Fusion proteins comprising human Lefty A protein variants and uses thereof

Abstract

The present invention relates to a human Lefty A protein variant having improved productivity and stability, a fusion protein comprising the protein variant, and a composition for preventing and / or treating neuromuscular diseases, the composition comprising the protein variant or the fusion protein. According to the present invention, a human Lefty A protein variant and a fusion protein comprising this variant are constructed, which have better stability than the naturally occurring human Lefty A protein, and thus are highly expressed and produced in high yields in animal cells. In addition, administration of the constructed human Lefty A protein variant or fusion protein can restore the nerve and motor functions of an animal model of peripheral neuropathy. Therefore, the use of the human Lefty A protein variant or fusion protein can effectively prevent or treat various nerve and muscle diseases.

Description

Technical Field

[0001] The present invention relates to a human Lefty A protein variant having improved productivity and stability, a fusion protein comprising the protein variant, and a composition for preventing and / or treating neuromuscular diseases, the composition comprising the protein variant or the fusion protein. Background Art

[0002] Lefty (left-right determination factor) is a morphogenetic factor belonging to the transforming growth factor-beta (TGF-β) superfamily and plays a key role in the differentiation and development of embryonic cells by binding to other TGF-β ligands (Shiratori H et al., Semin Cell Dev Biol. 2014; 32: 80-4).

[0003] The human LEFTY gene contains two genes (LEFTY2 encoding Lefty A protein and LEFTY1 encoding Lefty B protein) generated by independent duplication (Gharib WH, Robinson-Rechavi M, Brief Bioinform. 2011; 12(5): 436-41). The deduced amino acid sequences of Lefty A and Lefty B show 96% identity.

[0004] The human LEFTY gene translates a polypeptide consisting of 366 amino acids. The 21-amino acid amino-terminal signal peptide is cleaved during translocation to the endoplasmic reticulum (ER), and finally the extracellular release is a 42-kilodalton (kDa) pro-protein. The released 42 kDa Lefty protein can be processed by protease cleavage at its amino-terminal cleavage site (RXXR) to remove the propeptide. Unlike other members of the TGF-β family, Lefty has two instead of one putative cleavage site (RXXRs) and does not have the conserved cysteine required for the formation of homodimers (Juan et al., Genes to Cells. 2001; 6: 923-30).

[0005] The Lefty polypeptide can be cleaved at the carboxyl terminus of the second arginine at each cleavage site by proprotein convertase and processed into polypeptides of 34 kDa and 28 kDa. Proteolytic processing of the Lefty polypeptide can be blocked by mutating the two arginine residues at the first cleavage site (RGKR) to glycine or the first arginine at the second cleavage site (RHGR) to glycine (Ulloa L et al., J Biol Chem. 2001;276:217387-96).

[0006] It has been reported that the 42 kDa Lefty precursor protein and the 28 kDa mature protein can induce MAPK activation (Ulloa L et al., J Biol Chem. 2001;276:217387-96).

[0007] As an endogenous inhibitor of Nodal (a TGF-β family protein), Lefty plays an important role in early embryonic development during vertebrate development, such as inducing mesoderm and endoderm differentiation and controlling left-right asymmetry (Schier AF, Annu Rev Cell Biol. 2003;19:589-621). The exact mechanism of action by which Lefty inhibits Nodal is still unclear, but it is known that Lefty can bind to Nodal, thereby preventing Nodal from binding to the Activin receptor (such as ActRI or ActRII) and the co-receptor EGF-CFC (Branford et al., Current Biol. 2004;14:341-3 and Chen et al. Current Biol. 2004;14:618-24). In addition, it has been reported that Lefty inhibits TGF-β or BMP signaling (Ulloa L et al., J Biol Chem. 2001;276(24):2139-**404**).

[0008] Note: There seems to be a typo in the original text where "21397-404" in should probably be "21397-404" (the hyphenated range). Also, the last number in the citation in the translated text has been corrected to "404" as it seems to be an error in the original Chinese text which might have been mis-transcribed. If this is not an error, please let me know and I'll adjust accordingly.There are many TGF-β superfamily ligands, such as BMP, GDF, Nodal, activin, and TGF-β. They play important roles in regulating embryonic development and the functions of many cells and organs after birth. For example, TGF-β family members are involved in neural differentiation and regulate multiple parts of the central nervous system throughout development from early embryo to adult (Myers EA and Kessler JA, Cold Spring Harb Perspect Biol. 2017;9(8)). TGF-β inhibits the survival of Schwann cells during the process of neural differentiation (Parkinson et al., J Neurosci. 2001;21:8572-85). BMP7 inhibits the myelination process of Schwann cells by activating p38 in the peripheral nerves (Liu X et al., Sci Rep. 2016;6:31049). GDF8 (myostatin) inhibits skeletal muscle growth, and the expression of GDF8 induces skeletal muscle atrophy (Elkina Y et al., J Cachexia Sarcopenia Muscle. 2001;2:143-51).

[0009] Charcot-Marie-Tooth disease (CMT) is one of the most common hereditary peripheral neuropathies, with a prevalence of 1 in 2,500. More than 80 genes have been identified to cause CMT. Patients with CMT develop slow progressive muscular atrophy and sensory loss starting from the lower limbs.

[0010] CMT is an expressive and genetically heterogeneous disease, and is classified into a demyelinating type (CMT1) and an axonal type (CMT2) according to the motor nerve conduction velocities (MCV) of the upper limbs. Among these types, the demyelinating type (CMT1) accounts for about 70% of all CMT patients. The demyelinating type (CMT1) is caused by mutations or overexpression of myelin proteins such as PMP22 and MPZ. Although the pathological mechanism has not been elucidated, it has been proposed that protein lipid overexpression or mutation induces endoplasmic reticulum stress and is accompanied by apoptosis of Schwann cells as one of the mechanisms. Since no drug has been approved for the treatment of CMT, the interest in developing effective therapies for CMT is increasing.

[0011] In this technical background, the inventors of the present invention constructed variants with improved productivity and stability by introducing mutations into the human Lefty A protein, so that the Lefty A protein can be solubilized for the treatment of neuropathy or muscle diseases, and thus completed the present invention.

[0012] The information disclosed in the prior art is only used to strengthen the understanding of the background of the present invention, and therefore may not include information on the prior art known to those of ordinary skill in the art. Summary of the Invention

[0013] Technical Problem

[0014] One object of the present invention is to provide a variant of human Lefty A protein with improved productivity and stability.

[0015] Another object of the present invention is to provide a fusion protein comprising a variant of human Lefty A protein.

[0016] Yet another object of the present invention is to provide a nucleic acid molecule encoding the fusion protein, an expression vector comprising the nucleic acid molecule, a recombinant cell into which the expression vector is introduced, and a method for producing the fusion protein using the recombinant cell.

[0017] Still another object of the present invention is to provide a composition for preventing and / or treating neuromuscular diseases, comprising the protein variant or the fusion protein.

[0018] A further object of the present invention is to provide a method for preventing and / or treating neuromuscular diseases, comprising administering the protein variant or the fusion protein to a subject.

[0019] Another further object of the present invention is to provide the use of the protein variant or the fusion protein for the prevention and / or treatment of neuromuscular diseases.

[0020] The present invention further has an object to provide the use of a protein variant or a fusion protein for manufacturing a drug for treating neuromuscular diseases.

[0021] Solution

[0022] To achieve the above object, the present invention provides a human Lefty A protein variant, comprising the amino acid sequence from L22 to P366 of the human Lefty A protein, the human Lefty A protein having the amino acid sequence of SEQ ID NO: 131, wherein the human Lefty A protein variant comprises: (1) substitution of one or more amino acid residues at the processing sites (R74 to R77 and R132 to R135); and (2) substitution of one or more amino acid residues in the propeptide domain (L22 to S73).

[0023] The present invention also provides a fusion protein comprising the human Lefty A protein variant.

[0024] The present invention also provides a nucleic acid molecule encoding a fusion protein comprising the human Lefty A protein variant, an expression vector comprising the nucleic acid molecule, a recombinant cell into which the expression vector is introduced, and a method for manufacturing a fusion protein comprising the human Lefty A protein variant using the recombinant cell.

[0025] The present invention also provides a composition for preventing and / or treating neuromuscular diseases, comprising the human Lefty A protein variant or a fusion protein comprising the human Lefty A protein variant.

[0026] The present invention also provides a method for preventing and / or treating neuromuscular diseases, comprising administering the human Lefty A protein variant or a fusion protein comprising the human Lefty A protein variant to a subject.

[0027] The present invention also provides the use of a human Lefty A protein variant or a fusion protein comprising the human Lefty A protein variant for the prevention and / or treatment of neuromuscular diseases.

[0028] The present invention also provides the use of a human Lefty A protein variant or a fusion protein comprising the human Lefty A protein variant for manufacturing a drug for treating neuromuscular diseases. Description of the Drawings

[0029] Figure 1 Schematic diagram of the human Lefty A fusion protein.

[0030] Figure 2Western blot analysis of the culture supernatant was performed to evaluate the expression of human Lefty A protein variants fused with human serum albumin (HSA-fused).

[0031] Figure 3 SDS-PAGE analysis of 42LFc protein after ProA Affinity purification was shown.

[0032] Figure 4 SDS-PAGE analysis and size exclusion chromatography (SEC) analysis of Lefty A protein variants (combination) after ProA Affinity purification were shown.

[0033] Figure 5 Cell viability assay and Western blot analysis using anti-PARP antibody were shown to evaluate the effect on thapsigargin-induced apoptosis of S16 Schwann cells co-cultured with human Wharton's jelly-derived human mesenchymal stem cell (MSC).

[0034] Figure 6 Electrophysiological assessment of the therapeutic effect of human Lefty A fusion protein variants in Tr-J mice was shown.

[0035] Figure 7 Western blot analysis was shown to evaluate the expression of (A) Krox20 and MBP or (B), (C) Krox20 and phosphorylated / unphosphorylated p35 in RT4 Schwann cells treated with Nodal or / and human LeftyA fusion protein variants.

[0036] Figure 8 Electrophysiological assessment of the therapeutic effect of intraperitoneal injection of human Lefty A fusion protein variants in C22 mice was shown.

[0037] Figure 9Show the (A) rotarod test and (B) hindlimb grip strength test of C22 mice intraperitoneally injected with human Lefty A fusion protein variants.

[0038] Figure 10 Show the magnetic resonance imaging analysis of the gastrocnemius muscle of C22 mice intraperitoneally injected with human Lefty A fusion protein variants.

[0039] Figure 11 Show the gait analysis of wild-type and C22 mice intraperitoneally injected with human Lefty A fusion protein variants.

[0040] Figure 12 Show the electrophysiological evaluation of the therapeutic effect of intraperitoneal injection of human Lefty A fusion protein variants in C22 mice.

[0041] Figure 13 Show the (A) hindlimb grip strength test and (B) rotarod test of C22 mice intraperitoneally injected with human Lefty A fusion protein variants.

[0042] Figure 14 Show the gait analysis of C22 mice intraperitoneally injected with human Lefty A fusion protein variants.

[0043] Figure 15 Show the electrophysiological analysis of C22 mice subcutaneously injected with human Lefty A fusion protein variants for 4 weeks starting from 3 weeks of age.

[0044] Figure 16 Show the hindlimb grip strength test of C22 mice subcutaneously injected with human Lefty A fusion protein variants for 4 weeks starting from 3 weeks of age.

[0045] Figure 17 Show the luciferase reporter assay based on A204 cells to evaluate the effect of human Lefty A fusion protein variants on the signal transduction through myostatin.

[0046] Figure 18 Show the Western blot analysis of the sciatic nerve lysates of C22 mice injected with human Lefty A fusion protein variants.

[0047] Figure 19Electrophysiological analysis of C22 mice subcutaneously injected with human Lefty A fusion protein variant for 4 weeks starting at 5 weeks of age is shown.

[0048] Figure 20 (A) Rotarod test and (B) whole limb grip strength test of C22 mice subcutaneously injected with human Lefty A fusion protein variant for 4 weeks starting at 5 weeks of age are shown.

[0049] Figure 21 Western blot analysis to evaluate the effect of human Lefty A fusion protein variant on Nodal signaling in P19 cells is shown.

[0050] Figures 22 and 23 show the amino acid sequences of the human Lefty A fusion protein variant according to the present invention. Detailed Description

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature used herein and the experimental methods described below are well known and commonly employed in the art.

[0052] In the present invention, a fusion protein, compared with wild-type (WT) protein and previously reported human Lefty A variants, improves productivity and stability in Chinese hamster ovary cells (CHO cells), can also bind to human Nodal and inhibit Nodal-mediated signaling, and helps to improve the symptoms of peripheral neuropathy. The fusion protein comprises a human Lefty A (Uniprot No. 000292; NCBI DB NM_003240) protein variant with enhanced productivity and stability. Therefore, the fusion protein can be effectively used alone or in combination with conventional pharmaceutically acceptable carriers, neuropathy treatment reagents, muscle disease treatment reagents, etc. as a composition for preventing or treating nerve and muscle diseases.

[0053] Accordingly, in one aspect, the present invention relates to a human Lefty A protein variant comprising the amino acid sequence of L22 to P366 of human Lefty A protein, wherein the human Lefty A protein has the amino acid sequence of SEQ ID NO:131, and the human Lefty A protein variant comprises: (1) substitution of one or more amino acid residues at processing sites (R74 to R77 and R132 to R135); and (2) substitution of one or more amino acid residues in the propeptide domain (L22 to S73).

[0054] M1 to A21 in the human Lefty A protein having the amino acid sequence of SEQ ID NO:131 correspond to the signal peptide.

[0055] The human Lefty A protein variant according to the present invention also refers to a variant in which the amino acid residues at specific amino acid residue positions are conservatively substituted.

[0056] The term "conservative substitution" as used in the present invention refers to the processing of a Lefty A protein variant, which involves substituting one or more amino acids with amino acids having similar biochemical properties, and the amino acids having similar biochemical properties do not result in the loss of the biological or biochemical function of the Lefty A protein variant.

[0057] "Conservative amino acid substitution" is the replacement of an amino acid residue with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined and are well known in the art to which the present invention pertains. These families include amino acids having basic side chains (such as lysine, arginine, and histidine), amino acids having acidic side chains (such as aspartic acid and glutamic acid), amino acids having uncharged polar side chains (such as glycine, asparagine, glutamine, serine, threonine, tyrosine, and cysteine), amino acids having nonpolar side chains (such as alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan), amino acids having β-branched side chains (such as threonine, valine, and isoleucine), and amino acids having aromatic side chains (such as tyrosine, phenylalanine, tryptophan, and histidine).

[0058] It is conceivable that although the Lefty A protein variant of the present invention has conservative amino acid substitutions, it may still retain its activity.

[0059] In addition, the human Lefty A protein variants of the present invention are interpreted to include Lefty A protein variants having functions and / or effects substantially the same as those of the Lefty A protein variants of the present invention, and having at least 80% or 85% amino acid sequence homology with the Lefty A protein variants according to the present invention, preferably at least 90%, more preferably at least 95%, and most preferably at least 99%.

[0060] Preferably, the human Lefty A protein variants according to the present invention may include the amino acid sequence of L22 to P366 selected from any one of the group consisting of SEQ ID NO:86 to SEQ ID NO:111, or the amino acid sequence of L23 to P367 selected from any one of the group consisting of SEQ ID NO:112 to 129 and 133, but not limited thereto.

[0061] It should be understood that the Lefty A protein variants according to the present invention also include those having substantially the same efficacy as the Lefty A protein variants according to the present invention, even if the amino-terminal, carboxyl-terminal or internal amino acid sequences thereof are cleaved or substituted.

[0062] There is no mutation in wild-type Lefty A, that is, there is no substitution of amino acid residues at positions R74 to R77. Therefore, when protease-induced cleavage occurs in the corresponding region, a 34 kDa Lefty A protein fragment is obtained. In addition, there is no mutation at positions R132 to R135. Therefore, when protease-induced cleavage occurs in the corresponding region, a 28 kDa Lefty A protein fragment is obtained.

[0063] Therefore, the human Lefty A protein variants of the present invention preferably include substitutions of one or more amino acids at the processing sites, which are composed of R74 to R77 and R132 to R135.

[0064] In the present invention, the substitution of amino acid residues in the propeptide domain (L22 to S73) may be substitution of amino acid residues at one or more positions selected from the group consisting of E24, L27, R33, S38, V40, V42, R45, M48, K50, A55, V63, R66, R67, G70 and D71, but not limited thereto.

[0065] Preferably, the substitution of amino acid residues in the propeptide domain (L22 to S73) may be substitution of one or more amino acid residues selected from the group consisting of E24G, S38K, V42T, K50E, A55T, V63A and R66Q.

[0066] Most preferably, substitution of amino acid residues in the propeptide domain (L22 to S73) may include V63A, and may further include one or more amino acid residue substitutions selected from the group consisting of E24G, S38K, V42T, K50E, A55T, and R66Q.

