathogen surface. Activated MASP1 / MASP2 activates C4, leading to the successive activation of complement components C2, C3, and C5 and the formation of MAC. Unlike the classical and lectin pathways, the alternative pathway does not involve the activation of C4 and C2. Instead, it is triggered by the pathogen surface, leading to the activation of C3 via factors B, D, and properdin, followed by the activation of C5 and the formation of MAC. C3 and C5 can also be activated by proteins in the coagulation cascade.

[0009] Complement receptor type 1 (CR1) is a major regulator of complement activation. CR1 (also known as the C3b / C4b receptor) is a membrane-bound protein found on erythrocytes, macrophages / monocytes, granulocytes, B cells, some T cells, splenic follicular dendritic cells, and glomerular podocytes. A small amount of soluble CR1 (sCR1) is derived from the cleavage of cell surface CR1; a recombinant form of this soluble molecule, called TP10, has previously been produced. CR1 is a negative regulator of C3 activation; therefore, sCR1 can inhibit each of the classical, lectin, and alternative pathways.

[0010] sCR1 has a relatively short half-life of about 70 hours (3 days) (Zimmerman et al., 2000 Crit CareMed 28:3149-3154). sCR1 variants with one or more amino acid substitutions and / or truncated sCR1 variants that retain complement-inhibiting activity have been previously described (e.g., WO1994000571).

[0011] However, it will be apparent to those skilled in the art that there is a persistent need in the art for sCR1 variants with improved activity (e.g., increased complement inhibitory activity) and / or increased half-life. Summary of the Invention

[0012] This disclosure is based on the inventors’ identification of a type 1 soluble complement receptor (sCR1) variant in subjects that exhibits increased inhibitory complement activity.

[0013] In the process of generating this disclosure, the inventors produced truncated variants of sCR1 comprising defined amino acid sequences corresponding to one or more long homologous repeat (LHR) regions (i.e., LHR-A, LHR-B, LHR-C, and / or LHR-D). The inventors investigated the effect of each sCR1 variant on complement inhibitory activity. The truncated variants of sCR1 of this disclosure exhibit improved or increased inhibitory activity in all three complement pathways. The inventors have determined that sCR1 variants comprising residues 42 to 939 and / or residues 490 to 1392 of SEQ ID NO: 1 have increased inhibitory activity compared to another form of sCR1.

[0014] The inventors' discovery provides a basis for methods to inhibit complement activity in subjects, including administering an sCR1 variant to the subject. The inventors' discovery also provides a basis for methods to treat or prevent conditions in subjects, such as complement-mediated conditions.

[0015] This disclosure provides a method for inhibiting complement activity in a subject, the method comprising administering to the subject a type 1 soluble complement receptor (sCR1) variant comprising an amino acid sequence selected from:

[0016] (i) The amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1; and

[0017] (ii) The amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1.

[0018] In one instance, the sCR1 variant includes:

[0019] (i) The amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., amino acid residues 1393 to 1971 of SEQ ID NO: 1 are missing);

[0020] (ii) The amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1 (e.g., lacking amino acid residues 940 to 1971 of SEQ ID NO: 1);

[0021] (iii) The amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1 to 489 and 1393 to 1971 of SEQ ID NO: 1); or

[0022] (iv) The amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1 (e.g., amino acid residues 1 to 489 of SEQ ID NO: 1 are missing).

[0023] In one instance, the sCR1 variant contains the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1).

[0024] In one instance, the sCR1 variant contains the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1 (e.g., lacking amino acid residues 940 to 1971 of SEQ ID NO: 1).

[0025] In one instance, the sCR1 variant contains the amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1 to 489 and 1393 to 1971 of SEQ ID NO: 1).

[0026] In one instance, the sCR1 variant contains the amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1 to 489 of SEQ ID NO: 1).

[0027] In one instance, the sCR1 variant includes:

[0028] (i) The amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1;

[0029] (ii) The amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1;

[0030] (iii) The amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1; or

[0031] (iv) The amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1.

[0032] In one instance, the sCR1 variant consists of or contains the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1). The inventors have shown that such sCR1 variants exhibit improved complement-inhibiting activity compared to sCR1 variants containing amino acids 42 to 1971 of SEQ ID NO: 1. This finding is unexpected because the CR1 region in amino acids 1393 to 1971 binds to C1q and mannose-binding lectin (MBL), and its removal could reasonably be expected to be detrimental or ineffective for complement-inhibiting activity.

[0033] In one instance, the sCR1 variant consists of an amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1.

[0034] In one instance, the sCR1 variant consists of an amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1.

[0035] In one instance, the sCR1 variant consists of an amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1.

[0036] In one instance, the sCR1 variant does not consist of or contain the amino acid sequence corresponding to amino acids 1 to 1971 of SEQ ID NO: 1.

[0037] In one instance, the sCR1 variant does not consist of or contain the amino acid sequence corresponding to amino acids 42 to 1971 of SEQ ID NO: 1.

[0038] In one instance, the sCR1 variant of this disclosure optionally includes one or more amino acid substitutions, deletions, or insertions of any sequence disclosed herein. Amino acid substitutions applicable to this disclosure will be apparent to those skilled in the art and include both naturally occurring substitutions and engineered substitutions.

[0039] In one instance, the sCR1 variant of this disclosure contains one or more conserved amino acid substitutions compared to the sequence disclosed herein. In some instances, the sCR1 variant contains 10 or fewer, such as 9 or 8 or 7 or 6 or 5 or 4 or 3 or 2 or 1 conserved amino acid substitutions.

[0040] In one instance, the sCR1 variant of this disclosure includes one or more non-conservative amino acid alterations. For example, non-conservative amino acid substitutions increase the half-life of the sCR1 variant of this disclosure, decrease immunogenicity, and / or increase inhibitory activity. In one instance, the sCR1 variant includes fewer than 6, 5, 4, 3, 2, or 1 non-conservative amino acid substitutions.

[0041] In one instance, the sCR1 variant of this disclosure contains at least about 85% or about 90% or about 95% or about 97% or about 98% or about 99% of the sequence disclosed herein.

[0042] In one example, the sCR1 variant comprises at least about 85%, or about 90%, or about 97%, or about 98%, or about 99% of the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1). For example, the sCR1 variant comprises about 85% of the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1. In another example, the sCR1 variant comprises about 90% of the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1. In another example, the sCR1 variant comprises about 95% of the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1. In yet another example, the sCR1 variant comprises about 97% of the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1. In one instance, the sCR1 variant contains approximately 98% identical amino acid sequences to amino acids 42 to 1392 corresponding to SEQ ID NO: 1. In another instance, the sCR1 variant contains approximately 99% identical amino acid sequences to amino acids 42 to 1392 corresponding to SEQ ID NO: 1.

[0043] In one example, the sCR1 variant comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1. For example, the sCR1 variant contains about 85% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1. In another example, the sCR1 variant contains about 90% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1. In another example, the sCR1 variant contains about 95% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1. In yet another example, the sCR1 variant contains about 97% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1. In one example, the sCR1 variant contains about 98% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1. In another instance, the sCR1 variant contains approximately 99% of the same amino acid sequence as amino acids 42 to 939 corresponding to SEQ ID NO: 1.

[0044] In one example, the sCR1 variant comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1. For example, the sCR1 variant contains about 85% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1. In another example, the sCR1 variant contains about 90% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1. In yet another example, the sCR1 variant contains about 95% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1. In yet another example, the sCR1 variant contains about 97% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1. In one instance, the sCR1 variant contains approximately 98% of the same amino acid sequence as amino acids 490 to 1392 corresponding to SEQ ID NO: 1. In another instance, the sCR1 variant contains approximately 99% of the same amino acid sequence as amino acids 490 to 1392 corresponding to SEQ ID NO: 1.

[0045] In one example, the sCR1 variant comprises an amino acid sequence that is at least about 85%, about 90%, about 95%, about 97%, about 98%, or about 99% identical to the amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1. For example, the sCR1 variant contains about 85% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1. In another example, the sCR1 variant contains about 90% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1. In yet another example, the sCR1 variant contains about 95% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1. In yet another example, the sCR1 variant contains about 97% of the amino acid sequence that is identical to the amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1. In one instance, the sCR1 variant contains approximately 98% of the same amino acid sequence as amino acids 490 to 1971 corresponding to SEQ ID NO: 1. In another instance, the sCR1 variant contains approximately 99% of the same amino acid sequence as amino acids 490 to 1971 corresponding to SEQ ID NO: 1.

[0046] In one instance, the sCR1 variant of this disclosure exhibits increased inhibitory activity compared to sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the complement inhibitory activity of the sCR1 variant of this disclosure is increased by at least about 1.5 times, or about 2 times, or about 3 times, or about 3.5 times, or about 4 times, or about 5 times, or about 6 times, or about 8 times, or about 10 times compared to sCR1 containing the sequence shown in SEQ ID NO: 2.

[0047] Methods for determining the inhibitory activity of sCR1 variants are obvious to those skilled in the art and / or described herein. In one example, in vitro assays are used to determine complement inhibitory activity. For example, complement activity is determined using enzyme immunoassays (e.g., immunoassays measuring complement activation, such as...). The complement assay kit is used to determine the complement inhibitory activity. For example, a labeled antibody specific to the antigen or epitope generated during complement activation (e.g., an epitope present in C5b-9 or C5b-C9) is used to determine complement inhibitory activity. In one example, the wells of a microtiter plate are coated with a specific activator of the classical, lectin, or alternative pathways. In one example, the sCR1 variant is incubated with normal human serum and an appropriate assay diluent (i.e., a diluent containing appropriate blocking components to ensure specific activation of the classical, lectin, or alternative pathways) and added to the wells of a microtiter plate coated with a specific activator of the classical, lectin, or alternative pathways. The amount of C5b-9 complex formed is detected using an antibody labeled with an alkaline phosphatase specific for C5b-9. In one example, the amount of complement activation product (i.e., C5b-9) generated is proportional to the functional activity of the complement pathway. In one example, the half-maximal inhibitory concentration (i.e., IC50) is determined. 50 For example, determining the IC50 of sCR1 variants. 50 And it is compared with an IC containing sCR1, which contains the sequence shown in SEQ ID NO: 2. 50 Comparisons were made. In another instance, complement inhibitory activity was determined using hemolysis assays (e.g., classical pathway (i.e., CH50) and alternative pathway (ApH50) inhibition assays).

[0048] In one instance, the sCR1 variant exhibited increased inhibitory activity in the classical pathway, lectin pathway, and / or complement bypass pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2.

[0049] In one instance, the sCR1 variant exhibits increased inhibitory activity in the classical complement pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the sCR1 variant of this disclosure exhibits at least a 1.25-fold, or about 1.5-fold, or about 1.75-fold, or about 2-fold, or about 2.5-fold, or about 3-fold, or about 3.5-fold, or about 4-fold, or about 5-fold increase in inhibitory activity in the classical complement pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2.

[0050] In one instance, the sCR1 variant of this disclosure showed an IC50 value in a classic complement assay (e.g., the Wieslab complement assay). 50 Less than sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the sCR1 variant of this disclosure has an IC50 value in classical complement assays (e.g., Wieslab complement assay). 50 Less than about 1.0 nM, for example, about 0.95 nM, about 0.90 nM, about 0.85 nM, about 0.80 nM, about 0.75 nM, or about 0.70 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value of less than 1.0 nM in a classic complement assay (e.g., Wieslab complement assay). 50 The concentration is approximately 0.85 nM to 0.90 nM, for example, approximately 0.88 nM. In one instance, the IC50 of the sCR1 variant of this disclosure in a classic complement assay (e.g., Wieslab complement assay) is... 50 Less than about 0.65 nM, or about 0.60 nM, or about 0.55 nM, or about 0.50 nM, or about 0.45 nM, or about 0.40 nM, or about 0.35 nM, or about 0.30 nM, or about 0.25 nM, or about 0.20 nM, or about 0.15 nM, or about 0.10 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value of less than about 0.65 nM, or about 0.60 nM, or about 0.55 nM, or about 0.50 nM, or about 0.25 nM, or about 0.20 nM, or about 0.15 nM, or about 0.10 nM. 50 It ranges from about 0.35 nM to 0.45 nM, for example, about 0.40 nM.

[0051] In one instance, the sCR1 variant exhibits increased inhibitory activity in the lectin complement pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the sCR1 variant of this disclosure exhibits at least a 1.25-fold, or about 1.5-fold, or about 1.75-fold, or about 2-fold, or about 2.5-fold, or about 3-fold, or about 3.5-fold, or about 4-fold, or about 5-fold increase in inhibitory activity in the lectin complement pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2.

[0052] In one instance, the sCR1 variant of this disclosure showed an IC50 score in a lectin complement assay (e.g., the Wieslab complement assay). 50 Smaller than sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the sCR1 variant of this disclosure has an IC50 value in lectin complement assays (e.g., Wieslab complement assays). 50 Less than about 0.60 nM, or about 0.55 nM, or about 0.50 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value of less than 0.60 nM in a lectin complement assay (e.g., Wieslab complement assay). 50 The concentration is approximately 0.50 nM to 0.60 nM, for example, approximately 0.547 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value of approximately 0.547 nM in a lectin complement assay (e.g., Wieslab complement assay). 50 Less than about 0.50 nM, or about 0.45 nM, or about 0.40 nM, or about 0.35 nM, or about 0.30 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value of less than 0.50 nM in a lectin complement assay (e.g., Wieslab complement assay). 50 It ranges from about 0.40 nM to 0.45 nM, for example, about 0.43 nM.

[0053] In one instance, the sCR1 variant exhibits increased inhibitory activity in the complement bypass pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the sCR1 variant of this disclosure exhibits at least a 1.25-fold, or about 1.5-fold, or about 1.75-fold, or about 2-fold, or about 2.5-fold, or about 3-fold, or about 3.5-fold, or about 4-fold, or about 5-fold increase in inhibitory activity in the complement bypass pathway compared to the sCR1 containing the sequence shown in SEQ ID NO: 2.

[0054] In one instance, the sCR1 variant of this disclosure showed an IC50 value in a complement bypass assay (e.g., a Wieslab complement assay). 50 Smaller than sCR1 containing the sequence shown in SEQ ID NO: 2. For example, the sCR1 variant of this disclosure has an IC50 value in complement bypass assays (e.g., Wieslab complement assays). 50 Less than about 0.75 nM, or about 0.70 nM, or about 0.65 nM, or about 0.60 nM, or about 0.55 nM, or about 0.50 nM, or about 0.45 nM, or about 0.40 nM, or about 0.35 nM, or about 0.30 nM, or about 0.25 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value in a complement bypass assay (e.g., Wieslab complement assay). 50The concentration is approximately 0.35 nM to approximately 0.40 nM, for example, approximately 0.38 nM. In one instance, the sCR1 variant of this disclosure showed an IC50 value of approximately 0.35 nM in a complement bypass assay (e.g., Wieslab complement assay). 50 It ranges from about 0.25 nM to about 0.30 nM, for example, about 0.27 nM.

[0055] In one instance, the sCR1 variant of this disclosure contains a long homologous repeat (LHR) region selected from the following:

[0056] (i) LHR-A and LHR-B;

[0057] (ii) LHR-A, LHR-B and LHR-C;

[0058] (iii) LHR-B and LHR-C; and

[0059] (iv) LHR-B, LHR-C and LHR-D.

[0060] In one instance, the sCR1 variant of this disclosure includes an LHR region consisting of LHR-A and LHR-B but lacking LHR-C and LHR-D.

[0061] In one instance, the sCR1 variant of this disclosure includes an LHR region consisting of LHR-A, LHR-B, and LHR-C but lacking LHR-D.

[0062] In one instance, the sCR1 variant of this disclosure includes an LHR region consisting of LHR-B and LHR-C but lacking LHR-A and LHR-D.

[0063] In one instance, the sCR1 variant of this disclosure includes an LHR region consisting of LHR-B, LHR-C, and LHR-D but lacking LHR-A.

[0064] In one example, the LHR region LHR-A contains the amino acid sequence corresponding to amino acids 42 to 489 of SEQ ID NO: 1. For example, the LHR-A region contains the amino acid sequence shown in SEQ ID NO: 13. In one example, the LHR region LHR-A contains short common repeat (SCR) sequences 1 to 7. For example, SCR sequences 1 to 3 (i.e., site 1) are capable of binding C4b.

[0065] In one example, the LHR region LHR-B contains the amino acid sequence corresponding to amino acids 490 to 939 of SEQ ID NO: 1. For example, the LHR-B region contains the amino acid sequence shown in SEQ ID NO: 14. In one example, the LHR region LHR-B contains SCR sequences 8 to 14. For example, SCR sequences 8 to 10 (i.e., site 2) are capable of binding C3b and C4b.

[0066] In one example, the LHR region LHR-C contains the amino acid sequence corresponding to amino acids 940 to 1392 of SEQ ID NO: 1. For example, the LHR-C region contains the amino acid sequence shown in SEQ ID NO: 15. In one example, the LHR region LHR-C contains SCR sequences 15 to 21. For example, SCR sequences 15 to 17 are capable of binding C3b and C4b.

[0067] In one example, the LHR region LHR-D contains the amino acid sequence corresponding to amino acids 1393 to 1971 of SEQ ID NO: 1. For example, the LHR-D region contains the amino acid sequence shown in SEQ ID NO: 16. In one example, the LHR region LHR-D contains SCR sequences 22 to 28. For example, SCR sequences 22 to 28 are capable of binding C1q and MBL.

[0068] In one instance, the sCR1 variant of this disclosure comprises (or consists of) an SCR sequence selected from the following:

[0069] (i) SCR-1 to SCR-14 (e.g., missing SCR-15 to SCR-28);

[0070] (ii) SCR-1 to SCR-21 (e.g., missing SCR-22 to SCR-28);

[0071] (iii) SCR-8 to SCR-21 (e.g., missing SCR-1 to SCR-7 and SCR-22 to SCR-28); and

[0072] (iv) SCR-8 to SCR-28 (e.g., SCR-1 to SCR-7 are missing).

[0073] In one instance, the sCR1 variant of this disclosure includes SCR sequences SCR-1 through SCR-14 (e.g., lacking SCR-15 through SCR-28).

[0074] In one instance, the sCR1 variant of this disclosure includes SCR sequences SCR-1 to SCR-21 (e.g., lacking SCR-22 to SCR-28).

[0075] In one instance, the sCR1 variant of this disclosure includes SCR sequences SCR-8 to SCR-21 (e.g., lacking SCR-1 to SCR-7 and SCR-22 to SCR-28).

[0076] In one instance, the sCR1 variant of this disclosure includes SCR sequences SCR-8 to SCR-28 (e.g., lacking SCR-1 to SCR-7).

[0077] In one instance, the sCR1 variant is a monolithic variant (i.e., a single copy of the sCR1 variant).

[0078] In one instance, the sCR1 variant is a dimer or dimerized (i.e., two copies of the sCR1 variant are linked together in the fusion protein).

[0079] In one instance, the sCR1 variant is a multimer or polymerized (i.e., multiple copies of the sCR1 variant are linked in the fusion protein).

[0080] In one instance, two or more identical sCR1 variants are fused (i.e., expressed as a fusion protein).

[0081] In one instance, two or more different sCR1 variants are fused (i.e., expressed as a fusion protein).

[0082] In one instance, dimerized or multimerized sCR1 variants contain a connector between sCR1 variants.

[0083] In one instance, this disclosure provides a multimeric protein comprising two or more sCR1 variants containing multimerized domains, wherein the multimerized domains interact to form the multimeric protein.

[0084] In one instance, each sCR1 variant in the multimeric protein contains one sCR1 variant. In another instance, one or more sCR1 variants in the multimeric protein contain two or more sCR1 variants, for example, sCR1 variants are linked in a fusion protein.

[0085] In one instance, the polymerized domain contains an immunoglobulin hinge domain.

[0086] In one instance, the polymerizing domain is a leucine zipper domain, a cysteine ​​knot, or an antibody Fc region.

[0087] In one instance, the polymerized sCR1 variant is linear.

[0088] In one instance, the polymerized sCR1 variant is cyclic.

[0089] This disclosure provides sCR1 variants conjugated to a half-life-extending moiety or another soluble complement inhibitor, as described herein in any instance (e.g., the description of sCR1 variants relating to methods of inhibiting complement activity should apply to the following description of the sCR1 variants themselves). In one instance, the sCR1 variant is chemically conjugated to a half-life-extending moiety or another soluble complement inhibitor. In another instance, the sCR1 variant is fused to a half-life-extending moiety or another soluble complement inhibitor, for example, expressed as a fusion protein. In one instance, the half-life-extending moiety or another soluble complement inhibitor is conjugated to the C-terminus of the sCR1 variant. In one instance, the half-life-extending moiety or another soluble complement inhibitor is conjugated to the N-terminus of the sCR1 variant.

[0090] In one instance, a variant of the sCR1 disclosed herein is conjugated to the half-life extension portion. For example, the half-life extension portion is selected from: albumin or a functional fragment or variant thereof, human serum albumin or a functional fragment or variant thereof, immunoglobulin or a functional fragment thereof, afamin, alpha-fetoprotein, vitamin D-binding protein, and polymers.

[0091] In one instance, the immunoglobulin or a functional fragment thereof is an antibody fragment that binds to albumin. For example, the half-life-extending portion is an antibody Fc region (i.e., a monomeric or dimer immunoglobulin Fc region), such as a human IgG1 Fc region, a human IgG4 Fc region, or a stable human IgG4 Fc region. For example, the Fc region is a human IgG4 Fc region. In one instance, the antibody Fc region is modified to prevent dimerization (e.g., as discussed herein). For example, the antibody Fc region is a monomeric Fc region. In one instance, the Fc fragment and / or its variants contain one or more amino acid substitutions, deletions, or insertions. Amino acid substitutions suitable for this disclosure will be apparent to those skilled in the art and include naturally occurring substitutions and engineered substitutions, such as those described in WO2000042072, WO2002060919, WO2004035752, and WO2006053301.

[0092] In one instance, the sCR1 variant is fused to the antibody Fc region at its C-terminus. For example, the sCR1 variant consists of or contains an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1), and is fused to the antibody Fc region at its C-terminus. The inventors found that this fusion protein exhibits greater complement-inhibiting activity than expected compared to proteins lacking the Fc region. Considering Fc region dimerization, an approximately two-fold increase in activity could be expected, but the inventors observed an improvement of up to approximately eight-fold.

[0093] In one instance, the conjugated sCR1 variant of this disclosure has a longer serum half-life compared to sCR1 variant conjugates containing the sCR1 shown in SEQ ID NO: 2. Examples of increased serum half-life and the assays for determining serum half-life are described herein and should be adapted as necessary to apply to this instance of the disclosure.

[0094] In one instance, the extended half-life portion is albumin, its functional fragment, or a variant thereof. In one instance, albumin, its functional fragment, or a variant thereof is serum albumin, such as human serum albumin. In one instance, albumin, its functional fragment, or a variant thereof contains one or more amino acid substitutions, deletions, or insertions, for example, no more than 5, 4, 3, 2, or 1 substitution. Amino acid substitutions suitable for use in this disclosure will be apparent to those skilled in the art and include both naturally occurring substitutions and engineered substitutions, such as those described in, for example, WO2011051489, WO2014072481, WO2011103076, WO2012112188, WO2013075066, WO2015063611, WO2014179657, and WO2019075519.

[0095] In one instance, the sCR1 variant is fused at its C-terminus to albumin, a functional fragment thereof, or a variant thereof. For example, the sCR1 variant consists of or contains an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino residues 1393 to 1971 of SEQ ID NO: 1), and is fused at its C-terminus to albumin (e.g., serum albumin), a functional fragment thereof, or a variant thereof.

[0096] In one instance, other proteins that are structurally or evolutionarily related to albumin can be used as the extended half-life component, including but not limited to alpha-fetoprotein (WO 2005024044; Beattie and Dugaiczyk, 20 Gene 415-422, 1982), afamin (Lichenstein et al. 269(27) J. Biol. Chem. 18149-18154, 1994) and vitamin D-binding protein (Cooke and David, 76 J. Clin. Invest. 2420-2424, 1985).

[0097] In one instance, the extended half-life portion is alpha-fetoprotein.

[0098] In one instance, the extended half-life portion is afamin.

[0099] In one instance, the extended half-life portion is due to vitamin D-binding proteins.

[0100] In one instance, the extended half-life portion is an immunoglobulin or a functional fragment thereof. In one instance, the immunoglobulin contains an Fc region. For example, Ig is an Fc domain or an Fc fragment and / or a variant thereof. In one instance, Ig is part of an immunoglobulin constant domain. In one instance, the immunoglobulin is an antibody fragment that binds to albumin.

[0101] In one instance, the extended half-life portion is a polymer. Polymers suitable for use in this disclosure will be apparent to those skilled in the art, and include, for example, polyethylene glycol. In one instance, the polymer comprises mono- or poly(e.g., 2-4)-polyethylene glycol (PEG). In one instance, the polymer is PEG.

[0102] In one instance, the sCR1 variant of this disclosure is conjugated with another soluble complement inhibitor. For example, the other soluble complement inhibitor is selected from: C1 inhibitors (C1-INH), factor I (fI), factor H (fH), complement factor H-associated protein (CFHR), C4b-binding protein (C4bp), soluble CD55 (decay-promoting factor (DAF)), soluble CD46 (membrane cofactor protein (MCP)), soluble CD59 (protectin), soluble complement receptor 2 (sCR2), TT30 (CR2-fH), and cobra venom factor (CVF).

[0103] In one instance, another soluble complement inhibitor is the C1-inhibitor (C1-INH).

[0104] In one instance, another soluble complement inhibitor is factor I (fI).

[0105] In one instance, another soluble complement inhibitor is factor H (fH). In another instance, another soluble complement inhibitor is complement factor H-related protein (CFHR). For example, complement factor H-related protein is selected from CFHR1, CFHR2, CFHR3, CFHR4, and CFHR5.

[0106] In one instance, another soluble complement inhibitor is C4b-binding protein (C4bp).

[0107] In one instance, another soluble complement inhibitor is soluble CD55 (decay promoter (DAF)).

[0108] In one instance, another soluble complement inhibitor is soluble CD46 (membrane cofactor protein (MCP)).

[0109] In one instance, another soluble complement inhibitor is soluble CD59 (Protein).

[0110] In one instance, another soluble complement inhibitor is soluble complement receptor 2 (sCR2).

[0111] In one instance, another soluble complement inhibitor is TT30 (CR2-fH).

[0112] In one instance, another soluble complement inhibitor is cobra venom factor (CVF).

[0113] In one instance, the sCR1 variant of this disclosure is an sCR1 variant glycoform. For example, the sCR1 variant glycoform is a sialylated sCR1 variant glycoform. In one instance, the sCR1 variant glycoform in the composition comprises a sialylated glycan. For example, the sialylated sCR1 variant glycoform comprises at least one sialylated glycan (e.g., mono-, di-, tri-, or tetra-sialylated glycan). In one instance, the sialylated sCR1 variant glycoform is an sCR1 variant glycoform comprising one sialylated glycan (i.e., the sCR1 variant is monosialylated). In one instance, the sialylated sCR1 variant glycoform comprises at least two sialylated glycans (e.g., di-, tri-, or tetra-sialylated). In one instance, the sialylated sCR1 variant glycoform is an sCR1 variant glycoform comprising two sialylated glycans (i.e., the sCR1 variant is disialylated). In one instance, the sCR1 variant is a sCR1 variant glycoform containing three sialylated glycans (i.e., the sCR1 variant is trisialylated). In another instance, the sCR1 variant is a sCR1 variant glycoform containing four sialylated glycans (i.e., the sCR1 variant is tetrasialylated).

[0114] In one instance, compared to sCR1 containing the sequence shown in SEQ ID NO: 2, the sCR1 variant glycoform (i.e., containing at least two sialylated glycans) exhibits increased inhibitory activity in the classical pathway, lectin pathway, and / or complement bypass pathway.

[0115] This disclosure provides sCR1 variant glycoforms (i.e., comprising at least two sialylated glycans) conjugated with a half-life-extending portion or an additional soluble complement inhibitor. In one instance, the conjugated sCR1 variant glycoform of this disclosure has a longer serum half-life compared to sCR1 variant conjugates containing the sCR1 shown in SEQ ID NO: 2. Examples of increased serum half-life and the assays for determining serum half-life are described herein and should be adapted as necessary to apply to this example of the disclosure.

[0116] This disclosure also provides compositions comprising sCR1 variants and drug carriers and / or excipients. In one example, this disclosure also provides compositions comprising sCR1 variant glycoforms. For example, the composition comprises sialylated sCR1 variant glycoforms. In one example, at least 30% of the sCR1 variant glycoform in the composition comprises sialylated glycans. For example, at least 30% of the sialylated sCR1 variant glycoform comprises mono-, di-, tri-, and / or tetra-sialylated glycans. For example, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, or about 75% of the sCR1 variant glycoform in the composition comprises sialylated glycans (e.g., mono-, di-, tri-, and / or tetra-sialylated).

