CN107793479B - anti-CD 19 antibody and preparation method and application thereof - Google Patents

anti-CD 19 antibody and preparation method and application thereof Download PDF

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CN107793479B
CN107793479B CN201710793590.1A CN201710793590A CN107793479B CN 107793479 B CN107793479 B CN 107793479B CN 201710793590 A CN201710793590 A CN 201710793590A CN 107793479 B CN107793479 B CN 107793479B
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CN107793479A (en
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牟男
张云
马泽龙
袁纪军
曹跃琼
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Shanghai Genbase Biotechnology Co Ltd
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    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0646—Natural killers cells [NK], NKT cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5158—Antigen-pulsed cells, e.g. T-cells
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
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    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
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    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
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    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
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    • C07—ORGANIC CHEMISTRY
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    • C07K2319/00—Fusion polypeptide
    • C07K2319/32—Fusion polypeptide fusions with soluble part of a cell surface receptor, "decoy receptors"
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
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    • C12N2510/00—Genetically modified cells

Abstract

The invention relates to the field of biotechnology, in particular to an anti-CD 19 antibody, a preparation method and application thereof. The invention provides an anti-CD 19 antibody, which comprises a heavy chain variable region and a light chain variable region, wherein the complementarity determining region of the heavy chain variable region comprises CDR-H1 with an amino acid sequence shown as SEQ ID No.1, CDR-H2 with an amino acid sequence shown as SEQ ID No.2 and CDR-H3 with an amino acid sequence shown as SEQ ID No.3, and the complementarity determining region of the light chain variable region comprises CDR-L1 with an amino acid sequence shown as SEQ ID No.4, CDR-L2 with an amino acid sequence shown as SEQ ID No.5, SEQ ID No.6 or SEQ ID No.7 and CDR-L3 with an amino acid sequence shown as SEQ ID No. 8. The present inventors performed affinity maturation screening of FMC63 scFv using phage display technology, resulting in a high affinity single chain antibody to CD 19.