[0067] In the present invention, substitution of one or more amino acid residues at the processing sites (R74 to R77 and R132 to R135) may be one or more amino acid residue substitutions selected from the group consisting of R74G, R77G, R77V, R132G, and R135G.

[0068] The amino acid sequence at positions R74 to R77 may be RGKR, GGKG, RGKA, RGKV, or RHGG, and the amino acid sequence at positions R132 to R135 may be RHGR, GHGR, RHGG, RHER, GHGG, RHGA, or RHGV, which are derived from substitution of one or more amino acid residues at the processing sites (R74 to R77 and R132 to R135).

[0069] In the present invention, the human Lefty A protein variant may further include substitution of one or more amino acid residues at the thrombin cleavage site (L311, P313, R314, L359, P361, or R362), but is not limited thereto.

[0070] Preferably, the amino acid residues at one or more positions selected from the group consisting of thrombin cleavage sites L311, P313, R314, L359, P361, and R362 may be substituted with amino acid residues selected from the group consisting of aspartic acid (D), glutamic acid (E), serine (S), lysine (K), and glutamine (Q).

[0071] In the present invention, the human Lefty A protein variant may further include substitution of one or more amino acid residues at the fragmentation site (S202 or S223) with amino acid residues other than serine (S) and cysteine (C).

[0072] In the present invention, the human Lefty A protein variant may further include a signal peptide at the amino terminus.

[0073] In the content disclosed by the present invention, the signal peptide and the signal sequence have the same meaning. The signal peptide is a short amino acid sequence, and the short amino acid sequence plays an important role in delivering the newly expressed polypeptide to the endoplasmic reticulum and entering the secretory pathway. When a protein is synthesized in a cell and leaves the cell, the signal sequence will be cleaved, and the amino terminus of the Lefty A protein variant starts from L22. The signal sequence used in the present invention can be the signal sequence (M1 to A21) derived from the Lefty A protein, but is not limited thereto. A signal sequence derived from a protein other than the Lefty A protein can also be used. For example, it can be an antibody-derived sequence, such as MDMRVPAQLLGLLLLWFPGSRC (UniProt: A0A0C4DH73; SEQ ID NO: 132), but is not limited thereto.

[0074] On the other hand, the present invention relates to a fusion protein comprising a human Lefty A protein variant.

[0075] In the present invention, "a fusion protein comprising a human Lefty A protein variant" and "a Lefty A fusion protein variant" have the same meaning.

[0076] In the present invention, a fusion protein can be produced by fusing a human Lefty A protein variant with Fc or albumin, but is not limited thereto.

[0077] Preferably, the fusion protein of the present invention can be produced by fusing Fc or albumin to the amino terminus or carboxyl terminus of the human Lefty A protein variant, preferably the carboxyl terminus. In addition, the fusion protein of the present invention can be produced by fusing the human Lefty A protein variant with Fc or albumin through a linker.

[0078] The Fc in the present invention refers to the Fc (fragment crystallizable) region of an antibody. The Fc can be the Fc of an antibody selected from the group consisting of IgG, IgA, IgM, and IgE. Preferably, the Fc can be the Fc of an antibody selected from the group consisting of human IgG, IgA, IgM, and IgE, but is not limited thereto.

[0079] More preferably, the Fc can be one selected from IgG1, IgG2, IgG3, and IgG4 derived from human IgG. Most preferably, the Fc can be a human IgG1-derived Fc, but is not limited thereto. In addition, the Fc in the present invention can be a wild-type Fc or an amino acid sequence variant thereof.

[0080] In addition, the albumin of the present invention refers to all animal-derived albumins. Preferably, human albumin can be used, but the scope of the present invention is not limited thereto.

[0081] In the present invention, the fusion protein may have any amino acid sequence selected from the group consisting of the amino acid sequences listed in SEQ ID NOs: 134 to 178, but is not limited thereto.

[0082] In the present invention, the amino acid sequences listed in SEQ ID NOs: 134 to 159 respectively refer to the amino acid sequences of L22 to K614 except for the signal sequence in the amino acid sequences listed in SEQ ID NOs: 86 to 111. The amino acid sequences listed in SEQ ID NOs: 160 to 178 respectively refer to the amino acid sequences of L23 to K615 except for the signal sequence in the amino acid sequences listed in SEQ ID NOs: 112 to 129 and SEQ ID NO: 133 ( Figures 22a to 22j and Figures 23a to 23m ).

[0083] The fusion protein according to the present invention may further comprise a signal peptide located at the amino terminus. The signal peptide capable of binding to the amino terminus of the fusion protein may be the signal sequence (M1 to A21) derived from the Lefty A protein, similar to the signal sequence of the human Lefty A protein variant, but is not limited thereto. The signal peptide used in the present invention may be the signal sequence derived from a protein other than the Lefty A protein. For example, it may be an antibody-derived sequence such as MDMRVPAQLLGLLLLWFPGSRC (UniProt: A0A0C4DH73; SEQ ID NO: 132), but is not limited thereto.

[0084] On the other hand, the present invention relates to a nucleic acid molecule encoding a fusion protein, an expression vector containing the nucleic acid molecule, a recombinant cell into which the expression vector is introduced, and a method for producing a fusion protein containing a human Lefty A protein variant using the recombinant cell.

[0085] The term "nucleic acid molecule" used in the present invention refers to comprehensively including DNA (gDNA and cDNA) and RNA molecules, and nucleotides are the basic units of nucleic acid molecules. Nucleotides also include sugar- or base-modified analogs as well as natural nucleotides (Scheit, Nucleotide Analogs, John Wiley, New York (1980); Uhlman and Peyman, Chemical Reviews. 1990; 90: 543-84). The nucleic acid molecule can encode the human Lefty A fusion protein variant of the present invention, and the sequence of this nucleic acid molecule can be processed. Processing includes insertion, deletion, or non-conservative substitution or conservative substitution of nucleotides.

[0086] The term "vector" used in the present invention includes plasmid vectors, cosmid vectors, bacteriophage vectors, and viral vectors, such as adenovirus vectors, retroviral vectors, and adeno-associated viral vectors, as a means for expressing a target gene in a host cell. Preferably, the vector may include a plasmid vector, but is not limited thereto.

[0087] In the vector of the present invention, a nucleic acid molecule encoding a fusion protein comprising a human Lefty A protein variant may be operably linked to a promoter.

[0088] The term "operably linked" used in the present invention refers to a functional linkage between a nucleic acid expression control sequence (such as a series of promoters, signal sequences, or transcription regulatory factor binding sites) and another nucleic acid sequence, such that the control sequence controls the transcription and / or translation of the other nucleic acid sequence.

[0089] The recombinant vector system of the present invention can be constructed by various methods known in the relevant technical field. The detailed methods are disclosed in Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (2001), which is incorporated herein by reference.

[0090] Vectors can generally be constructed as vectors for cloning or for expression. Vectors can be constructed as vectors using prokaryotic cells or eukaryotic cells as hosts.

[0091] For example, when the vector of the present invention is an expression vector and a prokaryotic cell is used as the host cell, it usually contains a strong promoter capable of initiating transcription (such as tac promoter, lac promoter, lacUV5 promoter, lpp promoter, pLλ promoter, pRλ promoter, rac5 promoter, amp promoter, recA promoter, SP6 promoter, trp promoter, T7 promoter, etc.), a ribosome-binding site for translation initiation, and a transcription / translation termination sequence. As the host cell, when using Escherichia coli such as HB101, BL21, DH5α, etc., the operator and promoter for tryptophan biosynthesis in Escherichia coli (Yanofsky, C., J. Bacteriol., (1984) 158: 1018-1024) and the phage λ left promoter (pLλ promoter, Herskowitz, I. and Hagen, D., Ann. Rev. Genet., (1980) 14: 399-445) can be used as regulatory sequences. When using bacilli as the host cell, the promoter for the toxin (protein) gene derived from Bacillus thuringiensis or any promoter that can be expressed in bacilli can be used as the regulatory sequence.

[0092] Meanwhile, the expression vector of the present invention can be constructed by manipulating plasmids commonly used in the art (such as pCL, pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8 / 9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14, pGEX series, pET series, pUC19, etc.), phages (such as λgt·4λB, λ-Charon, λΔz1, M13, etc.) or viruses (such as SV40, etc.). For example, the expression vector of the present invention can be constructed by manipulating the pCL expression vector, specifically the pCLS05 (Korean Patent Registration No. 10-1420274) expression vector, but not limited thereto.

[0093] In addition, when the vector of the present invention is an expression vector and eukaryotic cells are used as host cells, promoters derived from the mammalian cell genome can be used, such as the metallothionein promoter, β-actin promoter, human hemoglobin promoter, and human muscle creatinine promoter, or promoters derived from mammalian viruses (such as the adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, cytomegalovirus (CMV) promoter, tk promoter of HSV, promoter of mouse mammary tumor virus (MMTV), LTR promoter of HIV, promoter of moloney virus, promoter of Epstein-Barr Virus (EBV), and promoter of Rous Sarcoma Virus (RSV)). Moreover, the vector usually contains a polyadenylation sequence as a transcription termination sequence. Specifically, the recombinant vector of the present invention contains a CMV promoter.

[0094] The recombinant vector of the present invention can be fused with other sequences to facilitate the purification of the recombinant protein expressed by the recombinant vector. The fusion sequences include, for example, glutathione S-transferase (Pharmacia, USA), maltose-binding protein (NEB, USA), FLAG tag (IBI, USA), 6xHis (hexahistidine) tag (Quiagen, USA), etc. In addition, since Fc is fused to the protein expressed by the vector of the present invention, the expressed protein can be easily purified through a protein A column, etc., without the need for additional sequences for purification.

[0095] Meanwhile, the recombinant vector of the present invention contains antibiotic resistance genes commonly used in the art as a selective marker, and the recombinant vector of the present invention may contain, for example, genes resistant to ampicillin, gentamicin, carbenicillin, chloramphenicol, streptomycin, kanamycin, geneticin, neomycin, and tetracycline.

[0096] Any host cell known in the art can be used as a recombinant cell capable of stably and continuously cloning and expressing the vector of the present invention. Host cells include prokaryotic host cells, for example, strains belonging to the genus Bacillus (such as Escherichia coli, Bacillus subtilis, and Bacillus thuringiensis), Streptomyces, Pseudomonas (such as Pseudomonas putida), Proteus mirabilis, and Staphylococcus (such as Staphylococcus carnosus), but are not limited thereto.

[0097] Eukaryotic host cells suitable for use with this vector include fungi (such as Aspergillus sp.) and yeasts (such as Pichia pastoris, Saccharomyces cerevisiae, Schizosaccharomyces, and Neurospora crassa) and other lower eukaryotic cells, as well as higher eukaryotic cells, such as insect-derived cells and cells derived from plants and mammals.

[0098] Specifically, the host cell may be monkey kidney cells (COS7), NSO cells, SP2 / 0, Chinese hamster ovary (CHO) cells, W138, baby hamster kidney (BHK) cells, MDCK, myeloma cells, HuT 78 cells, or HEK293 cells.

[0099] Compared with animal cells and the like, microorganisms such as Escherichia coli are highly productive, but due to problems such as the formation of disulfide bonds or glycosylation, they are not preferred for protein production. However, microorganisms can be used in production for the purpose of improving the in vivo stability of drugs through pegylation or the like.

[0100] In the present invention, transfection or transformation into a host cell includes any method capable of introducing nucleic acids into an organism, cell, tissue, or organ, and transfection or transformation into a host cell can be carried out by selecting a suitable standard technique according to the type of host cell, as known in the art. These methods include electroporation, protoplast fusion, calcium phosphate precipitation, calcium chloride precipitation, agitation with silicon carbide fiber, and agrobacterium-mediated, PEG-mediated, dextran sulfate-mediated, lipofectamine-mediated, and desiccation / inhibition-mediated transformation, but are not limited thereto.

[0101] The present invention provides a method for producing a recombinant protein using a recombinant cell. Specifically, this method can be a method for producing a human Lefty A fusion protein variant, comprising the following steps: (a) culturing a recombinant cell transformed with the recombinant vector of the present invention; and (b) expressing the recombinant protein in the recombinant cell.

[0102] The culturing steps for producing recombinant proteins can be carried out using suitable culture media and culture conditions known in the art. Those skilled in the art can adjust the culture conditions without difficulty according to the specific strain selected. These culturing methods are disclosed in many documents (such as James M. Lee, Biochemical Engineering, Prentice-Hall International Editions, 138 - 176). The methods for culturing cells can be classified into suspension culture and adherent culture based on the cell growth mode, and can be classified into batch method, fed-batch method, and continuous method according to the culture mode. The culture medium used for culturing should be appropriately selected to meet the conditions required by the specific strain used.

[0103] In animal cells, the culture medium contains carbon sources, nitrogen sources, and trace element components. Examples of carbon sources that can be used in the present invention include carbohydrates, fats, fatty acids, alcohols, and organic acids; carbohydrates, such as glucose, sucrose, lactose, fructose, maltose, starch, and cellulose; fats, such as soybean oil, sunflower oil, castor oil, and coconut oil; fatty acids, such as palmitic acid, stearic acid, and linoleic acid; alcohols, such as glycerol and ethanol; organic acids, such as acetic acid. These carbon sources can be used alone or in combination of two or more.

[0104] Nitrogen sources that can be used in the present invention include, for example, organic nitrogen sources and inorganic nitrogen sources; organic nitrogen sources, such as peptone, yeast extract, gravy, malt extract, corn steep liquor (CSL), and soybean meal powder; inorganic nitrogen sources, such as urea, ammonium sulfide, ammonium chloride, ammonium phosphate, ammonium carbonate, and ammonium nitrate. These nitrogen sources can be used alone or in combination of two or more. The culture medium can contain potassium dihydrogen phosphate, dipotassium hydrogen phosphate, and their corresponding sodium-containing salts as phosphate sources. In addition, metal salts, such as magnesium sulfate or iron sulfate, can be included in the culture medium as sulfate sources. In addition, amino acids, vitamins, and appropriate precursors can be included in the culture medium.

[0105] During the process of culturing cells, compounds such as ammonium hydroxide, potassium hydroxide, ammonia, phosphoric acid, and sulfuric acid can be added to the culture medium in a suitable manner to adjust the pH of the cell culture medium. In addition, during the culturing process, an antifoaming agent such as fatty acid polyglycol ester can be used to inhibit the generation of bubbles. Furthermore, oxygen or an oxygen-containing gas (such as air) is injected into the culture medium to keep the culture containing oxygen. The culturing temperature is generally 20°C to 45°C, preferably 25°C to 40°C.

[0106] The recombinant protein obtained by culturing the transformed recombinant cells can be used without purification, or can be used after being purified to a high purity by various conventional methods, such as dialysis, salt precipitation, and chromatography. Among these methods, chromatography-based methods are most commonly used, and the type and order of chromatography can be selected from ion exchange chromatography, size exclusion chromatography, or affinity chromatography, etc., according to the properties of the recombinant protein, the culturing method, and the like.

[0107] On the other hand, the present invention relates to a composition for preventing and / or treating neuromuscular diseases, comprising a human Lefty A protein variant or a fusion protein.

[0108] On another aspect, the present invention relates to a method for preventing and / or treating neuromuscular diseases, comprising administering a human Lefty A protein variant or a fusion protein comprising a human Lefty A protein variant to a subject.

[0109] On yet another aspect, the present invention relates to the use of a human Lefty A protein variant or a fusion protein comprising a human Lefty A protein variant for the prevention and / or treatment of neuromuscular diseases.

[0110] On still another aspect, the present invention relates to the use of a human Lefty A protein variant or a fusion protein comprising a human Lefty A protein variant for manufacturing a medicament for treating neuromuscular diseases.

[0111] The term "prevention" as used in the present invention refers to any action of inhibiting or delaying the progression of a disease such as a neuromuscular disease by administering the composition according to the present invention, and "treatment" refers to inhibiting the development, alleviating, or eliminating a disease such as a neuromuscular disease.

[0112] In one embodiment of the present invention, it is confirmed that a fusion protein comprising a human Lefty A protein variant according to the present invention can bind to human Nodal and inhibit Nodal-induced signal transduction, thus improving parameters related to peripheral neuropathy. In addition, in one embodiment of the present invention, it is confirmed that the human Lefty A fusion protein variant can inhibit myostatin signal transduction in a dose-dependent manner. Furthermore, it is found that the human Lefty A fusion protein variant can block p38 signal transduction, where p38 is a negative regulator of myelination, and thus can be used as a reagent for treating peripheral neuropathy, especially neuropathy caused by demyelination.

[0113] Therefore, in the present invention, a neuromuscular disease can be a disease occurring in the peripheral nerves or muscles, preferably a disease related to Nodal and / or myostatin signal transduction, but not limited thereto. A disease related to Nodal and / or myostatin signal transduction can be myopathy, peripheral neuropathy, or rigid spine syndrome.