[0117] In one example, the composition comprises a sialylated sCR1 variant glycoform containing at least two sialylated glycans (e.g., di-, tri-, or tetra-sialylated glycans). In one example, at least about 30% of the sCR1 variant glycoform in the composition comprises at least two sialylated glycans (e.g., di-, tri-, or tetra-sialylated glycans). In one example, at least about 30% of the sCR1 variant glycoform in the composition comprises di-, tri-, and / or tetra-sialylated glycans. For example, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, or about 75% of the sCR1 variant glycoform in the composition comprises at least two sialylated glycans (e.g., di-, tri-, or tetra-sialylated glycans). For example, about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the sCR1 variant glycoform in the composition comprises di, tri, and / or tetrasialylated glycans. In one example, the sCR1 variant consists of or contains an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1).

[0118] In one example, the composition comprises an sCR1 variant glycoform containing monosialotetrahexans. In one example, about 20% of the sCR1 variant glycoform in the composition comprises monosialotetrahexans. For example, about 20%, about 25%, about 30%, about 35%, about 40%, or about 45% of the sCR1 variant glycoform in the composition comprises monosialotetrahexans. In one example, about 22.5% to 25% of the sCR1 variant glycoform in the composition comprises monosialotetrahexans. In one example, about 35% to about 40% of the sCR1 variant glycoform in the composition comprises monosialotetrahexans. In one example, about 40% to about 45% of the sCR1 variant glycoform in the composition comprises monosialotetrahexans.

[0119] In one example, the composition comprises an sCR1 variant glycoform containing disialylated glycans. In one example, at least about 15% of the sCR1 variant glycoform in the composition comprises disialylated glycans. For example, about 15%, about 17.5%, about 20%, about 22.5%, or about 25% of the sCR1 variant glycoform in the composition comprises disialylated glycans. In one example, at least about 25% of the sCR1 variant glycoform in the composition comprises disialylated glycans. For example, about 25%, or about 27.5%, or about 30%, or about 35%, or about 40%, or about 45%, or about 50%, or about 55%, or about 60% of the sCR1 variant glycoform in the composition comprises disialylated glycans. In one example, about 17.5% to about 20% of the sCR1 variant glycoform in the composition comprises disialylated glycans. In one example, about 25% to about 30% of the sCR1 variant glycoform in the composition comprises disialialyzed glycans. In one example, about 40% to about 45% of the sCR1 variant glycoform in the composition comprises disialialyzed glycans. In one example, the sCR1 variant consists of or contains the amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1).

[0120] In one example, the composition comprises an sCR1 variant glycoform containing trisialyl glycan. In one example, at least about 1% of the sCR1 variant glycoform in the composition comprises trisialyl glycan. For example, about 1%, or about 2%, or about 3%, or about 4%, or about 5%, or about 6%, or about 7%, or about 8%, or about 9%, or about 10%, or about 11%, or about 12%, or about 13%, or about 14%, or about 15% of the sCR1 variant glycoform in the composition comprises trisialyl glycan. In one example, about 3.5% to about 4% of the sCR1 variant glycoform in the composition comprises trisialyl glycan. In one example, about 8% to about 8.5% of the sCR1 variant glycoform in the composition comprises trisialyl glycan. In one example, about 9% to about 9.5% of the sCR1 variant glycoform in the composition comprises trisialyl glycan. In one instance, the sCR1 variant consists of or contains an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1).

[0121] In one example, the composition comprises an sCR1 variant glycoform containing tetrasialyl glycan. In one example, at least about 0.5% of the sCR1 variant glycoform in the composition comprises tetrasialyl glycan. For example, about 0.5%, or about 0.75%, or about 1%, or about 1.25%, or about 1.5%, or about 1.75%, or about 2%, or about 2.25%, or about 2.5%, or about 2.75%, or about 3%, or about 4%, or about 5%, or about 6%, or about 7%, or about 8%, or about 9%, or about 10% of the sCR1 variant glycoform in the composition comprises tetrasialyl glycan. In one example, about 0.5% of the sCR1 variant glycoform in the composition comprises tetrasialyl glycan. In one example, about 1.5% to about 2% of the sCR1 variant glycoform in the composition comprises tetrasialyl glycan. In one example, about 2% to about 2.5% of the sCR1 variant glycoform in the composition comprises tetrasialylated glycan. In one example, the sCR1 variant consists of or contains an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1 (e.g., lacking amino acid residues 1393 to 1971 of SEQ ID NO: 1).

[0122] The methods for analyzing the complex carbohydrate structures of the sCR1 variants of this disclosure are obvious to those skilled in the art and / or described herein.

[0123] Methods for producing the sCR1 variant glycoform compositions of this disclosure will be apparent to those skilled in the art, and include, for example, expressing the sCR1 variant in cells that recombinantly express and / or overexpress sialyltransferases. For instance, expressing the sCR1 variant in cells that recombinantly express and / or overexpress human ST3GAL3 (ST3β-galactosyl α-2,3-sialyltransferase 3) and / or human B4GALT1 (human β1,4-galactosyltransferase). In another example, the sCR1 variant may be modified to include additional glycosylation sites to increase the level of glycosylation.

[0124] In one instance, the sCR1 variant of this disclosure is expressed in mammalian cell lines. For example, the sCR1 variant is expressed in cells selected from Chinese hamster ovary (CHO) cells, human embryonic kidney cells (e.g., HEK293), juvenile hamster kidney cells (e.g., BHK-21) cells, mouse myeloma cells (e.g., NS0, Sp2), and amniotic fluid cell-derived cells, such as… It is expressed in mammalian cell lines (e.g., CAP-Go.1, CAP-Go.2).

[0125] In one instance, the sCR1 variant was expressed in Chinese hamster ovary (CHO) cells.

[0126] In one instance, the sCR1 variant is expressed in human embryonic kidney cells (e.g., HEK293).

[0127] In another instance, the sCR1 variant is expressed in amniotic fluid cell-derived cells, such as human amniotic fluid cell-derived cells, for example... Cells, for example or Cells. Any method applicable to this disclosure. Cellular descriptions are found, for example, in Wissing et al. (2015; BMC Proc., 9(Suppl 9): P12).

[0128] This disclosure also provides compositions comprising sCR1 conjugates and drug carriers and / or excipients that include this disclosure.

[0129] In one instance, the composition has an increased serum half-life compared to a composition comprising an sCR1 conjugate containing the sCR1 sequence shown in SEQ ID NO: 2.

[0130] This disclosure provides a method for inhibiting complement activity in a subject, the method comprising administering an sCR1 variant conjugate of this disclosure or a composition containing an sCR1 variant.

[0131] This disclosure also provides a method for treating or preventing a disease or condition in a subject, the method comprising administering an sCR1 variant conjugate or a composition comprising an sCR1 variant of the disclosure.

[0132] In one instance, this disclosure provides an sCR1 variant or an sCR1 variant conjugate or a composition containing an sCR1 variant for inhibiting complement activity in a subject.

[0133] In one instance, this disclosure provides an sCR1 variant or sCR1 variant conjugate or a composition containing an sCR1 variant for the treatment or prevention of a disease or condition in a subject.

[0134] In one instance, this disclosure provides the use of an sCR1 variant or an sCR1 variant conjugate or a composition comprising an sCR1 variant of this disclosure in the preparation of a medicament for inhibiting complement activity in a subject.

[0135] In one instance, this disclosure provides the use of an sCR1 variant or an sCR1 variant conjugate or a composition comprising an sCR1 variant of this disclosure in the preparation of a medicament for treating or preventing a disease or condition in a subject.

[0136] In one instance, the subject was required to be treated with the sCR1 variant of this disclosure (i.e., there was a need for it).

[0137] In one instance, the disease or condition is a complement-mediated condition. For example, the subject has a complement-mediated condition or is at risk of developing a complement-mediated condition.

[0138] In one instance, the subject has a complement-mediated condition. In another instance, the subject has been diagnosed with a complement-mediated condition. In yet another instance, the subject is receiving treatment for a complement-mediated condition.

[0139] In one instance of any of the methods described herein, the sCR1 variant conjugate of this disclosure or a composition containing an sCR1 variant is administered before or after the onset of complement-mediated symptoms. In one instance of any of the methods described herein, the sCR1 variant conjugate of this disclosure or a composition containing an sCR1 variant is administered before the onset of complement-mediated symptoms. In one instance of any of the methods described herein, the sCR1 variant conjugate of this disclosure or a composition containing an sCR1 variant is administered after the onset of complement-mediated symptoms.

[0140] In one instance, the subject was at risk of developing complement-mediated symptoms.

[0141] In one instance, an sCR1 variant conjugate or a composition containing an sCR1 variant is administered before or after the onset of symptoms of a complement-mediated condition. In another instance, an sCR1 variant conjugate or a composition containing an sCR1 variant is administered before the onset of symptoms of a complement-mediated condition. In yet another instance, an sCR1 variant conjugate or a composition containing an sCR1 variant is administered after the onset of symptoms of a complement-mediated condition. In yet another instance, an sCR1 variant conjugate or a composition containing an sCR1 variant of this disclosure is administered at a dose that relieves or reduces one or more symptoms of a complement-mediated condition.

[0142] The symptoms of complement-mediated disorders will be apparent to a technician and will depend on the specific condition. Exemplary symptoms of complement-mediated disorders include, for example:

[0143] • Recurrent infections;

[0144] • Joint inflammation;

[0145] • Myasthenia gravis;

[0146] • Rash or skin discoloration;

[0147] • Edema, especially in the extremities (e.g., feet, hands, legs, or arms) or around the eyes;

[0148] ·stomach ache;

[0149] • Difficulty breathing;

[0150] ·nausea;

[0151] ·fatigue;

[0152] ·hematuria;

[0153] Partial or complete paralysis; and

[0154] • Poor cognitive ability.

[0155] In one instance, complement-mediated disorders are caused by primary dysregulation of the complement system, autoimmune diseases, acute injury, and / or inflammation. Examples of complement-mediated disorders include: hereditary angioedema, paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), thrombocytopenic purpura (TTP), thrombotic microangiopathy, C3 glomerulonephropathy, membranous proliferative glomerulonephritis (including anti-Thy 1 glomerulonephritis, anti-conA diffuse proliferative glomerulonephritis, and / or passive Heymann nephritis), transplant rejection (including lung transplantation (including graft salvage or antibody-mediated rejection) and / or solid organ transplantation (e.g., kidney transplantation (including antibody-mediated rejection)), neuromyelitis optica, multiple sclerosis, and Guillain-Barré syndrome. Myasthenia gravis (including autoimmune myasthenia gravis, demyelinating allergic encephalomyelitis, IgG immune complex alveolitis, and reversed passive Arthus reaction), lupus nephritis (including acute or chronic lupus nephritis), systemic lupus erythematosus (SLE), IgA nephropathy, rheumatoid arthritis, Crohn's disease, ulcerative colitis, autoimmune hemolytic anemia, pemphigus (including pemphigus vulgaris), bullous pemphigoid (including bullous bullous pemphigoid), antiphospholipid syndrome, and other related conditions. Traumatic trauma, neurological injury, hemodialysis, post-infectious HUS, macular degeneration, uveitis, ANCA-associated vasculitis, atherosclerosis, mood disorders, asthma, chronic obstructive pulmonary disease (COPD), chronic inflammatory demyelinating polyneuropathy (CIDP), allergic reactions, sepsis, cerebral malaria, psoriatic arthritis, dermatomyositis, osteoarthritis, dementia, glaucoma, diabetic angiopathy, myocardial infarction, ischemic stroke (with or without reperfusion), hemorrhagic stroke, post-bypass surgery, anti-glomerular basement membrane (GBM) nephritis (or Goodpasture syndrome), autoimmune epilepsy, herpetic dermatitis, eosinophilic granuloma with polyangiitis (EGPA; or Churg-Strauss syndrome), traumatic brain injury, physical trauma, hidradenitis suppurativa, Sjögren's syndrome ( Sjögren's syndrome, Sjögren's syndrome vasculitis, trauma (including glycogen-induced peritonitis, thermal trauma, nerve crush injury and / or closed head injury), local ischemia-reperfusion injury (IRI; including myocardial IRI, intestinal IRI, hepatic IRI and / or pancreatic IRI) and acute respiratory distress syndrome (or acute lung injury).

[0156] In one instance, complement-mediated conditions were selected from transplant rejection (e.g., antibody-mediated rejection), ischemia-reperfusion injury before, during, or after transplantation (including lung and / or kidney transplantation), delayed graft function (including lung and / or kidney transplantation), solid organ transplantation, neuromyelitis optica, myasthenia gravis, glomerular pathology, lupus nephritis, IgA nephropathy, bullous pemphigoid, antiphospholipid syndrome, uveitis, neurological disorders, Parkinson's disease, Huntington's disease, cerebral infarction, motor neuron disease, autoimmune hemolytic anemia, ANCA-associated vasculitis, chronic inflammatory demyelinating polyneuropathy (CIDP), and antiglomerular basement membrane (GBM) nephritis. In one instance, the subject had a condition requiring prophylactic treatment.

[0157] In one instance, complement-mediated conditions are selected from: transplant rejection (including delayed graft function, graft salvage, and antibody-mediated rejection), solid organ transplantation, nephropathy, ischemia-reperfusion injury, neuromyelitis optica, myasthenia gravis, glomerular pathology, lupus nephritis (acute and chronic), IgA nephropathy, bullous pemphigoid, antiphospholipid syndrome, uveitis, neurological disorders, Parkinson's disease, Huntington's disease, cerebral infarction, motor neuron disease, autoimmune hemolytic anemia, ANCA-associated vasculitis, chronic inflammatory demyelinating polyneuropathy, ischemic stroke (with and without reperfusion), traumatic brain injury, physical trauma, and antiglomerular basement membrane (GBM) nephritis.

[0158] In one instance, complement-mediated disease is transplant rejection (e.g., antibody-mediated rejection).

[0159] In one instance, complement-mediated disease is solid organ transplantation.

[0160] In one instance, complement-mediated pathology is ischemia-reperfusion injury before, during, or after transplantation (including lung and / or kidney transplantation).

[0161] In one instance, complement-mediated symptom is delayed graft function (including lung and / or kidney transplants).

[0162] In one instance, complement-mediated disease is neuromyelitis optica.

[0163] In one instance, complement-mediated disease is myasthenia gravis. Examples of myasthenia gravis include autoimmune myasthenia gravis, demyelinating allergic encephalomyelitis, IgG immune complex alveolitis, or reversed passive arthus response.

[0164] In one instance, complement-mediated disease is glomerular pathology.

[0165] In one instance, complement-mediated disease is lupus nephritis. For example, lupus nephritis can be acute or chronic.

[0166] In one instance, complement-mediated disease is systemic lupus erythematosus (SLE).

[0167] In one instance, complement-mediated disease is IgA nephropathy.

[0168] In one instance, complement-mediated disease is bullous pemphigoid. For example, bullous pemphigoid is a type of bullous pemphigoid.

[0169] In one instance, complement-mediated disease is antiphospholipid syndrome.

[0170] In one instance, complement-mediated disease was uveitis.

[0171] In one instance, complement-mediated disorders are neurological disorders.

[0172] In one example, complement-mediated disease is Parkinson's disease.

[0173] In one example, complement-mediated disease is Huntington's disease.

[0174] In one instance, complement-mediated disease was cerebral infarction.

[0175] In one instance, complement-mediated disease is motor neuron disease.

[0176] In one instance, the complement-mediated condition was autoimmune hemolytic anemia.

[0177] In one instance, the complement-mediated condition was ANCA-associated vasculitis.

[0178] In one instance, the complement-mediated condition was chronic inflammatory demyelinating polyneuropathy.

[0179] In one instance, the complement-mediated condition was hereditary angioedema.

[0180] In one instance, the complement-mediated condition was paroxysmal nocturnal hemoglobinuria (PNH).

[0181] In one instance, the complement-mediated condition was atypical hemolytic uremic syndrome (aHUS).

[0182] In one instance, complement-mediated disease is thrombocytopenic purpura (TTP).

[0183] In one instance, complement-mediated disease is thrombotic microangiopathy.

[0184] In one instance, complement-mediated disease is C3 glomerulonephropathy.

[0185] In one instance, complement-mediated disease is membranous proliferative glomerulonephritis. Examples of glomerulonephritis include anti-Thy 1 glomerulonephritis, anti-conA diffuse proliferative glomerulonephritis, and / or passive Heymann nephritis.

[0186] In one instance, complement-mediated disease is transplant rejection. Examples include lung transplants (including graft salvage or antibody-mediated rejection) and / or kidney transplants (including antibody-mediated rejection).

[0187] In one instance, complement-mediated disease is multiple sclerosis.

[0188] In one instance, complement-mediated syndrome is Guillain-Barré syndrome.

[0189] In one instance, complement-mediated disease is rheumatoid arthritis.

[0190] In one instance, complement-mediated disease is inflammatory bowel disease. Examples of inflammatory bowel disease include Crohn's disease or ulcerative colitis.

[0191] In one instance, complement-mediated disease is pemphigus. For example, pemphigus is pemphigus vulgaris.

[0192] In one instance, complement-mediated symptoms were multiple traumatic events.

[0193] In one instance, complement-mediated symptom was neurotrauma.

[0194] In one instance, complement-mediated disease is hemodialysis.

[0195] In one instance, complement-mediated symptoms were post-infectious HUS.

[0196] In one instance, complement-mediated disease is macular degeneration.

[0197] In one instance, complement-mediated disease is atherosclerosis.

[0198] In one instance, complement-mediated symptoms include mood disorders.

[0199] In one instance, complement-mediated disease is asthma.

[0200] In one instance, complement-mediated disease is chronic obstructive pulmonary disease (COPD).

[0201] In one instance, the complement-mediated condition was chronic inflammatory demyelinating polyneuropathy (CIDP).

[0202] In one instance, the complement-mediated condition was an allergic reaction.

[0203] In one instance, complement-mediated disease was sepsis.

[0204] In one instance, complement-mediated disease was cerebral malaria.

[0205] In one instance, complement-mediated disease is psoriatic arthritis.

[0206] In one instance, complement-mediated disease was dermatomyositis.

[0207] In one instance, complement-mediated disease is osteoarthritis.

[0208] In one instance, complement-mediated symptom is dementia.

[0209] In one instance, complement-mediated disease is glaucoma.

[0210] In one instance, complement-mediated disease is diabetic angiopathy.

[0211] In one instance, complement-mediated disease was myocardial infarction.

[0212] In one instance, complement-mediated disease is stroke. For example, the stroke is ischemic stroke (with or without reperfusion). In another example, the stroke is hemorrhagic stroke.

[0213] In one instance, the complement-mediated condition occurred after bypass surgery.

[0214] In one instance, complement-mediated disease is antiglomerular basement membrane (GBM) nephritis (or Goodpasture syndrome).

[0215] In one instance, complement-mediated syndrome is autoimmune epilepsy.

[0216] In one instance, complement-mediated symptoms include herpetic dermatitis.

[0217] In one instance, complement-mediated disease is eosinophilic granulomatosis (EGPA; or Churg-Strauss syndrome) with polyangiitis.

[0218] In one instance, complement-mediated symptom was traumatic brain injury.

[0219] In one instance, the complement-mediated condition is trauma. For example, the trauma is physical trauma. In one instance, the trauma is glycogen-induced peritonitis. In another instance, the trauma is thermal trauma. In yet another instance, the trauma is nerve crush injury and / or closed head injury.

[0220] In one instance, complement-mediated disease was hidradenitis suppurativa.

[0221] In one example, complement-mediated disease is Sjögren's syndrome. For instance, Sjögren's syndrome is Sjögren's vasculitis.

[0222] In one instance, complement-mediated pathology is ischemia-reperfusion injury (IRI). Examples of IRI include myocardial IRI, intestinal IRI, hepatic IRI, and / or pancreatic IRI.

[0223] In one instance, complement-mediated symptoms include acute respiratory distress syndrome (or acute lung injury).

[0224] In one instance, the sCR1 variant conjugate or a composition containing the sCR1 variant of this disclosure is administered to a subject in an amount that reduces the severity of complement-mediated symptoms in the subject.

[0225] In one instance of any of the methods described herein, the subject is a mammal, such as a primate, like a human.

[0226] The treatment methods described in this article may additionally include the administration of other compounds to reduce, treat, or prevent complement-mediated symptoms.

[0227] This disclosure provides a kit comprising at least one sCR1 conjugate of this disclosure or a composition comprising an sCR1 variant, the kit being packaged with instructions for inhibiting complement activity in a subject. Optionally, the kit may additionally comprise other therapeutically active compounds or drugs.

[0228] This disclosure further provides a kit comprising at least one sCR1 conjugate of this disclosure or a composition comprising an sCR1 variant, said kit being packaged with instructions for treating or preventing complement-mediated conditions in subjects. Optionally, the kit may additionally comprise other therapeutically active compounds or drugs.

[0229] This disclosure also provides a kit comprising at least one sCR1 variant conjugate of this disclosure, optionally in combination with other therapeutically active compounds or drugs, or a composition containing an sCR1 variant, the kit being packaged with instructions for administering the conjugate or composition to a subject who has a complement-mediated condition or is at risk of having a complement-mediated condition.

[0230] This document describes exemplary applications of sCR1 variant conjugates or compositions of this disclosure, and applies them, with necessary modifications, to the examples of this disclosure shown in the preceding four paragraphs. Attached Figure Description

[0231] Figure 1 This is a diagram showing the effect of sialylation of sCR1(1392)-8His on plasma half-life.

[0232] Figure 2 This is a diagram showing the effect of sCR1(1392)-8His treatment in an in vivo model of anti-GBM glomerulonephritis.

[0233] Figure 3 It displays sCR1(1392)-8His SIA Illustration of the role of treatment in an in vivo model of anti-GBM glomerulonephritis.

[0234] Key information in the sequence list

[0235] SEQ ID NO:1 Amino acid sequence of soluble complement receptor 1 (sCR1) with N-terminal endogenous human CR1 signal peptide

[0236] SEQ ID NO:2 The amino acid sequence of mature soluble complement receptor 1 (sCR1(1971)) lacking the N-terminal endogenous human CR1 signal peptide.

[0237] SEQ ID NO:3 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(1392)) lacking the N-terminal endogenous human CR1 signal peptide.

[0238] SEQ ID NO:4 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(939)) lacking the N-terminal endogenous human CR1 signal peptide.

[0239] SEQ ID NO:5 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(490-1392))

[0240] SEQ ID NO:6 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(490-1971))

[0241] SEQ ID NO:7 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(234)) lacking the N-terminal endogenous human CR1 signal peptide.

[0242] SEQ ID NO:8 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(489)) lacking the N-terminal endogenous human CR1 signal peptide.

[0243] SEQ ID NO:9 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(940-971))

[0244] SEQ ID NO:10 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(490-939))

[0245] SEQ ID NO:11 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(940-1392))

[0246] SEQ ID NO:12 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(1393-1971))

[0247] Amino acid sequence of SEQ ID NO:13 sCR1 LHR-A

[0248] Amino acid sequence of SEQ ID NO:14 sCR1 LHR-B

[0249] Amino acid sequence of SEQ ID NO:15 sCR1 LHR-C

[0250] Amino acid sequence of SEQ ID NO:16 sCR1 LHR-D

[0251] SEQ ID NO:17 8xHis tag

[0252] Amino acid sequence of endogenous signal peptide SEQ ID NO:18

[0253] Amino acid sequence of exogenous signal peptide SEQ ID NO:19

[0254] SEQ ID NO:20 Amino acid sequence of a His-tagged soluble complement receptor 1 (sCR1(1971)-8His) with an N-terminal endogenous signal peptide.

[0255] SEQ ID NO:21 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(1392)-8His) with an N-terminal endogenous signal peptide.

[0256] SEQ ID NO:22 Amino acid sequence of a truncated mature soluble complement receptor 1 (sCR1(939)-8His) with an N-terminal endogenous signal peptide.

[0257] SEQ ID NO:23 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(490-1392)-8His) with an N-terminal exogenous signal peptide.

[0258] SEQ ID NO:24 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(490-1971)-8His) with an N-terminal exogenous signal peptide.

[0259] SEQ ID NO:25 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(234)-8His) with an N-terminal endogenous signal peptide.

[0260] SEQ ID NO:26 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(489)-8His) with an N-terminal endogenous signal peptide.

[0261] SEQ ID NO:27 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(940-1971)-8His) with an N-terminal exogenous signal peptide.

[0262] SEQ ID NO:28 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(490-939)-8His) with an N-terminal exogenous signal peptide.

[0263] SEQ ID NO:29 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(940-1392)-8His) with an N-terminal exogenous signal peptide.

[0264] SEQ ID NO:30 Amino acid sequence of a His-tagged truncated soluble complement receptor 1 (sCR1(1393-1971)-8His) with an N-terminal exogenous signal peptide.

[0265] SEQ ID NO:31 GS13 connector

[0266] Amino acid sequence of mature human serum albumin (SEQ ID NO:32)

[0267] Amino acid sequence of SEQ ID NO:33 IgG1 Fc

[0268] Amino acid sequence of SEQ ID NO:34 IgG4 Fc

[0269] SEQ ID NO:35 GS30 connector

[0270] Amino acid sequence of exogenous HSA signal peptide SEQ ID NO:36

[0271] Amino acid sequence of exogenous signal peptide SEQ ID NO:37

[0272] SEQ ID NO:38 Amino acid sequence of soluble complement receptor 1 (sCR1(1971)-GS13-HSA) conjugated to HSA and possessing an N-terminal endogenous signal peptide.

[0273] SEQ ID NO:39 Amino acid sequence of an HSA-conjugated soluble complement receptor 1 (HSA-GS13-sCR1(1971)) having an N-terminal HSA signal peptide and a propeptide.

[0274] SEQ ID NO:40 Amino acid sequence of soluble complement receptor 1 (sCR1(1971)-IgG4 Fc) conjugated to IgG4 Fc with an N-terminal endogenous signal peptide.

[0275] SEQ ID NO:41 Amino acid sequence of soluble complement receptor 1 (IgG4Fc-sCR1(1971)) conjugated to IgG4 Fc with an N-terminal exogenous signal peptide.

[0276] Amino acid sequence of SEQ ID NO:42 HSA precursor

[0277] SEQ ID NO:43 Amino acid sequence of a truncated mature soluble complement receptor 1 (sCR1(1392)-GS13-HSA) conjugated to HSA with an N-terminal endogenous signal peptide.

[0278] SEQ ID NO:44 Amino acid sequence of a truncated mature soluble complement receptor 1 (HSA-GS13-sCR1(1392)) conjugated to HSA and having an N-terminal exogenous signal peptide.

[0279] SEQ ID NO:45 Amino acid sequence of a truncated mature soluble complement receptor 1 (sCR1(1392)-IgG1 Fc) conjugated to IgG1 Fc with an N-terminal endogenous signal peptide.

[0280] SEQ ID NO:46 Amino acid sequence of a truncated mature soluble complement receptor 1 (sCR1(1392)-IgG4 Fc) conjugated to IgG4 Fc with an N-terminal endogenous signal peptide.

[0281] SEQ ID NO:47 Amino acid sequence of a truncated mature soluble complement receptor 1 (IgG4 Fc-sCR1(1392)) conjugated to IgG4 Fc with an N-terminal exogenous signal peptide.

[0282] SEQ ID NO:48 Amino acid sequence of a truncated mature soluble complement receptor 1 (sCR1(939)-GS13-HSA) conjugated to HSA and possessing an N-terminal endogenous signal peptide.

[0283] SEQ ID NO:49 Amino acid sequence of a truncated mature soluble complement receptor 1 (sCR1(939)-IgG4 Fc) conjugated to IgG4 Fc with an N-terminal endogenous signal peptide.

[0284] SEQ ID NO:50 Amino acid sequence of a truncated mature soluble complement receptor 1 (IgG4 Fc-sCR1(939)) conjugated to IgG4 Fc with an N-terminal exogenous signal peptide.

[0285] SEQ ID NO:51 Amino acid sequence of truncated mature soluble complement receptor 1 (sCR1(1392)-HSA) conjugated to HSA. Detailed Implementation

[0286] General description

[0287] Throughout this specification, unless otherwise expressly stated or required by the context, references to a single step, a composition of substances, a group of steps, or a group of compositions of substances shall be construed as including one or more (i.e., more than one) of those steps, compositions of substances, groups of steps, or groups of compositions of substances.

[0288] Those skilled in the art will recognize that variations and modifications may be made to this disclosure beyond what has been specifically described. It should be understood that this disclosure includes all such variations and modifications. This disclosure also includes, individually or collectively, all steps, features, compositions, and compounds mentioned or indicated in this specification, as well as any and all combinations or any two or more of said steps or features.

[0289] This disclosure is not limited to the specific examples described herein, which are intended for illustrative purposes only. Functionally equivalent products, compositions, and methods are clearly within the scope of this disclosure.

[0290] Unless otherwise expressly stated, any instance of this disclosure herein should be adapted to any other instance of this disclosure. In other words, any particular instance of this disclosure may be combined with any other particular instance of this disclosure (unless mutually exclusive).

[0291] Any instance of this disclosure that discloses a specific feature or feature group or method or method step shall be understood as providing explicit support for exempting a specific feature or feature group or method or method step.