[0114] Myopathy can be selected from the group consisting of sarcopenia, muscular dystrophy, myasthenia gravis, amyotrophic lateral sclerosis (or Lou Gehrig's disease), primary lateral sclerosis, progressive muscular atrophy, Kennedy's disease (or spinobulbar muscular atrophy), spinal muscular atrophy, and distal myopathy.

[0115] The peripheral neuropathy may be selected from the group consisting of Charcot-Marie-Tooth disease, chronic inflammatory demyelinating polyneuropathy, carpal tunnel syndrome, diabetic peripheral neuropathy, and Guillain-Barre syndrome.

[0116] This composition may be a pharmaceutical composition, a quasi-pharmaceutical composition, or a health functional food composition.

[0117] The composition for preventing or treating a disease according to the present invention may comprise a pharmaceutically acceptable carrier.

[0118] The term "pharmaceutically acceptable carrier" used in the present invention refers to a carrier or diluent that does not impair the biological activity and properties of the administered compound and does not stimulate the organism. As the pharmaceutically acceptable carrier in the composition, it can be prepared as a liquid solution using a sterile and biocompatible carrier. The pharmaceutically acceptable carrier may be physiological saline, sterile water, buffered saline, albumin injection solution, glucose solution, maltodextrin solution, glycerol, or a mixture of two or more thereof. In addition, if necessary, the composition of the present invention may contain other known additives, including antioxidants, buffers, and bacteriostatic agents. Furthermore, the composition of the present invention can be prepared in an injectable form, such as an aqueous solution, suspension, or emulsion, with the help of diluents, dispersants, surfactants, binders, and lubricants. In addition, the composition according to the present invention can be prepared into pills, capsules, granules, or tablets.

[0119] The pharmaceutical composition according to the present invention can be formulated into oral or parenteral dosage forms. The pharmaceutical composition according to the present invention can be prepared using common diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants or surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations are prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple methods, lubricants such as magnesium stearate, talc, etc. can also be added. Liquid preparations for oral administration, such as suspensions, internal solutions, emulsions, syrups, etc., may contain simple diluents and various excipients, such as water and liquid paraffin for diluents, and wetting agents, sweeteners, flavoring agents, preservatives, etc. for excipients. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized products and suppositories. Non-aqueous solvents and suspensions can use propylene glycol, polyethylene glycol, vegetable oils such as olive oil or injectable esters such as ethyl oleate. As the base for suppositories, Witepsol, Macrogol, Tween 61, cacao fat, laurin fat, glycerogelatin, etc. can be used.

[0120] Pharmaceutically acceptable carriers and formulations are disclosed in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0121] The pharmaceutical composition of the present invention can be administered orally or parenterally. Parenteral administration can be achieved through intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, intrapulmonary administration, rectal administration, etc. For oral administration, considering that peptides and proteins will be digested in the stomach, the active ingredient in the composition needs to be made into a coated dosage form, or a dosage form that can protect the active ingredient from being digested in the stomach. Alternatively, the composition of the present invention can be administered through any device that can transfer the active ingredient to the target cells.

[0122] The appropriate dosage form of the composition for preventing or treating diseases according to the present invention can vary depending on different factors, such as the preparation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and sensitivity. Therefore, a medical practitioner with ordinary knowledge can easily judge and formulate an effective dose for the target disease to be treated or prevented.

[0123] According to an embodiment of the present invention, the daily dose of the pharmaceutical composition according to the present invention can be from 0.001 milligrams per kilogram (mg / kg) to 100 mg / kg. The term "pharmaceutically effective amount" used herein refers to an amount sufficient to prevent, treat, and diagnose diseases such as neuromuscular diseases.

[0124] The composition for preventing or treating according to the present invention can be formulated using pharmaceutically acceptable carriers and / or excipients according to methods that can be easily achieved by those with ordinary knowledge in the field to which the present invention pertains, to provide a unit dosage form or be contained in a multi-dose vial. Herein, the formulation can be in the form of a solution, suspension, or emulsion in an oil or water medium, or in the form of an extract, grain, suppository, powder, granule, tablet, or capsule, and the formulation can additionally contain dispersants or stabilizers.

[0125] Preferably, the composition of the present invention can be made into an injectable form. In this case, the injectable preparation can be a reconstituted lyophilized formulation or a liquid formulation provided in a ready-to-inject (RTI) form, but is not limited thereto.

[0126] The composition of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents. The composition of the present invention can be administered sequentially or simultaneously with conventional therapeutic agents.

[0127] Examples

[0128] The present invention will be described in more detail below with reference to examples. It will be apparent to those skilled in the art that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention.

[0129] Example 1: Construction of Human Lefty A Fusion Protein

[0130] Example 1-1: Construction of Human Lefty A Protein Variants

[0131] The amino acid sequence of wild-type human Lefty A protein is as follows:

[0132] MWPLWLCWALWVLPLAGPGAALTEEQLLGSLLRQLQLSEV PVLDRADMEKLVIPAHVRAQYVVLLRRSHGDRSRGKRFSQSFREVAGRFLASEASTHLLVFGMEQRLPPNSELVQAVLRLFQEPVPKAALHRHGRLSPRS AQARVTVEWL RVRDDGSNRTSLIDSRLVSV HESGWKAFDV TEAVNFWQQLSRPRQPLLLQVSVQREHLGP LASGAHKLVR FASQGAPAGL GEPQLELHTLDLRDYGAQGD CDPEAPMTEGTRCCRQEMYI DLQGMKWAKNWVLEPPGFLAYECVGTCQQP PEALAFNWPF LGPRQCIASE TASLPMIVSIKEGGRTRPQV VSLPNMRVQK CSCASDGALV PRRLQP(SEQ ID NO:131)

[0133] In this amino acid sequence, M1 to A21 correspond to the signal peptide sequence, L22 to S73 correspond to the propeptide domain, and R74 to R77 and R132 to R135 correspond to the processing sites.

[0134] The wild-type human Lefty A protein before processing is in the 42 kDa form, but is processed into 34 kDa or 28 kDa forms when the region containing the propeptide domain is removed by proprotein convertase. To elucidate the functions of the protein fragments in these three forms, expression vectors for the 42 kDa, 34 kDa, and 28 kDa proteins were constructed.

[0135] For the 42 kDa and 34 kDa forms of the protein, processing site mutations (42: R74G / R77G / R132G, 34: R132G) were introduced during construction to prevent additional cleavage (THE JOURNAL OF BIOLOGICAL CHEMISTRY, Vol.276, No.24, Issue of June 15, pp.21387–21396, 2001). Genes for the 42 fragment (L22 to P366, R74G / R77G / R132G), 34 fragment (F78 to P366, R132G), and 28 fragment (L136 to P366) of human Lefty A were synthesized by Bioneer Co., Ltd. (Korea) and used as PCR templates. Using the primer pair of L1_F (SEQ ID NO:13) and L2_R (SEQ ID NO:14) and the ProFlex system (Applied Biosystems, USA), the amplification process was carried out using Ex taq (Takara, Japan) according to the manufacturer's manual under the following conditions to obtain the 42, 34, and 28 fragments of human Lefty A protein. Conditions: 22 cycles, each cycle consisting of 60 seconds of denaturation at 95°C, 60 seconds of primer annealing at 58°C, and 60 seconds of extension at 72°C.

[0136] Example 1-2: Construction of carboxyl-terminal Fc fusion proteins

[0137] Three expression vectors for carboxyl-terminal Fc fusion proteins (42Fc (SEQ ID NO:1), 34Fc (SEQ ID NO:2), and 28Fc (SEQ ID NO:3)) were constructed by linking human IgG1 Fc to the human Lefty A protein variants of Example 1. 28Fc, 34Fc, and 42Fc refer to Fc fused to the carboxyl terminus of each 28 kDa, 34 kDa, and 42 kDa Lefty A protein.

[0138] Using DNA encoding the human IgG1 sequence (Uniprot: P01857) as a template and the primer pair of L3_F (SEQ ID NO: 15) and L4_R (SEQ ID NO: 16), amplification was carried out under the same conditions as the PCR amplification conditions of the human Lefty A fragment to obtain the human IgG1 fragment. The obtained PCR product was separated by 1.5% agarose gel electrophoresis, and then an assembly PCR reaction was carried out under the same conditions. The obtained reaction product was separated and purified by 1.5% agarose gel electrophoresis, and then digested with the restriction enzymes Hind III and Xho I (NEB, USA). Each digested product was ligated with the pCLS05 vector digested with the restriction enzymes Hind III and Xho I (Korean Patent Application No. 2011-0056685) at 25°C for 60 minutes, and then the ligation product was transformed into Escherichia coli DH5α. The transformed cells were cultured overnight in LB medium containing 100 micrograms / milliliter (μg / ml) of ampicillin, and the plasmid was extracted from the resulting colonies and then sequenced by the service of Cosmo Genetech Co., Ltd. (Korea), and then it was determined that three carboxyl-terminal Fc fusion protein expression vectors were constructed.

[0139] Example 1-3: Construction of carboxyl-terminal linker Fc fusion proteins

[0140] Three expression vectors of carboxyl-terminal linker Fc fusion proteins (42LFc (SEQ ID NO: 4), 34LFc (SEQ ID NO: 5), and 28LFc (SEQ ID NO: 6)) were constructed by linking human IgG1 Fc to human Lefty A through the SGGGGSGGGGSGGGGS linker (SEQ ID NO: 130). 28LFc, 34LFc, and 42LFc refer to Fc fused to the carboxyl terminus of each 28 kDa, 34 kDa, and 42 kDa Lefty A protein through the linker.

[0141] The method for obtaining the fragments of human Lefty A protein at positions 42, 34, and 28 can be the same as the method described for constructing the carboxyl-terminal Fc fusion protein expression vector. For the linker Fc fragment, DNA encoding the human IgG1 sequence (Uniprot: P01857) is used as a template, and the primer pair of L5_F (SEQ ID NO: 17) and L4_R (SEQ ID NO: 16) is used for PCR amplification under the same conditions to obtain the human linker IgG1 fragment. The subsequent procedures are the same as those described above for the construction of the carboxyl-terminal Fc fusion protein.

[0142] Example 1-4: Construction of Amino-Terminal Fc Fusion Proteins

[0143] Construct three expression vectors for amino-terminal Fc fusion proteins (Fc 42 (SEQ ID NO: 7), Fc 34 (SEQ ID NO: 8), and Fc 28 (SEQ ID NO: 9)). Fc 28, Fc 34, and Fc 42 refer to Fc fused to the amino-terminals of the 28 kDa, 34 kDa, and 42 kDa Lefty A proteins, respectively.

[0144] Using the primer pairs of L6_F (SEQ ID NO: 18), L7_F (SEQ ID NO: 19), L8_F (SEQ ID NO: 20), and L9_R (SEQ ID NO: 21), the fragments of human Lefty A protein at positions 42, 34, and 28 are obtained. Each of the obtained reaction products is separated and purified by 1.5% agarose gel electrophoresis, and then using the HD Cloning Kit (Clontech, 639650) at 50 °C for 15 minutes, each of the obtained reaction products is ligated to the pCLS05 vector encoding human IgG1 Fc digested with the restriction enzyme Xho I. The subsequent procedures are the same as those described above for the construction of the carboxyl-terminal Fc fusion protein.

[0145] Example 1-5: Construction of Carboxyl-Terminal HSA Fusion Proteins

[0146] When expressed in animal cells, human IgG1 Fc fusion proteins form homodimers. To this end, to construct monomeric fusion proteins, proteins fused with human serum albumin (HSA) were constructed. Three expression vectors of carboxyl-terminal HSA fusion proteins (42HSA (SEQ ID NO:10), 34HSA (SEQ ID NO:11), and 28HSA (SEQ ID NO:12)) were constructed. 28HSA, 34HSA, and 42HSA refer to HSA fused to the carboxyl terminus of each 28 kDa, 34 kDa, and 42 kDa Lefty A protein.

[0147] Using the primer pair of L1_F (SEQ ID NO:13) and L9_R (SEQ ID NO:21), 42, 34, and 28 fragments of the Lefty A protein were obtained in the same manner as in Example 1. Each fragment was cut with restriction enzymes Hind III and Xho I and ligated to the pCLS05 vector cut with restriction enzymes Hind III and Xho I at 25 °C for 60 minutes. The ligation product was transformed into Escherichia coli DH5α. The transformed cells were cultured overnight in LB medium containing 100 micrograms per milliliter (μg / ml) of ampicillin, and plasmids were extracted from the resulting colonies and then sequenced to determine the first-step cloning.

[0148] In the second-step cloning, using the human serum albumin gene synthesized by Bioneer as a template and the primer pair of L10_F (SEQ ID NO:22) and L11_R (SEQ ID NO:23), a fragment containing human serum albumin was obtained under the PCR conditions used in Example 1. The obtained PCR product was separated and purified by 1.5% agarose gel electrophoresis, and then the HD Cloning Kit (Clontech, 639650) was used to react at 50 °C for 15 minutes to ligate the obtained PCR product with the DNA cut with the restriction enzyme Xho I (encoding the 42, 34, and 28 fragments of the human Lefty A protein obtained in the first-step cloning). The subsequent procedures were the same as those for the construction of the carboxyl-terminal Fc fusion protein described above.

[0149] Schematic diagrams of the expression vectors constructed in Examples 1-1 to 1-5 are shown in Figure 1 , and their amino acid sequences are shown in Table 1 below, and the primers used for the construction of the fusion proteins are shown in Table 2 below.

[0150] [Table 1] Amino acid sequences of human Lefty A fusion proteins

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159] [Table 2] Primers Used for the Construction of Human Lefty A Fusion Protein

[0160]

[0161]

[0162] Examples 1 - 6: Transient Expression, Purification and Analysis of Human Lefty A Fusion Protein

[0163] The human Lefty A and its variant proteins were expressed using the FreestyleTM MAX CHO Expression system. Specifically, FreestyleTM CHO-S cells (Invitrogen, USA) were transfected with the expression vector as described above, and the cells were grown in CHO serum-free medium (Invitrogen, USA) for 5 days. The medium was centrifuged, the supernatant was recovered and filtered, and then used later for the analysis of the protein expression level and purification of Lefty A in the medium.

[0164] The fusion protein from the culture was purified by Protein A-based affinity chromatography using MabSelect Protein A resin (GE Healthcare, USA). The column packed with Protein A resin was equilibrated with phosphate-buffered saline (PBS, pH 7.4), and then the filtered cell culture supernatant was loaded onto the column. After washing with 10 column volumes (CV) of PBS, the protein was eluted with 5 column volumes of elution buffer (0.1 M Sodium Citrate), and the eluate was neutralized by adding 200 μl of 1 M Tris-HCl (pH 8.0). To exchange the buffer of the eluted recombinant protein aliquot with PBS (pH 7.4) buffer, an Amicon Ultra-15 Centrifugal filter (MWCO: 30000) (Millipore, Cat. No. UFC903096) was used. The purified protein aliquot mixed with the sample was placed and centrifuged at 4,000 rpm for 10 minutes at 4 °C, diluted 10-fold, and then centrifuged three more times to achieve a buffer exchange of 1,000-fold or more. The purified recombinant protein was filtered through a 25 mm PES syringe filter (0.22 μm; Nalgene, Cat. No. NAL-194-2520) in a clean bench while minimizing sample loss, then sealed and stored at 4 °C without exposure to the outside.

[0165] Purify the HSA fusion protein using the CaptureSelect™ HSA affinity matrix (ThermoFisher Scientific, USA) or the HiTrap Blue HP resin (HiTrap Blue HP resin) that specifically binds to HSA. Pass the filtered culture supernatant through a CaptureSelect affinity column filled with PBS, wash it with 20 mM Tris-Cl containing 1 M sodium chloride, and then elute it with 20 mM Tris-Cl containing 1 M arginine and 1 M sodium chloride. Alternatively, pass the prepared culture supernatant through a HiTrap Blue HP column filled with 20 mM sodium phosphate (pH 7.0) as the binding buffer so that only the desired protein binds to the resin, and then elute the protein with 20 mM sodium phosphate (pH 7.0) and 2 M sodium chloride. After completion of the purification, measure the protein concentration of each fraction at a wavelength of 280 nanometers (nm) using a NANODROP 2000 spectrophotometer (Thermo Scientific, Cat. No. ND-2000). To measure whether each fraction contains the desired protein, use a 1200 series HPLC (1200 series HPLC) (Agilent Technologies) with a SWxl Guard column (TOSOH, Cat. No. 08543) and a TSKgel G3000SWxl (TOSOH, Cat. No. 08541) as size exclusion HPLC.