[0292] Unless otherwise expressly defined, all technical and scientific terms used herein shall have the same meaning as commonly understood by one of ordinary skill in the art (e.g., in cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0293] Unless otherwise stated, the recombinant proteins, cell cultures and immunological techniques used in this disclosure are standard procedures well known to those skilled in the art. Such technical descriptions and explanations are found throughout the literature, such as J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984); J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989); TA Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991); DM Glover and BDHames (editors), DNA Cloning: A Practical Approach, Volumes 1-4, IRL Press (1995 and 1996); and FMAusubel et al. (eds.), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present); Ed Harlow and David Lane (eds.), Antibodies: A Laboratory Manual, Cold Spring Harbour. Laboratory, (1988), and J. E. Corpan et al. (eds.) Current Protocols in Immunology, John Wiley & Sons (including all updates to date).

[0294] The descriptions and definitions of variable regions and their portions, immunoglobulins, antibodies and their fragments in this article can be further clarified by the discussions in Kabat Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., 1987 and 1991, Bork et al., J Mol. Biol. 242, 309-320, 1994, Chothia and Lesk J. Mol Biol. 196: 901-917, 1987, Chothia et al., Nature 342, 877-883, 1989 and / or Al-Lazikani et al., J Mol Biol 273, 927-948, 1997.

[0295] The term “and / or”, such as “X and / or Y”, should be understood to mean “X and Y” or “X or Y”, and should be understood to provide explicit support for both meanings or either meaning.

[0296] Throughout this specification, the word “comprising” or variations such as “including” or “containing” shall be understood to mean including the elements, integers or steps or groups of elements, integers or steps stated therein, but not excluding any other elements, integers or steps or groups of elements, integers or steps.

[0297] As used herein, the term “from” should be understood to indicate that the specified integer can be obtained from a particular source, although not necessarily directly from that source.

[0298] Selected definition

[0299] The type 1 complement receptor (CR1) (also known as the C3b / C4b receptor or CD35) is a member of the complement activation regulator family. CR1 is present on the membranes of erythrocytes, monocytes / macrophages, granulocytes, B cells, some T cells, splenic follicular dendritic cells, and glomerular podocytes, and mediates the binding of cells to granules and immune complexes with activated complement. The encoded protein has a 41-amino acid signal peptide, a 1930-residue extracellular domain, a 25-residue transmembrane domain, and a 43-amino acid C-terminal cytoplasmic region. An exemplary sequence of human CR1 is shown in GenBank accession number NP_000564 for nomenclature and not limitation purposes only.

[0300] Soluble complement receptor type 1 (sCR1) is naturally produced through the cleavage of CR1 on the cell surface and plays a role in controlling complement activation at sites of inflammation. It should be understood that references to “sCR1” refer to a truncated CR1 lacking the transmembrane and cytoplasmic domains. An exemplary sequence of human sCR1 is listed in SEQ ID NO: 1 for nomenclature purposes only and not for limitation. In this document, amino acid positions are referred to with reference to the 1971-amino acid sCR1 protein (e.g., as shown in SEQ ID NO: 1). Full-length sCR1 contains four long homologous repeat (LHR) regions: LHR-A, B, C, and D. The LHR regions can be defined with reference to human sCR1 (as shown in SEQ ID NO: 1). For example, LHR-A contains amino acids 42 to 489 of SEQ ID NO: 1, LHR-B contains amino acids 490 to 939 of SEQ ID NO: 1, LHR-C contains amino acids 940 to 1392 of SEQ ID NO: 1, and LHR-D contains amino acids 1393 to 1971 of SEQ ID NO: 1. Each LHR contains short common repeat (SCR) sequences, totaling 30 SCR sequences, each containing 60 to 70 amino acids. For example, LHR-A contains SCRs 1 to 7 (corresponding to amino acids 42 to 489 of SEQ ID NO: 1), LHR-B contains SCRs 8 to 14 (corresponding to amino acids 491 to 939 of SEQ ID NO: 1), LHR-C contains SCRs 15 to 21 (corresponding to amino acids 941 to 1389 of SEQ ID NO: 1), and LHR-D contains SCRs 22 to 28 (corresponding to amino acids 1394 to 1842 of SEQ ID NO: 1) and SCRs 29 to 30 (corresponding to amino acids 1846 to 1967 of SEQ ID NO: 1). The sequence of mature human sCR1 lacks the N-terminal signal peptide corresponding to amino acids 1 to 41 of SEQ ID NO: 1. For example, the sequence of mature human sCR1 (i.e., lacking the N-terminal signal peptide) is shown in SEQ ID NO: 2.

[0301] The sCR1 sequence of other species can be determined using the sequences provided herein and / or sequences in publicly available databases, and / or using standard techniques (e.g., described in Ausubel et al., (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates to date) or Sambrook et al., Molecular Cloning: A Laboratory Manual, ColdSpring Harbor Laboratory Press (1989)).

[0302] As used herein, the phrase “corresponding to” regarding the amino acid position in SEQ ID NO: 1 should be understood as referring to an amino acid residue or position in the sCR1 sequence, and not necessarily to the sequence containing SEQ ID NO: 1. For example, in a truncated sCR1 sequence containing 41 amino acids (i.e., the mature sCR1), mentioning “corresponding to positions 42 to 939 of SEQ ID NO: 1” would necessarily refer to amino acids at positions 1 to 898. In one instance, sCR1 is contained in the sequence shown in SEQ ID NO: 1.

[0303] As used herein, the term "variant" refers to sCR1 that has undergone deletion or truncation of one or more amino acids using well-known techniques.

[0304] As used herein, in the context of complement activity, the terms “inhibition” or “suppression” should be understood to mean the level at which the sCR1 variant of this disclosure reduces or diminishes complement activity. It will be apparent from the foregoing that the sCR1 variant of this disclosure does not need to completely inhibit complement activity, but only needs to reduce activity by a statistically significant amount, such as at least about 10%, or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 95%. Methods for determining the inhibition of complement activity are known in the art and / or described herein.

[0305] As used herein, the term "complement inhibitor" should be understood to mean a compound that directly or indirectly reduces or diminishes the level of complement activity (including, for example, by inhibiting, blocking, degrading, and consuming complement compounds). In one instance of any of the methods described herein, the complement inhibitor directly reduces the level of complement activity. It will be apparent from the foregoing that the complement inhibitors of this disclosure do not need to completely inhibit complement activity, but only need to reduce the activity by a statistically significant amount, such as at least about 10%, or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 95%. Methods for determining the inhibition of complement activity are known in the art and / or described herein.

[0306] As used herein, the term "half-life extension portion" should be understood to refer to a peptide fusion chaperone that can increase the half-life of the sCR1 variant of this disclosure in a subject. Exemplary half-life extension portions include albumin, antibody Fc regions, and polymers.

[0307] As used herein, in the context of this disclosure, the term "serum half-life" or "plasma half-life" refers to the time required for a 50% (i.e., half) reduction in the concentration or amount of sCR1 in serum (e.g., due to degradation and / or clearance or sequestration by natural mechanisms). Those skilled in the art will recognize that the serum half-life of sCR1 in a subject depends on various physiological conditions (e.g., health status, body size / weight). In healthy human subjects, the serum half-life of sCR1 is approximately 70 hours (3 days). Methods for determining the serum half-life of sCR1 are known in the art and include, for example, pharmacokinetic analysis. For the purposes of this disclosure, an "increased" or "enhanced" serum half-life refers to an increase or acceleration in the time taken for a 50% reduction in the serum concentration of a variant of sCR1 compared to the sCR1 shown in SEQ ID NO: 2.

[0308] The term "recombination" should be understood to refer to the product of artificial genetic recombination. When recombinant proteins are expressed in cells, tissues, or subjects (e.g., therein), it also includes proteins expressed through artificial recombination.

[0309] The term "protein" should be considered to include a single polypeptide chain, that is, a series of consecutive amino acids linked by peptide bonds, or a series of polypeptide chains (i.e., polypeptide complexes) linked covalently or nonvalently to each other. For example, a series of polypeptide chains can be covalently linked using suitable chemical bonds or disulfide bonds. Examples of nonvalent bonds include hydrogen bonds, ionic bonds, van der Waals forces, and hydrophobic interactions.

[0310] From the preceding paragraphs, you will understand that the term "polypeptide" or "polypeptide chain" refers to a series of consecutive amino acids linked by peptide bonds.

[0311] The phrase "conservative amino acid substitution" refers to the replacement or substitution of an amino acid residue with an amino acid residue having similar side chains and / or hydropathicity and / or hydrophilicity. Families of amino acid residues with similar side chains are defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Hydropathic indices are described, for example, by Kyte and Doolittle J.Mol.Biol., 157:105-132, 1982, and hydrophilicity index is described, for example, by US4554101.

[0312] As used herein, the term “condition” refers to a disruption or interference with normal functioning and is not limited to any particular condition, and will include diseases or symptoms.

[0313] As used herein, a subject who is “at risk” of developing a disease or condition or of its recurrence or relapse may or may not have detectable symptoms of the disease or condition, and may or may not have exhibited detectable symptoms of the disease or condition prior to treatment in accordance with this disclosure. “At risk” means that the subject has one or more risk factors, which are measurable parameters associated with the development of a disease or condition, as known in the art and / or described herein.

[0314] As used herein, the terms “treatment,” “curing,” or “therapeutic treatment” include the administration of the serum albumin variant conjugates described herein to thereby reduce or eliminate at least one symptom of a particular disease or condition or to slow the progression of the disease or condition.

[0315] As used herein, the terms “prevention,” “avoidance,” or “prevention” include providing prevention against the onset or recurrence of a specific disease or condition in an individual. An individual may be susceptible to a disease or recurrence of a disease, or at risk of developing a disease or recurrence of a disease, but has not yet been diagnosed with the disease or recurrence of a disease.

[0316] As used herein, the term "subject" should be understood to mean any animal, including humans, such as mammals. Exemplary subjects include, but are not limited to, humans and non-human primates. For example, a subject is a human.

[0317] Inhibit complement activity

[0318] This disclosure provides, for example, a method for inhibiting complement activity in a subject, which includes administering a type 1 soluble complement receptor (sCR1) variant of this disclosure to the subject.

[0319] This disclosure also provides methods for treating or preventing a disease or condition in a subject, the method comprising administering to the subject an sCR1 variant or sCR1 variant conjugate or a composition comprising an sCR1 variant of the disclosure. In one instance, this disclosure provides a method for treating a disease or condition in a subject in need.

[0320] This disclosure also provides for the use of the sCR1 variant or sCR1 variant conjugate or composition comprising the sCR1 variant for the treatment or prevention of a disease or condition in a subject. In one instance, this disclosure provides for the use of the serum albumin conjugate of this disclosure for the treatment of a disease or condition in a subject in need.

[0321] In one instance, the method includes inhibiting activity in the classical pathway, the lectin pathway, and / or the complement alternative pathway. For example, the method includes administering an sCR1 variant of the present disclosure to inhibit activation of the classical complement pathway. In another instance, the method includes administering an sCR1 variant of the present disclosure to inhibit activation of the lectin pathway. In yet another instance, the method includes administering an sCR1 variant of the present disclosure to inhibit activation of the complement alternative pathway.

[0322] In one instance, the method includes inhibiting activity in the extrinsic complement pathway. For example, the method includes administering an sCR1 variant of the present disclosure to inhibit activation of the extrinsic complement pathway.

[0323] In one instance, the disease or condition is a complement-mediated symptom.

[0324] In one instance, the subject had a complement-mediated condition. Complement-mediated conditions can be genetic or acquired.

[0325] In one instance, complement-mediated conditions are selected from: transplant rejection (including delayed graft function, graft salvage, and antibody-mediated rejection), solid organ transplantation, nephropathy, ischemia-reperfusion injury, neuromyelitis optica, myasthenia gravis, glomerular pathology, lupus nephritis (acute and chronic), IgA nephropathy, bullous pemphigoid, antiphospholipid syndrome, uveitis, neurological disorders, Parkinson's disease, Huntington's disease, cerebral infarction, motor neuron disease, autoimmune hemolytic anemia, ANCA-associated vasculitis, chronic inflammatory demyelinating polyneuropathy, ischemic stroke (with and without reperfusion), traumatic brain injury, physical trauma, and antiglomerular basement membrane (GBM) nephritis.

[0326] In one instance, complement-mediated conditions are primary dysregulations, such as hereditary angioedema, paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), thrombotic microangiopathy, C3 glomerulonephropathy, membranous proliferative glomerulonephritis, or transplant rejection (including graft function delay, graft salvage, and antibody-mediated rejection).

[0327] In one instance, complement-mediated conditions are autoimmune conditions such as neuromyelitis optica, multiple sclerosis, myasthenia gravis, Guillain-Barré syndrome, lupus nephritis (acute and chronic), IgA nephropathy, rheumatoid arthritis, Crohn's disease, ulcerative colitis, autoimmune hemolytic anemia, pemphigus, bullous pemphigoid (including bullous bullous pemphigoid), chronic inflammatory demyelinating polyneuropathy (CIDP), antiglomerular basement membrane (GBM) nephritis, or antiphospholipid syndrome.

[0328] In one instance, complement-mediated conditions are acute injuries, such as multiple traumas, neurotrauma, hemodialysis, traumatic brain injury, physical trauma, or post-infectious HUS.

[0329] In one instance, complement-mediated conditions are inflammatory diseases, such as macular degeneration, uveitis, ANCA-associated vasculitis, atherosclerosis, asthma, COPD, sepsis, acute respiratory distress syndrome, cerebral malaria, psoriatic arthritis, or dermatomyositis.

[0330] In one instance, complement-mediated conditions are degenerative conditions such as osteoarthritis, dementia, glaucoma, neurological disorders, Parkinson's disease, Huntington's disease, motor neuron disease, or diabetic vascular disease.

[0331] In one instance, complement-mediated disease is ischemia-reperfusion condition / injury, such as that occurring during or after organ transplantation, surgery, or after stroke or myocardial infarction.

[0332] Methods for diagnosing complement-mediated disorders will readily be apparent to those skilled in the art, and include, for example, the hemolytic classical complement pathway (CH-50) test, the hemolytic alternative complement pathway (AP-50) test, screening for immune complex disorders, antinuclear serology for testing lupus, urinalysis, and complete blood count (CBC).

[0333] In one instance, a subject is at risk of developing a complement-mediated disease. A subject is considered at risk if they have a higher risk of developing a complement-mediated disease than a control group. A control group may include one or more subjects randomly selected from a general population (e.g., matched for age, sex, race, and / or ethnicity) who do not have a complement-mediated disease or a family history of it. A subject may be considered at risk of developing a complement-mediated disease if a “risk factor” associated with the disease is found to be relevant to them. Risk factors may include any activity, trait, event, or nature associated with a given disease (e.g., through statistical or epidemiological studies of a subject population). Therefore, a subject may be classified as at risk of developing a complement-mediated disease even if the study identifying potential risk factors did not explicitly include them.

[0334] In one instance, the subject is at risk of developing complement-mediated disease, and the sCR1 variant is administered before or after the onset of symptoms of complement-mediated disease. In one instance, the sCR1 variant is administered before the onset of symptoms of complement-mediated disease. In one instance, sCR1 is administered after the onset of symptoms of complement-mediated disease. In one instance, the sCR1 variant of this disclosure is administered at a dose that alleviates or reduces one or more symptoms of complement-mediated disease in the subject at risk.

[0335] The methods disclosed herein can be readily applied to any form of complement-mediated condition in subjects.

[0336] In one instance, the methods of this disclosure alleviated any symptoms of complement-mediated conditions known in the art and / or described herein.

[0337] It will be apparent to those skilled in the art that the “reduction” of symptoms of complement-mediated disease in a subject will be compared with that of another subject who also has complement-mediated disease but has not received treatment using the methods described herein. This does not necessarily require comparing two subjects simultaneously. Instead, it can rely on population data. For example, assess a population of subjects with complement-mediated disease who have not received treatment using the methods described herein (optionally, a population of subjects similar to the treated subjects (e.g., in age, weight, ethnicity)) and compare the mean to the results of subjects or a population of subjects treated using the methods described herein.

[0338] In complement-mediated conditions resulting from or related to organ transplantation-induced or organ transplantation-associated ischemia-reperfusion injury, the sCR1 variants or compositions comprising the sCR1 variants of this disclosure may be administered before, during, or after transplantation. In some instances, the sCR1 variants or compositions are administered to organ transplant donors. In other instances, the sCR1 variants or compositions are administered to subjects, where the subjects are organ transplant recipients. In one instance, the sCR1 variants or compositions are administered ex vivo to harvested organs prior to organ transplantation. For example, prior to transplantation, harvested organs may be perfused or infused with a solution comprising the sCR1 variants or compositions.

[0339] In one instance, organ transplantation is a solid organ transplant. For example, a solid organ transplant is a lung transplant.

[0340] It will be apparent to those skilled in the art from the foregoing that this disclosure provides methods for organ transplantation or for improving the outcome of organ transplantation or improving the function of transplanted organs or for preventing delays in graft function, the methods comprising administering an sCR1 variant or composition to an organ transplant donor prior to organ collection; collecting the organ; and transplanting the organ into an organ transplant recipient.

[0341] This disclosure also provides a method for preparing transplant organs from organ donors to improve organ function in organ transplant recipients, the method comprising administering an sCR1 variant or composition to the organ donor prior to organ collection.

[0342] This disclosure also provides a method for preventing organ transplant rejection, which includes administering an sCR1 variant or composition to an organ donor prior to organ collection, collecting the organ, and transplanting the organ into an organ transplant recipient.

[0343] In some instances, the method further includes administering an sCR1 variant or composition to the organ transplant recipient. For example, the sCR1 variant or composition may be administered to the organ transplant recipient before or during organ transplantation (i.e., during transplantation).

[0344] This disclosure also provides methods for organ transplantation or for improving organ transplantation outcomes or improving the function of transplanted organs or for preventing graft function delay, the method comprising administering an sCR1 variant or composition to an organ transplant recipient prior to organ transplantation, and then transplanting the organ into the organ transplant recipient.

[0345] In one instance, the organ transplant donor was brain dead. For example, the organ donor was alive with life support but was brain dead.

[0346] In one example of this disclosure, the sCR1 variant or composition is administered prior to reperfusion, for example in the case of organ transplantation, where the sCR1 variant or composition is administered to the organ transplant recipient prior to reperfusion of the transplanted organ (e.g., the sCR1 variant or composition is administered before or during transplantation but prior to reperfusion).

[0347] In cases of administration to brain-dead donors, the sCR1 variant or composition can be administered at any time between brain death and organ harvesting. In some instances, the sCR1 variant or composition is administered ex vivo to the harvested organ prior to organ transplantation. For example, the harvested organ can be perfused or infused with a solution containing the sCR1 variant or composition prior to transplantation.

[0348] Type 1 soluble complement receptor variant

[0349] This disclosure provides sCR1 variants for any of the methods described herein.

[0350] In one instance, this disclosure provides an sCR1 variant that has improved or increased complement-inhibiting activity compared to the sequence shown in SEQ ID NO: 2. The inventors have determined that sCR1 variants comprising residues 42 to 939 and / or residues 490 to 1392 of SEQ ID NO: 1 have improved and / or increased complement-inhibiting activity.

[0351] This disclosure provides a method for inhibiting complement activity in a subject, the method comprising administering to the subject a type 1 soluble complement receptor (sCR1) variant comprising an amino acid sequence selected from:

[0352] (i) The amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1; and

[0353] (ii) The amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1.

[0354] For example, the inventors have identified amino acid residues in the sequence shown in SEQ ID NO: 1 that can be deleted without loss of function or resulting in functional improvement. In one instance, the sCR1 variant contains deletions of 489 to 1073 amino acids compared to the sequence shown in SEQ ID NO: 1. For example, the sCR1 variant contains deletions of 489, 620, 1068, or 1073 amino acids compared to the sequence shown in SEQ ID NO: 1.

[0355] In one instance, this disclosure provides a truncated sCR1 containing 898 to 1482 amino acids compared to the sequence shown in SEQ ID NO: 1. For example, the truncated sCR1 contains 898, 903, 1351, or 1482 amino acids compared to the sequence shown in SEQ ID NO: 1.

[0356] In one instance, the sCR1 variant of this disclosure comprises a variant of the sequence shown in SEQ ID NO: 1, wherein the variant sequence comprises an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO: 1.

[0357] In one instance, the sCR1 variant of this disclosure comprises a variant of the sequence shown in SEQ ID NO: 1, wherein the variant sequence comprises an amino acid sequence corresponding to amino acids 42 to 939 of SEQ ID NO: 1.

[0358] In one instance, the sCR1 variant of this disclosure comprises a variant of the sequence shown in SEQ ID NO: 1, wherein the variant sequence comprises an amino acid sequence corresponding to amino acids 490 to 1392 of SEQ ID NO: 1.

[0359] In one instance, the sCR1 variant of this disclosure comprises a variant of the sequence shown in SEQ ID NO: 1, wherein the variant sequence comprises an amino acid sequence corresponding to amino acids 490 to 1971 of SEQ ID NO: 1.

[0360] In one instance, the sCR1 variant of this disclosure does not contain or consist of the sequences shown in SEQ ID NO: 1 and / or SEQ ID NO: 2.

[0361] In one instance, the sCR1 variant of this disclosure does not contain the amino acid sequence corresponding to amino acids 1 to 41 of SEQ ID NO: 1.

[0362] In one instance, the sCR1 variant of this disclosure does not contain the amino acid sequence corresponding to amino acids 940 to 1971 of SEQ ID NO: 1.

[0363] In one instance, the sCR1 variant of this disclosure does not contain the amino acid sequence corresponding to amino acids 1393 to 1971 of SEQ ID NO: 1.

[0364] In one instance, the sCR1 variant of this disclosure does not contain the amino acid sequence corresponding to amino acids 1 to 489 of SEQ ID NO: 1.

[0365] In one instance, the sCR1 variant is a monolithic variant (i.e., a single copy of the sCR1 variant).

[0366] In one instance, the sCR1 variant is a dimer or dimerized (i.e., two copies of the sCR1 variant are linked together in the fusion protein).

[0367] In one instance, the sCR1 variant is a multimer or polymerized (i.e., multiple copies of the sCR1 variant are linked in the fusion protein).

[0368] Methods for achieving dimerization or polymerization of sCR1 variants are known in the art and / or described herein, and include, for example, direct or indirect conjugation between two or more sCR1 variants (e.g., by means of a linker between two or more sCR1 variants). In one instance, dimerization or polymerization is formed by chemical conjugation (e.g., by disulfide bonds or cystine knots) or by genetic fusion.

[0369] In one instance, two or more identical sCR1 variants are fused (i.e., expressed as a fusion protein).

[0370] In one instance, two or more different sCR1 variants are fused (i.e., expressed as a fusion protein).

[0371] In one instance, dimerized or multimerized sCR1 variants contain a connector between sCR1 variants.

[0372] In one instance, this disclosure provides a multimeric protein comprising two or more sCR1 variants containing multimerizing domains, wherein the multimerizing domains interact to form the multimeric protein.

[0373] In one instance, each sCR1 variant in the multimeric protein contains one sCR1 variant. In another instance, one or more sCR1 variants in the multimeric protein contain two or more sCR1 variants, for example, variants linked in a fusion protein.

[0374] In one instance, the polymerized domain contains an immunoglobulin hinge domain.

[0375] In one instance, the polymerizing domain is a leucine zipper domain, a cysteine ​​knot, or an antibody Fc region. For example, the polymerizing domain is a leucine zipper domain. Suitable leucine zipper peptides are known in the art and include c-Jun and c-Fos leucine zipper domains. Leucine zipper fusions are described in Riley et al., Protein Eng. (1996), which is incorporated herein by reference. In another instance, the polymerizing domain is a cysteine ​​knot. For example, a cysteine ​​knot may contain up to 60 amino acids in length and include a core domain with three or more intertwined disulfide bonds. In yet another instance, the polymerizing domain is an antibody Fc region (e.g., as described herein).

[0376] In one instance, the polymerized sCR1 variant is linear.

[0377] In one instance, the polymerized sCR1 variant is circular. For example, the polymerized sCR1 variant may contain sorting enzyme cleavage sites, as described in Popp, MW et al., PNAS (2011), which is incorporated herein by reference.

[0378] In one instance, the sCR1 variant used in this disclosure comprises at least two sialylated glycans (e.g., di-, tri-, or tetra-sialylated glycans). For example, a composition used in any of the methods described herein comprises a sialylated sCR1 variant glycoform. In one instance, the sialylated sCR1 variant glycoform used in any of the methods described herein comprises a di-, tri-, or tetra-sialylated glycoform. Methods for producing variant sCR1 glycoforms comprising at least two sialylated glycans (e.g., di-, tri-, or tetra-sialylated glycans) are obvious to those skilled in the art and / or described herein.

[0379] Exemplary methods for determining the biological activity of the sCR1 variants of this disclosure are obvious to those skilled in the art and / or described herein. For example, methods for determining inhibitory activity of the classical pathway, lectin pathway, and / or alternative pathway are described herein.

[0380] Conjugate

[0381] This disclosure provides sCR1 variant conjugates for use in any of the methods described herein. Methods for conjugating sCR1 variants will be obvious to those skilled in the art and / or described herein. This disclosure considers all forms and methods of conjugation (i.e., binding), including, for example, direct or indirect conjugation between an sCR1 variant and another compound / part described herein (e.g., by means of a linker between the sCR1 variant and other compounds / parts). In one instance, the conjugate is formed by chemical conjugation (e.g., via an amine bond or disulfide bond) or by genetic fusion.

[0382] In one instance, the sCR1 variant of this disclosure is conjugated to a half-life extension portion or another soluble complement inhibitor.

[0383] In one instance, the sCR1 variant of this disclosure is conjugated to a half-life-extended portion or another soluble complement inhibitor that binds directly or indirectly to the sCR1 variant. The half-life-extended portion or the other soluble complement inhibitor may bind directly or indirectly to the sCR1 variant (e.g., in the case of indirect binding, it may contain a linker).

[0384] In one instance, the sCR1 variant is conjugated via an amine bond to either a half-life-extending portion or an additional soluble complement inhibitor.

[0385] In one instance, this disclosure provides a fusion protein comprising an sCR1 variant and an extended half-life portion or an additional soluble complement inhibitor. For example, the extended half-life portion or the additional soluble complement inhibitor is located at the N-terminus of the sCR1 variant, the C-terminus of the sCR1 variant, or any combination thereof.

[0386] In one instance, the sCR1 variant conjugates to a half-life-extending portion or another soluble complement inhibitor via a linker. For example, the linker is a peptide linker.

[0387] In one instance, the linker is a flexible linker. A “flexible” linker is an amino acid sequence that does not have a fixed structure (secondary or tertiary structure) in solution. Therefore, such flexible linkers are free to adopt various conformations. Flexible linkers suitable for this disclosure are known in the art. Examples of flexible linkers used in this disclosure are linker sequences SGGGGS / GGGGS / GGGGS or (Gly4Ser)3. Flexible linkers are also disclosed in WO1999045132.

[0388] The linker can contain any amino acid sequence that does not substantially impede the interaction between the binding region and its target. Preferred amino acid residues for flexible linker sequences include, but are not limited to, glycine, alanine, serine, threonine, proline, lysine, arginine, glutamine, and glutamic acid.

[0389] The linker sequence between the binding regions preferably comprises five or more amino acid residues. The flexible linker sequence according to this disclosure consists of five or more residues (preferably 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, 25, or 30 or more residues). In a highly preferred embodiment of the invention, the flexible linker sequence consists of 5, 7, 10, 13, 16, or 30 residues.

[0390] In one instance, the flexible connector has the amino acid sequence according to SEQ ID NO: 31, namely GSGGSGGSGGSGS(GS13).

[0391] In one example, the flexible connector has the amino acid sequence according to SEQ ID NO: 35, namely, SGGSGGSGGSGGSGGSGGSGGSGGSGGSGS(GS30).

[0392] Exemplary compounds that can be conjugated with the sCR1 variants of this disclosure, and methods for such conjugation, are known in the art and are described herein.

[0393] Extended half-life portion

[0394] In one instance, an sCR1 variant is conjugated to a half-life extension portion. Half-life extension portions suitable for this disclosure will be apparent to those skilled in the art, and include, but are not limited to, those described herein. For example, half-life extension portions are selected from human serum albumin or functional fragments thereof, immunoglobulin Fc regions or functional fragments thereof, afamin, alpha-fetoprotein, vitamin D-binding protein, albumin-bound antibody fragments, and polymers.

[0395] In one instance, the extended half-life portion is human serum albumin or a functional fragment thereof.

[0396] In one instance, the extended half-life portion is the Fc region of an immunoglobulin or a functional fragment thereof.

[0397] In one instance, the extended half-life portion is afamin.

[0398] In one instance, the extended half-life portion is alpha-fetoprotein.

[0399] In one instance, the extended half-life portion is due to vitamin D-binding proteins.

[0400] In one instance, the extended half-life portion is an antibody fragment that binds to albumin.

[0401] In one instance, the extended half-life portion is a polymer.

[0402] Albumin and its variants

[0403] In one instance, the extended half-life portion is albumin or a functional fragment or variant thereof.

[0404] Serum albumin (or blood albumin) is the most abundant blood protein and acts as a carrier protein for steroids, fatty acids and thyroid hormones in the blood, and plays a major role in stabilizing extracellular fluid volume.