[0166] Analyze each purified fusion protein by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography (SEC). Perform SDS-PAGE on a 4-12% acrylamide gel of Tris / glycine (Tris / glycine 4-12% acrylamide gel) (Invitrogen, USA). Stain the gel used for analysis with Coomassie blue and image it by digital scanning. Analyze each purified protein by size exclusion chromatography (SEC) using a TSK-GEL G300 SWXL column (7.8 x 300 mm, Tosohaas, USA) equilibrated with phosphate buffer containing 0.02% sodium azide (NaN3) at a flow rate of 0.5 ml / min to determine the protein expression level and purity. The results are shown in Table 3 below.

[0167] [Table 3] Purification Yield and Purity of Human Lefty A Fusion Protein

[0168] Protein Name Yield (μg / L) Purity SEC (%) 42Fc 63.8 45.9 34Fc Not Expressed - 28Fc Not Expressed - 42LFc 125 46.2 34LFc Not Expressed - 28LFc Not Expressed - Fc 42 7.2 - Fc 34 Not Expressed - Fc 28 Not Expressed -

[0169] Regardless of the Fc fusion site, neither the 34- or 28-Fc fusion protein was expressed. In the case of the "42" fragment, carboxyl-terminal fusion ("42Fc") had a significantly higher protein expression efficiency compared to amino-terminal fusion ("Fc42"), especially in the case of linker Fc fusion ("42LFc"), where the protein expression efficiency was greatly enhanced. Similarly, in the case of the HSA fusion protein, only 42HSA was expressed, while 34HSA or 28HSA was not expressed. This was confirmed by Western blot analysis of the culture.

[0170] Based on the expression data and in vivo efficacy described in Examples 1-6, 42Fc or 42LFc will be used as a template for variant construction later ( Figure 2 ).

[0171] Example 2: Construction of Variants of Human Lefty A Fusion Protein with Improved Productivity and Stability

[0172] Based on the expression vector described in Example 1, a Lefty A fusion protein variant with improved protein stability and expression in animal cells compared to 42LFc was constructed.

[0173] Example 2-1: Construction of the Expression Vector for Human Lefty A Fusion Protein Variant

[0174] Most of the fusion proteins with the pre-peptide removed constructed in Example 1 were not expressed. Even the 42LFc protein was poorly expressed in host cells, with poor purity and low yield of the purified protein. As Figure 3 shown, after affinity purification of the 42LFc protein, additional smaller bands appeared on the SDS-PAGE gel, which may be due to cleavage at the processing site or non-specific cleavage caused by protein instability ( Figure 3)。In addition, the ExPASy PeptideCutter DB (https: / / web.expasy.org / peptide_cutter / ) was used to predict the possible cleavage sites cleaved by various enzymes and chemicals, and it was predicted that the leucine residues at positions 311 and 359 might be cleaved by thrombin. Therefore, in order to increase the expression level, stability, and purification purity of the human Lefty A fusion protein variant, mutations were introduced into the propeptide domain, processing sites, and positions L311, P313, R314, L359, P361, and R362. Using the 42LFc gene as a template and the primer pairs shown in Table 4 below, each fragment was obtained by PCR under the same conditions as described in Example 1, and then each fragment was subjected to an assembly PCR reaction. Each obtained reaction product was separated and purified by 1.5% agarose gel electrophoresis, and then the HD Cloning Kit (Clontech, 639650) was reacted at 50 °C for 15 minutes, and each obtained reaction product was ligated to the pCLS05 vector digested with restriction enzymes Hind III and Xho I. The subsequent procedures were the same as those for the construction of the carboxyl-terminal Fc fusion protein described above.

[0175] [Table 4] Primers used for the construction of human Lefty A variants

[0176]

[0177]

[0178]

[0179]

[0180] The amino acid sequences of the variants are shown in Table 5 below, and substitutions of amino acid residues in the sequence of 42LFc in the propeptide domain (L22 to S73) occurred in the variants.

[0181] [Table 5] Amino acid sequences of human Lefty A fusion protein variants (propeptide domain variants)

[0182]

[0183]

[0184]

[0185]

[0186] LFB 42LFc is a variant in which amino acid residue substitutions of E24G, S38K, V42T, K50E, A55T, V63A and R66Q occur in the sequence of 42LFc.

[0187] The amino acid sequence of the variant is shown in Table 6 below, in which amino acid residue substitutions occur at the processing sites (R74 to R77 and R132 to R135) in the sequence of 42LFc.

[0188] [Table 6] Amino acid sequence of human Lefty A fusion protein variant (processing site variant)

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198] 42LFc V1 is a variant in which there is an amino acid residue substitution of G132R in the sequence of 42LFc; 42LFc V2 is a variant in which there are amino acid residue substitutions of G132R and R135G in the sequence of 42LFc; 42LFc V3 is a variant in which there are amino acid residue substitutions of G132R and G134E in the sequence of 42LFc. 42LFc V4 is a variant in which there is an amino acid residue substitution of R135G in the sequence of 42LFc; 42LFc V5 is a variant in which there are amino acid residue substitutions of G74R and G77A in the sequence of 42LFc; 42LFc V6 is a variant in which there are amino acid residue substitutions of G74R, G77A, G132R and R135A in the sequence of 42LFc; 42LFc V7 is a variant in which there are amino acid residue substitutions of G74R, G77A, G132R and R135V in the sequence of 42LFc; 42LFc V8 is a variant in which there are amino acid residue substitutions of G74R and G77V in the sequence of 42LFc; 42LFc V9 is a variant in which there are amino acid residue substitutions of G74R, G77V, G132R and R135A in the sequence of 42LFc; 42LFc V10 is a variant in which there are amino acid residue substitutions of G74R, G77V, G132R and R135V in the sequence of 42LFc; 42LFc V11 is a variant in which there are amino acid residue substitutions of G74R, G75H, K76G, G132R and R135G in the sequence of 42LFc; 42LFc V12 is a variant in which there are amino acid residue substitutions of G74R, G132R and R135G in the sequence of 42LFc; 42LFc V13 is a variant in which there are amino acid residue substitutions of G74R, G77A, G132R and R135G in the sequence of 42LFc.

[0199] Table 7 below reveals the amino acid sequences of the variants in which the following amino acid residue substitutions occur: substitutions of amino acid residues in the propeptide domain (L22 to S73) as shown in Table 5 above; substitutions of amino acid residues at the processing sites (R74 to R77 and R132 to R135) as shown in Table 6 above; and additional substitutions of amino acid residues at the cleavage sites (S202 or S223).

[0200] [Table 7] Amino acid sequences of Lefty A fusion protein variants (combinations)

[0201]

[0202]

[0203]

[0204]

[0205]

[0206] CX196 is a variant in which amino acid residue substitutions of E24G, S38K, V42T, K50E, A55T, V63A, R66Q, G132R, and R135G occur in the sequence of 42LFc; CX197 is a variant in which amino acid residue substitutions of E24G, S38K, V42T, K50E, A55T, V63A, R66Q, G132R, and R135A occur in the sequence of 42LFc.

[0207] CX201 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, and R135A occur in the sequence of 42LFc; CX203 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, and G134E occur in the sequence of 42LFc.

[0208] CX206 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, R135G, and S202T occur in the sequence of 42LFc; CX207 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, R135G, and S223G occur in the sequence of 42LFc; CX208 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, R135G, S202T, and S223G occur in the sequence of 42LFc.

[0209] Table 8 below reveals the amino acid sequences of the variants in which amino acid residue substitutions occur at the thrombin cleavage site (L311, P313, R314, L359, P361, or R362) in the sequence of CX201, and CX201 is as shown in Table 7 above.

[0210] [Table 8] Amino Acid Sequences of Human Lefty A Fusion Protein Variants (Thrombin Cleavage Site)

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223] For the thrombin cleavage site, the antibody-derived MDMRVPAQLLGLLLLWFPGSRC (UniProt: A0A0C4DH73; SEQ ID NO: 132) sequence was used as the signal sequence.

[0224] Example 2-2: Transient Expression and Purification

[0225] The constructed expression vector was transfected, and the protein was expressed and purified in the same manner as in Example 1, and then the expression level and purity of the protein were analyzed. The results are shown in Tables 9 to 12 below.

[0226] After the first affinity purification, the 42-LFc fusion protein used as the basis for variant construction showed an expression level of approximately 2.8 μg / ml and an average protein purity of approximately 64.3% as measured by SE-HPLC. The Lefty A variant obtained by replacing the propeptide domain with LFB showed an approximately 7.5-fold increase in expression and an average protein purity of approximately 83% as measured by SE-HPLC. The substitution of V63A in the propeptide domain of human Lefty A increased the expression level by approximately 2-fold, and the double substitution of V63A / R66Q increased the expression level by approximately 2.3-fold (Table 9).

[0227] [Table 9] Expression Levels and Purities of Human Lefty A Fusion Protein Variants (Propeptide Domain Variants)

[0228]

[0229]

[0230] Sequence mutations were introduced at the processing site. As a result, the protein purity of 42LFc V2 measured by SE-HPLC was 84.1%, and the expression level of 42LFc V2 was similar to that before the introduction of the mutation. 42LFc V6 showed the highest protein purity of 90.9%, but the expression level of 42LFc V6 decreased to about 60% compared to that before the introduction of the mutation (Table 10).

[0231] [Table 10] Expression levels and purities of human Lefty A fusion protein variants (processing site variants)

[0232] Analysis of human Lefty A fusion protein variants (combinations) showed that CX196 obtained using the LFB propeptide showed an approximately 1.8-fold increase in expression level and an increase in protein purity to 94.1%; CX201 obtained using the human propeptide showed an approximately 1.9-fold increase in expression level and an increase in protein purity to 91.3% (Table 11 and Figure 4 ).

[0233] [Table 11] Expression levels and purities of human Lefty A fusion protein variants (combinations)

[0234]

[0235] Analysis of human Lefty A fusion protein variants (thrombin cleavage site) showed that a single mutation of P361S showed a 1.5-fold increase in expression level. Double mutations of L311D / R362Q, L311E / R362Q, triple mutations of L311D / P361D / R362Q and L311E / P361D / R362Q all showed a protein purity of approximately 95% (corresponding to an approximately 9% increase) and a 1.5-fold increase in expression level (Table 12).

[0236] [Table 12] Expression levels and purities of human Lefty A fusion protein variants (thrombin cleavage site)

[0237]

[0238] Example 3: Improvement in stability in serum

[0239] Using the human Lefty A fusion protein variant (thrombin cleavage site) constructed in Example 2, an in vitro serum stability test was conducted. Each protein variant was diluted in mouse serum or plasma to a final concentration of 10 μg / ml, incubated at 37 °C for 4 hours, and then analyzed by sandwich enzyme-linked immunosorbent assay (Sandwich ELISA). The stability in mouse serum was analyzed by measuring the relative remaining amount of each human Lefty A variant in serum under in vitro conditions. The relative stability of each variant relative to CX201 is shown in Table 13 below. It was confirmed that when putative thrombin cleavage site mutations such as L311D, L311E, P361D, or R362Q occurred, the stability of the variant in serum was enhanced.

[0240] [Table 13] Relative stability in serum after incubation at 37 °C for 4 hours

[0241]

[0242]

[0243] Example 4: Measurement of the binding affinity for Nodal

[0244] The binding affinity between human Nodal protein and human Lefty A fusion protein variants was measured using BIAcore.

[0245] Specifically, 50 RU of human Nodal (R&D systems, 3218-ND-025 / CF) protein was immobilized on a CM5 sensor chip. The human Lefty A fusion protein and its variants were diluted to concentrations of 500, 250, 125, 62.5, and 31.25 nM, and injected sequentially starting from the lower concentration. Then, each dilution was allowed to associate for 3 minutes at a flow rate of 30 μl / min, and each dilution was dissociated for 5 minutes using a running buffer. The chip was regenerated using 15 μl of 50 mM sodium hydroxide. The association and dissociation of each cycle were evaluated using the "Bivalent analyte" mode of BIAevaluation software version 4.1, and the BIAcore data is summarized in Table 14 below.

[0246] [Table 14] Affinity of Lefty A fusion protein and its variants for human Nodal

[0247] Example 5: Prevention of apoptosis of Schwann cells induced by endoplasmic reticulum stress by human Lefty A protein

[0248] In this study, to identify substances for treating hereditary peripheral neuropathy, a Schwann cell model was developed using an endoplasmic reticulum stress-inducing drug, and the Schwann cell model was used for drug identification.

[0249] First, when mesenchymal stem cells were co-cultured with the endoplasmic reticulum stress cell model, cell death was effectively inhibited and endoplasmic reticulum stress was reduced. Apoptosis of Schwann cells induced by endoplasmic reticulum stress could be prevented when co-cultured with mesenchymal stem cells. Specifically, thapsigargin-induced endoplasmic reticulum stress was induced in S16 Schwann cells in a transwell chamber, and then umbilical cord-derived mesenchymal stem cells were co-cultured in the upper chamber, and the death of S16 cells was analyzed. At this time, co-culture of umbilical cord-derived mesenchymal stem cells inhibited the death of S16 Schwann cells by about 30% or more ( Figure 5 ).

[0250] From this result, it can be inferred that paracrine factors secreted by umbilical cord stem cells inhibit the death of Schwann cells. Furthermore, a cytokine antibody array was performed to identify therapeutic proteins secreted from mesenchymal stem cells. In the case where Schwann cells were increased by 2-fold or more, proteins secreted from umbilical cord stem cells during co-culture with Schwann cells were analyzed. In particular, the secretion of Lefty A protein was effectively increased.

[0251] Example 6: Improvement of nerve function by human Lefty A fusion protein in Tr-J mice

[0252] To test whether human Lefty A fusion protein improves nerve function, electrophysiological studies were performed using Trembler-J (Tr-J) mice, an animal model for CMT1 (Meekins et al., J Peripher Nerv Syst 9(3):177-82(2004)). Tr-J mice are spontaneously mutated mice with an L16P mutation in the PMP22 gene. Tr-J mice have an expression pattern of peripheral nerve demyelination and reduced nerve conduction velocity and compound muscle action potential (CMAP) expression, as seen in CMT1 patients (Henry et al., J Neuropathol Exp Neurol 2(6):688-706(1983); Valentijin et al., Nat Genet 2(4):288-911992(1992)).

[0253] From postnatal day 6 (p6) to postnatal day 20 (p20), Tr-J mice were intraperitoneally injected with PBS or human Lefty A fusion protein (1 μg / kg) every two days for a total of 8 injections. One day before the nerve conduction study, the hair covering the hindlimbs was removed and depilated. The next day, the mice were anesthetized, an active recording needle electrode was placed in the gastrocnemius muscle, and a reference electrode was placed under the recording electrode to evaluate nerve conduction in the sciatic nerve of the mice. The stimulus cathode was placed in the hip region 6 mm from the recording electrode, and the amplitude of the compound muscle action potential (CMAP) and the motor nerve conduction velocity (MNCV) were measured using a Nicolet VikingQuest (Natus Medical, San Carlos, CA) device.

[0254] As Figure 6 and Table 15 below show, the nerve conduction study showed that the 42LFc fusion protein improved nerve function in Tr-J mice.

[0255] [Table 15] Electrophysiological effects of Lefty A fusion protein on nerve conduction in Tr-J mice

[0256] Drug Distal CMAP (mV) Proximal CMAP (mV) MNCV (m / s) PBS 13.7±1.1 11.8±1.2 13.9±0.9 42-LFc <![CDATA[16.9±1.1 * > <![CDATA[15.6±1.1 * > 16.2±1.7 42-HSA 11.6±0.8 10.1±0.7 14.7±0.8

[0257] *p < 0.05

[0258] At a dose of 1 μg / kg, compared with PBS or HSA fusion protein, the LFc-fused human Lefty A protein significantly increased the compound muscle action potential (CMAP) and motor nerve conduction velocity (MNCV) in Tr-J mice. These data indicate that the human Lefty A fusion protein can be used to improve the nerve function of neuropathy patients.

[0259] Example 7: Promotion of Schwann cell myelination by human Lefty A fusion protein variants

[0260] Treatment of Lefty A fusion protein variants (42LFc; X-42) in RT4-D6P2T Schwann cells resulted in increased expression of Krox20 and myelin basic protein (MBP) Figure 7 A), where Krox20 is an important transcriptional regulator of Schwann cell myelination and myelin basic protein is induced during the process of myelination.

[0261] On the other hand, treatment of Nodal in RT4-D6P2T Schwann cells resulted in decreased expression of Krox20, and the inhibition of myelination caused by Nodal could be restored by Lefty A fusion protein variants Figure 7 B and Figure 7 C). These results indicate that Lefty A fusion protein variants can promote Schwann cell myelination, which can be achieved by inhibiting Nodal, a negative regulator of Schwann cell myelination.