[0405] In one instance, albumin, its functional fragments, or variants are serum albumin, such as human serum albumin. An exemplary sequence of mature human serum albumin is listed in NCBI GenBank accession number AEE60908 and SEQ ID NO: 32 for nomenclature and not limitation purposes only.

[0406] In one instance, albumin, its functional fragments, or variants contain one or more amino acid substitutions, deletions, or insertions. Amino acid substitutions suitable for use in this disclosure will be apparent to those skilled in the art and include both naturally occurring substitutions and engineered substitutions, such as those described in, for example, WO2011051489, WO2014072481, WO2011103076, WO2012112188, WO2013075066, WO2015063611, WO2014179657, and WO2019075519.

[0407] In one instance, this disclosure provides sCR1 variants conjugated to albumin family proteins (e.g., proteins structurally or evolutionarily related to albumin). For example, sCR1 variants conjugated to afamin, alpha-fetoprotein, or vitamin D-binding proteins.

[0408] In another instance, the sCR1 variant is fused with an albumin family protein (e.g., a protein structurally or evolutionarily related to albumin), for example, expressed as a fusion protein. For instance, the sCR1 variant is fused with afamin, alpha-fetoprotein, or a vitamin D-binding protein, for example, as a fusion protein.

[0409] Immunoglobulin Fc region and its fragments

[0410] In one instance, the extended half-life portion is an immunoglobulin or a functional fragment thereof. For example, an immunoglobulin contains an Fc region, such as an Fc domain or Fc fragment and / or variants thereof. In one instance, Ig is part of an immunoglobulin constant domain. An exemplary sequence of human IgG1 Fc is set in SEQ ID NO: 33 for nomenclature and not limitation only. An exemplary sequence of human IgG4 Fc is set in SEQ ID NO: 34 for nomenclature and not limitation only.

[0411] In one instance, the Fc fragment and / or its variants contain one or more amino acid substitutions, deletions, or insertions. Amino acid substitutions applicable to this disclosure will be apparent to those skilled in the art and include both naturally occurring substitutions and engineered substitutions, such as those described in WO2000042072, WO2002060919, WO2004035752, and WO2006053301.

[0412] In one instance, the immunoglobulin or fragment thereof used in this disclosure contains an IgG4 constant region or a stable IgG4 constant region. For example, a stable IgG4 constant region contains proline at hinge region position 241 according to the Kabat system (Kabat et al., Sequences of Proteins of Immunological Interest, Washington DC, United States Department of Health and Human Services, 1987 and / or 1991) or at hinge region position 228 according to the EU numbering system (Edelman, GM et al., Proc. Natl. Acad. USA, 63, 78-85 (1969)).

[0413] In one instance, the Fc domain is modified to prevent it from dimerizing. For example, the Fc region is a monomeric Fc region.

[0414] Methods for generating haptens are known in the art, and exemplary methods are described herein.

[0415] In one instance, a hapten can be secreted by introducing a gene containing a protein with an Fc domain for expression into a cell. In one instance, the constant region (e.g., the IgG4 Fc domain) contains a "bond or pore" (or "bump or hole") mutation to prevent heterodimer formation. In one instance, the constant region (e.g., the IgG4 Fc domain) contains a T366W mutation (or a bump). In another instance, the constant region (e.g., the IgG4 Fc domain) contains T366S, L368A, and Y407V mutations (or holes). In another instance, the Fc domain contains T350V, T366L, K392L, and T394W mutations (bumps). In yet another instance, the constant region contains T350V, L351Y, F405A, and Y407V mutations (holes). Exemplary amino acid substitutions in the constant region are numbered according to the EU numbering system.

[0416] For example, the Fc domain is the IgG4 Fc domain, which contains the sequence shown in SEQ ID NO: 34 with the following substitutions:

[0417] • Position 228, arginine replaces proline;

[0418] • Position 351, phenylalanine replaces leucine;

[0419] • Position 366, arginine replaces threonine;

[0420] • Position 395, lysine replaces proline;

[0421] • Position 405, arginine replaces phenylalanine; and

[0422] • Position 407, glutamic acid replaces tyrosine.

[0423] In one instance, this disclosure provides sCR1 variants that conjugate to immunoglobulins or functional fragments thereof, such as Fc regions, Fc domains, or Fc fragments and / or variants thereof. For example, sCR1 variants conjugate to human IgG4 Fc.

[0424] In another instance, the sCR1 variant is fused with an immunoglobulin or a functional fragment thereof (e.g., an Fc region, such as an Fc domain or Fc fragment and / or a variant thereof), for example, expressed as a fusion protein. For instance, the sCR1 variant is fused with human IgG4 Fc, for example, as a fusion protein.

[0425] Antibodies and their fragments

[0426] In one instance, an immunoglobulin is an antibody or antigen-binding fragment that binds to albumin. Exemplary antibody or antigen-binding fragments are known in the art and are described, for example, in Kang et al., Immunol Lett.; 169:33-40, 2016; Protein Eng Des Sel. 21(5):283-8, 2008; and Holt et al., MAbs. 8(7):1336-1346, 2016.

[0427] Other exemplary antibodies or antigen-binding fragments thereof used in this disclosure are described herein or known in the art, including:

[0428] • Humanized antibodies or fragments thereof, such as proteins containing human-like variable regions, including CDRs of antibodies from non-human species (e.g., mice or rats or non-human primates) transplanted into or inserted into the framework region (FR) of human antibodies (e.g., generated by the methods described in US5225539, US6054297, US7566771 or US5585089);

[0429] • Human antibodies or fragments thereof, such as antibodies having variable antibody regions and optional constant antibody regions found in humans (e.g., in human germline or somatic cells or in libraries produced using such regions). “Human” antibodies may include amino acid residues not encoded by human sequences, for example, mutations introduced by random or site-directed mutagenesis in vitro (e.g., generated by the methods described in US5565332), and affinity-matured forms of such antibodies;

[0430] • Synthesize humanized antibodies or fragments thereof, such as antibodies containing variable regions comprising FR from New World primate antibody variable regions and CDR from non-New World primate antibody variable regions (e.g., generated by the method described in WO2007019620);

[0431] • Primate-modified antibodies or fragments thereof, such as antibodies containing variable regions of antibodies produced after immunization from non-human primates (e.g., cynomolgus monkeys) (e.g., produced by the method described in US6113898);

[0432] • Chimeric antibodies or chimeric antigen-binding fragments, such as antibodies or fragments in which one or more variable domains are derived from a specific species (e.g., rodents, such as mice or rats) or belong to a specific antibody class or subclass, while the remainder of the antibody or fragment is derived from another species (e.g., humans or non-human primates) or belongs to another antibody class or subclass (e.g., generated by the methods described in US6331415, US5807715, US4816567, and US4816397);

[0433] • Deimmunizing antibodies or their antigen-binding fragments, such as antibodies and fragments that remove (i.e. mutate) one or more epitopes (e.g., B-cell epitopes or T-cell epitopes), thereby reducing the likelihood that a subject will elicit an immune response against the antibody or protein (e.g., as described in WO2000034317 and WO2004108158).

[0434] • Bispecific antibodies or fragments thereof, for example, antibodies comprising two types of antibodies or antibody fragments (e.g., two half-antibodies) that are specific to different antigens or epitopes (e.g., as described in US5731168).

[0435] Other exemplary antibody fragments used in this disclosure are described herein or are known in the art, and include:

[0436] • Single-domain antibodies (domain antibodies or dAbs), for example, a single polypeptide chain containing all or part of the variable domain of the antibody heavy chain;

[0437] • Biantibodies, triantibodies, tetraantibodies or higher-order protein complexes (e.g., as described in WO98 / 044001 and / or WO94 / 007921);

[0438] • Single-chain Fv (scFv) fragments, for example, V contained in a polypeptide chain H and V L District and V H and V L The fragment of the polypeptide linker between the peptides allows scFv to form the desired antigen-binding structure (i.e., V in a polypeptide chain). H and V L They combine to form Fv);

[0439] • Half-antibody or half-molecule, such as a protein containing a heavy chain and a light chain.

[0440] This disclosure also considers other antibodies and antibody fragments, such as:

[0441] (i) Microantibodies, such as those described in US5837821;

[0442] (ii) Heteroconjugated proteins, for example, as described in US4676980;

[0443] (iii) Heteroconjugated proteins produced using chemical cross-linking agents, such as those described in US4676980; and

[0444] (iv) Fab3 (e.g., as described in EP19930302894).

[0445] polymer

[0446] In one instance, this disclosure provides sCR1 variants conjugated to polymers. Suitable polymers for use in this disclosure will be obvious to those skilled in the art and / or are described herein.

[0447] In one example, the sCR1 variant is conjugated to polyethylene glycol (PEG). For example, the polymer contains a mono- or poly(e.g., 2-4)-polyethylene glycol (PEG) moiety. For example, the mono- or poly(e.g., 2-4)-polyethylene glycol (PEG) moiety prolongs the in vivo half-life of the sCR1 variant.

[0448] PEGylation can be carried out by any available PEGylation reaction. Exemplary methods for preparing PEGylated protein products typically include: (a) reacting a polypeptide with polyethylene glycol (e.g., a reactive ester or aldehyde derivative of PEG) under conditions in which the protein is linked to one or more PEG groups; and (b) obtaining the reaction product.

[0449] Those skilled in the art will recognize various PEG attachment methods, including but not limited to those described in, for example, EP 0 401 384; Malik et al., Exp. Hematol., 20:1028-1035 (1992); Francis, Focus on Growth Factors, 3(2):4-10 (1992); EP 0 154 316; EP 0 401 384; WO 92 / 16221; WO 95 / 34326; and U.S. Patent No. 5,252,714.

[0450] Soluble complement inhibitors

[0451] This disclosure provides sCR1 variants conjugated to other soluble complement inhibitors.

[0452] In one instance, an sCR1 variant was conjugated to a soluble complement inhibitor or a modified (i.e., variant) form thereof.

[0453] Suitable complement inhibitors for use in this disclosure will be apparent to those skilled in the art, and include, for example, factor I (fI), factor H (fH), complement factor H-associated protein (CFHR), C4b-binding protein (C4bp), soluble CD55 (decay-promoting factor (DAF)), soluble CD46 (membrane cofactor protein (MCP)), soluble CD59 (protectin), soluble complement receptor 2 (sCR2), TT30 (CR2-fH), cobra venom factor (CVF)), and functional fragments or variants thereof.

[0454] Assay activity of sCR1 variant

[0455] For example, as described below, the biological activity of the sCR1 variants of this disclosure can be easily screened.

[0456] Measuring complement activity

[0457] In one instance, complement activity was assessed using an enzyme immunoassay (e.g., The complement assay kit measures the complement inhibitory activity. For example, a labeled antibody specific to an antigen or epitope generated during complement activation (e.g., C5b-9 or an epitope present in C5b-9) is used to determine complement inhibitory activity. In one example, the wells of a microtiter plate are coated with a specific activator of the classical, lectin, or alternative pathways. In another example, the sCR1 variant is incubated with normal human serum and an appropriate assay diluent (i.e., a diluent containing appropriate components to ensure specific activation of the classical, lectin, or alternative pathways) and added to the wells of a microtiter plate coated with a specific activator of the classical, lectin, or alternative pathways. The amount of C5b-9 complex formed is detected using an antibody labeled with a specific alkaline phosphatase against C5b-9. In one example, the amount of complement activation product (i.e., C5b-9) generated is proportional to the functional activity of the complement pathway. In one example, the half-maximal inhibitory concentration (i.e., IC50) is determined. 50 For example, determining the IC of the sCR1 variant. 50 And it is compared with an IC containing sCR1, which contains the sequence shown in SEQ ID NO: 2. 50 Compare them.

[0458] In another example, hemolysis assays (e.g., classical pathway (i.e., CH50) and alternative pathway (ApH50) inhibition assays) are used to determine complement inhibitory activity. The CH50 assay is a method for measuring total classical complement activity in serum. This test is a lysis assay that uses antibody-sensitized erythrocytes as activators of the classical complement pathway, with human serum as the complement source. The percentage of hemolysis can be determined, for example, using a spectrophotometer. The CH50 assay indirectly measures the formation of the terminal complement complex (TCC), as the TCC itself is directly responsible for the hemolysis being measured. This assay is well-known. In short, to assess inhibition of the classical complement pathway, pre-diluted human serum is pre-incubated in micro-assay wells with serially diluted sCR1 variants. Next, antibody-sensitized erythrocytes (e.g., sheep erythrocytes sensitized with rabbit anti-sheep antibodies) are added. After centrifugation, free hemoglobin in the supernatant is measured using a spectrophotometer. The reduction in free hemoglobin reflected the inhibition of TCC-mediated erythrocyte lysis. The sCR1-mediated inhibition was then calculated relative to erythrocytes incubated only with human serum (100% lysed sample).

[0459] Complement inhibition can also be assessed using any method known in the art, including, for example, in vitro yeast polysaccharide assays, erythrocyte lysis assays, antibody or immune complex activation assays, bypass pathway activation assays, and lectin pathway activation assays.

[0460] Pharmaceutical compositions and treatment methods

[0461] Suitably, in compositions or methods for administering the sCR1 variant of this disclosure to a subject, the sCR1 variant conjugate of this disclosure (i.e., the sCR1 variant conjugated with a half-life-extending portion or another soluble complement inhibitor) is combined with a pharmaceutically acceptable carrier as understood in the art. Thus, one example of this disclosure provides a composition (e.g., a pharmaceutical composition) comprising the sCR1 variant of this disclosure combined with a pharmaceutically acceptable carrier. Another example of this disclosure provides a composition (e.g., a pharmaceutical composition) comprising the sCR1 variant conjugate of this disclosure combined with a pharmaceutically acceptable carrier.

[0462] Generally speaking, a "carrier" refers to a solid or liquid filler, binder, diluent, encapsulating substance, emulsifier, wetting agent, solvent, suspending agent, coating, or lubricant that can be safely applied to any subject (e.g., a human). Depending on the specific route of application, a variety of acceptable carriers known in the art may be used, such as those described in Remington's Pharmaceutical Sciences (Mack Publishing Co., NJUSA, 1991).

[0463] The sCR1 variants or sCR1 variant conjugates disclosed herein may be used for prevention or treatment via parenteral, topical, oral, local, aerosol, intrathecal, or transdermal administration. In one instance, the sCR1 variant or sCR1 variant conjugate is administered parenterally, such as subcutaneously or intravenously. For example, intravenous administration of the sCR1 variant or sCR1 variant conjugate.

[0464] The formulation of the sCR1 variant to sCR1 variant conjugate to be administered will vary depending on the route of administration and the chosen formulation (e.g., solution, emulsion, capsule). Suitable pharmaceutical compositions to be administered can be prepared in physiologically acceptable carriers. For solutions or emulsions, suitable carriers include, for example, aqueous or alcohol / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral media may include sodium chloride solution, Ringer's glucose, glucose and sodium chloride, lactated Ringer's solution, or non-volatile oils. Various suitable aqueous carriers are known to those skilled in the art, including water, buffered water, buffered saline, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), glucose solutions, and amino acids, including, for example, glycine, proline, lysine, histidine, methionine, arginine, alanine, valine, serine, asparagine, phenylalanine, tyrosine, cysteine, threonine, leucine, tryptophan, glutamine, isoleucine, glutamic acid, and combinations thereof. Intravenous mediators may include various additives, preservatives, or liquids, nutrients, or electrolyte supplements (see Remington's Pharmaceutical Science, 16th edition, Mack, 1980). The composition may optionally contain pharmaceutically acceptable adjuvants required to approximate physiological conditions, such as pH adjusters and buffers, and toxicity modifiers, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, and sodium lactate. The composition may be stored in a liquid state or may be lyophilized for storage and reconstituted in a suitable carrier prior to use according to lyophilization and reconstitution techniques known in the art.

[0465] The method of this disclosure may also include the co-administration of an sCR1 variant or sCR1 variant conjugate according to this disclosure and the administration of another therapeutically effective agent for inhibiting complement activity or for preventing or treating complement-mediated conditions.

[0466] In one instance, the sCR1 variant or conjugate of this disclosure is used in combination with at least one other known compound or therapeutic protein that is currently in use or under development to inhibit complement activity or to prevent or treat complement-mediated conditions. Compounds currently used to treat complement-mediated conditions are known in the art and include antibodies against C5 and its activated form (C5a), such as eculizumab, Berinert human C1 esterase inhibitor, human C1 esterase inhibitor, Ruconest recombinant C1 esterase inhibitor, Cinryze human C1 esterase inhibitor, anti-human MASP-2 monoclonal antibody, APL-2C3 inhibitory peptide, lamparazine, and TNT009 anti-C1s antibody. Further compounds are described in Reis et al., Clin Immunol. Dec; 161(2):225–240, 2015.

[0467] As will be apparent from the foregoing, this disclosure provides a method of concomitant therapeutic treatment of a subject, comprising administering an effective amount of a first agent and a second agent to a subject in need, wherein the first agent is an sCR1 variant or sCR1 variant conjugate of this disclosure, and the second agent is also used to inhibit complement activity or to prevent or treat complement-mediated conditions.

[0468] As used herein, the term "companionate" in the phrase "companionate treatment" includes administering a first drug in the presence of a second drug. A companion treatment method includes a method in which a first, second, third, or additional drug is administered concurrently. A companion treatment method also includes a method in which a first or additional drug is administered in the presence of a second or additional drug, wherein, for example, the second or additional drug may have been previously administered. Companionate treatment can be performed sequentially by different participants. For example, one participant may administer a first drug to a subject, and a second participant may administer a second drug to said subject, and the administration steps may be performed simultaneously, nearly simultaneously, or at distant times, provided that the first drug (and / or the additional drug) is administered subsequently in the presence of the second drug (and / or the additional drug). The participant and the subject may be the same entity (e.g., a person).

[0469] The optimal concentration of the active ingredient in the selected medium can be determined empirically using methods known to the technician, depending on the desired final pharmaceutical formulation.

[0470] The dose range used to administer the sCR1 variant of this disclosure is a dose range that is sufficiently large to produce the desired effect. For example, the composition contains an effective amount of the sCR1 variant or sCR1 variant conjugate. In one example, the composition contains a therapeutically effective amount of the sCR1 variant or sCR1 variant conjugate. In another example, the composition contains a preventatively effective amount of the sCR1 variant or sCR1 variant conjugate.

[0471] The dosage should not be too high to avoid adverse side effects. Generally, the dosage will vary depending on the patient's age, condition, sex, and severity of the disease, and can be determined by a person skilled in the art. In the event of any complications, the dosage may be adjusted by an individual physician.

[0472] The dosage can vary from about 0.1 mg / kg to about 300 mg / kg, for example from about 0.2 mg / kg to about 200 mg / kg, for example from about 0.5 mg / kg to about 20 mg / kg, and is administered once or more daily for one to several days.

[0473] In some instances, the sCR1 variant or sCR1 variant conjugate is administered at an initial (or loading) dose higher than the subsequent (maintenance dose). For example, the sCR1 variant or sCR1 variant conjugate is administered at an initial dose of about 10 mg / kg to about 30 mg / kg. The sCR1 variant or sCR1 variant conjugate is then administered at a maintenance dose of about 0.0001 mg / kg to about 30 mg / kg. The maintenance dose may be administered every 2 to 30 days, for example, every 2 or 3 days or 6 or 9 days or 12 or 15 days or 18 days or 21 days or 24 days or 27 days or 30 days.

[0474] In some instances, a dose-escalation regimen is used, where the sCR1 variant or sCR1 variant conjugate is initially administered at a lower dose than is used in subsequent doses. This dosage regimen is available in cases where the subject initially experiences an adverse event.

[0475] If a subject does not respond adequately to treatment, multiple doses may be administered within a week. Alternatively, or additionally, increased doses may be administered.

[0476] Subjects can be retreated with sCR1 variants or sCR1 variant conjugates by administering one or more exposures or a series of doses, such as at least two exposures, such as about 2 to 60 exposures, more particularly about 2 to 40 exposures, and most particularly about 2 to 20 exposures.

[0477] In one instance, any retreatment can be performed when signs or symptoms of the disease, such as a bacterial infection, recur.

[0478] In another instance, any retreatment can be administered at defined intervals. For example, subsequent exposures can be administered at various intervals, such as approximately 24–28 weeks, 48–56 weeks, or longer. For example, the exposure can be administered at intervals of approximately 24–26 weeks, approximately 38–42 weeks, or approximately 50–54 weeks.

[0479] If a subject does not respond adequately to treatment, multiple doses may be administered within a week. Alternatively, or additionally, increased doses may be administered.

[0480] In another instance, for subjects experiencing adverse reactions, the initial (or loading) dose may be administered separately over multiple days within a week or over multiple consecutive days.

[0481] According to the methods of this disclosure, the administration of sCR1 variants or sCR1 variant conjugates can be continuous or intermittent, depending on factors such as the recipient's physiological condition, whether the purpose of administration is therapeutic or prophylactic, and other factors known to a skilled practitioner. The administration can be substantially continuous over a predetermined period of time, or it can be a series of intervals of doses, such as during or after the progression of the condition.

[0482] Kit and other substance combinations

[0483] Another example of this disclosure provides a kit containing an sCR1 variant or sCR1 variant conjugate of this disclosure, which, as described above, can be used to inhibit complement activity or to treat or prevent complement-mediated conditions.

[0484] In one instance, the kit includes (a) a container containing an sCR1 variant or sCR1 variant conjugate in a pharmaceutically acceptable carrier or diluent; and (b) a packaging insert with instructions for inhibiting complement activity in a subject or for treating or preventing complement-mediated conditions.

[0485] In one instance, the kit comprises (a) at least one sCR1 variant or sCR1 variant conjugate in a pharmaceutically acceptable carrier or diluent; (b) instructions for using the kit in a subject to inhibit complement activity or to treat or prevent complement-mediated conditions; and (c) optionally, at least one other therapeutically active compound or drug.

[0486] According to this example of the disclosure, a packaging insert is on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, etc. The container can be formed from a variety of materials, such as glass or plastic. The container contains or contains a composition that effectively inhibits complement activity or treats or prevents complement-mediated conditions and may have a sterile inlet (e.g., the container may be an intravenous bag or vial with a stopper that can be punctured by a hypodermic needle). At least one active agent in the composition is an sCR1 variant. The label or packaging insert indicates that the composition is intended for the treatment of eligible subjects, such as subjects with or susceptible to complement-mediated conditions, and provides specific guidance on the dosage and interval of the sCR1 variant and any other medications. The kit may further include additional containers containing pharmaceutically acceptable diluent buffers, such as sterile water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and / or glucose solution. The kit may further include other materials desired from a commercial and user perspective, including additional buffers, diluents, filters, needles, and syringes.

[0487] The kit optionally further includes a container containing a second drug, wherein the sCR1 variant or sCR1 variant conjugate is the first drug, and the product also includes instructions on a packaging insert regarding the treatment of the subject with the second drug in an effective amount. The second drug may be the therapeutic protein described above.

[0488] This disclosure includes the following non-limiting embodiments.

[0489] Example

[0490] Example 1: Generation of sCR1 variants

[0491] The human complement receptor type 1 (CR1) cDNA (GenBank accession number NP_000564) was codon-optimized for human expression and derived by... (Invitrogen TM Synthesized by Thermo Fisher Scientific. Full-length and truncated soluble CR1 (sCR1) variants were generated using standard PCR-based mutagenesis techniques. cDNA was generated with the Kozak concordant sequence (GCCACC) immediately upstream of the starting methionine (+1), then digested with NheI and XhoI and ligated into pcDNA3.1 (Invitrogen). TM (Thermo Fisher Scientific). The sCR1 variant cDNA was cloned with the C-terminal 8x histidine tag conforming to the reading frame. See Table 1 for a list of sCR1-8His variants.

[0492] Large-scale preparation of plasmid DNA was performed using the QIAGEN Plasmid Giga Kit according to the manufacturer's instructions. This was achieved using BigDye. TM Terminator version 3.1 Easy Reactive Cyclic Sequencing (BigDye) TM Terminator Version3.1Ready Reaction Cycle Sequencing, Invitrogen TM Thermo Fisher Scientific and Applied Biosystems 3130xl Genetic Analyzer validated the nucleotide sequences of all plasmid constructs by sequencing both strands.

[0493] According to the manufacturer's recommendation (Invitrogen) TM Thermo Fisher Scientific, using Expi293 TMThe expression system was tested using the sCR1 expression plasmid on Expi293F. TM Transient transfection of cells. All cell culture media were supplemented with antibiotics and antifungal agents. Thermo Fisher Scientific) and kept the cells in an incubator at 37°C and an atmosphere of 8% CO2.

[0494] Purification of the sCR1-8His peptide. In brief, to purify the sCR1 protein with a 6-histidine tag, culture supernatant was directly loaded onto NiGAgarose Excel affinity resin (GE Healthcare) pre-equilibrated with 20 mM NaH2PO4, 500 mM NaCl, and 10 mM imidazole (pH 7.4). After loading, the resin was washed with 20 mM NaH2PO4, 500 mM NaCl, and 25 mM imidazole (pH 7.4). The resin-bound sCR1 was blocked and eluted with 20 mM NaH2PO4, 500 mM NaCl, and 500 mM imidazole (pH 7.4), and the eluted protein was collected based on absorbance at 280 nm. The collected proteins were loaded onto a HiLoad 26 / 60 Superdex 200 preparative-grade column (GE Healthcare) pre-equilibrated in mt-PBS (137 mM NaCl, 27 mM KCl, 8.1 mM Na2HPO4, 1.15 mM KH2PO4, pH 7.4) to remove any contaminating proteins and buffer exchanged to the desired buffer. The purified proteins were concentrated to the desired concentration using an Amicon ultracentrifuge filter with a 50 kDa MWCO, sterilely filtered, and stored at -80°C. Due to intracellular processing, the mature sCR1-8His variant lacks the N-terminal 41 amino acid human CR1 signal peptide.

[0495] Table 1: sCR1-8His variants

[0496]

[0497] Example 2: The sCR1-8His variant exhibits complement-inhibiting activity in vitro.

[0498] To assess complement inhibitory activity, according to the manufacturer's instructions, in The sCR1-8His variant was tested in a complement assay (EuroDiagnostica). Briefly, the sCR1-8His variant protein was serially diluted with PBS in a 96-well plate. 50 μl of each diluted sCR1-8His variant sample or PBS alone was added to 202.5 μl of pre-diluted human serum (classical / lectin pathway 1:101) or 220 μl of diluted serum (bypass pathway 1:18) (using the appropriate assay diluent for each complement pathway (according to the manufacturer's instructions)) and incubated at room temperature (RT) for 30 min. Once added to the pre-diluted serum, the final starting concentration for each protein was 40 nM. 100 μl samples were transferred in duplicate to the assay plate and incubated at 37 °C (CO2-free) for 1 h. The wells were drained and washed three times with 300 μl / well of 1x wash buffer (according to the manufacturer's instructions). The C5b-9 terminal complex was detected using 100 μl / well of alkaline phosphatase-conjugated anti-C5b-9 specific monoclonal antibody and incubated at room temperature for 30 minutes. Unbound antibody was discarded, and the wells were washed three times with 300 μl / well of 1x wash buffer. Bound antibody was detected using 100 μl / well of alkaline phosphatase substrate solution and incubated at room temperature for 30 minutes. The absorbance was read at 405 nm using an Envision plate reader.

[0499] Raw values ​​are expressed as the percentage of C5b-9 formation in the control group (i.e., 100% C5b-9 formation) containing only serum and PBS. Results were analyzed in Graph Pad Prism using a logarithm (inhibitor) fitted to a response-variable slope (four parameters) to determine the IC50. 50 Values. The bottom and top are limited to values ​​of 0 and 100, respectively.

[0500] Except for sCR1(490-939)-8His, sCR1(940-1392)-8His, and sCR1(1393-1971)-8His, all sCR1-8His variants are functionally active in the classical, lectin, and alternative pathways. sCR1(490-939)-8His is functionally active only in the alternative pathway, while sCR1(940-1392)-8His is functionally active in both the lectin and alternative pathways. sCR1(1939-1971)-8His showed no activity in any of the classical, lectin, or alternative pathways.

[0501] As shown in Table 2 below, in Wieslab assays, sCR1(1392)-8His exhibited increased inhibitory activity in all three complement pathways (i.e., the classical pathway, the lectin pathway, and the bypass pathway) compared to the full-length sCR1(1971)-8His and other sCR1 fragments.

[0502] The functional activity of the sCR1-8His variant was also tested using hemolysis assays (e.g., classical pathway (i.e., CH50) and alternative pathway (ApH50) inhibition assays).

[0503] To evaluate the inhibitory effect of the sCR1 variant on the classical complement pathway (CH50), sheep erythrocytes (Siemens) were sensitized with rabbit anti-sheep antibody (Ambozeptor 6000; Siemens) and diluted to 4 x 10⁻⁶. 8 cells / mL GVB ++ (GVB, 0.15 mM CaCl2, 0.5 mM MgCl2). The sCR1 variant was pre-incubated in 1 / 40 diluted NHS (room temperature, 30 min), then added to erythrocytes at a 1 / 1 (v / v) ratio and incubated at 37 °C in a microtiter plate shaker for 1 h. Hemolysis in the supernatant was determined by measuring the absorbance of the released hemoglobin at 412 nm after adding ice-cold GVBE (GVB, 10 mM EDTA) and centrifuging (4 °C, 1250 x g, 5 min). Cells incubated with NHS and buffer alone served as a 100% lysis control. The inhibition of lysis by the sCR1 variant was calculated relative to the control.