[0262] Example 8: Improvement of nerve and motor function by intraperitoneal injection (IP) of human Lefty A fusion protein variant (X-42) in C22 mice (p6 mice)

[0263] C22 [Strain origin: (C57BL / 6J x CBA / CA)F1] mice are transgenic mice carrying 7 copies of the human PMP22 gene and having an expression pattern of severe peripheral nerve demyelination. C22 mice have been widely studied as an animal model of Charcot-Marie-Tooth disease type 1A (CMT1A) (Robertson et al., J Anat 200(4):377-90 (2002); Norreel et al., Neuroscience 116(3):695-703 (2003)). From postnatal day 6 (p6) to postnatal day 24 (p24), a dose of 10 μg / kg of the human Lefty A fusion protein variant (42LFc; X-42) was intraperitoneally administered to the mice every two days for a total of 10 times to evaluate nerve conduction and muscle movement expression.

[0264] Example 8-1: Nerve Conduction Study

[0265] One day before the nerve conduction study, the hair covering the hindlimbs was removed and depilated. The next day, the mice were anesthetized, an active recording needle electrode was placed in the gastrocnemius muscle, and a reference electrode was placed under the recording electrode to evaluate nerve conduction in the sciatic nerve of the mice. The stimulus cathode was placed in the hip region 6 mm away from the recording electrode, and a Nicolet VikingQuest (Natus Medical, San Carlos, CA) device was used to measure the amplitude of the compound muscle action potential (CMAP) and the motor nerve conduction velocity (MNCV). Compared with wild-type mice, C22 mice showed a significant decrease in motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP). On the other hand, compared with the vehicle-administered group, C22 mice administered with the human Lefty A fusion protein variant showed a significant increase in motor nerve transmission velocity (MNCV) and compound muscle action potential (CMAP) ( Figure 8 ).

[0266] Example 8-2: Behavioral Analysis

[0267] To evaluate motor function, a rotarod test and a hindlimb grip strength analysis were performed.

[0268] Specifically, the rotarod test was performed by placing the mice on a 3-cm horizontal rotating rod (2 m / min) and then measuring the retention time. The mice were trained for 3 days before the test and tested four times a week. The latency to fall from the rotarod was recorded, with a maximum limit of 5 minutes. As Figure 9 shown, the locomotor performance of C22 mice was much lower than that of wild-type (WT) mice. In the rotarod test, administration of the human Lefty A fusion protein variant significantly improved the locomotor ability of C22 mice ( Figure 9 A).

[0269] The hindlimb grip strength of 3- to 4-week-old C22 mice was evaluated using a grip strength meter. The mice were placed with their backs to the meter and their hindlimbs on an angled mesh assembly. Once the mice grasped the metal grid or triangular bar with their hind toes, they were pulled directly by the tail towards the meter at a fixed speed. The peak force was recorded in grams by the device. Compared with wild-type mice, C22 mice had reduced hindlimb grip strength, and treatment with the human Lefty A protein variant showed a significant therapeutic effect on grip strength ( Figure 9 B). The hindlimb grip strength of C22 mice administered the human Lefty A fusion protein variant increased approximately 3-fold ( Figure 9 B).

[0270] Example 9: Intraperitoneal injection (IP) of the human Lefty A fusion protein variant (X-42) increased muscle mass in C22 mice (p6 mice)

[0271] Six-day-old (p6) wild-type (WT) or C22 mice were used to test for changes in muscle mass caused by the human Lefty A fusion protein variant. From postnatal day 6 (p6) to postnatal day 24 (p24), PBS or the human Lefty A fusion protein variant (10 μg / kg) was administered intraperitoneally to the mice every two days for a total of 10 times, and then the gastrocnemius region was measured using magnetic resonance imaging. Administration of the human Lefty A fusion protein variant showed a significant increase in muscle mass in CMT1 model mice as well as wild-type mice ( Figure 10 ).

[0272] Example 10: Improvement of gait in C22 mice (p6 mice) by intraperitoneal injection (IP) of human Lefty A fusion protein variant (X-42)

[0273] To elucidate the effect of the Lefty A fusion protein variant on the gait of C22 mice, PBS or the human Lefty A fusion protein variant (10 μg / kg) was intraperitoneally administered to 6-day-old mice, 10 times in total (once every two days). After allowing the mice to pass through a restricted path, the gait was monitored, and the hip, knee, and ankle joint angles, as well as two parameters (stride length and base of support (BOS)), were analyzed ( Figure 11 ). The stride length was calculated by tracking the right and left hind paws, and the BOS value was determined by calculating the left and right of the stride length. As a result, an improvement in the pelvic stride length was observed in the group of mice administered the Lefty A fusion protein variant ( Figure 11 right figure).

[0274] Compared to wild-type mice, C22 mice showed significant differences in the hip, knee, and ankle joint angles due to gait abnormalities. However, when the Lefty A fusion protein was administered to C22 mice, the change in angle in C22 mice decreased ( Figure 11 lower left figure). This result indicates that neuromuscular function can be improved to a certain extent, but not to the level of wild-type mice.

[0275] Example 11: Improvement of nerve and motor function in C22 mice (p6 mice) by intraperitoneal injection (IP) of human Lefty A fusion protein variant (CX201)

[0276] C22 mice were intraperitoneally injected with the human Lefty A fusion protein variant CX201 at a dose of 0.2 mg / kg once every two days for 10 times. The injection started as early as on the 6th day after birth, even when myelination has not yet started under normal circumstances. Then, as described in Example 8 above, nerve conduction studies and behavioral analyses were performed.

[0277] Compared to the vehicle-administered group, C22 mice injected with the human Lefty A fusion protein variant CX201 showed a significant increase in the motor nerve conduction velocity (MNCV) and compound muscle action potential (CMAP) ( Figure 12 ).

[0278] In addition, as Figure 13As shown, compared with the vehicle control group, C22 mice injected with CX201 showed statistically significant improvement in hindlimb grip strength and motor performance on the rotarod.

[0279] Example 12: Effect of Intraperitoneal Injection (IP) of Human Lefty A Fusion Protein Variant (CX201) on Gait in C22 Mice (p6 Mice)

[0280] To elucidate the effect of Lefty A fusion protein variant CX201 on the gait of C22 mice, PBS or human Lefty A fusion protein variant (200 μg / kg) was intraperitoneally administered to 6-day-old mice 10 times (once every two days), and then gait analysis was performed as described in Example 10.

[0281] In particular, analysis of the step length from one heel to the next showed that the gait of C22 mice administered with the Lefty A fusion protein variant was significantly improved ( Figure 14 ).

[0282] Example 13: Improvement of Neural and Motor Functions by Subcutaneous Injection of Human Lefty A Fusion Protein Variant in C22 Mice (p21 Mice)

[0283] On postnatal day 21 (p21), when myelination development mainly occurs, human Lefty A fusion protein variant CX201 was subcutaneously injected into C22 mice, and then nerve conduction studies and behavioral analysis were performed as described in Example 8 above.

[0284] Compared with the group administered with the vehicle, C22 mice administered with CX201 for 4 weeks showed a significant increase in both motor nerve conduction velocity and compound muscle action potential ( Figure 15 ). In particular, the hindlimb grip strength of male mice increased, with statistical significance ( Figure 16 ). These data show that administration of the fusion protein variant CX201 in the late stage of myelination differentiation and early postpartum can improve neural and motor functions.

[0285] Example 14: Inhibition of Myostatin Signaling by Human Lefty A Fusion Protein Variant

[0286] Use a reporter gene to evaluate whether the human Lefty A fusion protein variant can inhibit myostatin signaling. Transfect rhabdomyosarcoma A204 cells with the Smad2 / 3-reactive luciferase reporter vector pGL4.48 (Promega, USA), and then screen through antibiotics to select cells stably transfected with the vector. Pre-incubate myostatin with increasing concentrations of the human Lefty A fusion protein variant for 45 minutes. After adding the medium, culture the cells for 6 hours, and then detect the luciferase activity induced by myostatin using the bio-glo luciferase assay reagent (Promega, USA). Myostatin induces strong luciferase expression in the cell line stably transfected with the vector, and the human Lefty A fusion protein variant inhibits myostatin signaling in a dose-dependent manner ( Figure 17 ).

[0287] Example 15: Inhibition of p38 Signaling by the Human Lefty A Fusion Protein Variant

[0288] It is known that the TGF-β family member BMP7 inhibits the expression of myelin genes and delays peripheral myelination through the phosphorylation of p38 (Liu X et al., Sci Rep 6:31049 (2016)). Therefore, investigate whether the human Lefty A fusion protein variant can inhibit p38 phosphorylation. Specifically, as described in Example 7, administer the human Lefty A fusion protein variant to C22 mice, and analyze phosphorylated p38 in the sciatic nerve using Western blotting.

[0289] As Figure 18 shown, in the sciatic nerve of C22 mice administered with the human Lefty A fusion protein variant, the phosphorylation of p38 is inhibited, and the expression of the MBP protein increases. These data indicate that the human Lefty A fusion protein variant can promote myelination by blocking p38 signaling of the negative regulator of myelination. Therefore, the human Lefty A fusion protein can be used as a reagent for treating peripheral neuropathy, such as neurodegenerative diseases caused by demyelination.

[0290] Example 16: Construction of a Cell Line for Producing the Human Lefty A Fusion Protein Variant

[0291] In a 125 mL conical flask, CHO-S (cGMP-banked) cells were inoculated into 30 mL of CD-FortiCHO medium at a density of 1 x 10 6 cells / mL. 50 μg of an expression vector inserted with the gene of the human Lefty A fusion protein variant (CX201; containing the antibody-derived MDMRVPAQLLGLLLLWFPGSRC sequence as a signal sequence in the CX201 fusion protein; human Lefty A was linked to human IgG1 Fc through the SGGGGSGGGGSGGGGS linker; SEQ ID NO: 133 as shown in Table 16) was placed into a 50 mL conical tube, OptiPRO SFM was added to a final volume of 1.5 mL, and then vortexed. Freestyle MAX solution was added to the DNA solution, and then the solution was left standing at room temperature for 10 minutes. After 10 minutes, the conical flask containing the cells was treated with the DNA-Freestyle MAX reagent (DNA-Freestyle MAX reagent complex) to transform the cells.

[0292] The first screening and the second screening were sequentially performed to induce gene amplification. CX201 of the production cell line was selected from the pool showing the highest expression level among the pools obtained after the first and second screenings using ClonePix. The selected clone was seed-cultured for 6 days and then suspended in the cryopreservation medium (90% CD-FortiCHO medium + 10% DMSO) to a concentration of 1.0 x 10 7 cells / mL, and 1 mL of the suspension was dispensed into each cryotube to prepare the research cell bank (RCD).

[0293] [Table 16] Amino acid sequence of the human Lefty A fusion protein variant

[0294]

[0295] CX201 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, and R135A occur in the sequence of 42LFc; CX203 is a variant in which amino acid residue substitutions of V63A, R66Q, G132R, and G134E occur in the sequence of 42LFc

[0296] Example 17: Improvement of Neuromotor Function by Subcutaneous Injection of Human Lefty A Fusion Protein Variants in C22 Mice (p35 Mice)

[0297] Five-week-old C22 mice (p35) undergoing peripheral nerve myelination were subcutaneously administered with the human Lefty A fusion protein variant CX201 once or twice a week at a dose of 5 mg / kg or once a week at a dose of 10 mg / kg. As described in Example 8, electrophysiological evaluation of nerve function was performed. Compared with the vehicle-administered group, C22 mice administered with CX201 showed significant improvement in both nerve conduction velocity (NCV) and compound muscle action potential (CMAP) ( Figure 19 ).

[0298] The rotarod test and grip strength analysis were performed to evaluate motor function. For the rotarod test, mice were placed on a rod rotating at 12 rpm, and the latency to fall off the rod was recorded, with a maximum limit of 200 seconds. As Figure 19 shown, compared with the vehicle control group, the motor ability of C22 mice injected with CX201 on the rotarod was significantly improved. In addition, the whole-limb grip strength of mice was measured using a grip strength meter to evaluate the neuromuscular function of mice. When C22 mice were injected with CX201 at a dose of 5 mg / kg twice a week, the whole-limb grip strength was significantly improved statistically ( Figure 20 ).

[0299] In summary, subcutaneous injection of the human Lefty A fusion protein variant CX201 into 5-week-old C22 mice for four weeks or longer improved the nerve conduction and motor function of C22 mice. In particular, administration of CX201 at a dose of 5 mg / kg twice a week was the most effective.

[0300] Example 18: Inhibition of Nodal Signaling by Human Lefty A Fusion Protein Variants

[0301] Nodal is a member of TGF-β, and Nodal activates Smad signaling. Once Nodal binds to the activin receptor, Smad2 and Smad3 are phosphorylated and bind to Smad4 and move into the nucleus, and then regulate the transcription of various genes. The Nodal-responsive P19 mouse embryonic cancer cell line was used to evaluate the effect of human Lefty A fusion protein variants on Nodal-induced Smad signaling.

[0302] Nodal was pre-incubated with various concentrations of the human Lefty A fusion protein variant CX201 for 30 minutes. Cells were treated with Nodal alone or with the Nodal-Lefty A fusion protein variant for 1 hour, and then Smad3 phosphorylation induced by Nodal was evaluated by Western blot analysis using cell lysates. Treatment of cells with Nodal protein alone induced strong Smad3 phosphorylation, and the Smad3 phosphorylation was inhibited in a dose-dependent manner by the human Lefty A fusion protein variant ( Figure 21 ).

[0303] Industrial Applicability

[0304] According to the present invention, a human Lefty A protein variant and a fusion protein containing the variant are constructed, which have better stability than the naturally occurring human Lefty A protein, and thus are highly expressed and produced in high yield in animal cells. In addition, administration of the constructed human Lefty A protein variant or fusion protein can restore the nerve and motor functions of an animal model of peripheral neuropathy. Therefore, the use of the human Lefty A protein variant or fusion protein can effectively prevent or treat various nerve and muscle diseases.

[0305] Although the present invention has been described in detail with reference to the drawings, it will be apparent to those skilled in the art that these descriptions are only for the preferred embodiments and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the claims and their equivalents.