[0504] To assess the inhibitory effect of the sCR1 variant on the complement system bypass pathway (i.e., ApH50), rabbit erythrocytes (Jackson Laboratories) were washed and diluted to 2 x 10⁻⁶. 8 Cells / mL GVB / MgEGTA (GVB, 5 mM MgEGTA). The sCR1 variant was pre-incubated in 1 / 6 diluted NHS (at room temperature for 30 min), then added to erythrocytes at a 2 / 1 (v / v) ratio and incubated at 37 °C in a microtiter plate shaker for 1 h. Hemolysis in the supernatant was determined by measuring the absorbance of released hemoglobin at 412 nm after adding ice-cold GVBE and centrifuging (1250 x g, 10 min). Cells incubated with NHS and buffer alone served as a 100% lysis control. The inhibition of lysis by the sCR1 variant was calculated relative to the control.

[0505] Except for sCR1(1393-1971)-8His, all variants showed functional activity in both CH50 and ApH50 assays. As shown in Table 3, sCR1(1392)-8His exhibited increased activity compared to sCR1(1971)-8His in both assays.

[0506] Table 2: Relative in vitro activity of sCR1 variants as determined by Wieslab

[0507]

[0508]

[0509] Table 3: Relative in vitro activity of sCR1 variants in hemolysis assays

[0510]

[0511] Example 3: Compared with sCR1(1971)-8His, the sCR1(1392)-8His variant showed increased stability.

[0512] To assess the stability of sCR1(1392)-8His under different buffer conditions, differential scanning fluorometry (DSF) was performed to measure the thermal stability of sCR1(1392)-8His protein compared to the full-length sCR1(1971)-8His protein. Protein stability was assessed under a range of salts (NaCl 0 mM, 50 mM, 150 mM, and 500 mM) and pH conditions for the following buffers: citrate, HEPES, sodium acetate, phosphate, glycine, histidine, TRIS, and proline.

[0513] In short, aliquot 5 μl of 4x buffer concentrate into two 384-well plates. Dilute sCR1(1392)-8His and sCR1(1971)-8His proteins to 0.13 mg / ml in MT-PBS, then incorporate 1 / 20 of the dye stock solution. Orange; Sigma), diluted with water to a final dilution of 1 / 400 for each assay reaction. Then, 15 μl of the protein / dye mixture was dispensed into each well of a 384-well plate containing the buffer concentrate. The plate was sealed with optical adhesive caps and centrifuged at 3220 g for 1 minute, then processed using QuantStudio. TM The experiment was conducted on a real-time PCR instrument (Applied Biosystems). Melting curves were generated by cooling and holding the temperature at 20.0 °C for 1 minute, followed by increasing the temperature from 20.0 °C to 99.0 °C at a rate of 0.05 °C / s. The transition midpoint (Tm) value was calculated from each melting curve using the first derivative function and Protein Thermal Shift software (Applied Biosystems). Contour plots were generated using JMP13 to graphically display how the Tm value changed relative to NaCl concentration (x-axis) and pH (y-axis).

[0514] sCR1(1392)-8His is stable under the following buffer conditions: phosphate (pH 6.0-8.0; NaCl 0-500mM); citrate phosphate (pH 6.0-8.0; NaCl 0-500mM); Tris (pH 7.0-9.0; NaCl 0-500mM); glycine (pH 9.0-10.0; NaCl 0-500mM); HEPES (pH 6.5-8.5; NaCl 0-500mM); and histidine (pH 6.0-7.0; NaCl 0-500mM). The maximum Tm value measured for sCR1(1392)-8His is 61.4℃, and the maximum Tm value measured for sCR1(1971)-8His is 61.7℃.

[0515] Based on buffer screening, sCR1(1392)-8His is more stable than sCR1(1971)-8His.

[0516] Example 4: Sialidized sCR1(1392)-8His has an improved in vivo half-life

[0517] To assess whether the in vivo half-life of sCR1(1392)-8His could be prolonged, a sialylated version of sCR1(1392)-8His (sCR1(1392)-8His) was prepared. SIA In short, the sialylated substance was produced by co-transfecting Expi293F cells with cDNA encoding sCR1(1392)-8His, cDNA encoding human ST3GAL3 (ST3β-galactosyl α-2,3-sialyltransferase 3, GenBank accession number NP_006270), and cDNA encoding human B4GALT1 (human β1,4-galactosyltransferase, GenBank accession number NP_001488.2) in a ratio of 94:3:3.

[0518] As shown in Table 4, sCR1(1392)-8His generated in cells transfected with ST3GAL3 / B4GALT1 SIA The substance has a higher proportion of sialylated glycans. In particular, sialylated sCR1(1392)-8His SIA The substance has a higher proportion of di, tri, and tetra sialylated glycans.

[0519] Table 4: Proportion of polysaccharides in sialylated sCR1(1392)-8His

[0520]

[0521] In human FcRn transgenic mice (B6.Cg-Fcgrt)tm1Dcr Tg(FCGRT)32Dcr / DcrJ; The Jackson Laboratory stock number 014565) tested sCR1(1392)-8His and its sialylated form (sCR1(1392)-8His) SIA The in vivo half-life of sCR1(1392)-8His was determined. Mice were administered 30 mg / kg sCR1(1392)-8His intravenously (a single bolus injection into the tail vein). SIA Plasma was collected at different time points (Group A: 5 min and 4 h, n=3); Group B: 0.5 h and 8 h, n=3; Group C: 1 h and 16 h, n=3; Group D: 2 h and 48 h, n=3). Blood was mixed with citrate buffer at a ratio of 8 parts blood to 2 parts citrate buffer. Following the manufacturer's instructions with the following modifications: a standard curve (range 3-250 ng / mL) was generated using each test sample. The assay buffer used was 1% BSA heat shock fraction, protease-free (Sigma catalog number A3059). The wash buffer was PBS + 0.05% v / v Tween-20. Human sCR1 plasma levels were measured in an anti-human CD35 ELISA (RayBiotech, catalog number ELH CD35). Mean retention time (MRT) and area under the curve (AUC) were calculated using standard statistical formulas.

[0522] like Figure 1 As shown, compared with sCR1(1392)-8His, sCR1(1392)-8His SIA With improved in vivo retention, MRT increased 25-fold (14.7 hours vs. 35 minutes), while AUC increased 8-fold (AUC = 516.5 vs. 65.74).

[0523] Example 5: sCR1(1392)-8His alleviates anti-GBM glomerulonephritis

[0524] The efficacy of sCR1(1392)-8His treatment was evaluated in an in vivo model of anti-glomerular basement membrane (GBM) glomerulonephritis. Briefly, anti-GBM glomerulonephritis IgG was induced in C57BL / 6 mice by intravenous injection of 1 mg polyclonal rabbit anti-GBM antiserum (IgG fraction) on day 0, followed by intraperitoneal injection of 2 mg mouse monoclonal anti-rabbit IgG (MsαRb IgG generated from hybridoma CRL-1753 (ATCC)) on day 6. Mice were intraperitoneally injected with PBS or 60 mg / kg sCR1(1392)-8His, sCR1(1971)-8His, or anti-mouse C5 mAb (muBB5.1-mIgG1κ; Rother R et al., Nat Biotechnol. 2007; Wang Y et al., Proc Natl Acad Sci 1995). Another group of mice were treated with 60 mg / kg sCR1(1392)-8His only on day 6, not on day 5 (sCR1(1392)-8His, once). On day 6, the drug (i.e., sCR1 variant or BB5.1) or PBS control was administered approximately one hour before the injection of MsαRb IgG mAb. After MsαRb injection, mice were placed individually in metabolic cages, and urine was collected over 24 hours. Urinary albumin levels were measured using an ELISA kit (Bethyl Laboratories), and albuminuria per mouse was plotted in μg / 24h.

[0525] The sCR1(1392)-8His and sCR1(1971)-8His substances used in this experiment were relatively unsialylated and were produced in Expi293F cells that were not co-transfected with ST3GAL3 and B4GALT1 cDNA.

[0526] like Figure 2 As shown, urinary albumin levels were significantly reduced in mice treated with sCR1(1392)-8His (PBS vs. sCR1(1392)-8His, 2 times: p = 0.0147; PBS vs. sCR1(1392)-8His, 1 time: p = 0.1526; PBS vs. BB5.1: p = 0.0078).

[0527] Example 6: Sialidized sCR1(1392)-8His alleviates anti-GBM glomerulonephritis

[0528] As described in Example 4, sCR1(1392)-8His was evaluated in an in vivo model of anti-glomerular basement membrane (GBM) glomerulonephritis. SIAThe efficacy of treatment. On day 6, mice were injected intraperitoneally with PBS or 10 mg / kg, 30 mg / kg, or 60 mg / kg sCR1(1392)-8His SIA .

[0529] On day 6, approximately one hour before the injection of MsαRb IgG mAb, sCR1(1392)-8His was administered. SIA Alternatively, a PBS control was used. After MsαRb injection, mice were placed individually in metabolic cages, and urine was collected over a 24-hour period. Urinary albumin levels were measured using an ELISA kit (BethylLaboratories), and albuminuria for each mouse was plotted in μg / 24h.

[0530] like Figure 3 As shown, compared with control (i.e., PBS) treated mice, mice treated with 10 mg / kg, 30 mg / kg and 60 mg / kg sCR1(1392)-8His SIA The urinary albumin level decreased in the treated mice.

[0531] Example 7: Sialidified sCR1(1392)-8His protects against renal ischemia-reperfusion injury

[0532] sCR1(1392)-8His SIA The therapeutic effect was evaluated in a warm in vivo model of renal ischemia-reperfusion (IR) injury. Male C57BL / 6 mice aged 10–20 weeks were anesthetized and underwent right nephrectomy and 22-minute left renal ischemia or right-only nephrectomy (Sham) at 37°C. Briefly, a midline abdominal incision was made and the renal pedicle was bluntly dissected. After right nephrectomy, the left renal pedicle was clamped with a microvascular clamp for 22 min while the animals were held at 37°C. After ischemia, the microvascular clamp was removed, and the kidney was observed to confirm complete reperfusion. One hour before ischemia, the mice were administered 60 mg / kg sCR1(1392)-8His intraperitoneally. SIA Mice were treated with either a mediator control (n=14) or a mediator control (n=8). Mice were sacrificed 24 hours after reperfusion, and serum and plasma were collected to assess renal function (creatinine, urea) and complement activation (C3b, C5a ELISA). Kidneys were removed, and complement C9 deposition and immune cell infiltration were analyzed by immunofluorescence / confocal microscopy.

[0533] As shown in Table 5 below, compared with Sham, mice treated with the vector showed significantly increased IR-induced severe kidney damage, as indicated by a marked increase in serum creatinine and urea, plasma C3b and C5a, tissue C9 deposition, and neutrophil and macrophage infiltration.

[0534] Using sCR1(1392)-8His SIATreatment significantly protected against IR-induced damage, manifested as markedly reduced renal function (i.e., serum creatinine and urea), complement activation and deposition (i.e., plasma C3b, C5a, and tissue C9 deposition), and cellular infiltration (i.e., neutrophil and macrophage infiltration) (see Table 5).

[0535] The results show that in this model, sCR1(1392)-8His SIA It protects against IR-mediated kidney damage, accompanied by a significant reduction in renal function loss (indicated by serum creatinine and urea) and decreased plasma complement activation products, complement tissue deposition, and infiltration of innate immune cells.

[0536] Table 5: sCR1(1392)-8His SIA Effects on IR-mediated renal damage

[0537]

[0538] Example 8: Effect of sialylated sCR1(1392)-8His dose alteration on renal ischemia-reperfusion injury

[0539] To evaluate sCR1(1392)-8His SIA Dose-response relationships in an in vivo model of warm renal ischemia-reperfusion (IR) injury, where mice were administered 10 mg / kg, 30 mg / kg, or 60 mg / kg sCR1(1392)-8His intraperitoneally 1 hour before ischemia. SIA Treatment may be administered as a mediator control. Induce IR as described above, and assess IR-mediated renal damage as previously described.

[0540] By administering 60 mg / kg sCR1(1392)-8His 1 hour before ischemia SIA Or, as a mediator control, administer 60 mg / kg sCR1(1392)-8His at 1 hour and / or 2 hours after ischemia. SIA Or use a mediator control to assess the maintenance of elevated levels of sCR1(1392)-8His SIA The effects were as follows: IR was induced as described above, and IR-mediated kidney damage was assessed as previously described.

[0541] Example 9: Generation of sCR1 variant fusion

[0542] Recombinant sCR1 fusion proteins were generated by fusing sCR1 variants at the N-terminus or C-terminus of the sCR1 sequence with human serum albumin (HSA) (GenBank accession number NP_000468), human IgG1 Fc (GenBank accession number P01857), or human IgG4 Fc (aa99-327; GenBank accession number P01861) (Table 6). Recombinant fusion proteins were prepared using standard cloning techniques. In the case of HSA fusion, a GS13 adapter (GSGGSGGSGGSGS) was used to link the sCR1 and HSA sequences. In the case of IgG4 Fc and IgG1 Fc fusion, no adapter was used. For some constructs, ceruloplasmin signal peptide (GenBank accession number NP_000087) was used. All fusion proteins were analyzed at Expi 293F. TM The sCR1 protein was expressed in cells and purified as described above.

[0543] To purify the human serum albumin (HSA)-labeled sCR1 fusion, culture supernatant was directly loaded onto Capture Select Human Albumin Affinity Matrix affinity resin (GE Healthcare) pre-equilibrated with 20 mM Tris, pH 7.4. After loading all supernatant, the resin was washed with 20 mM Tris (pH 7.4). The resin-bound sCR1 was eluted with 20 mM Tris and 2 M MgCl2 (pH 7.4), and the eluted protein was collected based on absorbance at 280 nm. The eluted protein was loaded onto a HiLoad 26 / 60 Superdex 200 preparative-grade column (GE Healthcare) pre-equilibrated in mt-PBS (137 mM NaCl, 27 mM KCl, 8.1 mM Na2HPO4, 1.15 mM KH2PO4, pH 7.4) to remove any contaminating proteins and buffer exchanged to the required buffer. The purified protein was concentrated to the desired concentration using an Amicon ultracentrifuge filter with a 50 kDa MWCO, sterile filtered, and stored at -80°C.

[0544] To purify the Fc-fused sCR1 variant, culture supernatant was directly loaded onto MabSelect SuRe affinity resin (GE Healthcare) pre-equilibrated with mt-PBS (137 mM NaCl, 27 mM KCl, 8.1 mM Na2HPO4, 1.15 mM KH2PO4, pH 7.4). After loading all supernatant, the resin was washed with mt-PBS at pH 7.4. Weakly bound non-target proteins were eluted with 0.1 M sodium citrate at pH 5.0. Resin-bound sCR1 was eluted with 0.1 M sodium citrate (pH 3.0), and the eluted protein was collected based on absorbance at 280 nm. The eluted protein was loaded onto a HiLoad 26 / 60 Superdex 200 preparative-grade column (GE Healthcare) pre-equilibrated in mt-PBS to remove any contaminating proteins and buffer exchange to the desired buffer. The purified protein was concentrated to the desired concentration using an Amicon ultracentrifuge filter with a 50 kDa MWCO, sterile filtered, and stored at -80°C.

[0545] Table 6: sCR1 Variant Fusions

[0546]

[0547]

[0548] As previously described, the Wieslab assay was used to determine that all sCR1 variants conjugated to HSA, IgG1 Fc, or IgG4 Fc at the N- or C-terminus exhibited complement inhibitory activity in the classical, lectin, and alternative pathways.

[0549] As shown in Tables 7 and 8 below, in Wieslab assays and hemolysis (i.e., CH50 and ApH50) inhibition assays, sCR1(1392)-GS13-HSA and sCR1(1392)-IgG4 Fc exhibited increased complement inhibitory activity compared to sCR1(1392)-8His in all three complement pathways (i.e., classical pathway, lectin pathway, and alternative pathway). Compared to sCR1(1392)-8His, the sCR1(1392)-IgG4 Fc fusion complex showed approximately a 2-fold increase in complement inhibitory activity in the classical pathway (mean 2.31 ± 0.16) and the lectin pathway (mean 2.37 ± 0.44), and a 7-8 fold increase in complement inhibitory activity in the alternative pathway (mean 7.61 ± 2.52). Fusion at the C-terminus with HSA and IgG4 Fc did not adversely affect the complement inhibitory activity of sCR1(1392).

[0550] Table 7: Relative in vitro activity of sCR1 variant fusions as determined by Wieslab

[0551]

[0552] Table 8: Relative in vitro activity of sCR1 variant fusion bodies in hemolysis assay

[0553]

[0554]

[0555] As described above, a recombinant sCR1-HSA fusion (sCR1(1392)-HSA; SEQ ID NO: 51) was generated, and in CHO The compound was expressed in cells and purified as described above. Complement activity in the classical and lectin pathways was measured using Wieslab assays, confirming its origin from CHO. The substance and the substance derived from Expi293F TM It has similar activity to other substances.

[0556] Example 10: Compared with the monomeric Fc sCR1 variant fusion, the dimer Fc sCR1 variant fusion has increased inhibitory activity.

[0557] As previously described, a recombinant sCR1 fusion is generated, which is conjugated at the C-terminus to a dimer IgG1 Fc, a dimer IgG4 Fc, or a monomeric IgG4 Fc.

[0558] As shown in Tables 9 and 10 below, in the Wieslab assay and the hemolysis (i.e., CH50 and ApH50) inhibition assay, sCR1(1392)-IgG1 Fc and sCR1(1392)-IgG4 Fc exhibited increased complement inhibitory activity compared to sCR1(1392)-8His when measured in all three complement pathways (i.e., the classical pathway, the lectin pathway, and the alternative pathway). In particular, compared to sCR1(1392)-8His and the sCR1(1392)-monomer IgG1 Fc fusion, the sCR1(1392)-dimer IgG4 Fc fusion showed approximately 2-fold increased complement inhibitory activity in the classical and lectin pathways, and 4-fold increased complement inhibitory activity in the alternative pathway. These results also indicate that N-terminal fusion has an adverse effect on the complement-inhibiting activity of sCR1(1392) in the bypass pathway.

[0559] Table 9: Relative in vitro activity of sCR1 variant fusions as determined by Wieslab

[0560]

[0561] Table 10: Relative in vitro activity of sCR1 variant fusion bodies in hemolysis assay