[0306] Sequence Listing editable text

[0307] Attached in the electronic file. Sequence Listing <110> Junggonang Co., Ltd. Samsung Life Welfare Foundation, a social welfare corporation <120> Fusion Protein Containing Human Lefty A Protein Variant and Its Use <130> PF-B2272 <140> PCT / KR2019 / 017917 <141> 2019-12-17 <150> KR 10-2018-0166802 <151> 2018-12-20 <160> 178 <170> PatentIn version 3.5 <210> 1 <211> 598 <212> PRT <213> Artificial Sequence <220> <223> 42 Fc <400> 1 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Glu Pro 355 360 365 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 370 375 380 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 385 390 395 400 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 405 410 415 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 420 425 430 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 435 440 445 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 450 455 460 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 465 470 475 480 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 485 490 495 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 500 505 510 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 515 520 525 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 530 535 540 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 545 550 555 560 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 565 570 575 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 580 585 590 Ser Leu Ser Pro Gly Lys 595 <210> 2 <211> 542 <212> PRT <213> Artificial Sequence <220> <223> 34 Fc <400> 2 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Phe Ser Gln Ser Phe Arg Glu Val Ala Gly Arg 20 25 30 Phe Leu Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe Gly Met Glu 35 40 45 Gln Arg Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val Leu Arg Leu 50 55 60 Phe Gln Glu Pro Val Pro Lys Ala Ala Leu His Gly His Gly Arg Leu 65 70 75 80 Ser Pro Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp Leu Arg Val 85 90 95 Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser Arg Leu Val 100 105 110 Ser Val His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr Glu Ala Val 115 120 125 Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg Gln Pro Leu Leu Leu Gln 130 135 140 Val Ser Val Gln Arg Glu His Leu Gly Pro Leu Ala Ser Gly Ala His 145 150 155 160 Lys Leu Val Arg Phe Ala Ser Gln Gly Ala Pro Ala Gly Leu Gly Glu 165 170 175 Pro Gln Leu Glu Leu His Thr Leu Asp Leu Arg Asp Tyr Gly Ala Gln 180 185 190 Gly Asp Cys Asp Pro Glu Ala Pro Met Thr Glu Gly Thr Arg Cys Cys 195 200 205 Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly Met Lys Trp Ala Lys Asn 210 215 220 Trp Val Leu Glu Pro Pro Gly Phe Leu Ala Tyr Glu Cys Val Gly Thr 225 230 235 240 Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe Asn Trp Pro Phe Leu Gly 245 250 255 Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala Ser Leu Pro Met Ile Val 260 265 270 Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro Gln Val Val Ser Leu Pro 275 280 285 Asn Met Arg Val Gln Lys Cys Ser Cys Ala Ser Asp Gly Ala Leu Val 290 295 300 Pro Arg Arg Leu Gln Pro Glu Pro Lys Ser Cys Asp Lys Thr His Thr 305 310 315 320 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 325 330 335 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 340 345 350 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 355 360 365 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 370 375 380 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 385 390 395 400 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 405 410 415 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 420 425 430 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 435 440 445 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 450 455 460 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 465 470 475 480 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 485 490 495 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 500 505 510 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 515 520 525 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 530 535 540 <210> 3 <211> 484 <212> PRT <213> Artificial Sequence <220> <223> 28 Fc <400> 3 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr 20 25 30 Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu 35 40 45 Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe 50 55 60 Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg 65 70 75 80 Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu Gly Pro 85 90 95 Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln Gly Ala 100 105 110 Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu Asp Leu 115 120 125 Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro Met Thr 130 135 140 Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly 145 150 155 160 Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe Leu Ala 165 170 175 Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe 180 185 190 Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala 195 200 205 Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro 210 215 220 Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser Cys Ala 225 230 235 240 Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Glu Pro Lys Ser 245 250 255 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 260 265 270 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 275 280 285 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 290 295 300 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 305 310 315 320 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 325 330 335 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 340 345 350 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 355 360 365 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 370 375 380 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 385 390 395 400 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 405 410 415 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 420 425 430 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 435 440 445 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 450 455 460 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 465 470 475 480 Ser Pro Gly Lys <210> 4 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc <400> 4 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 5 <211> 558 <212> PRT <213> Artificial Sequence <220> <223> 34 LFc <400> 5 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Phe Ser Gln Ser Phe Arg Glu Val Ala Gly Arg 20 25 30 Phe Leu Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe Gly Met Glu 35 40 45 Gln Arg Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val Leu Arg Leu 50 55 60 Phe Gln Glu Pro Val Pro Lys Ala Ala Leu His Gly His Gly Arg Leu 65 70 75 80 Ser Pro Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp Leu Arg Val 85 90 95 Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser Arg Leu Val 100 105 110 Ser Val His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr Glu Ala Val 115 120 125 Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg Gln Pro Leu Leu Leu Gln 130 135 140 Val Ser Val Gln Arg Glu His Leu Gly Pro Leu Ala Ser Gly Ala His 145 150 155 160 Lys Leu Val Arg Phe Ala Ser Gln Gly Ala Pro Ala Gly Leu Gly Glu 165 170 175 Pro Gln Leu Glu Leu His Thr Leu Asp Leu Arg Asp Tyr Gly Ala Gln 180 185 190 Gly Asp Cys Asp Pro Glu Ala Pro Met Thr Glu Gly Thr Arg Cys Cys 195 200 205 Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly Met Lys Trp Ala Lys Asn 210 215 220 Trp Val Leu Glu Pro Pro Gly Phe Leu Ala Tyr Glu Cys Val Gly Thr 225 230 235 240 Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe Asn Trp Pro Phe Leu Gly 245 250 255 Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala Ser Leu Pro Met Ile Val 260 265 270 Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro Gln Val Val Ser Leu Pro 275 280 285 Asn Met Arg Val Gln Lys Cys Ser Cys Ala Ser Asp Gly Ala Leu Val 290 295 300 Pro Arg Arg Leu Gln Pro Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 305 310 315 320 Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp Lys Thr His Thr 325 330 335 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 340 345 350 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 355 360 365 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 370 375 380 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 385 390 395 400 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 405 410 415 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 420 425 430 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 435 440 445 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 450 455 460 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 465 470 475 480 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 485 490 495 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 500 505 510 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 515 520 525 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 530 535 540 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 545 550 555 <210> 6 <211> 500 <212> PRT <213> Artificial Sequence <220> <223> 28 LFc <400> 6 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr 20 25 30 Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu 35 40 45 Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe 50 55 60 Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg 65 70 75 80 Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu Gly Pro 85 90 95 Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln Gly Ala 100 105 110 Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu Asp Leu 115 120 125 Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro Met Thr 130 135 140 Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly 145 150 155 160 Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe Leu Ala 165 170 175 Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe 180 185 190 Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala 195 200 205 Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro 210 215 220 Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser Cys Ala 225 230 235 240 Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly Gly Gly 245 250 255 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser 260 265 270 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 275 280 285 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 290 295 300 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 305 310 315 320 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 325 330 335 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 340 345 350 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 355 360 365 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 370 375 380 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 385 390 395 400 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 405 410 415 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 420 425 430 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 435 440 445 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 450 455 460 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 465 470 475 480 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 485 490 495 Ser Pro Gly Lys 500 <210> 7 <211> 596 <212> PRT <213> Artificial Sequence <220> <223> Fc 42 <400> 7 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro 20 25 30 Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 35 40 45 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 50 55 60 Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 65 70 75 80 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 85 90 95 Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 100 105 110 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 115 120 125 Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys 130 135 140 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp 145 150 155 160 Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 165 170 175 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 180 185 190 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 195 200 205 Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 210 215 220 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 225 230 235 240 Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Thr Glu Glu Gln 245 250 255 Leu Leu Gly Ser Leu Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val 260 265 270 Leu Asp Arg Ala Asp Met Glu Lys Leu Val Ile Pro Ala His Val Arg 275 280 285 Ala Gln Tyr Val Val Leu Leu Arg Arg Ser His Gly Asp Arg Ser Arg 290 295 300 Gly Lys Arg Phe Ser Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu 305 310 315 320 Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe Gly Met Glu Gln Arg 325 330 335 Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln 340 345 350 Glu Pro Val Pro Lys Ala Ala Leu His Arg His Gly Arg Leu Ser Pro 355 360 365 Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp 370 375 380 Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val 385 390 395 400 His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe 405 410 415 Trp Gln Gln Leu Ser Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser 420 425 430 Val Gln Arg Glu His Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu 435 440 445 Val Arg Phe Ala Ser Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln 450 455 460 Leu Glu Leu His Thr Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp 465 470 475 480 Cys Asp Pro Glu Ala Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln 485 490 495 Glu Met Tyr Ile Asp Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val 500 505 510 Leu Glu Pro Pro Gly Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln 515 520 525 Gln Pro Pro Glu Ala Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg 530 535 540 Gln Cys Ile Ala Ser Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile 545 550 555 560 Lys Glu Gly Gly Arg Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met 565 570 575 Arg Val Gln Lys Cys Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg 580 585 590 Arg Leu Gln Pro 595 <210> 8 <211> 540 <212> PRT <213> Artificial Sequence <220> <223> Fc 34 <400> 8 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro 20 25 30 Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 35 40 45 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 50 55 60 Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 65 70 75 80 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 85 90 95 Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 100 105 110 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 115 120 125 Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys 130 135 140 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp 145 150 155 160 Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 165 170 175 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 180 185 190 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 195 200 205 Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 210 215 220 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 225 230 235 240 Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Phe Ser Gln Ser Phe 245 250 255 Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr His Leu 260 265 270 Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu Leu Val 275 280 285 Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala Ala Leu 290 295 300 His Arg His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr 305 310 315 320 Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu 325 330 335 Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe 340 345 350 Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg 355 360 365 Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu Gly Pro 370 375 380 Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln Gly Ala 385 390 395 400 Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu Asp Leu 405 410 415 Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro Met Thr 420 425 430 Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly 435 440 445 Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe Leu Ala 450 455 460 Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe 465 470 475 480 Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala 485 490 495 Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro 500 505 510 Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser Cys Ala 515 520 525 Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro 530 535 540 <210> 9 <211> 482 <212> PRT <213> Artificial Sequence <220> <223> Fc 28 <400> 9 Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly 1 5 10 15 Val His Ser Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro 20 25 30 Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 35 40 45 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 50 55 60 Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 65 70 75 80 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 85 90 95 Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 100 105 110 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 115 120 125 Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys 130 135 140 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp 145 150 155 160 Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 165 170 175 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 180 185 190 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 195 200 205 Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 210 215 220 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 225 230 235 240 Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Leu Ser Pro Arg Ser 245 250 255 Ala Gln Ala Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly 260 265 270 Ser Asn Arg Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu 275 280 285 Ser Gly Trp Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln 290 295 300 Gln Leu Ser Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln 305 310 315 320 Arg Glu His Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg 325 330 335 Phe Ala Ser Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu 340 345 350 Leu His Thr Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp 355 360 365 Pro Glu Ala Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met 370 375 380 Tyr Ile Asp Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu 385 390 395 400 Pro Pro Gly Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro 405 410 415 Pro Glu Ala Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys 420 425 430 Ile Ala Ser Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu 435 440 445 Gly Gly Arg Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val 450 455 460 Gln Lys Cys Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu 465 470 475 480 Gln Pro <210> 10 <211> 951 <212> PRT <213> Artificial Sequence <220> <223> 42 HSA <400> 10 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Asp Ala 355 360 365 His Lys Ser Glu Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn 370 375 380 Phe Lys Ala Leu Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys 385 390 395 400 Pro Phe Glu Asp His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala 405 410 415 Lys Thr Cys Val Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu 420 425 430 His Thr Leu Phe Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu 435 440 445 Thr Tyr Gly Glu Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg 450 455 460 Asn Glu Cys Phe Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg 465 470 475 480 Leu Val Arg Pro Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn 485 490 495 Glu Glu Thr Phe Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His 500 505 510 Pro Tyr Phe Tyr Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys 515 520 525 Ala Ala Phe Thr Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu 530 535 540 Leu Pro Lys Leu Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala 545 550 555 560 Lys Gln Arg Leu Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala 565 570 575 Phe Lys Ala Trp Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala 580 585 590 Glu Phe Ala Glu Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His 595 600 605 Thr Glu Cys Cys His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala 610 615 620 Asp Leu Ala Lys Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys 625 630 635 640 Leu Lys Glu Cys Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile 645 650 655 Ala Glu Val Glu Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala 660 665 670 Ala Asp Phe Val Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala 675 680 685 Lys Asp Val Phe Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His 690 695 700 Pro Asp Tyr Ser Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu 705 710 715 720 Thr Thr Leu Glu Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr 725 730 735 Ala Lys Val Phe Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn 740 745 750 Leu Ile Lys Gln Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys 755 760 765 Phe Gln Asn Ala Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val 770 775 780 Ser Thr Pro Thr Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly 785 790 795 800 Ser Lys Cys Cys Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu 805 810 815 Asp Tyr Leu Ser Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys 820 825 830 Thr Pro Val Ser Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val 835 840 845 Asn Arg Arg Pro Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val 850 855 860 Pro Lys Glu Phe Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys 865 870 875 880 Thr Leu Ser Glu Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val 885 890 895 Glu Leu Val Lys His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala 900 905 910 Val Met Asp Asp Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp 915 920 925 Asp Lys Glu Thr Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala 930 935 940 Ser Gln Ala Ala Leu Gly Leu 945 950 <210> 11 <211> 895 <212> PRT <213> Artificial Sequence <220> <223> 34 HSA <400> 11 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Phe Ser Gln Ser Phe Arg Glu Val Ala Gly Arg 20 25 30 Phe Leu Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe Gly Met Glu 35 40 45 Gln Arg Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val Leu Arg Leu 50 55 60 Phe Gln Glu Pro Val Pro Lys Ala Ala Leu His Gly His Gly Arg Leu 65 70 75 80 Ser Pro Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp Leu Arg Val 85 90 95 Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser Arg Leu Val 100 105 110 Ser Val His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr Glu Ala Val 115 120 125 Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg Gln Pro Leu Leu Leu Gln 130 135 140 Val Ser Val Gln Arg Glu His Leu Gly Pro Leu Ala Ser Gly Ala His 145 150 155 160 Lys Leu Val Arg Phe Ala Ser Gln Gly Ala Pro Ala Gly Leu Gly Glu 165 170 175 Pro Gln Leu Glu Leu His Thr Leu Asp Leu Arg Asp Tyr Gly Ala Gln 180 185 190 Gly Asp Cys Asp Pro Glu Ala Pro Met Thr Glu Gly Thr Arg Cys Cys 195 200 205 Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly Met Lys Trp Ala Lys Asn 210 215 220 Trp Val Leu Glu Pro Pro Gly Phe Leu Ala Tyr Glu Cys Val Gly Thr 225 230 235 240 Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe Asn Trp Pro Phe Leu Gly 245 250 255 Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala Ser Leu Pro Met Ile Val 260 265 270 Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro Gln Val Val Ser Leu Pro 275 280 285 Asn Met Arg Val Gln Lys Cys Ser Cys Ala Ser Asp Gly Ala Leu Val 290 295 300 Pro Arg Arg Leu Gln Pro Asp Ala His Lys Ser Glu Val Ala His Arg 305 310 315 320 Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu Val Leu Ile Ala 325 330 335 Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp His Val Lys Leu 340 345 350 Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val Ala Asp Glu Ser 355 360 365 Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe Gly Asp Lys Leu 370 375 380 Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu Met Ala Asp Cys 385 390 395 400 Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe Leu Gln His Lys 405 410 415 Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro Glu Val Asp Val 420 425 430 Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe Leu Lys Lys Tyr 435 440 445 Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr Ala Pro Glu Leu 450 455 460 Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr Glu Cys Cys Gln 465 470 475 480 Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu Asp Glu Leu Arg 485 490 495 Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu Lys Cys Ala Ser 500 505 510 Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp Ala Val Ala Arg 515 520 525 Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu Val Ser Lys Leu 530 535 540 Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys His Gly Asp Leu 545 550 555 560 Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys Tyr Ile Cys Glu 565 570 575 Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys Cys Glu Lys Pro 580 585 590 Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu Asn Asp Glu Met 595 600 605 Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val Glu Ser Lys Asp 610 615 620 Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe Leu Gly Met Phe 625 630 635 640 Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser Val Val Leu Leu 645 650 655 Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr Leu Glu Lys Cys Cys Ala 660 665 670 Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe Asp Glu Phe Lys 675 680 685 Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln Asn Cys Glu Leu 690 695 700 Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala Leu Leu Val Arg 705 710 715 720 Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr Leu Val Glu Val 725 730 735 Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys Lys His Pro Glu 740 745 750 Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser Val Val Leu Asn 755 760 765 Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Asp Arg Val Thr 770 775 780 Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro Cys Phe Ser Ala 785 790 795 800 Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe Asn Ala Glu Thr 805 810 815 Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu Lys Glu Arg Gln 820 825 830 Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys His Lys Pro Lys 835 840 845 Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp Phe Ala Ala Phe 850 855 860 Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr Cys Phe Ala Glu 865 870 875 880 Glu Gly Lys Lys Leu Val Ala Ala Ser Gln Ala Ala Leu Gly Leu 885 890 895 <210> 12 <211> 837 <212> PRT <213> Artificial Sequence <220> <223> 28 HSA <400> 12 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr 20 25 30 Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu 35 40 45 Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe 50 55 60 Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg 65 70 75 80 Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu Gly Pro 85 90 95 Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln Gly Ala 100 105 110 Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu Asp Leu 115 120 125 Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro Met Thr 130 135 140 Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly 145 150 155 160 Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe Leu Ala 165 170 175 Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe 180 185 190 Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala 195 200 205 Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro 210 215 220 Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser Cys Ala 225 230 235 240 Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Asp Ala His Lys 245 250 255 Ser Glu Val Ala His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys 260 265 270 Ala Leu Val Leu Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe 275 280 285 Glu Asp His Val Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr 290 295 300 Cys Val Ala Asp Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr 305 310 315 320 Leu Phe Gly Asp Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr 325 330 335 Gly Glu Met Ala Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu 340 345 350 Cys Phe Leu Gln His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val 355 360 365 Arg Pro Glu Val Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu 370 375 380 Thr Phe Leu Lys Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr 385 390 395 400 Phe Tyr Ala Pro Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala 405 410 415 Phe Thr Glu Cys Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro 420 425 430 Lys Leu Asp Glu Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln 435 440 445 Arg Leu Lys Cys Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys 450 455 460 Ala Trp Ala Val Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe 465 470 475 480 Ala Glu Val Ser Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu 485 490 495 Cys Cys His Gly Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu 500 505 510 Ala Lys Tyr Ile Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys 515 520 525 Glu Cys Cys Glu Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu 530 535 540 Val Glu Asn Asp Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp 545 550 555 560 Phe Val Glu Ser Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp 565 570 575 Val Phe Leu Gly Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp 580 585 590 Tyr Ser Val Val Leu Leu Leu Arg Leu Ala Lys Thr Tyr Glu Thr Thr 595 600 605 Leu Glu Lys Cys Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys 610 615 620 Val Phe Asp Glu Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile 625 630 635 640 Lys Gln Asn Cys Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln 645 650 655 Asn Ala Leu Leu Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr 660 665 670 Pro Thr Leu Val Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys 675 680 685 Cys Cys Lys His Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr 690 695 700 Leu Ser Val Val Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro 705 710 715 720 Val Ser Asp Arg Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg 725 730 735 Arg Pro Cys Phe Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys 740 745 750 Glu Phe Asn Ala Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu 755 760 765 Ser Glu Lys Glu Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu 770 775 780 Val Lys His Lys Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met 785 790 795 800 Asp Asp Phe Ala Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys 805 810 815 Glu Thr Cys Phe Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Gln 820 825 830 Ala Ala Leu Gly Leu 835 <210> 13 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L1_F <400> 13 aattaagctt gccaccatgt ggcctctgtg gctctg 36 <210> 14 <211> 24 <212> DNA <213> Artificial sequence <220> <223> L2_R <400> 14 aggttgcaat cgtcttggga caag 24 <210> 15 <211> 45 <212> DNA <213> Artificial sequence <220> <223> L3_F <400> 15 cttgtcccaa gacgattgca acctgagccc aaatcttgtg acaaa 45 <210> 16 <211> 31 <212> DNA <213> Artificial sequence <220> <223> L4_R <400> 16 aattctcgag tcatttaccc ggagacaggg a 31 <210> 17 <211> 87 <212> DNA <213> Artificial Sequence <220> <223> L5_F <400> 17 gtcccaagac gattgcaacc tagcggcggt ggcggttctg gcggtggtgg aagtggcggt 60 ggcgggtctg agcccaaatc ttgtgac 87 <210> 18 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L6_F <400> 18 ctctccctgt ctccgggtaa actcaccgaa gagcagttgt tg 42 <210> 19 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L7_F <400> 19 ctctccctgt ctccgggtaa attcagccaa tcattccgtg ag 42 <210> 20 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L8_F <400> 20 ctctccctgt ctccgggtaa actctccccc cgaagcgccc ag 42 <210> 21 <211> 43 <212> DNA <213> Artificial Sequence <220> <223> L9_R <400> 21 gaaggcacag ctcgagtcaa ggttgcaatc gtcttgggac aag 43 <210> 22 <211> 45 <212> DNA <213> Artificial sequence <220> <223> L10_F <400> 22 gtcccaagac gattgcaacc tgacgctcac aagtctgaag ttgct 45 <210> 23 <211> 48 <212> DNA <213> Artificial sequence <220> <223> L11_R <400> 23 agatccacgc ggaaccagct cgagtcacag ccccaaagcg gcctgtga 48 <210> 24 <211> 21 <212> DNA <213> Artificial sequence <220> <223> L12_F <400> 24 actctagagg atcgaaccct t 21 <210> 25 <211> 54 <212> DNA <213> Artificial sequence <220> <223> L12_R <400> 25 gctggcaact agaaggcaca gctcgagtca tttacccgga gacagggaga ggct 54 <210> 26 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L13_F <400> 26 ggggccgctc tcacagrgga gcagktgctc gggtcattgt tg 42 <210> 27 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L13_R <400> 27 caacaatgac ccgagcamct gctccyctgt gagagcggcc cc 42 <210> 28 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L14_F <400> 28 ctcgggtcat tgttgaggca gcttcagctt tcc 33 <210> 29 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L14_R <400> 29 ggaaagctga agctgcctca acaatgaccc gag 33 <210> 30 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L15_F <400> 30 cttcagcttt ccgaagtgcc cgtcctggac aag 33 <210> 31 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L15_R <400> 31 cttgtccagg acgggcactt cggaaagctg aag 33 <210> 32 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L16_F <400> 32 ccacccgtcc tggacarggc cgatrtggag gggctcgtta tc 42 <210> 33 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L16_R <400> 33 gataacgagc ccctccayat cggccytgtc caggacgggt gg 42 <210> 34 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L17_F <400> 34 aaggccgatg tcgagaagct cgttatccca tcc 33 <210> 35 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L17_R <400> 35 ggatgggata acgagcttct cgacatcggc ctt 33 <210> 36 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L18_F <400> 36 gggctcgtta tcccagccca tgtccgggct cag 33 <210> 37 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L18_R <400> 37 ctgagcccgg acatgggctg ggataacgag ccc 33 <210> 38 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L19_F <400> 38 cgggctcagt acgttgtgtt gctgcaacat tca 33 <210> 39 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L19_R <400> 39 tgaatgttgc agcaacacaa cgtactgagc ccg 33 <210> 40 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L20_F <400> 40 tacgttgcct tgctgagacg gtcacacgct agtaga 36 <210> 41 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L20_R <400> 41 tctactagcg tgtgaccgtc tcagcaaggc aacgta 36 <210> 42 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L21_F <400> 42 ctgcaacatt cacacgsakm cagaagtgga ggcaag 36 <210> 43 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L21_R <400> 43 cttgcctcca cttctgkmts cgtgtgaatg ttgcag 36 <210> 44 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L22_F <400> 44 tacgttgcct tgctgcrgcr ctcacacgct agtaga 36 <210> 45 <211> 36 <212> DNA <213> Artificial sequence <220> <223> L22_R <400> 45 tctactagcg tgtgagygcy gcagcaaggc aacgta 36 <210> 46 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L23_F <400> 46 ggggccgccc tcaccggcga gcagttgttg ggc 33 <210> 47 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L23_R <400> 47 gcccaacaac tgctcgccgg tgagggcggc ccc 33 <210> 48 <211> 42 <212> DNA <213> Artificial sequence <220> <223> L24_F <400> 48 cgcgcccaat atgtggycct gctccrgcga tctcacggag at 42 <210> 49 <211> 42 <212> DNA <213> Artificial sequence <220> <223> L24_R <400> 49 atctccgtga gatcgcygga gcaggrccac atattgggcg cg 42 <210> 50 <211> 45 <212> DNA <213> Artificial sequence <220> <223> L25_F <400> 50 cgcgcccaat atgtggccct gctccrgcac tctcacggag atagg 45 <210> 51 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> L25_R <400> 51 cctatctccg tgagagtgcy ggagcagggc cacatattgg gcgcg 45 <210> 52 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> L26_F <400> 52 aaggccgatg tcgagaagct cgttatccca gcccatgtcc gggctcag 48 <210> 53 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> L26_R <400> 53 ctgagcccgg acatgggctg ggataacgag cttctcgaca tcggcctt 48 <210> 54 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L27_F <400> 54 aaggctgccc tgcaccggca tggtcgtctc tccccccgaa gc 42 <210> 55 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L27_R <400> 55 gcttcggggg gagagacgac catgccggtg cagggcagcc tt 42 <210> 56 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L28_F <400> 56 ctgcacggcc atggtggcct ctccccccga agc 33 <210> 57 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L28_R <400> 57 gcttcggggg gagaggccac catggccgtg cag 33 <210> 58 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L29_F <400> 58 aggagcaggg gcaaggyctt cagccaatca ttc 33 <210> 59 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L29_R <400> 59 gaatgattgg ctgaagrcct tgcccctgct cct 33 <210> 60 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L30_F <400> 60 ctgcaccggc atggtgycct ctccccccga agc 33 <210> 61 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L30_R <400> 61 gcttcggggg gagaggrcac catgccggtg cag 33 <210> 62 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L31_F <400> 62 ctgcaccggc atggtggcct ctccccccga agc 33 <210> 63 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L31_R <400> 63 gcttcggggg gagaggccac catgccggtg cag 33 <210> 64 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> L32_F <220> <221> misc_feature <222> (16)..(17) <223> n is a, c, g, or t <400> 64 ctccttctgc aagtgnnkgt ccagcgcgaa cac 33 <210> 65 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L32_R <220> <221> misc_feature <222> (17)..(18) <223> n is a, c, g, or t <400> 65 gtgttcgcgc tggacmnnca cttgcagaag gag 33 <210> 66 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L33_F <220> <221> misc_feature <222> (16)..(17) <223> n is a, c, g, or t <400> 66 ctggtgcgat tcgctnnkca gggtgcacca gcc 33 <210> 67 <211> 33 <212> DNA <213> Artificial sequence <220> <223> L33_R <220> <221> misc_feature <222> (17)..(18) <223> n is a, c, g, or t <400> 67 ggctggtgca ccctgmnnag cgaatcgcac cag 33 <210> 68 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L34_F <400> 68 ttcaactggc cctttgasgg acctcggcaa tgtattgcca gc 42 <210> 69 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L34_R <400> 69 gctggcaata cattgccgag gtccstcaaa gggccagttg aa 42 <210> 70 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L35_F <400> 70 ttcaactggc cctttctcgg agascggcaa tgtattgcca gc 42 <210> 71 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L35_R <400> 71 gctggcaata cattgccgst ctccgagaaa gggccagttg aa 42 <210> 72 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L36_F <400> 72 ttcaactggc cctttctcgg atcccggcaa tgtattgcca gc 42 <210> 73 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L36_R <400> 73 gctggcaata cattgccggg atccgagaaa gggccagttg aa 42 <210> 74 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L37_F <400> 74 ttcaactggc cctttctcgg acctmagcaa tgtattgcca gc 42 <210> 75 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L37_R <400> 75 gctggcaata cattgctkag gtccgagaaa gggccagttg aa 42 <210> 76 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L38_F <400> 76 gcctctgacg gggctgasgt cccaagacga ttgcaacctt ct 42 <210> 77 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L38_R <400> 77 agaaggttgc aatcgtcttg ggacstcagc cccgtcagag gc 42 <210> 78 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L39_F <400> 78 gcctctgacg gggctcttgt cgasagacga ttgcaacctt ct 42 <210> 79 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L39_R <400> 79 agaaggttgc aatcgtctst cgacaagagc cccgtcagag gc 42 <210> 80 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L40_F <400> 80 gcctctgacg gggctcttgt ctccagacga ttgcaacctt ct 42 <210> 81 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L40_R <400> 81 agaaggttgc aatcgtctgg agacaagagc cccgtcagag gc 42 <210> 82 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L41_F <400> 82 gcctctgacg gggctcttgt cccamagcga ttgcaacctt ct 42 <210> 83 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L41_R <400> 83 agaaggttgc aatcgctktg ggacaagagc cccgtcagag gc 42 <210> 84 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L42_F <400> 84 gcctctgacg gggctcttgt cgaccagcga ttgcaacctt ct 42 <210> 85 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> L42_R <400> 85 agaaggttgc aatcgctggt cgacaagagc cccgtcagag gc 42 <210> 86 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(V63A) <400> 86 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 87 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(R66Q) <400> 87 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 88 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(V63A / R66Q) <400> 88 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 89 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(V63A / R67H) <400> 89 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Arg His Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 90 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(V63A / R66Q / R67H) <400> 90 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln His Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Methionine, Threonine, Glutamic acid, Glycine, Threonine, Arginine, Cysteine, Cysteine, Arginine, Glutamine, Glutamic acid, Methionine, Tyrosine, Isoleucine, Aspartic acid, Leucine 260 265 270 Glutamine, Glycine, Methionine, Lysine, Tryptophan, Alanine, Lysine, Asparagine, Tryptophan, Valine, Leucine, Glutamic acid, Proline, Proline, Glycine, Phenylalanine 275 280 285 Leucine, Alanine, Tyrosine, Glutamic acid, Cysteine, Valine, Glycine, Threonine, Cysteine, Glutamine, Glutamine, Proline, Proline, Glutamic acid, Alanine, Leucine 290 295 300 Alanine, Phenylalanine, Asparagine, Tryptophan, Proline, Phenylalanine, Leucine, Glycine, Proline, Arginine, Glutamine, Cysteine, Isoleucine, Alanine, Serine, Glutamic acid 305 310 315 320 Threonine, Alanine, Serine, Leucine, Proline, Methionine, Isoleucine, Valine, Serine, Isoleucine, Lysine, Glutamic acid, Glycine, Glycine, Arginine, Threonine 325 330 335 Arginine, Proline, Glutamine, Valine, Valine, Serine, Leucine, Proline, Asparagine, Methionine, Arginine, Valine, Glutamine, Lysine, Cysteine, Serine 340 345 350 Cysteine, Alanine, Serine, Aspartic acid, Glycine, Alanine, Leucine, Valine, Proline, Arginine, Arginine, Leucine, Glutamine, Proline, Serine, Glycine 355 360 365 Glycine, Glycine, Glycine, Serine, Glycine, Glycine, Glycine, Glycine, Serine, Glycine, Glycine, Glycine, Glycine, Serine, Glutamic acid, Proline 370 375 380 Lysine, Serine, Cysteine, Aspartic acid, Lysine, Threonine, Histidine, Threonine, Cysteine, Proline, Proline, Cysteine, Proline, Alanine, Proline, Glutamic acid 385 390 395 400 Leucine, Leucine, Glycine, Glycine, Proline, Serine, Valine, Phenylalanine, Leucine, Phenylalanine, Proline, Proline, Lysine, Proline, Lysine, Aspartic acid 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 91 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> LFB 42 LFc <400> 91 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Gly Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Lys Glu Val Pro Thr Leu Asp Arg Ala Asp Met 35 40 45 Glu Glu Leu Val Ile Pro Thr His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 92 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V1 <400> 92 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 93 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V2 <400> 93 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leucine Arginine Arginine Serine Histidine Glycine Aspartic acid Arginine Serine Glycine Glycine Lysine Glycine Phenylalanine Serine Glutamine 65 70 75 80 Serine Phenylalanine Arginine Glutamic acid Valine Alanine Glycine Arginine Phenylalanine Leucine Alanine Serine Glutamic acid Alanine Serine Threonine 85 90 95 Histidine Leucine Leucine Valine Phenylalanine Glycine Methionine Glutamic acid Glutamine Arginine Leucine Proline Proline Asparagine Serine Glutamic acid 100 105 110 Leucine Valine Glutamine Alanine Valine Leucine Arginine Leucine Phenylalanine Glutamine Glutamic acid Proline Valine Proline Lysine Alanine 115 120 125 Alanine Leucine Histidine Arginine Histidine Glycine Glycine Leucine Serine Proline Arginine Serine Alanine Glutamine Alanine Arginine 130 135 140 Valine Threonine Valine Glutamic acid Tryptophan Leucine Arginine Valine Arginine Aspartic acid Aspartic acid Glycine Serine Asparagine Arginine Threonine 145 150 155 160 Serine Leucine Isoleucine Aspartic acid Serine Arginine Leucine Valine Serine Valine Histidine Glutamic acid Serine Glycine Tryptophan Lysine 165 170 175 Alanine Phenylalanine Aspartic acid Valine Threonine Glutamic acid Alanine Valine Asparagine Phenylalanine Tryptophan Glutamine Glutamine Leucine Serine Arginine 180 185 190 Proline Arginine Glutamine Proline Leucine Leucine Leucine Glutamine Valine Serine Valine Glutamine Arginine Glutamic acid Histidine Leucine 195 200 205 Glycine Proline Leucine Alanine Serine Glycine Alanine Histidine Lysine Leucine Valine Arginine Phenylalanine Alanine Serine Glutamine 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 94 <211> 614 <212> PRT <213> Artificial sequence <220> <223> 42 LFc V3 <400> 94 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Glu Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 95 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V4 <400> 95 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 96 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V5 <400> 96 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Ala Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 97 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V6 <400> 97 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Ala Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Ala Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 98 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V7 <400> 98 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Ala Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Val Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 99 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V8 <400> 99 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Val Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Gly His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 100 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V9 <400> 100 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Val Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Ala Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 101 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V10 <400> 101 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Val Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Val Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu[[ID=z8]] 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 102 <211> 614 <212> PRT <213> Artificial sequence <220> <223> 42 LFc V11 <400> 102 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg His Gly Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 103 <211> 614 <212> PRT <213> Artificial sequence <220> <223> 42 LFc V12 <400> 103 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 104 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc V13 <400> 104 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Ala Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 105 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> CX196 <400> 105 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Gly Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Lys Glu Val Pro Thr Leu Asp Arg Ala Asp