[0562] sCR1(1392)-8His 427 956 <![CDATA[sCR1(1392)-IgG4Fc]]> 151 165 <![CDATA[IgG4Fc-sCR1(1392)]]> 178 2061 <![CDATA[sCR1(1392)-monomeric IgG4Fc]]> 259 1060 sequence list <110> Jet Ltd. <120> Type 1 soluble complement receptor variants and their uses <130> 524790PCT <150> 2018901703 <151> 2018-05-16 <160> 51 <170> PatentIn version 3.5 <210> 1 <211> 1971 <212> PRT <213> Artificial sequence <220> <223> sCR1 <400> 1 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly Gln Cys Asn Ala Pro Glu Trp 35 40 45 Leu Pro Phe Ala Arg Pro Thr Asn Leu Thr Asp Glu Phe Glu Phe Pro 50 55 60 Ile Gly Thr Tyr Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Ser Gly Arg 65 70 75 80 Pro Phe Ser Ile Ile Cys Leu Lys Asn Ser Val Trp Thr Gly Ala Lys 85 90 95 Asp Arg Cys Arg Arg Lys Ser Cys Arg Asn Pro Pro Asp Pro Val Asn 100 105 110 Gly Met Val His Val Ile Lys Gly Ile Gln Phe Gly Ser Gln Ile Lys 115 120 125 Tyr Ser Cys Thr Lys Gly Tyr Arg Leu Ile Gly Ser Ser Ser Ala Thr 130 135 140 Cys Ile Ile Ser Gly Asp Thr Val Ile Trp Asp Asn Glu Thr Pro Ile 145 150 155 160 Cys Asp Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Thr Asn Gly Asp 165 170 175 Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr 180 185 190 Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val 195 200 205 Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 210 215 220 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro 225 230 235 240 Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe 245 250 255 Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met 260 265 270 Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 275 280 285 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu 290 295 300 His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln 305 310 315 320 Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala 325 330 335 Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr 340 345 350 Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly 355 360 365 Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe 370 375 380 Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys 385 390 395 400 Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys 405 410 415 Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His 420 425 430 Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Thr Val Asn Tyr 435 440 445 Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly 450 455 460 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 465 470 475 480 Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln Ala Pro 485 490 495 Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp 500 505 510 Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr 515 520 525 Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp Ser Ser 530 535 540 Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro 545 550 555 560 Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val Gly Ser Arg 565 570 575 Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile Gly His Ser Ser 580 585 590 Ala Glu Cys Ile Leu Ser Gly Asn Ala Ala His Trp Ser Thr Lys Pro 595 600 605 Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn 610 615 620 Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val 625 630 635 640 Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu 645 650 655 Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val 660 665 670 Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys 675 680 685 Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser 690 695 700 Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe 705 710 715 720 Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp 725 730 735 Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp 740 745 750 Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro 755 760 765 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly 770 775 780 Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala 785 790 795 800 Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu 805 810 815 Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val 820 825 830 Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser 835 840 845 Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro 850 855 860 Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly 865 870 875 880 Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Ala Val 885 890 895 Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu 900 905 910 Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly 915 920 925 Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln 930 935 940 Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala 945 950 955 960 Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu 965 970 975 Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp 980 985 990 Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro 995 1000 1005 Asp Pro Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val 1010 1015 1020 Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile 1025 1030 1035 Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Thr Ala His 1040 1045 1050 Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu 1055 1060 1065 Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 1070 1075 1080 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu Gly 1085 1090 1095 Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 1100 1105 1110 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 1115 1120 1125 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val 1130 1135 1140 Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu 1145 1150 1155 Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys 1160 1165 1170 Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 1175 1180 1185 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile 1190 1195 1200 Leu His Gly Glu His Thr Pro Ser His Gln Asp Asn Phe Ser Pro 1205 1210 1215 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg 1220 1225 1230 Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly Asp Trp Ser Pro 1235 1240 1245 Glu Ala Pro Arg Cys Ala Val Lys Ser Cys Asp Asp Phe Leu Gly 1250 1255 1260 Gln Leu Pro His Gly Arg Val Leu Phe Pro Leu Asn Leu Gln Leu 1265 1270 1275 Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg Leu Lys 1280 1285 1290 Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met Arg Ser Leu 1295 1300 1305 Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys Pro Asn 1310 1315 1320 Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser Gly 1325 1330 1335 Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 1340 1345 1350 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile 1355 1360 1365 Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro 1370 1375 1380 Ala Pro Arg Cys Glu Leu Ser Val Arg Ala Gly His Cys Lys Thr 1385 1390 1395 Pro Glu Gln Phe Pro Phe Ala Ser Pro Thr Ile Pro Ile Asn Asp 1400 1405 1410 Phe Glu Phe Pro Val Gly Thr Ser Leu Asn Tyr Glu Cys Arg Pro 1415 1420 1425 Gly Tyr Phe Gly Lys Met Phe Ser Ile Ser Cys Leu Glu Asn Leu 1430 1435 1440 Val Trp Ser Ser Val Glu Asp Asn Cys Arg Arg Lys Ser Cys Gly 1445 1450 1455 Pro Pro Pro Glu Pro Phe Asn Gly Met Val His Ile Asn Thr Asp 1460 1465 1470 Thr Gln Phe Gly Ser Thr Val Asn Tyr Ser Cys Asn Glu Gly Phe 1475 1480 1485 Arg Leu Ile Gly Ser Pro Ser Thr Thr Cys Leu Val Ser Gly Asn 1490 1495 1500 Asn Val Thr Trp Asp Lys Lys Ala Pro Ile Cys Glu Ile Ile Ser 1505 1510 1515 Cys Glu Pro Pro Pro Thr Ile Ser Asn Gly Asp Phe Tyr Ser Asn 1520 1525 1530 Asn Arg Thr Ser Phe His Asn Gly Thr Val Val Thr Tyr Gln Cys 1535 1540 1545 His Thr Gly Pro Asp Gly Glu Gln Leu Phe Glu Leu Val Gly Glu 1550 1555 1560 Arg Ser Ile Tyr Cys Thr Ser Lys Asp Asp Gln Val Gly Val Trp 1565 1570 1575 Ser Ser Pro Pro Pro Arg Cys Ile Ser Thr Asn Lys Cys Thr Ala 1580 1585 1590 Pro Glu Val Glu Asn Ala Ile Arg Val Pro Gly Asn Arg Ser Phe 1595 1600 1605 Phe Ser Leu Thr Glu Ile Ile Arg Phe Arg Cys Gln Pro Gly Phe 1610 1615 1620 Val Met Val Gly Ser His Thr Val Gln Cys Gln Thr Asn Gly Arg 1625 1630 1635 Trp Gly Pro Lys Leu Pro His Cys Ser Arg Val Cys Gln Pro Pro 1640 1645 1650 Pro Glu Ile Leu His Gly Glu His Thr Leu Ser His Gln Asp Asn 1655 1660 1665 Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Ser Tyr 1670 1675 1680 Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly Asp 1685 1690 1695 Trp Ser Pro Glu Ala Pro Arg Cys Thr Val Lys Ser Cys Asp Asp 1700 1705 1710 Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Leu Pro Leu Asn 1715 1720 1725 Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe 1730 1735 1740 Arg Leu Lys Gly Arg Ser Ala Ser His Cys Val Leu Ala Gly Met 1745 1750 1755 Lys Ala Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe 1760 1765 1770 Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr 1775 1780 1785 Pro Phe Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Ala Cys 1790 1795 1800 Asp Thr His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu 1805 1810 1815 Ser Ser Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 1820 1825 1830 Ser Ser Pro Ala Pro Arg Cys Glu Leu Ser Val Pro Ala Ala Cys 1835 1840 1845 Pro His Pro Pro Lys Ile Gln Asn Gly His Tyr Ile Gly Gly His 1850 1855 1860 Val Ser Leu Tyr Leu Pro Gly Met Thr Ile Ser Tyr Ile Cys Asp 1865 1870 1875 Pro Gly Tyr Leu Leu Val Gly Lys Gly Phe Ile Phe Cys Thr Asp 1880 1885 1890 Gln Gly Ile Trp Ser Gln Leu Asp His Tyr Cys Lys Glu Val Asn 1895 1900 1905 Cys Ser Phe Pro Leu Phe Met Asn Gly Ile Ser Lys Glu Leu Glu 1910 1915 1920 Met Lys Lys Val Tyr His Tyr Gly Asp Tyr Val Thr Leu Lys Cys 1925 1930 1935 Glu Asp Gly Tyr Thr Leu Glu Gly Ser Pro Trp Ser Gln Cys Gln 1940 1945 1950 Ala Asp Asp Arg Trp Asp Pro Pro Leu Ala Lys Cys Thr Ser Arg 1955 1960 1965 Thr His Asp 1970 <210> 2 <211> 1930 <212> PRT <213> Artificial sequence <220> <223> sCR1(1971) <400> 2 Gln Cys Asn Ala Pro Glu Trp Leu Pro Phe Ala Arg Pro Thr Asn Leu 1 5 10 15 Thr Asp Glu Phe Glu Phe Pro Ile Gly Thr Tyr Leu Asn Tyr Glu Cys 20 25 30 Arg Pro Gly Tyr Ser Gly Arg Pro Phe Ser Ile Ile Cys Leu Lys Asn 35 40 45 Ser Val Trp Thr Gly Ala Lys Asp Arg Cys Arg Arg Lys Ser Cys Arg 50 55 60 Asn Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Lys Gly Ile 65 70 75 80 Gln Phe Gly Ser Gln Ile Lys Tyr Ser Cys Thr Lys Gly Tyr Arg Leu 85 90 95 Ile Gly Ser Ser Ser Ala Thr Cys Ile Ile Ser Gly Asp Thr Val Ile 100 105 110 Trp Asp Asn Glu Thr Pro Ile Cys Asp Arg Ile Pro Cys Gly Leu Pro 115 120 125 Pro Thr Ile Thr Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe 130 135 140 His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly 145 150 155 160 Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser 165 170 175 Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile 180 185 190 Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val 195 200 205 Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg 210 215 220 Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln 225 230 235 240 Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys 245 250 255 Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys 260 265 270 Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 275 280 285 Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp 290 295 300 Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe 305 310 315 320 Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln 325 330 335 Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys 340 345 350 Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp 355 360 365 Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro 370 375 380 Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro 385 390 395 400 Phe Gly Lys Thr Val Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly 405 410 415 Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp 420 425 430 Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile 435 440 445 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 450 455 460 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 465 470 475 480 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 485 490 495 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 500 505 510 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 515 520 525 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 530 535 540 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 545 550 555 560 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 565 570 575 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 580 585 590 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 595 600 605 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 610 615 620 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 625 630 635 640 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 645 650 655 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 660 665 670 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 675 680 685 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 690 695 700 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 705 710 715 720 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 725 730 735 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 740 745 750 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 755 760 765 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 770 775 780 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 785 790 795 800 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 805 810 815 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 820 825 830 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 835 840 845 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 850 855 860 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 865 870 875 880 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 885 890 895 Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys 900 905 910 Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu 915 920 925 Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr 930 935 940 Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg 945 950 955 960 Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val 965 970 975 Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr 980 985 990 Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly 995 1000 1005 Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 1010 1015 1020 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser 1025 1030 1035 Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg 1040 1045 1050 Cys Asn Leu Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly 1055 1060 1065 Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 1070 1075 1080 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr 1085 1090 1095 Pro Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser 1100 1105 1110 Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly 1115 1120 1125 Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn 1130 1135 1140 Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro 1145 1150 1155 Pro Pro Glu Ile Leu His Gly Glu His Thr Pro Ser His Gln Asp 1160 1165 1170 Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 1175 1180 1185 Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly 1190 1195 1200 Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser Cys Asp 1205 1210 1215 Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro Leu 1220 1225 1230 Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly 1235 1240 1245 Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly 1250 1255 1260 Met Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile 1265 1270 1275 Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly 1280 1285 1290 Thr Pro Ser Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr 1295 1300 1305 Cys Asp Pro His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly 1310 1315 1320 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val 1325 1330 1335 Trp Ser Ser Pro Ala Pro Arg Cys Glu Leu Ser Val Arg Ala Gly 1340 1345 1350 His Cys Lys Thr Pro Glu Gln Phe Pro Phe Ala Ser Pro Thr Ile 1355 1360 1365 Pro Ile Asn Asp Phe Glu Phe Pro Val Gly Thr Ser Leu Asn Tyr 1370 1375 1380 Glu Cys Arg Pro Gly Tyr Phe Gly Lys Met Phe Ser Ile Ser Cys 1385 1390 1395 Leu Glu Asn Leu Val Trp Ser Ser Val Glu Asp Asn Cys Arg Arg 1400 1405 1410 Lys Ser Cys Gly Pro Pro Pro Glu Pro Phe Asn Gly Met Val His 1415 1420 1425 Ile Asn Thr Asp Thr Gln Phe Gly Ser Thr Val Asn Tyr Ser Cys 1430 1435 1440 Asn Glu Gly Phe Arg Leu Ile Gly Ser Pro Ser Thr Thr Cys Leu 1445 1450 1455 Val Ser Gly Asn Asn Val Thr Trp Asp Lys Lys Ala Pro Ile Cys 1460 1465 1470 Glu Ile Ile Ser Cys Glu Pro Pro Pro Thr Ile Ser Asn Gly Asp 1475 1480 1485 Phe Tyr Ser Asn Asn Arg Thr Ser Phe His Asn Gly Thr Val Val 1490 1495 1500 Thr Tyr Gln Cys His Thr Gly Pro Asp Gly Glu Gln Leu Phe Glu 1505 1510 1515 Leu Val Gly Glu Arg Ser Ile Tyr Cys Thr Ser Lys Asp Asp Gln 1520 1525 1530 Val Gly Val Trp Ser Ser Pro Pro Pro Arg Cys Ile Ser Thr Asn 1535 1540 1545 Lys Cys Thr Ala Pro Glu Val Glu Asn Ala Ile Arg Val Pro Gly 1550 1555 1560 Asn Arg Ser Phe Phe Ser Leu Thr Glu Ile Ile Arg Phe Arg Cys 1565 1570 1575 Gln Pro Gly Phe Val Met Val Gly Ser His Thr Val Gln Cys Gln 1580 1585 1590 Thr Asn Gly Arg Trp Gly Pro Lys Leu Pro His Cys Ser Arg Val 1595 1600 1605 Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr Leu Ser 1610 1615 1620 His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys 1625 1630 1635 Glu Pro Ser Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr 1640 1645 1650 Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Thr Val Lys 1655 1660 1665 Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu 1670 1675 1680 Leu Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys 1685 1690 1695 Asp Glu Gly Phe Arg Leu Lys Gly Arg Ser Ala Ser His Cys Val 1700 1705 1710 Leu Ala Gly Met Lys Ala Leu Trp Asn Ser Ser Val Pro Val Cys 1715 1720 1725 Glu Gln Ile Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg 1730 1735 1740 His Thr Gly Thr Pro Phe Gly Asp Ile Pro Tyr Gly Lys Glu Ile 1745 1750 1755 Ser Tyr Ala Cys Asp Thr His Pro Asp Arg Gly Met Thr Phe Asn 1760 1765 1770 Leu Ile Gly Glu Ser Ser Ile Arg Cys Thr Ser Asp Pro Gln Gly 1775 1780 1785 Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Glu Leu Ser Val 1790 1795 1800 Pro Ala Ala Cys Pro His Pro Pro Lys Ile Gln Asn Gly His Tyr 1805 1810 1815 Ile Gly Gly His Val Ser Leu Tyr Leu Pro Gly Met Thr Ile Ser 1820 1825 1830 Tyr Ile Cys Asp Pro Gly Tyr Leu Leu Val Gly Lys Gly Phe Ile 1835 1840 1845 Phe Cys Thr Asp Gln Gly Ile Trp Ser Gln Leu Asp His Tyr Cys 1850 1855 1860 Lys Glu Val Asn Cys Ser Phe Pro Leu Phe Met Asn Gly Ile Ser 1865 1870 1875 Lys Glu Leu Glu Met Lys Lys Val Tyr His Tyr Gly Asp Tyr Val 1880 1885 1890 Thr Leu Lys Cys Glu Asp Gly Tyr Thr Leu Glu Gly Ser Pro Trp 1895 1900 1905 Ser Gln Cys Gln Ala Asp Asp Arg Trp Asp Pro Pro Leu Ala Lys 1910 1915 1920 Cys Thr Ser Arg Thr His Asp 1925 1930 <210> 3 <211> 1351 <212> PRT <213> Artificial sequence <220> <223> sCR1(1392) <400> 3 Gln Cys Asn Ala Pro Glu Trp Leu Pro Phe Ala Arg Pro Thr Asn Leu 1 5 10 15 Thr Asp Glu Phe Glu Phe Pro Ile Gly Thr Tyr Leu Asn Tyr Glu Cys 20 25 30 Arg Pro Gly Tyr Ser Gly Arg Pro Phe Ser Ile Ile Cys Leu Lys Asn 35 40 45 Ser Val Trp Thr Gly Ala Lys Asp Arg Cys Arg Arg Lys Ser Cys Arg 50 55 60 Asn Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Lys Gly Ile 65 70 75 80 Gln Phe Gly Ser Gln Ile Lys Tyr Ser Cys Thr Lys Gly Tyr Arg Leu 85 90 95 Ile Gly Ser Ser Ser Ala Thr Cys Ile Ile Ser Gly Asp Thr Val Ile 100 105 110 Trp Asp Asn Glu Thr Pro Ile Cys Asp Arg Ile Pro Cys Gly Leu Pro 115 120 125 Pro Thr Ile Thr Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe 130 135 140 His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly 145 150 155 160 Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser 165 170 175 Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile 180 185 190 Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val 195 200 205 Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg 210 215 220 Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln 225 230 235 240 Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys 245 250 255 Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys 260 265 270 Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 275 280 285 Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp 290 295 300 Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe 305 310 315 320 Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln 325 330 335 Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys 340 345 350 Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp 355 360 365 Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro 370 375 380 Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro 385 390 395 400 Phe Gly Lys Thr Val Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly 405 410 415 Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp 420 425 430 Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile 435 440 445 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 450 455 460 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 465 470 475 480 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 485 490 495 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 500 505 510 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 515 520 525 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 530 535 540 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 545 550 555 560 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 565 570 575 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 580 585 590 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 595 600 605 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 610 615 620 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 625 630 635 640 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 645 650 655 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 660 665 670 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 675 680 685 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 690 695 700 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 705 710 715 720 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 725 730 735 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 740 745 750 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 755 760 765 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 770 775 780 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 785 790 795 800 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 805 810 815 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 820 825 830 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 835 840 845 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 850 855 860 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 865 870 875 880 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 885 890 895 Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys 900 905 910 Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu 915 920 925 Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr 930 935 940 Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg 945 950 955 960 Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val 965 970 975 Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr 980 985 990 Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly 995 1000 1005 Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 1010 1015 1020 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser 1025 1030 1035 Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg 1040 1045 1050 Cys Asn Leu Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly 1055 1060 1065 Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 1070 1075 1080 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr 1085 1090 1095 Pro Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser 1100 1105 1110 Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly 1115 1120 1125 Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn 1130 1135 1140 Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro 1145 1150 1155 Pro Pro Glu Ile Leu His Gly Glu His Thr Pro Ser His Gln Asp 1160 1165 1170 Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 1175 1180 1185 Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly 1190 1195 1200 Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser Cys Asp 1205 1210 1215 Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro Leu 1220 1225 1230 Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly 1235 1240 1245 Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly 1250 1255 1260 Met Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile 1265 1270 1275 Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly 1280 1285 1290 Thr Pro Ser Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr 1295 1300 1305 Cys Asp Pro His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly 1310 1315 1320 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val 1325 1330 1335 Trp Ser Ser Pro Ala Pro Arg Cys Glu Leu Ser Val Arg 1340 1345 1350 <210> 4 <211> 898 <212> PRT <213> Artificial Sequence <220> <223> sCR1(939) <400> 4 Gln Cys Asn Ala Pro Glu Trp Leu Pro Phe Ala Arg Pro Thr Asn Leu 1 5 10 15 Thr Asp Glu Phe Glu Phe Pro Ile Gly Thr Tyr Leu Asn Tyr Glu Cys 20 25 30 Arg Pro Gly Tyr Ser Gly Arg Pro Phe Ser Ile Ile Cys Leu Lys Asn 35 40 45 Ser Val Trp Thr Gly Ala Lys Asp Arg Cys Arg Arg Lys Ser Cys Arg 50 55 60 Asn Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Lys Gly Ile 65 70 75 80 Gln Phe Gly Ser Gln Ile Lys Tyr Ser Cys Thr Lys Gly Tyr Arg Leu 85 90 95 Ile Gly Ser Ser Ser Ala Thr Cys Ile Ile Ser Gly Asp Thr Val Ile 100 105 110 Trp Asp Asn Glu Thr Pro Ile Cys Asp Arg Ile Pro Cys Gly Leu Pro 115 120 125 Pro Thr Ile Thr Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe 130 135 140 His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly 145 150 155 160 Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser 165 170 175 Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile 180 185 190 Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val 195 200 205 Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg 210 215 220 Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln 225 230 235 240 Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys 245 250 255 Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys 260 265 270 Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 275 280 285 Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp 290 295 300 Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe 305 310 315 320 Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln 325 330 335 Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys 340 345 350 Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp 355 360 365 Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro 370 375 380 Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro 385 390 395 400 Phe Gly Lys Thr Val Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly 405 410 415 Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp 420 425 430 Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile 435 440 445 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 450 455 460 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 465 470 475 480 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 485 490 495 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 500 505 510 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 515 520 525 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 530 535 540 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 545 550 555 560 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 565 570 575 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 580 585 590 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 595 600 605 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 610 615 620 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 625 630 635 640 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 645 650 655 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 660 665 670 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 675 680 685 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 690 695 700 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 705 710 715 720 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 725 730 735 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 740 745 750 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 755 760 765 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 770 775 780 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 785 790 795 800 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 805 810 815 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 820 825 830 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 835 840 845 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 850 855 860 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 865 870 875 880 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 885 890 895 Gly Ile <210> 5 <211> 903 <212> PRT <213> Artificial Sequence <220> <223> sCR1(490-1392) <400> 5 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 145 150 155 160 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 260 265 270 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 340 345 350 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 370 375 380 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 385 390 395 400 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys 450 455 460 Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu 465 470 475 480 Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr 485 490 495 Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg 500 505 510 Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val 515 520 525 Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr 530 535 540 Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly 545 550 555 560 Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro 565 570 575 Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn 580 585 590 Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu 595 600 605 Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 610 615 620 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala 625 630 635 640 Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn 645 650 655 Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val 660 665 670 Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg 675 680 685 Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys 690 695 700 Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr 705 710 715 720 Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser 725 730 735 Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr 740 745 750 Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser 755 760 765 Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro 770 775 780 Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly 785 790 795 800 Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met 805 810 815 Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys 820 825 830 Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser 835 840 845 Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 850 855 860 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile Arg 865 870 875 880 Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala Pro 885 890 895 Arg Cys Glu Leu Ser Val Arg 900 <210> 6 <211> 1482 <212> PRT <213> Artificial Sequence <220> <223> sCR1(490-1971) <400> 6 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 145 150 155 160 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 260 265 270 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 340 345 350 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 370 375 380 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 385 390 395 400 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys 450 455 460 Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu 465 470 475 480 Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr 485 490 495 Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg 500 505 510 Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val 515 520 525 Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr 530 535 540 Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly 545 550 555 560 Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro 565 570 575 Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn 580 585 590 Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu 595 600 605 Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 610 615 620 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala 625 630 635 640 Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn 645 650 655 Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val 660 665 670 Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg 675 680 685 Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys 690 695 700 Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr 705 710 715 720 Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser 725 730 735 Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr 740 745 750 Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser 755 760 765 Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro 770 775 780 Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly 785 790 795 800 Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met 805 810 815 Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys 820 825 830 Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser 835 840 845 Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 850 855 860 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile Arg 865 870 875 880 Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala Pro 885 890 895 Arg Cys Glu Leu Ser Val Arg Ala Gly His Cys Lys Thr Pro Glu Gln 900 905 910 Phe Pro Phe Ala Ser Pro Thr Ile Pro Ile Asn Asp Phe Glu Phe Pro 915 920 925 Val Gly Thr Ser Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Phe Gly Lys 930 935 940 Met Phe Ser Ile Ser Cys Leu Glu Asn Leu Val Trp Ser Ser Val Glu 945 950 955 960 Asp Asn Cys Arg Arg Lys Ser Cys Gly Pro Pro Pro Glu Pro Phe Asn 965 970 975 Gly Met Val His Ile Asn Thr Asp Thr Gln Phe Gly Ser Thr Val Asn 980 985 990 Tyr Ser Cys Asn Glu Gly Phe Arg Leu Ile Gly Ser Pro Ser Thr Thr 995 1000 1005 Cys Leu Val Ser Gly Asn Asn Val Thr Trp Asp Lys Lys Ala Pro 1010 1015 1020 Ile Cys Glu Ile Ile Ser Cys Glu Pro Pro Pro Thr Ile Ser Asn 1025 1030 1035 Gly Asp Phe Tyr Ser Asn Asn Arg Thr Ser Phe His Asn Gly Thr 1040 1045 1050 Val Val Thr Tyr Gln Cys His Thr Gly Pro Asp Gly Glu Gln Leu 1055 1060 1065 Phe Glu Leu Val Gly Glu Arg Ser Ile Tyr Cys Thr Ser Lys Asp 1070 1075 1080 Asp Gln Val Gly Val Trp Ser Ser Pro Pro Pro Arg Cys Ile Ser 1085 1090 1095 Thr Asn Lys Cys Thr Ala Pro Glu Val Glu Asn Ala Ile Arg Val 1100 1105 1110 Pro Gly Asn Arg Ser Phe Phe Ser Leu Thr Glu Ile Ile Arg Phe 1115 1120 1125 Arg Cys Gln Pro Gly Phe Val Met Val Gly Ser His Thr Val Gln 1130 1135 1140 Cys Gln Thr Asn Gly Arg Trp Gly Pro Lys Leu Pro His Cys Ser 1145 1150 1155 Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr 1160 1165 1170 Leu Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 1175 1180 1185 Ser Cys Glu Pro Ser Tyr Asp Leu Arg Gly Ala Ala Ser Leu His 1190 1195 1200 Cys Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Thr 1205 1210 1215 Val Lys Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg 1220 1225 1230 Val Leu Leu Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe 1235 1240 1245 Val Cys Asp Glu Gly Phe Arg Leu Lys Gly Arg Ser Ala Ser His 1250 1255 1260 Cys Val Leu Ala Gly Met Lys Ala Leu Trp Asn Ser Ser Val Pro 1265 1270 1275 Val Cys Glu Gln Ile Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn 1280 1285 1290 Gly Arg His Thr Gly Thr Pro Phe Gly Asp Ile Pro Tyr Gly Lys 1295 1300 1305 Glu Ile Ser Tyr Ala Cys Asp Thr His Pro Asp Arg Gly Met Thr 1310 1315 1320 Phe Asn Leu Ile Gly Glu Ser Ser Ile Arg Cys Thr Ser Asp Pro 1325 1330 1335 Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Glu Leu 1340 1345 1350 Ser Val Pro Ala Ala Cys Pro His Pro Pro Lys Ile Gln Asn Gly 1355 1360 1365 His Tyr Ile Gly Gly His Val Ser Leu Tyr Leu Pro Gly Met Thr 1370 1375 1380 Ile Ser Tyr Ile Cys Asp Pro Gly Tyr Leu Leu Val Gly Lys Gly 1385 1390 1395 Phe Ile Phe Cys Thr Asp Gln Gly Ile Trp Ser Gln Leu Asp His 1400 1405 1410 Tyr Cys Lys Glu Val Asn Cys Ser Phe Pro Leu Phe Met Asn Gly 1415 1420 1425 Ile Ser Lys Glu Leu Glu Met Lys Lys Val Tyr His Tyr Gly Asp 1430 1435 1440 Tyr Val Thr Leu Lys Cys Glu Asp Gly Tyr Thr Leu Glu Gly Ser 1445 1450 1455 Pro Trp Ser Gln Cys Gln Ala Asp Asp Arg Trp Asp Pro Pro Leu 1460 1465 1470 Ala Lys Cys Thr Ser Arg Thr His Asp 1475 1480 <210> 7 <211> 193 <212> PRT <213> Artificial Sequence <220> <223> sCR1(234) <400> 7 Gln Cys Asn Ala Pro Glu Trp Leu Pro Phe Ala Arg Pro Thr Asn Leu 1 5 10 15 Thr Asp Glu Phe Glu Phe Pro Ile Gly Thr Tyr Leu Asn Tyr Glu Cys 20 25 30 Arg Pro Gly Tyr Ser Gly Arg Pro Phe Ser Ile Ile Cys Leu Lys Asn 35 40 45 Ser Val Trp Thr Gly Ala Lys Asp Arg Cys Arg Arg Lys Ser Cys Arg 50 55 60 Asn Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Lys Gly Ile 65 70 75 80 Gln Phe Gly Ser Gln Ile Lys Tyr Ser Cys Thr Lys Gly Tyr Arg Leu 85 90 95 Ile Gly Ser Ser Ser Ala Thr Cys Ile Ile Ser Gly Asp Thr Val Ile 100 105 110 Trp Asp Asn Glu Thr Pro Ile Cys Asp Arg Ile Pro Cys Gly Leu Pro 115 120 125 Pro Thr Ile Thr Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe 130 135 140 His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly 145 150 155 160 Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser 165 170 175 Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile 180 185 190 Ile <210> 8 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> sCR1(489) <400> 8 Gln Cys Asn Ala Pro Glu Trp Leu Pro Phe Ala Arg Pro Thr Asn Leu 1 5 10 15 Thr Asp Glu Phe Glu Phe Pro Ile Gly Thr Tyr Leu Asn Tyr Glu Cys 20 25 30 Arg Pro Gly Tyr Ser Gly Arg Pro Phe Ser Ile Ile Cys Leu Lys Asn 35 40 45 Ser Val Trp Thr Gly Ala Lys Asp Arg Cys Arg Arg Lys Ser Cys Arg 50 55 60 Asn Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Lys Gly Ile 65 70 75 80 Gln Phe Gly Ser Gln Ile Lys Tyr Ser Cys Thr Lys Gly Tyr Arg Leu 85 90 95 Ile Gly Ser Ser Ser Ala Thr Cys Ile Ile Ser Gly Asp Thr Val Ile 100 105 110 Trp Asp Asn Glu Thr Pro Ile Cys Asp Arg Ile Pro Cys Gly Leu Pro 115 120 125 Pro Thr Ile Thr Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe 130 135 140 His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly 145 150 155 160 Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser 165 170 175 Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile 180 185 190 Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val 195 200 205 Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg 210 215 220 Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln 225 230 235 240 Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys 245 250 255 Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys 260 265 270 Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 275 280 285 Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp 290 295 300 Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe 305 310 315 320 Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln 325 330 335 Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys 340 345 350 Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp 355 360 365 Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro 370 375 380 Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro 385 390 395 400 Phe Gly Lys Thr Val Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly 405 410 415 Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp 420 425 430 Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile 435 440 445 <210> 9 <211> 1482 <212> PRT <213> Artificial sequence <220> sCR1(940-1971) <400> 9 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 145 150 155 160 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 260 265 270 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 340 345 350 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 370 375 380 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 385 390 395 400 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys 450 455 460 Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu 465 470 475 480 Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr 485 490 495 Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg 500 505 510 Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val 515 520 525 Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr 530 535 540 Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly 545 550 555 560 Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro 565 570 575 Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn 580 585 590 Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu 595 600 605 Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 610 615 620 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala 625 630 635 640 Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn 645 650 655 Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val 660 665 670 Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg 675 680 685 Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys 690 695 700 Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr 705 710 715 720 Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser 725 730 735 Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr 740 745 750 Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser 755 760 765 Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro 770 775 780 Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly 785 790 795 800 Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met 805 810 815 Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys 820 825 830 Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser 835 840 845 Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 850 855 860 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile Arg 865 870 875 880 Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala Pro 885 890 895 Arg Cys Glu Leu Ser Val Arg Ala Gly His Cys Lys Thr Pro Glu Gln 900 905 910 Phe Pro Phe Ala Ser Pro Thr Ile Pro Ile Asn Asp Phe Glu Phe Pro 915 920 925 Val Gly Thr Ser Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Phe Gly Lys 930 935 940 Met Phe Ser Ile Ser Cys Leu Glu Asn Leu Val Trp Ser Ser Val Glu 945 950 955 960 Asp Asn Cys Arg Arg Lys Ser Cys Gly Pro Pro Pro Glu Pro Phe Asn 965 970 975 Gly Met Val His Ile Asn Thr Asp Thr Gln Phe Gly Ser Thr Val Asn 980 985 990 Tyr Ser Cys Asn Glu Gly Phe Arg Leu Ile Gly Ser Pro Ser Thr Thr 995 1000 1005 Cys Leu Val Ser Gly Asn Asn Val Thr Trp Asp Lys Lys Ala Pro 1010 1015 1020 Ile Cys Glu Ile Ile Ser Cys Glu Pro Pro Pro Thr Ile Ser Asn 1025 1030 1035 Gly Asp Phe Tyr Ser Asn Asn Arg Thr Ser Phe His Asn Gly Thr 1040 1045 1050 Val Val Thr Tyr Gln Cys His Thr Gly Pro Asp Gly Glu Gln Leu 1055 1060 1065 Phe Glu Leu Val Gly Glu Arg Ser Ile Tyr Cys Thr Ser Lys Asp 1070 1075 1080 Asp Gln Val Gly Val Trp Ser Ser Pro Pro Pro Arg Cys Ile Ser 1085 1090 1095 Thr Asn Lys Cys Thr Ala Pro Glu Val Glu Asn Ala Ile Arg Val 1100 1105 1110 Pro Gly Asn Arg Ser Phe Phe Ser Leu Thr Glu Ile Ile Arg Phe 1115 1120 1125 Arg Cys Gln Pro Gly Phe Val Met Val Gly Ser His Thr Val Gln 1130 1135 1140 Cys Gln Thr Asn Gly Arg Trp Gly Pro Lys Leu Pro His Cys Ser 1145 1150 1155 Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr 1160 1165 1170 Leu Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 1175 1180 1185 Ser Cys Glu Pro Ser Tyr Asp Leu Arg Gly Ala Ala Ser Leu His 1190 1195 1200 Cys Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Thr 1205 1210 1215 Val Lys Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg 1220 1225 1230 Val Leu Leu Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe 1235 1240 1245 Val Cys Asp Glu Gly Phe Arg Leu Lys Gly Arg Ser Ala Ser His 1250 1255 1260 Cys Val Leu Ala Gly Met Lys Ala Leu Trp Asn Ser Ser Val Pro 1265 1270 1275 Val Cys Glu Gln Ile Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn 1280 1285 1290 Gly Arg His Thr Gly Thr Pro Phe Gly Asp Ile Pro Tyr Gly Lys 1295 1300 1305 Glu Ile Ser Tyr Ala Cys Asp Thr His Pro Asp Arg Gly Met Thr 1310 1315 1320 Phe Asn Leu Ile Gly Glu Ser Ser Ile Arg Cys Thr Ser Asp Pro 1325 1330 1335 Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Glu Leu 1340 1345 1350 Ser Val Pro Ala Ala Cys Pro His Pro Pro Lys Ile Gln Asn Gly 1355 1360 1365 His Tyr Ile Gly Gly His Val Ser Leu Tyr Leu Pro Gly Met Thr 1370 1375 1380 Ile Ser Tyr Ile Cys Asp Pro Gly Tyr Leu Leu Val Gly Lys Gly 1385 1390 1395 Phe Ile Phe Cys Thr Asp Gln Gly Ile Trp Ser Gln Leu Asp His 1400 1405 1410 Tyr Cys Lys Glu Val Asn Cys Ser Phe Pro Leu Phe Met Asn Gly 1415 1420 1425 Ile Ser Lys Glu Leu Glu Met Lys Lys Val Tyr His Tyr Gly Asp 1430 1435 1440 Tyr Val Thr Leu Lys Cys Glu Asp Gly Tyr Thr Leu Glu Gly Ser 1445 1450 1455 Pro Trp Ser Gln Cys Gln Ala Asp Asp Arg Trp Asp Pro Pro Leu 1460 1465 1470 Ala Lys Cys Thr Ser Arg Thr His Asp 1475 1480 <210> 10 <211> 450 <212> PRT <213> Artificial Sequence <220> <223> sCR1(490-939) <400> 10 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 145 150 155 160 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 260 265 270 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 340 345 350 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 370 375 380 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 385 390 395 400 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Gly Ile 450 <210> 11 <211> 903 <212> PRT <213> Artificial Sequence <220> <223> sCR1(940-1392) <400> 11 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 145 150 155 160 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 260 265 270 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 340 345 350 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 370 375 380 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 385 390 395 400 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys 450 455 460 Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu 465 470 475 480 Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr 485 490 495 Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg 500 505 510 Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val 515 520 525 Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr 530 535 540 Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly 545 550 555 560 Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro 565 570 575 Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn 580 585 590 Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu 595 600 605 Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 610 615 620 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala 625 630 635 640 Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn 645 650 655 Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val 660 665 670 Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg 675 680 685 Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys 690 695 700 Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr 705 710 715 720 Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser 725 730 735 Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr 740 745 750 Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser 755 760 765 Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro 770 775 780 Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly 785 790 795 800 Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met 805 810 815 Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys 820 825 830 Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser 835 840 845 Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 850 855 860 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile Arg 865 870 875 880 Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala Pro 885 890 895 Arg Cys Glu Leu Ser Val Arg 900 <210> 12 <211> 579 <212> PRT <213> Artificial Sequence <220> <223> sCR1(1393-1971) <400> 12 Ala Gly His Cys Lys Thr Pro Glu Gln Phe Pro Phe Ala Ser Pro Thr 1 5 10 15 Ile Pro Ile Asn Asp Phe Glu Phe Pro Val Gly Thr Ser Leu Asn Tyr 20 25 30 Glu Cys Arg Pro Gly Tyr Phe Gly Lys Met Phe Ser Ile Ser Cys Leu 35 40 45 Glu Asn Leu Val Trp Ser Ser Val Glu Asp Asn Cys Arg Arg Lys Ser 50 55 60 Cys Gly Pro Pro Pro Glu Pro Phe Asn Gly Met Val His Ile Asn Thr 65 70 75 80 Asp Thr Gln Phe Gly Ser Thr Val Asn Tyr Ser Cys Asn Glu Gly Phe 85 90 95 Arg Leu Ile Gly Ser Pro Ser Thr Thr Cys Leu Val Ser Gly Asn Asn 100 105 110 Val Thr Trp Asp Lys Lys Ala Pro Ile Cys Glu Ile Ile Ser Cys Glu 115 120 125 Pro Pro Pro Thr Ile Ser Asn Gly Asp Phe Tyr Ser Asn Asn Arg Thr 130 135 140 Ser Phe His Asn Gly Thr Val Val Thr Tyr Gln Cys His Thr Gly Pro 145 150 155 160 Asp Gly Glu Gln Leu Phe Glu Leu Val Gly Glu Arg Ser Ile Tyr Cys 165 170 175 Thr Ser Lys Asp Asp Gln Val Gly Val Trp Ser Ser Pro Pro Pro Arg 180 185 190 Cys Ile Ser Thr Asn Lys Cys Thr Ala Pro Glu Val Glu Asn Ala Ile 195 200 205 Arg Val Pro Gly Asn Arg Ser Phe Phe Ser Leu Thr Glu Ile Ile Arg 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Val Gly Ser His Thr Val Gln 225 230 235 240 Cys Gln Thr Asn Gly Arg Trp Gly Pro Lys Leu Pro His Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr Leu Ser 260 265 270 His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Ser Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Thr Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Leu Pro Leu Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg 340 345 350 Leu Lys Gly Arg Ser Ala Ser His Cys Val Leu Ala Gly Met Lys Ala 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Asn 370 375 380 Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Phe Gly Asp 385 390 395 400 Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Ala Cys Asp Thr His Pro Asp 405 410 415 Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Ser Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Glu Leu Ser Val Pro Ala Ala Cys Pro His Pro Pro Lys Ile Gln Asn 450 455 460 Gly His Tyr Ile Gly Gly His Val Ser Leu Tyr Leu Pro Gly Met Thr 465 470 475 480 Ile Ser Tyr Ile Cys Asp Pro Gly Tyr Leu Leu Val Gly Lys Gly Phe 485 490 495 Ile Phe Cys Thr Asp Gln Gly Ile Trp Ser Gln Leu Asp His Tyr Cys 500 505 510 Lys Glu Val Asn Cys Ser Phe Pro Leu Phe Met Asn Gly Ile Ser Lys 515 520 525 Glu Leu Glu Met Lys Lys Val Tyr His Tyr Gly Asp Tyr Val Thr Leu 530 535 540 Lys Cys Glu Asp Gly Tyr Thr Leu Glu Gly Ser Pro Trp Ser Gln Cys 545 550 555 560 Gln Ala Asp Asp Arg Trp Asp Pro Pro Leu Ala Lys Cys Thr Ser Arg 565 570 575 Thr His Asp <210> 13 <211> 448 <212> PRT <213> Artificial sequence <220> <223> sCR1 LHR-A <400> 13 Gln Cys Asn Ala Pro Glu Trp Leu Pro Phe Ala Arg Pro Thr Asn Leu 1 5 10 15 Thr Asp Glu Phe Glu Phe Pro Ile Gly Thr Tyr Leu Asn Tyr Glu Cys 20 25 30 Arg Pro Gly Tyr Ser Gly Arg Pro Phe Ser Ile Ile Cys Leu Lys Asn 35 40 45 Ser Val Trp Thr Gly Ala Lys Asp Arg Cys Arg Arg Lys Ser Cys Arg 50 55 60 Asn Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Lys Gly Ile 65 70 75 80 Gln Phe Gly Ser Gln Ile Lys Tyr Ser Cys Thr Lys Gly Tyr Arg Leu 85 90 95 Ile Gly Ser Ser Ser Ala Thr Cys Ile Ile Ser Gly Asp Thr Val Ile 100 105 110 Trp Asp Asn Glu Thr Pro Ile Cys Asp Arg Ile Pro Cys Gly Leu Pro 115 120 125 Pro Thr Ile Thr Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe 130 135 140 His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly 145 150 155 160 Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser 165 170 175 Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile 180 185 190 Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val 195 200 205 Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg 210 215 220 Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln 225 230 235 240 Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys 245 250 255 Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys 260 265 270 Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly 275 280 285 Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp 290 295 300 Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe 305 310 315 320 Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln 325 330 335 Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys 340 345 350 Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp 355 360 365 Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro 370 375 380 Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro 385 390 395 400 Phe Gly Lys Thr Val Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly 405 410 415 Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp 420 425 430 Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile 435 440 445 <210> 14 <211> 450 <212> PRT <213> Artificial Sequence <220> <223> sCR1 LHR-B <400> 14 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Ala 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Pro Gly Ser 145 150 155 160 Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg Thr Gln Arg 260 265 270 Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu Gly Phe Gln 340 345 350 Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly Met Glu Ser 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Ser 370 375 380 Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro Leu Glu Val 385 390 395 400 Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Gly Ile 450 <210> 15 <211> 453 <212> PRT <213> Artificial sequence <220> <223> sCR1 LHR-C <400> 15 Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys 1 5 10 15 Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr 20 25 30 Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu 35 40 45 Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser 50 55 60 Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His Val Ile Thr 65 70 75 80 Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His 85 90 95 Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Thr 100 105 110 Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly 115 120 125 Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 130 135 140 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu Gly Ser 145 150 155 160 Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile Tyr Cys 165 170 175 Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro Ala Pro Gln 180 185 190 Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu Asn Gly Ile 195 200 205 Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu Val Val Glu 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg Arg Val Lys 225 230 235 240 Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr Pro Ser 260 265 270 His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe Pro Leu Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg 340 345 350 Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met Arg Ser 355 360 365 Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys Pro Asn 370 375 380 Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser Gly Asp 385 390 395 400 Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His Pro Asp 405 410 415 Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile Arg Cys Thr 420 425 430 Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Glu Leu Ser Val Arg 450 <210> 16 <211> 579 <212> PRT <213> Artificial Sequence <220> <223> sCR1 LHR-D <400> 16 Ala Gly His Cys Lys Thr Pro Glu Gln Phe Pro Phe Ala Ser Pro Thr 1 5 10 15 Ile Pro Ile Asn Asp Phe Glu Phe Pro Val Gly Thr Ser Leu Asn Tyr 20 25 30 Glu Cys Arg Pro Gly Tyr Phe Gly Lys Met Phe Ser Ile Ser Cys Leu 35 40 45 Glu Asn Leu Val Trp Ser Ser Val Glu Asp Asn Cys Arg Arg Lys Ser 50 55 60 Cys Gly Pro Pro Pro Glu Pro Phe Asn Gly Met Val His Ile Asn Thr 65 70 75 80 Asp Thr Gln Phe Gly Ser Thr Val Asn Tyr Ser Cys Asn Glu Gly Phe 85 90 95 Arg Leu Ile Gly Ser Pro Ser Thr Thr Cys Leu Val Ser Gly Asn Asn 100 105 110 Val Thr Trp Asp Lys Lys Ala Pro Ile Cys Glu Ile Ile Ser Cys Glu 115 120 125 Pro Pro Pro Thr Ile Ser Asn Gly Asp Phe Tyr Ser Asn Asn Arg Thr 130 135 140 Ser Phe His Asn Gly Thr Val Val Thr Tyr Gln Cys His Thr Gly Pro 145 150 155 160 Asp Gly Glu Gln Leu Phe Glu Leu Val Gly Glu Arg Ser Ile Tyr Cys 165 170 175 Thr Ser Lys Asp Asp Gln Val Gly Val Trp Ser Ser Pro Pro Pro Arg 180 185 190 Cys Ile Ser Thr Asn Lys Cys Thr Ala Pro Glu Val Glu Asn Ala Ile 195 200 205 Arg Val Pro Gly Asn Arg Ser Phe Phe Ser Leu Thr Glu Ile Ile Arg 210 215 220 Phe Arg Cys Gln Pro Gly Phe Val Met Val Gly Ser His Thr Val Gln 225 230 235 240 Cys Gln Thr Asn Gly Arg Trp Gly Pro Lys Leu Pro His Cys Ser Arg 245 250 255 Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His Thr Leu Ser 260 265 270 His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu 275 280 285 Pro Ser Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln 290 295 300 Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Thr Val Lys Ser Cys Asp 305 310 315 320 Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Leu Pro Leu Asn 325 330 335 Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg 340 345 350 Leu Lys Gly Arg Ser Ala Ser His Cys Val Leu Ala Gly Met Lys Ala 355 360 365 Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Asn 370 375 380 Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Phe Gly Asp 385 390 395 400 Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Ala Cys Asp Thr His Pro Asp 405 410 415 Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Ser Ile Arg Cys Thr 420 425 430 Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro Arg Cys 435 440 445 Glu Leu Ser Val Pro Ala Ala Cys Pro His Pro Pro Lys Ile Gln Asn 450 455 460 Gly His Tyr Ile Gly Gly His Val Ser Leu Tyr Leu Pro Gly Met Thr 465 470 475 480 Ile Ser Tyr Ile Cys Asp Pro Gly Tyr Leu Leu Val Gly Lys Gly Phe 485 490 495 Ile Phe Cys Thr Asp Gln Gly Ile Trp Ser Gln Leu Asp His Tyr Cys 500 505 510 Lys Glu Val Asn Cys Ser Phe Pro Leu Phe Met Asn Gly Ile Ser Lys 515 520 525 Glu Leu Glu Met Lys Lys Val Tyr His Tyr Gly Asp Tyr Val Thr Leu 530 535 540 Lys Cys Glu Asp Gly Tyr Thr Leu Glu Gly Ser Pro Trp Ser Gln Cys 545 550 555 560 Gln Ala Asp Asp Arg Trp Asp Pro Pro Leu Ala Lys Cys Thr Ser Arg 565 570 575 Thr His Asp <210> 17 <211> 8 <212> PRT <213> Artificial sequence <220> <223> 8xHis tag <400> 17 His His His His His His 1 5 <210> 18 <211> 41 <212> PRT <213> Artificial sequence <220> <223> endogenous signal peptide <400> 18 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly 35 40 <210> 19 <211> 19 <212> PRT <213> Artificial sequence <220> <223> Exogenous signal peptides <400> 19 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala <210> 20 <211> 1979 <212> PRT <213> artificial sequence <220> <223> sCR1(1971)-8His <400> 20 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly Gln Cys Asn Ala Pro Glu Trp 35 40 45 Leu Pro Phe Ala Arg Pro Thr Asn Leu Thr Asp Glu Phe Glu Phe Pro 50 55 60 Ile Gly Thr Tyr Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Ser Gly Arg 65 70 75 80 Pro Phe Ser Ile Ile Cys Leu Lys Asn Ser Val Trp Thr Gly Ala Lys 85 90 95 Asp Arg Cys Arg Arg Lys Ser Cys Arg Asn Pro Pro Asp Pro Val Asn 100 105 110 Gly Met Val His Val Ile Lys Gly Ile Gln Phe Gly Ser Gln Ile Lys 115 120 125 Tyr Ser Cys Thr Lys Gly Tyr Arg Leu Ile Gly Ser Ser Ser Ala Thr 130 135 140 Cys Ile Ile Ser Gly Asp Thr Val Ile Trp Asp Asn Glu Thr Pro Ile 145 150 155 160 Cys Asp Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Thr Asn Gly Asp 165 170 175 Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr 180 185 190 Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val 195 200 205 Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 210 215 220 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro 225 230 235 240 Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe 245 250 255 Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met 260 265 270 Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 275 280 285 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu 290 295 300 His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln 305 310 315 320 Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala 325 330 335 Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr 340 345 350 Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly 355 360 365 Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe 370 375 380 Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys 385 390 395 400 Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys 405 410 415 Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His 420 425 430 Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Thr Val Asn Tyr 435 440 445 Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly 450 455 460 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 465 470 475 480 Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln Ala Pro 485 490 495 Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp 500 505 510 Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr 515 520 525 Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp Ser Ser 530 535 540 Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro 545 550 555 560 Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val Gly Ser Arg 565 570 575 Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile Gly His Ser Ser 580 585 590 Ala Glu Cys Ile Leu Ser Gly Asn Ala Ala His Trp Ser Thr Lys Pro 595 600 605 Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn 610 615 620 Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val 625 630 635 640 Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu 645 650 655 Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val 660 665 670 Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys 675 680 685 Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser 690 695 700 Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe 705 710 715 720 Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp 725 730 735 Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp 740 745 750 Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro 755 760 765 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly 770 775 780 Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala 785 790 795 800 Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu 805 810 815 Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val 820 825 830 Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser 835 840 845 Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro 850 855 860 Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly 865 870 875 880 Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Ala Val 885 890 895 Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu 900 905 910 Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly 915 920 925 Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln 930 935 940 Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala 945 950 955 960 Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu 965 970 975 Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp 980 985 990 Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro 995 1000 1005 Asp Pro Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val 1010 1015 1020 Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile 1025 1030 1035 Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Thr Ala His 1040 1045 1050 Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu 1055 1060 1065 Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 1070 1075 1080 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu Gly 1085 1090 1095 Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 1100 1105 1110 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 1115 1120 1125 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val 1130 1135 1140 Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu 1145 1150 1155 Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys 1160 1165 1170 Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 1175 1180 1185 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile 1190 1195 1200 Leu His Gly Glu His Thr Pro Ser His Gln Asp Asn Phe Ser Pro 1205 1210 1215 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg 1220 1225 1230 Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly Asp Trp Ser Pro 1235 1240 1245 Glu Ala Pro Arg Cys Ala Val Lys Ser Cys Asp Asp Phe Leu Gly 1250 1255 1260 Gln Leu Pro His Gly Arg Val Leu Phe Pro Leu Asn Leu Gln Leu 1265 1270 1275 Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg Leu Lys 1280 1285 1290 Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met Arg Ser Leu 1295 1300 1305 Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys Pro Asn 1310 1315 1320 Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser Gly 1325 1330 1335 Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 1340 1345 1350 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile 1355 1360 1365 Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro 1370 1375 1380 Ala Pro Arg Cys Glu Leu Ser Val Arg Ala Gly His Cys Lys Thr 1385 1390 1395 Pro Glu Gln Phe Pro Phe Ala Ser Pro Thr Ile Pro Ile Asn Asp 1400 1405 1410 Phe Glu Phe Pro Val Gly Thr Ser Leu Asn Tyr Glu Cys Arg Pro 1415 1420 1425 Gly Tyr Phe Gly Lys Met Phe Ser Ile Ser Cys Leu Glu Asn Leu 1430 1435 1440 Val Trp Ser Ser Val Glu Asp Asn Cys Arg Arg Lys Ser Cys Gly 1445 1450 1455 Pro Pro Pro Glu Pro Phe Asn Gly Met Val His Ile Asn Thr Asp 1460 1465 1470 Thr Gln Phe Gly Ser Thr Val Asn Tyr Ser Cys Asn Glu Gly Phe 1475 1480 1485 Arg Leu Ile Gly Ser Pro Ser Thr Thr Cys Leu Val Ser Gly Asn 1490 1495 1500 Asn Val Thr Trp Asp Lys Lys Ala Pro Ile Cys Glu Ile Ile Ser 1505 1510 1515 Cys Glu Pro Pro Pro Thr Ile Ser Asn Gly Asp Phe Tyr Ser Asn 1520 1525 1530 Asn Arg Thr Ser Phe His Asn Gly Thr Val Val Thr Tyr Gln Cys 1535 1540 1545 His Thr Gly Pro Asp Gly Glu Gln Leu Phe Glu Leu Val Gly Glu 1550 1555 1560 Arg Ser Ile Tyr Cys Thr Ser Lys Asp Asp Gln Val Gly Val Trp 1565 1570 1575 Ser Ser Pro Pro Pro Arg Cys Ile Ser Thr Asn Lys Cys Thr Ala 1580 1585 1590 Pro Glu Val Glu Asn Ala Ile Arg Val Pro Gly Asn Arg Ser Phe 1595 1600 1605 Phe Ser Leu Thr Glu Ile Ile Arg Phe Arg Cys Gln Pro Gly Phe 1610 1615 1620 Val Met Val Gly Ser His Thr Val Gln Cys Gln Thr Asn Gly Arg 1625 1630 1635 Trp Gly Pro Lys Leu Pro His Cys Ser Arg Val Cys Gln Pro Pro 1640 1645 1650 Pro Glu Ile Leu His Gly Glu His Thr Leu Ser His Gln Asp Asn 1655 1660 1665 Phe Ser Pro Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Ser Tyr 1670 1675 1680 Asp Leu Arg Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly Asp 1685 1690 1695 Trp Ser Pro Glu Ala Pro Arg Cys Thr Val Lys Ser Cys Asp Asp 1700 1705 1710 Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Leu Pro Leu Asn 1715 1720 1725 Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe 1730 1735 1740 Arg Leu Lys Gly Arg Ser Ala Ser His Cys Val Leu Ala Gly Met 1745 1750 1755 Lys Ala Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe 1760 1765 1770 Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr 1775 1780 1785 Pro Phe Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Ala Cys 1790 1795 1800 Asp Thr His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu 1805 1810 1815 Ser Ser Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 1820 1825 1830 Ser Ser Pro Ala Pro Arg Cys Glu Leu Ser Val Pro Ala Ala Cys 1835 1840 1845 Pro His Pro Pro Lys Ile Gln Asn Gly His Tyr Ile Gly Gly His 1850 1855 1860 Val Ser Leu Tyr Leu Pro Gly Met Thr Ile Ser Tyr Ile Cys Asp 1865 1870 1875 Pro Gly Tyr Leu Leu Val Gly Lys Gly Phe Ile Phe Cys Thr Asp 1880 1885 1890 Gln Gly Ile Trp Ser Gln Leu Asp His Tyr Cys Lys Glu Val Asn 1895 1900 1905 Cys Ser Phe Pro Leu Phe Met Asn Gly Ile Ser Lys Glu Leu Glu 1910 1915 1920 Met Lys Lys Val Tyr His Tyr Gly Asp Tyr Val Thr Leu Lys Cys 1925 1930 1935 Glu Asp Gly Tyr Thr Leu Glu Gly Ser Pro Trp Ser Gln Cys Gln 1940 1945 1950 Ala Asp Asp Arg Trp Asp Pro Pro Leu Ala Lys Cys Thr Ser Arg 1955 1960 1965 Thr His Asp His His His His His His His His 1970 1975 <210> 21 <211> 1400 <212> PRT <213> Artificial Sequence <220> <223> sCR1(1392)-8His <400> 21 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly Gln Cys Asn Ala Pro Glu Trp 35 40 45 Leu Pro Phe Ala Arg Pro Thr Asn Leu Thr Asp Glu Phe Glu Phe Pro 50 55 60 Ile Gly Thr Tyr Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Ser Gly Arg 65 70 75 80 Pro Phe Ser Ile Ile Cys Leu Lys Asn Ser Val Trp Thr Gly Ala Lys 85 90 95 Asp Arg Cys Arg Arg Lys Ser Cys Arg Asn Pro Pro Asp Pro Val Asn 100 105 110 Gly Met Val His Val Ile Lys Gly Ile Gln Phe Gly Ser Gln Ile Lys 115 120 125 Tyr Ser Cys Thr Lys Gly Tyr Arg Leu Ile Gly Ser Ser Ser Ala Thr 130 135 140 Cys Ile Ile Ser Gly Asp Thr Val Ile Trp Asp Asn Glu Thr Pro Ile 145 150 155 160 Cys Asp Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Thr Asn Gly Asp 165 170 175 Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr 180 185 190 Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val 195 200 205 Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 210 215 220 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro 225 230 235 240 Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe 245 250 255 Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met 260 265 270 Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 275 280 285 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu 290 295 300 His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln 305 310 315 320 Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala 325 330 335 Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr 340 345 350 Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly 355 360 365 Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe 370 375 380 Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys 385 390 395 400 Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys 405 410 415 Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His 420 425 430 Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Thr Val Asn Tyr 435 440 445 Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly 450 455 460 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 465 470 475 480 Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln Ala Pro 485 490 495 Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp 500 505 510 Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr 515 520 525 Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp Ser Ser 530 535 540 Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro 545 550 555 560 Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val Gly Ser Arg 565 570 575 Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile Gly His Ser Ser 580 585 590 Ala Glu Cys Ile Leu Ser Gly Asn Ala Ala His Trp Ser Thr Lys Pro 595 600 605 Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn 610 615 620 Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val 625 630 635 640 Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu 645 650 655 Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val 660 665 670 Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys 675 680 685 Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser 690 695 700 Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe 705 710 715 720 Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp 725 730 735 Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp 740 745 750 Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro 755 760 765 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly 770 775 780 Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala 785 790 795 800 Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu 805 810 815 Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val 820 825 830 Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser 835 840 845 Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro 850 855 860 Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly 865 870 875 880 Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Ala Val 885 890 895 Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu 900 905 910 Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly 915 920 925 Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln 930 935 940 Ala Pro Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala 945 950 955 960 Ser Asp Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu 965 970 975 Tyr Tyr Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp 980 985 990 Ser Ser Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro 995 1000 1005 Asp Pro Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val 1010 1015 1020 Gly Ser Arg Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile 1025 1030 1035 Gly His Ser Ser Ala Glu Cys Ile Leu Ser Gly Asn Thr Ala His 1040 1045 1050 Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu 1055 1060 1065 Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr Asn Arg Glu 1070 1075 1080 Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn Leu Gly 1085 1090 1095 Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser Ile 1100 1105 1110 Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 1115 1120 1125 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val 1130 1135 1140 Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu 1145 1150 1155 Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys 1160 1165 1170 Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 1175 1180 1185 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile 1190 1195 1200 Leu His Gly Glu His Thr Pro Ser His Gln Asp Asn Phe Ser Pro 1205 1210 1215 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg 1220 1225 1230 Gly Ala Ala Ser Leu His Cys Thr Pro Gln Gly Asp Trp Ser Pro 1235 1240 1245 Glu Ala Pro Arg Cys Ala Val Lys Ser Cys Asp Asp Phe Leu Gly 1250 1255 1260 Gln Leu Pro His Gly Arg Val Leu Phe Pro Leu Asn Leu Gln Leu 1265 1270 1275 Gly Ala Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg Leu Lys 1280 1285 1290 Gly Ser Ser Val Ser His Cys Val Leu Val Gly Met Arg Ser Leu 1295 1300 1305 Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe Cys Pro Asn 1310 1315 1320 Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Ser Gly 1325 1330 1335 Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro His 1340 1345 1350 Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile 1355 1360 1365 Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro 1370 1375 1380 Ala Pro Arg Cys Glu Leu Ser Val Arg His His His His His His 1385 1390 1395 His His 1400 <210> 22 <211> 947 <212> PRT <213> Artificial Sequence <220> <223> sCR1(939)-8His <400> 22 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly Gln Cys Asn Ala Pro Glu Trp 35 40 45 Leu Pro Phe Ala Arg Pro Thr Asn Leu Thr Asp Glu Phe Glu Phe Pro 50 55 60 Ile Gly Thr Tyr Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Ser Gly Arg 65 70 75 80 Pro Phe Ser Ile Ile Cys Leu Lys Asn Ser Val Trp Thr Gly Ala Lys 85 90 95 Asp Arg Cys Arg Arg Lys Ser Cys Arg Asn Pro Pro Asp Pro Val Asn 100 105 110 Gly Met Val His Val Ile Lys Gly Ile Gln Phe Gly Ser Gln Ile Lys 115 120 125 Tyr Ser Cys Thr Lys Gly Tyr Arg Leu Ile Gly Ser Ser Ser Ala Thr 130 135 140 Cys Ile Ile Ser Gly Asp Thr Val Ile Trp Asp Asn Glu Thr Pro Ile 145 150 155 160 Cys Asp Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Thr Asn Gly Asp 165 170 175 Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr 180 185 190 Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val 195 200 205 Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 210 215 220 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro 225 230 235 240 Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe 245 250 255 Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met 260 265 270 Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 275 280 285 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu 290 295 300 His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln 305 310 315 320 Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala 325 330 335 Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr 340 345 350 Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly 355 360 365 Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe 370 375 380 Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys 385 390 395 400 Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys 405 410 415 Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His 420 425 430 Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Thr Val Asn Tyr 435 440 445 Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly 450 455 460 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 465 470 475 480 Ser Ser Pro Ala Pro Arg Cys Gly Ile Leu Gly His Cys Gln Ala Pro 485 490 495 Asp His Phe Leu Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp 500 505 510 Phe Pro Ile Gly Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr 515 520 525 Gly Arg Pro Phe Ser Ile Thr Cys Leu Asp Asn Leu Val Trp Ser Ser 530 535 540 Pro Lys Asp Val Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro 545 550 555 560 Val Asn Gly Met Val His Val Ile Thr Asp Ile Gln Val Gly Ser Arg 565 570 575 Ile Asn Tyr Ser Cys Thr Thr Gly His Arg Leu Ile Gly His Ser Ser 580 585 590 Ala Glu Cys Ile Leu Ser Gly Asn Ala Ala His Trp Ser Thr Lys Pro 595 600 605 Pro Ile Cys Gln Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn 610 615 620 Gly Asp Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val 625 630 635 640 Val Thr Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu 645 650 655 Leu Val Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val 660 665 670 Gly Ile Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys 675 680 685 Thr Pro Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser 690 695 700 Leu Phe Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe 705 710 715 720 Val Met Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp 725 730 735 Glu Pro Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp 740 745 750 Val Leu His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro 755 760 765 Gly Gln Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly 770 775 780 Ala Ala Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala 785 790 795 800 Pro Thr Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu 805 810 815 Asn Gly Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val 820 825 830 Asp Phe Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser 835 840 845 Tyr Cys Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro 850 855 860 Val Cys Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly 865 870 875 880 Arg His Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Ala Val 885 890 895 Asn Tyr Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu 900 905 910 Ile Gly Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly 915 920 925 Val Trp Ser Ser Pro Ala Pro Arg Cys Gly Ile His His His His His 930 935 940 His His His 945 <210> 23 <211> 930 <212> PRT <213> Artificial Sequence <220> <223> sCR1(490-1392)-8His <400> 23 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala 20 25 30 Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser 35 40 45 Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile 50 55 60 Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys 65 70 75 80 Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His 85 90 95 Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr 100 105 110 Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser 115 120 125 Gly Asn Ala Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 130 135 140 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr 145 150 155 160 Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn 165 170 175 Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser 180 185 190 Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 195 200 205 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu 210 215 220 Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu 225 230 235 240 Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg 245 250 255 Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser 260 265 270 Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg 275 280 285 Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 290 295 300 Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys 305 310 315 320 Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys 325 330 335 Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe 340 345 350 Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu 355 360 365 Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly 370 375 380 Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe 385 390 395 400 Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro 405 410 415 Leu Glu Val Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro 420 425 430 His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile 435 440 445 Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala 450 455 460 Pro Arg Cys Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu 465 470 475 480 Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly 485 490 495 Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe 500 505 510 Ser Ile Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val 515 520 525 Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met 530 535 540 Val His Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser 545 550 555 560 Cys Thr Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile 565 570 575 Leu Ser Gly Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln 580 585 590 Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile 595 600 605 Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg 610 615 620 Cys Asn Leu Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu 625 630 635 640 Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser 645 650 655 Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn 660 665 670 Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu 675 680 685 Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly 690 695 700 Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu 705 710 715 720 Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly 725 730 735 Glu His Thr Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val 740 745 750 Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu 755 760 765 His Cys Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala 770 775 780 Val Lys Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val 785 790 795 800 Leu Phe Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys 805 810 815 Asp Glu Gly Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu 820 825 830 Val Gly Met Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His 835 840 845 Ile Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly 850 855 860 Thr Pro Ser Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys 865 870 875 880 Asp Pro His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser 885 890 895 Thr Ile Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser 900 905 910 Pro Ala Pro Arg Cys Glu Leu Ser Val Arg His His His His His His 915 920 925 His His 930 <210> 24 <211> 1509 <212> PRT <213> Artificial Sequence <220> <223> sCR1(490-1971)-8His <400> 24 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala 20 25 30 Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser 35 40 45 Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile 50 55 60 Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys 65 70 75 80 Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His 85 90 95 Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr 100 105 110 Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser 115 120 125 Gly Asn Ala Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 130 135 140 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr 145 150 155 160 Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn 165 170 175 Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser 180 185 190 Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 195 200 205 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu 210 215 220 Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu 225 230 235 240 Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg 245 250 255 Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser 260 265 270 Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg 275 280 285 Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 290 295 300 Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys 305 310 315 320 Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys 325 330 335 Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe 340 345 350 Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu 355 360 365 Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly 370 375 380 Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe 385 390 395 400 Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro 405 410 415 Leu Glu Val Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro 420 425 430 His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile 435 440 445 Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala 450 455 460 Pro Arg Cys Gly Ile Leu Gly His Cys Gln Ala Pro Asp His Phe Leu 465 470 475 480 Phe Ala Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly 485 490 495 Thr Ser Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe 500 505 510 Ser Ile Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val 515 520 525 Cys Lys Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met 530 535 540 Val His Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser 545 550 555 560 Cys Thr Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile 565 570 575 Leu Ser Gly Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln 580 585 590 Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile 595 600 605 Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg 610 615 620 Cys Asn Leu Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu 625 630 635 640 Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser 645 650 655 Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn 660 665 670 Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu 675 680 685 Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly 690 695 700 Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu 705 710 715 720 Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly 725 730 735 Glu His Thr Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val 740 745 750 Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu 755 760 765 His Cys Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala 770 775 780 Val Lys Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val 785 790 795 800 Leu Phe Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys 805 810 815 Asp Glu Gly Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu 820 825 830 Val Gly Met Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His 835 840 845 Ile Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly 850 855 860 Thr Pro Ser Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys 865 870 875 880 Asp Pro His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser 885 890 895 Thr Ile Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser 900 905 910 Pro Ala Pro Arg Cys Glu Leu Ser Val Arg Ala Gly His Cys Lys Thr 915 920 925 Pro Glu Gln Phe Pro Phe Ala Ser Pro Thr Ile Pro Ile Asn Asp Phe 930 935 940 Glu Phe Pro Val Gly Thr Ser Leu Asn Tyr Glu Cys Arg Pro Gly Tyr 945 950 955 960 Phe Gly Lys Met Phe Ser Ile Ser Cys Leu Glu Asn Leu Val Trp Ser 965 970 975 Ser Val Glu Asp Asn Cys Arg Arg Lys Ser Cys Gly Pro Pro Pro Glu 980 985 990 Pro Phe Asn Gly Met Val His Ile Asn Thr Asp Thr Gln Phe Gly Ser 995 1000 1005 Thr Val Asn Tyr Ser Cys Asn Glu Gly Phe Arg Leu Ile Gly Ser 1010 1015 1020 Pro Ser Thr Thr Cys Leu Val Ser Gly Asn Asn Val Thr Trp Asp 1025 1030 1035 Lys Lys Ala Pro Ile Cys Glu Ile Ile Ser Cys Glu Pro Pro Pro 1040 1045 1050 Thr Ile Ser Asn Gly Asp Phe Tyr Ser Asn Asn Arg Thr Ser Phe 1055 1060 1065 His Asn Gly Thr Val Val Thr Tyr Gln Cys His Thr Gly Pro Asp 1070 1075 1080 Gly Glu Gln Leu Phe Glu Leu Val Gly Glu Arg Ser Ile Tyr Cys 1085 1090 1095 Thr Ser Lys Asp Asp Gln Val Gly Val Trp Ser Ser Pro Pro Pro 1100 1105 1110 Arg Cys Ile Ser Thr Asn Lys Cys Thr Ala Pro Glu Val Glu Asn 1115 1120 1125 Ala Ile Arg Val Pro Gly Asn Arg Ser Phe Phe Ser Leu Thr Glu 1130 1135 1140 Ile Ile Arg Phe Arg Cys Gln Pro Gly Phe Val Met Val Gly Ser 1145 1150 1155 His Thr Val Gln Cys Gln Thr Asn Gly Arg Trp Gly Pro Lys Leu 1160 1165 1170 Pro His Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His 1175 1180 1185 Gly Glu His Thr Leu Ser His Gln Asp Asn Phe Ser Pro Gly Gln 1190 1195 1200 Glu Val Phe Tyr Ser Cys Glu Pro Ser Tyr Asp Leu Arg Gly Ala 1205 1210 1215 Ala Ser Leu His Cys Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala 1220 1225 1230 Pro Arg Cys Thr Val Lys Ser Cys Asp Asp Phe Leu Gly Gln Leu 1235 1240 1245 Pro His Gly Arg Val Leu Leu Pro Leu Asn Leu Gln Leu Gly Ala 1250 1255 1260 Lys Val Ser Phe Val Cys Asp Glu Gly Phe Arg Leu Lys Gly Arg 1265 1270 1275 Ser Ala Ser His Cys Val Leu Ala Gly Met Lys Ala Leu Trp Asn 1280 1285 1290 Ser Ser Val Pro Val Cys Glu Gln Ile Phe Cys Pro Asn Pro Pro 1295 1300 1305 Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro Phe Gly Asp Ile 1310 1315 1320 Pro Tyr Gly Lys Glu Ile Ser Tyr Ala Cys Asp Thr His Pro Asp 1325 1330 1335 Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Ser Ile Arg Cys 1340 1345 1350 Thr Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala Pro 1355 1360 1365 Arg Cys Glu Leu Ser Val Pro Ala Ala Cys Pro His Pro Pro Lys 1370 1375 1380 Ile Gln Asn Gly His Tyr Ile Gly Gly His Val Ser Leu Tyr Leu 1385 1390 1395 Pro Gly Met Thr Ile Ser Tyr Ile Cys Asp Pro Gly Tyr Leu Leu 1400 1405 1410 Val Gly Lys Gly Phe Ile Phe Cys Thr Asp Gln Gly Ile Trp Ser 1415 1420 1425 Gln Leu Asp His Tyr Cys Lys Glu Val Asn Cys Ser Phe Pro Leu 1430 1435 1440 Phe Met Asn Gly Ile Ser Lys Glu Leu Glu Met Lys Lys Val Tyr 1445 1450 1455 His Tyr Gly Asp Tyr Val Thr Leu Lys Cys Glu Asp Gly Tyr Thr 1460 1465 1470 Leu Glu Gly Ser Pro Trp Ser Gln Cys Gln Ala Asp Asp Arg Trp 1475 1480 1485 Asp Pro Pro Leu Ala Lys Cys Thr Ser Arg Thr His Asp His His 1490 1495 1500 His His His His His His 1505 <210> 25 <211> 242 <212> PRT <213> Artificial Sequence <220> <223> sCR1(234)-8His <400> 25 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly Gln Cys Asn Ala Pro Glu Trp 35 40 45 Leu Pro Phe Ala Arg Pro Thr Asn Leu Thr Asp Glu Phe Glu Phe Pro 50 55 60 Ile Gly Thr Tyr Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Ser Gly Arg 65 70 75 80 Pro Phe Ser Ile Ile Cys Leu Lys Asn Ser Val Trp Thr Gly Ala Lys 85 90 95 Asp Arg Cys Arg Arg Lys Ser Cys Arg Asn Pro Pro Asp Pro Val Asn 100 105 110 Gly Met Val His Val Ile Lys Gly Ile Gln Phe Gly Ser Gln Ile Lys 115 120 125 Tyr Ser Cys Thr Lys Gly Tyr Arg Leu Ile Gly Ser Ser Ser Ala Thr 130 135 140 Cys Ile Ile Ser Gly Asp Thr Val Ile Trp Asp Asn Glu Thr Pro Ile 145 150 155 160 Cys Asp Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Thr Asn Gly Asp 165 170 175 Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr 180 185 190 Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val 195 200 205 Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 210 215 220 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile His His His His His His 225 230 235 240 His His <210> 26 <211> 497 <212> PRT <213> Artificial Sequence <220> <223> sCR1(489)-8His <400> 26 Met Gly Ala Ser Ser Pro Arg Ser Pro Glu Pro Val Gly Pro Pro Ala 1 5 10 15 Pro Gly Leu Pro Phe Cys Cys Gly Gly Ser Leu Leu Ala Val Val Val 20 25 30 Leu Leu Ala Leu Pro Val Ala Trp Gly Gln Cys Asn Ala Pro Glu Trp 35 40 45 Leu Pro Phe Ala Arg Pro Thr Asn Leu Thr Asp Glu Phe Glu Phe Pro 50 55 60 Ile Gly Thr Tyr Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Ser Gly Arg 65 70 75 80 Pro Phe Ser Ile Ile Cys Leu Lys Asn Ser Val Trp Thr Gly Ala Lys 85 90 95 Asp Arg Cys Arg Arg Lys Ser Cys Arg Asn Pro Pro Asp Pro Val Asn 100 105 110 Gly Met Val His Val Ile Lys Gly Ile Gln Phe Gly Ser Gln Ile Lys 115 120 125 Tyr Ser Cys Thr Lys Gly Tyr Arg Leu Ile Gly Ser Ser Ser Ala Thr 130 135 140 Cys Ile Ile Ser Gly Asp Thr Val Ile Trp Asp Asn Glu Thr Pro Ile 145 150 155 160 Cys Asp Arg Ile Pro Cys Gly Leu Pro Pro Thr Ile Thr Asn Gly Asp 165 170 175 Phe Ile Ser Thr Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr 180 185 190 Tyr Arg Cys Asn Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val 195 200 205 Gly Glu Pro Ser Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile 210 215 220 Trp Ser Gly Pro Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro 225 230 235 240 Pro Asn Val Glu Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe 245 250 255 Ser Leu Asn Glu Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met 260 265 270 Lys Gly Pro Arg Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro 275 280 285 Glu Leu Pro Ser Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu 290 295 300 His Ala Glu Arg Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln 305 310 315 320 Glu Val Phe Tyr Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala 325 330 335 Ser Met Arg Cys Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr 340 345 350 Cys Glu Val Lys Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly 355 360 365 Arg Val Leu Phe Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe 370 375 380 Val Cys Asp Glu Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys 385 390 395 400 Val Leu Ala Gly Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys 405 410 415 Glu Gln Ile Phe Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His 420 425 430 Thr Gly Lys Pro Leu Glu Val Phe Pro Phe Gly Lys Thr Val Asn Tyr 435 440 445 Thr Cys Asp Pro His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly 450 455 460 Glu Ser Thr Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp 465 470 475 480 Ser Ser Pro Ala Pro Arg Cys Gly Ile His His His His His His His 485 490 495 His <210> 27 <211> 1059 <212> PRT <213> Artificial Sequence <220> <223> sCR1(940-1971)-8His <400> 27 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala 20 25 30 Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser 35 40 45 Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile 50 55 60 Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys 65 70 75 80 Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His 85 90 95 Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr 100 105 110 Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser 115 120 125 Gly Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 130 135 140 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr 145 150 155 160 Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn 165 170 175 Leu Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser 180 185 190 Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 195 200 205 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu 210 215 220 Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu 225 230 235 240 Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg 245 250 255 Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser 260 265 270 Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His 275 280 285 Thr Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 290 295 300 Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys 305 310 315 320 Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys 325 330 335 Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe 340 345 350 Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu 355 360 365 Gly Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly 370 375 380 Met Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe 385 390 395 400 Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro 405 410 415 Ser Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro 420 425 430 His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile 435 440 445 Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala 450 455 460 Pro Arg Cys Glu Leu Ser Val Arg Ala Gly His Cys Lys Thr Pro Glu 465 470 475 480 Gln Phe Pro Phe Ala Ser Pro Thr Ile Pro Ile Asn Asp Phe Glu Phe 485 490 495 Pro Val Gly Thr Ser Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Phe Gly 500 505 510 Lys Met Phe Ser Ile Ser Cys Leu Glu Asn Leu Val Trp Ser Ser Val 515 520 525 Glu Asp Asn Cys Arg Arg Lys Ser Cys Gly Pro Pro Pro Glu Pro Phe 530 535 540 Asn Gly Met Val His Ile Asn Thr Asp Thr Gln Phe Gly Ser Thr Val 545 550 555 560 Asn Tyr Ser Cys Asn Glu Gly Phe Arg Leu Ile Gly Ser Pro Ser Thr 565 570 575 Thr Cys Leu Val Ser Gly Asn Asn Val Thr Trp Asp Lys Lys Ala Pro 580 585 590 Ile Cys Glu Ile Ile Ser Cys Glu Pro Pro Pro Thr Ile Ser Asn Gly 595 600 605 Asp Phe Tyr Ser Asn Asn Arg Thr Ser Phe His Asn Gly Thr Val Val 610 615 620 Thr Tyr Gln Cys His Thr Gly Pro Asp Gly Glu Gln Leu Phe Glu Leu 625 630 635 640 Val Gly Glu Arg Ser Ile Tyr Cys Thr Ser Lys Asp Asp Gln Val Gly 645 650 655 Val Trp Ser Ser Pro Pro Pro Arg Cys Ile Ser Thr Asn Lys Cys Thr 660 665 670 Ala Pro Glu Val Glu Asn Ala Ile Arg Val Pro Gly Asn Arg Ser Phe 675 680 685 Phe Ser Leu Thr Glu Ile Ile Arg Phe Arg Cys Gln Pro Gly Phe Val 690 695 700 Met Val Gly Ser His Thr Val Gln Cys Gln Thr Asn Gly Arg Trp Gly 705 710 715 720 Pro Lys Leu Pro His Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile 725 730 735 Leu His Gly Glu His Thr Leu Ser His Gln Asp Asn Phe Ser Pro Gly 740 745 750 Gln Glu Val Phe Tyr Ser Cys Glu Pro Ser Tyr Asp Leu Arg Gly Ala 755 760 765 Ala Ser Leu His Cys Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro 770 775 780 Arg Cys Thr Val Lys Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His 785 790 795 800 Gly Arg Val Leu Leu Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser 805 810 815 Phe Val Cys Asp Glu Gly Phe Arg Leu Lys Gly Arg Ser Ala Ser His 820 825 830 Cys Val Leu Ala Gly Met Lys Ala Leu Trp Asn Ser Ser Val Pro Val 835 840 845 Cys Glu Gln Ile Phe Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg 850 855 860 His Thr Gly Thr Pro Phe Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser 865 870 875 880 Tyr Ala Cys Asp Thr His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile 885 890 895 Gly Glu Ser Ser Ile Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val 900 905 910 Trp Ser Ser Pro Ala Pro Arg Cys Glu Leu Ser Val Pro Ala Ala Cys 915 920 925 Pro His Pro Pro Lys Ile Gln Asn Gly His Tyr Ile Gly Gly His Val 930 935 940 Ser Leu Tyr Leu Pro Gly Met Thr Ile Ser Tyr Ile Cys Asp Pro Gly 945 950 955 960 Tyr Leu Leu Val Gly Lys Gly Phe Ile Phe Cys Thr Asp Gln Gly Ile 965 970 975 Trp Ser Gln Leu Asp His Tyr Cys Lys Glu Val Asn Cys Ser Phe Pro 980 985 990 Leu Phe Met Asn Gly Ile Ser Lys Glu Leu Glu Met Lys Lys Val Tyr 995 1000 1005 His Tyr Gly Asp Tyr Val Thr Leu Lys Cys Glu Asp Gly Tyr Thr 1010 1015 1020 Leu Glu Gly Ser Pro Trp Ser Gln Cys Gln Ala Asp Asp Arg Trp 1025 1030 1035 Asp Pro Pro Leu Ala Lys Cys Thr Ser Arg Thr His Asp His His 1040 1045 1050 His His His His His His 1055 <210> 28 <211> 477 <212> PRT <213> Artificial sequence <220> <223> sCR1(490-939)-8His <400> 28 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala 20 25 30 Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser 35 40 45 Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile 50 55 60 Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys 65 70 75 80 Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His 85 90 95 Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr 100 105 110 Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser 115 120 125 Gly Asn Ala Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 130 135 140 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr 145 150 155 160 Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn 165 170 175 Pro Gly Ser Gly Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser 180 185 190 Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 195 200 205 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu 210 215 220 Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu 225 230 235 240 Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg 245 250 255 Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser 260 265 270 Cys Ser Arg Val Cys Gln Pro Pro Pro Asp Val Leu His Ala Glu Arg 275 280 285 Thr Gln Arg Asp Lys Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 290 295 300 Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Met Arg Cys 305 310 315 320 Thr Pro Gln Gly Asp Trp Ser Pro Ala Ala Pro Thr Cys Glu Val Lys 325 330 335 Ser Cys Asp Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe 340 345 350 Pro Val Asn Leu Gln Leu Gly Ala Lys Val Asp Phe Val Cys Asp Glu 355 360 365 Gly Phe Gln Leu Lys Gly Ser Ser Ala Ser Tyr Cys Val Leu Ala Gly 370 375 380 Met Glu Ser Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe 385 390 395 400 Cys Pro Ser Pro Pro Val Ile Pro Asn Gly Arg His Thr Gly Lys Pro 405 410 415 Leu Glu Val Phe Pro Phe Gly Lys Ala Val Asn Tyr Thr Cys Asp Pro 420 425 430 His Pro Asp Arg Gly Thr Ser Phe Asp Leu Ile Gly Glu Ser Thr Ile 435 440 445 Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala 450 455 460 Pro Arg Cys Gly Ile His His His His His His His His 465 470 475 <210> 29 <211> 480 <212> PRT <213> Artificial Sequence <220> <223> sCR1(940-1392)-8His <400> 29 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala Leu Gly His Cys Gln Ala Pro Asp His Phe Leu Phe Ala 20 25 30 Lys Leu Lys Thr Gln Thr Asn Ala Ser Asp Phe Pro Ile Gly Thr Ser 35 40 45 Leu Lys Tyr Glu Cys Arg Pro Glu Tyr Tyr Gly Arg Pro Phe Ser Ile 50 55 60 Thr Cys Leu Asp Asn Leu Val Trp Ser Ser Pro Lys Asp Val Cys Lys 65 70 75 80 Arg Lys Ser Cys Lys Thr Pro Pro Asp Pro Val Asn Gly Met Val His 85 90 95 Val Ile Thr Asp Ile Gln Val Gly Ser Arg Ile Asn Tyr Ser Cys Thr 100 105 110 Thr Gly His Arg Leu Ile Gly His Ser Ser Ala Glu Cys Ile Leu Ser 115 120 125 Gly Asn Thr Ala His Trp Ser Thr Lys Pro Pro Ile Cys Gln Arg Ile 130 135 140 Pro Cys Gly Leu Pro Pro Thr Ile Ala Asn Gly Asp Phe Ile Ser Thr 145 150 155 160 Asn Arg Glu Asn Phe His Tyr Gly Ser Val Val Thr Tyr Arg Cys Asn 165 170 175 Leu Gly Ser Arg Gly Arg Lys Val Phe Glu Leu Val Gly Glu Pro Ser 180 185 190 Ile Tyr Cys Thr Ser Asn Asp Asp Gln Val Gly Ile Trp Ser Gly Pro 195 200 205 Ala Pro Gln Cys Ile Ile Pro Asn Lys Cys Thr Pro Pro Asn Val Glu 210 215 220 Asn Gly Ile Leu Val Ser Asp Asn Arg Ser Leu Phe Ser Leu Asn Glu 225 230 235 240 Val Val Glu Phe Arg Cys Gln Pro Gly Phe Val Met Lys Gly Pro Arg 245 250 255 Arg Val Lys Cys Gln Ala Leu Asn Lys Trp Glu Pro Glu Leu Pro Ser 260 265 270 Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His 275 280 285 Thr Pro Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 290 295 300 Ser Cys Glu Pro Gly Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys 305 310 315 320 Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Ala Val Lys 325 330 335 Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Phe 340 345 350 Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu 355 360 365 Gly Phe Arg Leu Lys Gly Ser Ser Val Ser His Cys Val Leu Val Gly 370 375 380 Met Arg Ser Leu Trp Asn Asn Ser Val Pro Val Cys Glu His Ile Phe 385 390 395 400 Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro 405 410 415 Ser Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Thr Cys Asp Pro 420 425 430 His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Thr Ile 435 440 445 Arg Cys Thr Ser Asp Pro His Gly Asn Gly Val Trp Ser Ser Pro Ala 450 455 460 Pro Arg Cys Glu Leu Ser Val Arg His His His His His His His His 465 470 475 480 <210> 30 <211> 606 <212> PRT <213> Artificial Sequence <220> <223> sCR1(1393-1971)-8His <400> 30 Met Lys Ile Leu Ile Leu Gly Ile Phe Leu Phe Leu Cys Ser Thr Pro 1 5 10 15 Ala Trp Ala Ala Gly His Cys Lys Thr Pro Glu Gln Phe Pro Phe Ala 20 25 30 Ser Pro Thr Ile Pro Ile Asn Asp Phe Glu Phe Pro Val Gly Thr Ser 35 40 45 Leu Asn Tyr Glu Cys Arg Pro Gly Tyr Phe Gly Lys Met Phe Ser Ile 50 55 60 Ser Cys Leu Glu Asn Leu Val Trp Ser Ser Val Glu Asp Asn Cys Arg 65 70 75 80 Arg Lys Ser Cys Gly Pro Pro Pro Glu Pro Phe Asn Gly Met Val His 85 90 95 Ile Asn Thr Asp Thr Gln Phe Gly Ser Thr Val Asn Tyr Ser Cys Asn 100 105 110 Glu Gly Phe Arg Leu Ile Gly Ser Pro Ser Thr Thr Cys Leu Val Ser 115 120 125 Gly Asn Asn Val Thr Trp Asp Lys Lys Ala Pro Ile Cys Glu Ile Ile 130 135 140 Ser Cys Glu Pro Pro Pro Thr Ile Ser Asn Gly Asp Phe Tyr Ser Asn 145 150 155 160 Asn Arg Thr Ser Phe His Asn Gly Thr Val Val Thr Tyr Gln Cys His 165 170 175 Thr Gly Pro Asp Gly Glu Gln Leu Phe Glu Leu Val Gly Glu Arg Ser 180 185 190 Ile Tyr Cys Thr Ser Lys Asp Asp Gln Val Gly Val Trp Ser Ser Pro 195 200 205 Pro Pro Arg Cys Ile Ser Thr Asn Lys Cys Thr Ala Pro Glu Val Glu 210 215 220 Asn Ala Ile Arg Val Pro Gly Asn Arg Ser Phe Phe Ser Leu Thr Glu 225 230 235 240 Ile Ile Arg Phe Arg Cys Gln Pro Gly Phe Val Met Val Gly Ser His 245 250 255 Thr Val Gln Cys Gln Thr Asn Gly Arg Trp Gly Pro Lys Leu Pro His 260 265 270 Cys Ser Arg Val Cys Gln Pro Pro Pro Glu Ile Leu His Gly Glu His 275 280 285 Thr Leu Ser His Gln Asp Asn Phe Ser Pro Gly Gln Glu Val Phe Tyr 290 295 300 Ser Cys Glu Pro Ser Tyr Asp Leu Arg Gly Ala Ala Ser Leu His Cys 305 310 315 320 Thr Pro Gln Gly Asp Trp Ser Pro Glu Ala Pro Arg Cys Thr Val Lys 325 330 335 Ser Cys Asp Asp Phe Leu Gly Gln Leu Pro His Gly Arg Val Leu Leu 340 345 350 Pro Leu Asn Leu Gln Leu Gly Ala Lys Val Ser Phe Val Cys Asp Glu 355 360 365 Gly Phe Arg Leu Lys Gly Arg Ser Ala Ser His Cys Val Leu Ala Gly 370 375 380 Met Lys Ala Leu Trp Asn Ser Ser Val Pro Val Cys Glu Gln Ile Phe 385 390 395 400 Cys Pro Asn Pro Pro Ala Ile Leu Asn Gly Arg His Thr Gly Thr Pro 405 410 415 Phe Gly Asp Ile Pro Tyr Gly Lys Glu Ile Ser Tyr Ala Cys Asp Thr 420 425 430 His Pro Asp Arg Gly Met Thr Phe Asn Leu Ile Gly Glu Ser Ser Ile 435 440 445 Arg Cys Thr Ser Asp Pro Gln Gly Asn Gly Val Trp Ser Ser Pro Ala 450 455 460 Pro Arg Cys Glu Leu Ser Val Pro Ala Ala Cys Pro His Pro Pro Lys 465 470 475 480 Ile Gln Asn Gly His Tyr Ile Gly Gly His Val Ser Leu Tyr Leu Pro 485 490 495 Gly Met Thr Ile Ser Tyr Ile Cys Asp Pro Gly Tyr Leu Leu Val Gly 500 505 510 Lys Gly Phe Ile Phe Cys Thr Asp Gln Gly Ile Trp Ser Gln Leu Asp 515 520 525 His Tyr Cys Lys Glu Val Asn Cys Ser Phe Pro Leu Phe Met Asn Gly 530 535 540 Ile Ser Lys Glu Leu Glu Met Lys Lys Val Tyr His Tyr Gly Asp Tyr 545 550 555 560 Val Thr Leu Lys Cys Glu Asp Gly Tyr Thr Leu Glu Gly Ser Pro Trp 565 570 575 Ser Gln Cys Gln Ala Asp Asp Arg Trp Asp Pro Pro Leu Ala Lys Cys 580 585 590 Thr Ser Arg Thr His Asp His His His His His His His His 595 600 605 <210> 31 <211> 13 <212> PRT <213> Artificial sequence <220> <223> connector <400> 31 Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Ser 1 5 10 <210> 32 <211> 585 <212> PRT <213> Artificial sequence <220> <223> Human serum albumin <400> 32 Asp Ala His Lys Ser Glu Val Ala His Arg Phe Lys Asp Leu Gly Glu 1 5 10 15 Glu Asn Phe Lys Ala Leu Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln 20 25 30 Gln Cys Pro Phe Glu Asp His Val Lys Leu Val Asn Glu Val Thr Glu 35 40 45 Phe Ala Lys Thr Cys Val Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys 50 55 60 Ser Leu His Thr Leu Phe Gly Asp Lys Leu Cys Thr Val Ala Thr Leu 65 70 75 80 Arg Glu Thr Tyr Gly Glu Met Ala Asp Cys Cys Ala Lys Gln Glu Pro 85 90 95 Glu Arg Asn Glu Cys Phe Leu Gln His Lys Asp Asp Asn Pro Asn Leu 100 105 110 Pro Arg Leu Val Arg Pro Glu Val Asp Val Met Cys Thr Ala Phe His 115 120 125 Asp Asn Glu Glu Thr Phe Leu Lys Tyr Leu Tyr Glu Ile Ala Arg 130 135 140 Arg His Pro Tyr Phe Tyr Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg 145 150 155 160 Tyr Lys Ala Phe Thr Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala 165 170 175 Cys Leu Leu Pro Lys Leu Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser 180 185 190 The Ser Al of Lys Gln Arg Leu Lys Cys Al Ser Leu Gln Lys Phe Gly Glu 195 200 205 Arg Ala Phe Lys Ala Trp Ala Val Ala Arg Leu Ser Gln Arg Phe Pro 210 215 220 Lys Ala Glu Phe Ala Glu Val Ser Leu Val Thr Asp Leu Thr Lys 225 230 235 240 Val His Thr Glu Cys His Gly Asp Leads To Glu Cys Ala Asp Asp 245 250 255 Arg Ala Asp Leu Ala Lys Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser 260 265 270 Ser Lys Leu Lys Glu Cys Cys Glu Lys Pro Leu Leu Glu Lys Ser His 275 280 285 Cys Ile Ala Glu Val Glu Asn Asp Glu Met Pro Ala Asp Leu Pro Ser 290,295,300 Leu Ala Ala Asp Phe Val Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala 305 310 315 320 Glu Ala Lys Asp Val Phe Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg 325 330 335 Arg His Pro Asp Tyr Ser Val Val Leu Leu Leu Arg Leu Ala Lys Thr 340 345 350 Tyr Glu Thr Thr Leu Glu Lys Cys Ala Ala Ala Asp Pro His Glu 355 360 365 Cys Tyr Ala Lys Val Phe Asp Glu Phe Lys Pro Leu Val Glu Glu Pro 370 375 380 Gln Asn With Lys Gln Asn Cys Glu To Phe Glu Gln To Gly Glu 385 390 395 400 Tyr Lys Phe Gln Asn Ala Leu Leu Val Arg Tyr Thr Lys Val Pro 405 410 415 Gln Val Ser Thr Pro Thr Leu Val Glu Val Ser Arg Asn Leu Gly Lys 420 425 430 Val Gly Ser Lys Cys Cys Lys His Pro Glu Ala Lys Arg Met Pro Cys 435 440 445 Ala Glu Asp Tyr Leu Ser Val Val Leu Asn Gln Leu Cys Val Leu His 450 455 460 Glu Lys Thr Pro Val Ser Asp Arg Val Thr Lys Cys Cys Thr Glu Ser 465 470 475 480 Leu Val Asn Arg Arg Pro Cys Phe Ser Ala Leu Glu Val Asp Glu Thr 485 490 495 Tyr Val Pro Lys Glu Phe Asn Ala Glu Thr Phe Thr Phe His Ala Asp 500 505 510 Ile Cys Thr Leu Ser Glu Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala 515 520 525 Leu Val Glu Leu Val Lys His Lys Pro Lys Ala Thr Lys Glu Gln Leu 530 535 540 Lys Ala Val Met Asp Asp Phe Ala Ala Phe Val Glu Lys Cys Cys Lys 545 550 555 560 Ala Asp Asp Lys Glu Thr Cys Phe Ala Glu Glu Gly Lys Lys Leu Val 565 570 575 Ala Ala Ser Gln Ala Ala Leu Gly Leu 580 585 <210> 33 <211> 229 <212> PRT <213> Artificial Sequence <220> <223> IgG1 Fc <400> 33 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe 1 5 10 15 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 65 70 75 80 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 85 90 95 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 130 135 140 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ala Ile 145 150 155 160 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu 180 185 190 Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 195 200 205 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 Leu Ser Leu Gly Lys 225 <210> 34 <211> 229 <212> PRT <213> Artificial Sequence <220> <223> IgG4 Fc <400> 34 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe 1 5 10 15 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 65 70 75 80 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His ...