Met 35 40 45 Glu Glu Leu Val Ile Pro Thr His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 106 <211> 614 <212> PRT <213> Artificial sequence <220> <223> CX197 <400> 106 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Gly Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Lys Glu Val Pro Thr Leu Asp Arg Ala Asp Met 35 40 45 Glu Glu Leu Val Ile Pro Thr His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Ala Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Methionine, Threonine, Glutamic acid, Glycine, Threonine, Arginine, Cysteine, Cysteine, Arginine, Glutamine, Glutamic acid, Methionine, Tyrosine, Isoleucine, Aspartic acid, Leucine 260 265 270 Glutamine, Glycine, Methionine, Lysine, Tryptophan, Alanine, Lysine, Asparagine, Tryptophan, Valine, Leucine, Glutamic acid, Proline, Proline, Glycine, Phenylalanine 275 280 285 Leucine, Alanine, Tyrosine, Glutamic acid, Cysteine, Valine, Glycine, Threonine, Cysteine, Glutamine, Glutamine, Proline, Proline, Glutamic acid, Alanine, Leucine 290 295 300 Alanine, Phenylalanine, Asparagine, Tryptophan, Proline, Phenylalanine, Leucine, Glycine, Proline, Arginine, Glutamine, Cysteine, Isoleucine, Alanine, Serine, Glutamic acid 305 310 315 320 Threonine, Alanine, Serine, Leucine, Proline, Methionine, Isoleucine, Valine, Serine, Isoleucine, Lysine, Glutamic acid, Glycine, Glycine, Arginine, Threonine 325 330 335 Arginine, Proline, Glutamine, Valine, Valine, Serine, Leucine, Proline, Asparagine, Methionine, Arginine, Valine, Glutamine, Lysine, Cysteine, Serine 340 345 350 Cysteine, Alanine, Serine, Aspartic acid, Glycine, Alanine, Leucine, Valine, Proline, Arginine, Arginine, Leucine, Glutamine, Proline, Serine, Glycine 355 360 365 Glycine, Glycine, Glycine, Serine, Glycine, Glycine, Glycine, Glycine, Serine, Glycine, Glycine, Glycine, Glycine, Serine, Glutamic acid, Proline 370 375 380 Lysine, Serine, Cysteine, Aspartic acid, Lysine, Threonine, Histidine, Threonine, Cysteine, Proline, Proline, Cysteine, Proline, Alanine, Proline, Glutamic acid 385 390 395 400 Leucine, Leucine, Glycine, Glycine, Proline, Serine, Valine, Phenylalanine, Leucine, Phenylalanine, Proline, Proline, Lysine, Proline, Lysine, Aspartic acid 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 107 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> CX201 <400> 107 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 108 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> CX203 <400> 108 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Glu Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 109 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> CX206 <400> 109 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Thr Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 110 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> CX207 <400> 110 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Gly Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 111 <211> 614 <212> PRT <213> Artificial Sequence <220> <223> CX208 <400> 111 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu 50 55 60 Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Thr Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Gly Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly 355 360 365 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro 370 375 380 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 385 390 395 400 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 405 410 415 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 420 425 430 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 435 440 445 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 450 455 460 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 465 470 475 480 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 485 490 495 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 500 505 510 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn 515 520 525 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 530 535 540 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 545 550 555 560 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 565 570 575 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 580 585 590 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 595 600 605 Ser Leu Ser Pro Gly Lys 610 <210> 112 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(L311D) <400> 112 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Asp Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 113 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(L311E) <400> 113 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Glu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 114 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(P313D) <400> 114 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Asp Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 115 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(P313E) <400> 115 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Glu Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 116 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(P313S) <400> 116 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Ser Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 117 <211> 615 <212> PRT <213> Artificial sequence <220> <223> CX201(R314K) <400> 117 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Lys Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 118 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(R314Q) <400> 118 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Gln Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 119 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(L359D) <400> 119 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Asp Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 120 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(L359E) <400> 120 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Glu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 121 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(P361D) <400> 121 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Asp Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 122 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(P361E) <400> 122 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Glu Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 123 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(P361S) <400> 123 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Methionine, Glutamic acid, Lysine, Leucine, Valine, Isoleucine, Proline, Alanine, Histidine, Valine, Arginine, Alanine, Glutamine, Tyrosine, Valine, Alanine 50 55 60 Leucine, Leucine, Glutamine, Arginine, Serine, Histidine, Glycine, Aspartic acid, Arginine, Serine, Glycine, Glycine, Lysine, Glycine, Phenylalanine, Serine 65 70 75 80 Glutamine, Serine, Phenylalanine, Arginine, Glutamic acid, Valine, Alanine, Glycine, Arginine, Phenylalanine, Leucine, Alanine, Serine, Glutamic acid, Alanine, Serine 85 90 95 Threonine, Histidine, Leucine, Leucine, Valine, Phenylalanine, Glycine, Methionine, Glutamic acid, Glutamine, Arginine, Leucine, Proline, Proline, Asparagine, Serine 100 105 110 Glutamic acid, Leucine, Valine, Glutamine, Alanine, Valine, Leucine, Arginine, Leucine, Phenylalanine, Glutamine, Glutamic acid, Proline, Valine, Proline, Lysine 115 120 125 Alanine, Alanine, Leucine, Histidine, Arginine, Histidine, Glycine, Glycine, Leucine, Serine, Proline, Arginine, Serine, Alanine, Glutamine, Alanine 130 135 140 Arginine, Valine, Threonine, Valine, Glutamic acid, Tryptophan, Leucine, Arginine, Valine, Arginine, Aspartic acid, Aspartic acid, Glycine, Serine, Asparagine, Arginine 145 150 155 160 Threonine, Serine, Leucine, Isoleucine, Aspartic acid, Serine, Arginine, Leucine, Valine, Serine, Valine, Histidine, Glutamic acid, Serine, Glycine, Tryptophan 165 170 175 Lysine, Alanine, Phenylalanine, Aspartic acid, Valine, Threonine, Glutamic acid, Alanine, Valine, Asparagine, Phenylalanine, Tryptophan, Glutamine, Glutamine, Leucine, Serine 180 185 190 Arginine, Proline, Arginine, Glutamine, Proline, Leucine, Leucine, Leucine, Glutamine, Valine, Serine, Valine, Glutamine, Arginine, Glutamic acid, Histidine 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Ser Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 124 <211> 615 <212> PRT <213> Artificial sequence <220> <223> CX201(R362K) <400> 124 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Lys Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 125 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(R362Q) <400> 125 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Gln Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 126 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(L311D / R362Q) <400> 126 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Asp Gly Pro Arg Gln Cys Ile Ala Ser ​ Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Gln Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 127 <211> 615 <212> PRT <213> Artificial Sequence <220> <223> CX201(L311E / R362Q) <400> 127 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leucine Alanine Phenylalanine Asparagine Tryptophan Proline Phenylalanine Glutamic acid Glycine Proline Arginine Glutamine Cysteine Isoleucine Alanine Serine 305 310 315 320 Glutamic acid Threonine Alanine Serine Leucine Proline Methionine Isoleucine Valine Serine Isoleucine Lysine Glutamic acid Glycine Glycine Arginine 325 330 335 Threonine Arginine Proline Glutamine Valine Valine Serine Leucine Proline Asparagine Methionine Arginine Valine Glutamine Lysine Cysteine 340 345 350 Serine Cysteine Alanine Serine Aspartic acid Glycine Alanine Leucine Valine Proline Glutamine Arginine Leucine Glutamine Proline Serine 355 360 365 Glycine Glycine Glycine Glycine Serine Glycine Glycine Glycine Serine Glycine Glycine Glycine Serine Glutamic acid 370 375 380 Proline Lysine Serine Cysteine Aspartic acid Lysine Threonine Histidine Threonine Cysteine Proline Proline Cysteine Proline Alanine Proline 385 390 395 400 Glutamic acid Leucine Leucine Glycine Glycine Proline Serine Valine Phenylalanine Leucine Phenylalanine Proline Proline Lysine Proline Lysine 405 410 415 Aspartic acid Threonine Leucine Methionine Isoleucine Serine Arginine Threonine Proline Glutamic acid Valine Threonine Cysteine Valine Valine Valine 420 425 430 Aspartic acid Valine Serine Histidine Glutamic acid Aspartic acid Proline Glutamic acid Valine Lysine Phenylalanine Asparagine Tryptophan Tyrosine Valine Aspartic acid 435 440 445 Glycine Valine Glutamic acid Valine Histidine Asparagine Alanine Lysine Threonine Lysine Proline Arginine Glutamic acid Glutamic acid Glutamine Tyrosine 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 128 <211> 615 <212> PRT <213> Artificial sequence <220> <223> CX201(L311D / P361D / R362Q) <400> 128 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Asp Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Asp Gln Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 129 <211> 615 <212> PRT <213> artificial sequence <220> <223> CX201(L311E / P361D / R362Q) <400> 129 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Glu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Asp Gln Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 130 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Linker <400> 130 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <210> 131 <211> 366 <212> PRT <213> Artificial sequence <220> <223> Human Lefty A <400> 131 Met Trp Pro Leu Trp Leu Cys Trp Ala Leu Trp Val Leu Pro Leu Ala 1 5 10 15 Gly Pro Gly Ala Ala Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu 20 25 30 Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met 35 40 45 Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Val Leu 50 55 60 Leu Arg Arg Ser His Gly Asp Arg Ser Arg Gly Lys Arg Phe Ser Gln 65 70 75 80 Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr 85 90 95 His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu 100 105 110 Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala 115 120 125 Ala Leu His Arg His Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg 130 135 140 Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr 145 150 155 160 Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp Lys 165 170 175 Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg 180 185 190 Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His Leu 195 200 205 Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln 210 215 220 Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu 225 230 235 240 Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro 245 250 255 Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu 260 265 270 Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe 275 280 285 Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu 290 295 300 Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu 305 310 315 320 Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr 325 330 335 Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser 340 345 350 Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro 355 360 365 <210> 132 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> Signal peptide <400> 132 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys 20 <210> 133 <211> 615 <212> PRT <213> Artificial sequence <220> <223> CX201s <400> 133 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ser Arg Cys Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu 20 25 30 Leu Arg Gln Leu Gln Leu Ser Glu Val Pro Val Leu Asp Arg Ala Asp 35 40 45 Met Glu Lys Leu Val Ile Pro Ala His Val Arg Ala Gln Tyr Val Ala 50 55 60 Leu Leu Gln Arg Ser His Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser 65 70 75 80 Gln Ser Phe Arg Glu Val Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser 85 90 95 Thr His Leu Leu Val Phe Gly Met Glu Gln Arg Leu Pro Pro Asn Ser 100 105 110 Glu Leu Val Gln Ala Val Leu Arg Leu Phe Gln Glu Pro Val Pro Lys 115 120 125 Ala Ala Leu His Arg His Gly Gly Leu Ser Pro Arg Ser Ala Gln Ala 130 135 140 Arg Val Thr Val Glu Trp Leu Arg Val Arg Asp Asp Gly Ser Asn Arg 145 150 155 160 Thr Ser Leu Ile Asp Ser Arg Leu Val Ser Val His Glu Ser Gly Trp 165 170 175 Lys Ala Phe Asp Val Thr Glu Ala Val Asn Phe Trp Gln Gln Leu Ser 180 185 190 Arg Pro Arg Gln Pro Leu Leu Leu Gln Val Ser Val Gln Arg Glu His 195 200 205 Leu Gly Pro Leu Ala Ser Gly Ala His Lys Leu Val Arg Phe Ala Ser 210 215 220 Gln Gly Ala Pro Ala Gly Leu Gly Glu Pro Gln Leu Glu Leu His Thr 225 230 235 240 Leu Asp Leu Arg Asp Tyr Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala 245 250 255 Pro Met Thr Glu Gly Thr Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp 260 265 270 Leu Gln Gly Met Lys Trp Ala Lys Asn Trp Val Leu Glu Pro Pro Gly 275 280 285 Phe Leu Ala Tyr Glu Cys Val Gly Thr Cys Gln Gln Pro Pro Glu Ala 290 295 300 Leu Ala Phe Asn Trp Pro Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser 305 310 315 320 Glu Thr Ala Ser Leu Pro Met Ile Val Ser Ile Lys Glu Gly Gly Arg 325 330 335 Thr Arg Pro Gln Val Val Ser Leu Pro Asn Met Arg Val Gln Lys Cys 340 345 350 Ser Cys Ala Ser Asp Gly Ala Leu Val Pro Arg Arg Leu Gln Pro Ser 355 360 365 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu 370 375 380 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 385 390 395 400 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 405 410 415 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 420 425 430 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 435 440 445 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 450 455 460 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 465 470 475 480 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 485 490 495 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 500 505 510 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 515 520 525 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 530 535 540 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 545 550 555 560 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 565 570 575 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 580 585 590 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 595 600 605 Leu Ser Leu Ser Pro Gly Lys 610 615 <210> 134 <211> 593 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(V63A) <400> 134 Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu Arg Gln Leu Gln Leu 1 5 10 15 Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met Glu Lys Leu Val Ile 20 25 30 Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu Leu Arg Arg Ser His 35 40 45 Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln Ser Phe Arg Glu Val 50 55 60 Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe 65 70 75 80 Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val 85 90 95 Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala Ala Leu His Gly His 100 105 110 Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp 115 120 125 Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser 130 135 140 Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr 145 150 155 160 Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg Gln Pro Leu 165 170 175 Leu Leu Gln Val Ser Val Gln Arg Glu His Leu Gly Pro Leu Ala Ser 180 185 190 Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln Gly Ala Pro Ala Gly 195 200 205 Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu Asp Leu Arg Asp Tyr 210 215 220 Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro Met Thr Glu Gly Thr 225 230 235 240 Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly Met Lys Trp 245 250 255 Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe Leu Ala Tyr Glu Cys 260 265 270 Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe Asn Trp Pro 275 280 285 Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala Ser Leu Pro 290 295 300 Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro Gln Val Val 305 310 315 320 Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser Cys Ala Ser Asp Gly 325 330 335 Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly Gly Gly Gly Ser Gly 340 345 350 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp Lys 355 360 365 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 370 375 380 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 385 390 395 400 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 405 410 415 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 420 425 430 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 435 440 445 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 450 455 460 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 465 470 475 480 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 485 490 495 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 500 505 510 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 515 520 525 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 530 535 540 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 545 550 555 560 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 565 570 575 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 580 585 590 Lys <210> 135 <211> 593 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(R66Q) <400> 135 Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu Arg Gln Leu Gln Leu 1 5 10 15 Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met Glu Lys Leu Val Ile 20 25 30 Pro Ala His Val Arg Ala Gln Tyr Val Val Leu Leu Gln Arg Ser His 35 40 45 Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln Ser Phe Arg Glu Val 50 55 60 Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe 65 70 75 80 Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val 85 90 95 Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala Ala Leu His Gly His 100 105 110 Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp 115 120 125 Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser 130 135 140 Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr 145 150 155 160 Glu Ala Val Asn Phe Trp Gln Gln Leu Ser Arg Pro Arg Gln Pro Leu 165 170 175 Leu Leu Gln Val Ser Val Gln Arg Glu His Leu Gly Pro Leu Ala Ser 180 185 190 Gly Ala His Lys Leu Val Arg Phe Ala Ser Gln Gly Ala Pro Ala Gly 195 200 205 Leu Gly Glu Pro Gln Leu Glu Leu His Thr Leu Asp Leu Arg Asp Tyr 210 215 220 Gly Ala Gln Gly Asp Cys Asp Pro Glu Ala Pro Met Thr Glu Gly Thr 225 230 235 240 Arg Cys Cys Arg Gln Glu Met Tyr Ile Asp Leu Gln Gly Met Lys Trp 245 250 255 Ala Lys Asn Trp Val Leu Glu Pro Pro Gly Phe Leu Ala Tyr Glu Cys 260 265 270 Val Gly Thr Cys Gln Gln Pro Pro Glu Ala Leu Ala Phe Asn Trp Pro 275 280 285 Phe Leu Gly Pro Arg Gln Cys Ile Ala Ser Glu Thr Ala Ser Leu Pro 290 295 300 Met Ile Val Ser Ile Lys Glu Gly Gly Arg Thr Arg Pro Gln Val Val 305 310 315 320 Ser Leu Pro Asn Met Arg Val Gln Lys Cys Ser Cys Ala Ser Asp Gly 325 330 335 Ala Leu Val Pro Arg Arg Leu Gln Pro Ser Gly Gly Gly Gly Ser Gly 340 345 350 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Cys Asp Lys 355 360 365 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 370 375 380 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 385 390 395 400 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 405 410 415 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 420 425 430 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 435 440 445 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 450 455 460 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 465 470 475 480 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 485 490 495 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 500 505 510 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 515 520 525 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 530 535 540 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 545 550 555 560 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 565 570 575 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 580 585 590 Lys <210> 136 <211> 593 <212> PRT <213> Artificial Sequence <220> <223> 42 LFc(V63A / R66Q) <400> 136 Leu Thr Glu Glu Gln Leu Leu Gly Ser Leu Leu Arg Gln Leu Gln Leu 1 5 10 15 Ser Glu Val Pro Val Leu Asp Arg Ala Asp Met Glu Lys Leu Val Ile 20 25 30 Pro Ala His Val Arg Ala Gln Tyr Val Ala Leu Leu Gln Arg Ser His 35 40 45 Gly Asp Arg Ser Gly Gly Lys Gly Phe Ser Gln Ser Phe Arg Glu Val 50 55 60 Ala Gly Arg Phe Leu Ala Ser Glu Ala Ser Thr His Leu Leu Val Phe 65 70 75 80 Gly Met Glu Gln Arg Leu Pro Pro Asn Ser Glu Leu Val Gln Ala Val 85 90 95 Leu Arg Leu Phe Gln Glu Pro Val Pro Lys Ala Ala Leu His Gly His 100 105 110 Gly Arg Leu Ser Pro Arg Ser Ala Gln Ala Arg Val Thr Val Glu Trp 115 120 125 Leu Arg Val Arg Asp Asp Gly Ser Asn Arg Thr Ser Leu Ile Asp Ser 130 135 140 Arg Leu Val Ser Val His Glu Ser Gly Trp Lys Ala Phe Asp Val Thr 145 150 155 160 Glu Ala Val Asn P...

Claims

1. A fusion protein comprising a human Lefty A protein variant, wherein the amino acid sequence of the fusion protein is selected from the group consisting of the amino acid sequences listed in SEQ ID NOs: 134 to 178.

2. A nucleic acid molecule encoding the fusion protein according to claim 1.

3. An expression vector comprising the nucleic acid molecule according to claim 2.

4. A recombinant cell into which the expression vector according to claim 3 is introduced. A method for producing a fusion protein comprising a human Lefty A protein variant using the recombinant cell according to claim 4 .

6. Use of a composition for preparing a medicament for preventing and / or treating a neuromuscular disease, the composition comprising a human Lefty A fusion protein variant, wherein the amino acid sequence of the human Lefty A fusion protein variant is SEQ ID NO: 133, and wherein the neuromuscular disease is Charcot-Marie-Stokes disease.

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