Claims

Use of a type 1.1 soluble complement receptor (sCR1) variant in the preparation of a medicament for the prevention or treatment of complement-mediated conditions in subjects of need, wherein the sCR1 variant comprises an amino acid sequence corresponding to amino acids 42 to 1392 of SEQ ID NO:

1. That is, the sCR1 variant consists of long homologous repeat (LHR) regions LHR-A, LHR-B, and LHR-C, but lacks LHR-D. The complement-mediated conditions mentioned above are selected from renal ischemia-reperfusion injury and antiglomerular basement membrane (GBM) nephritis.

2. The use according to claim 1, wherein the complement-mediated condition is antiglomerular basement membrane (GBM) nephritis.

3. The use according to claim 1, wherein the complement-mediated condition is renal ischemia-reperfusion injury.

4. The use according to claim 1, wherein the sCR1 variant has increased complement inhibitory activity compared to the sCR1 containing the sequence shown in SEQ ID NO:

2.

5. The use according to claim 1, wherein the sCR1 variant has increased complement inhibitory activity in the classical pathway, lectin pathway and / or complement bypass pathway compared to the sCR1 comprising the sequence shown in SEQ ID NO:

2.

6. The use according to claim 1, wherein the sCR1 variant is conjugated with a half-life-extending portion or an additional soluble complement inhibitor.

7. The use according to claim 6, wherein the extended half-life portion is selected from: human serum albumin or a functional fragment thereof, the Fc region of monomeric or dimeric immunoglobulin or a functional fragment thereof, afamin, alpha-fetoprotein, vitamin D-binding protein, antibody fragments bound to albumin, and polymers.

8. The use according to claim 6, wherein the additional soluble complement inhibitor is selected from: C1 inhibitor (C1-INH), factor I (fI), factor H (fH), complement factor H-related protein (CFHR), C4b binding protein (C4bp), soluble CD55, i.e., decay-promoting factor (DAF), soluble CD46, i.e., membrane cofactor protein (MCP), soluble CD59, i.e., protectin, soluble complement receptor 2 (sCR2), TT30, i.e., CR2-fH, and cobra venom factor (CVF).

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