Description

anti-CD 19 antibody and preparation method and application thereof
Technical Field
The invention relates to the field of biotechnology, in particular to an anti-CD 19 antibody, a preparation method and application thereof.
Background
The CD19 antigen (B lymphocyte antigen CD, CD 19) is a protein that is specifically expressed on the surface of human B cells. CD19 is widely expressed on the surface of B cells in various development stages and plays an important role, wherein the CD19 is taken as a co-receptor (co-receptor) of a B Cell Receptor (BCR) and can reduce the signal threshold value required by antigen-mediated B cell receptor-dependent activation (anti-receptor-dependent stimulation); b cell receptor activation relies on phosphorylation of the CD19 intracellular region followed by recruitment of Src kinase and PI3K kinase to fully activate B cells. Fully mature B cells are called plasma cells (plasmacytes), where they lose expression of CD19 antigen after full maturation.
B-cell malignant proliferation-related hematological tumors include: b-cell acute lymphoblastic leukemia (acute lymphoblastic leukemia, B-ALL), B-cell chronic lymphoblastic leukemia (chronic lymphoblastic leukemia, B-CLL), and B-cell lymphoma (B lymphoma). B-cell acute lymphoblastic leukemia and B-cell lymphoma are highly malignant cancers characterized by peripheral blood, bone marrow accompanied by high levels of B-cell malignant proliferation and formation of systemic B-cell solid tumors, severely interfering with the patient's blood circulation system. Today, treatment of B cell tumors mainly includes small molecule targeted drugs such as imatinib (BCR-Abl kinase inhibitor) and ibrutinib (bruton kinase inhibitor), antibody drugs such as rituximab (CD 20 anti) and bone marrow transplantation (bone marrow transplantation); in clinical application, the micromolecular medicaments and the antibody medicaments can obviously prolong the survival time of patients and strive for time for bone marrow transplantation, but partial patients can have drug resistance relapse, and the clinical medicaments are difficult to obtain curative effect for patients with drug resistance relapse.
The chimeric antigen receptor T cell therapy (CART) belongs to a novel treatment mode, is expanded in vitro based on T cells of a patient and is subjected to genetic engineering modification, the CART cells can identify tumor cell specific antigens (such as CD 19), and after activation, the tumor cells can be specifically lysed to achieve the purpose of killing tumors. Chimeric antigen receptor composition is single chain antibody (scFv) -hinge region (transmembrane domain) -transmembrane region (transmembrane domain) -co-stimulatory domain (co-stimulatory domain) -essential stimulatory domain (essential signaling domain), wherein having a highly avid single chain antibody to an antigen is the most important component of Chimeric Antigen Receptor (CAR), the affinity of scFv for an antigen determines whether CART cells can activate and kill tumor cells, and thus screening for affinity maturation of antigen-specific scFv is particularly important.
FMC63-mIgG2a was murine antibody against CD19 obtained by animal immunization in the last century. FMC63 scFv has been successfully applied to anti-CD 19 CAR constructs such as norvanis CTL019 and Juno Therapeutics JCAR015.CTL019 and JCAR015 gave good results in clinical trials of B-cell acute lymphoblastic leukemia, with a complete remission (complete remission) of >70%.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide an anti-CD 19 antibody, a method for preparing the same and use thereof, which are used for solving the problems of the prior art.
To achieve the above and other related objects, in one aspect, the present invention provides an anti-CD 19 antibody comprising a heavy chain variable region and a light chain variable region, the anti-CD 19 antibody having one or more of the following technical features:
<1> the complementarity determining region of the heavy chain variable region includes CDR-H1 having the amino acid sequence shown in SEQ ID No. 1;
<2> complementarity determining regions of the heavy chain variable region include CDR-H2 having the amino acid sequence shown in SEQ ID No. 2;
<3> complementarity determining regions of the heavy chain variable region include CDR-H3 having the amino acid sequence shown in SEQ ID No. 3;
<4> complementarity determining regions of the light chain variable region include CDR-L1 having the amino acid sequence shown in SEQ ID No. 4;
<5> the complementarity determining region of the light chain variable region comprises CDR-L2 having an amino acid sequence as shown in SEQ ID No.5, SEQ ID No.6 or SEQ ID No. 7;
<6> complementarity determining regions of the light chain variable region include CDR-L3 having the amino acid sequence shown in SEQ ID No. 8.
GWSLEDYGVS(SEQ ID No.1)
VIWGSETTYYNSALKS(SEQ ID No.2)
HX 1 YYGGSX 2 X 3 MDY, wherein X 1 ＝R、Y、W；X 2 ＝F、M；X 3 ＝D、E、A(SEQ ID No.3)
RASX 1 X 2 IX 3 X 4 YLN, where X 1 ＝Q、M；X 2 ＝D、P；X 3 ＝R、K；X 4 ＝R、K(SEQ ID No.4)
QTSRLAW(SEQ ID No.5)
QTDRKQS(SEQ ID No.6)
HTSRLHS(SEQ ID No.7)
QQGX 1 X 2 X 3 PX 4 T, where X 1 ＝N、K；X 2 ＝T、A、K、R；X 3 ＝L、A、G；X 4 ＝F、Y(SEQ ID No.8)
CDRs (complementarity determining regions, complementarity determining region) generally refer to regions of an antibody that can be spatially complementary to an epitope. Variability in antibodies is typically not evenly distributed throughout the variable regions of the antibody, and the heavy and light chain variable regions of a monoclonal antibody typically each have 3 hypervariable regions (hypervariable region, HVR) which are typically complementary to an epitope in spatial structure, so that the hypervariable regions are also referred to as complementarity determining regions (complementarity determining region, CDRs), i.e., the heavy chain variable regions typically comprise three complementarity determining regions, i.e., HCDR1, HCDR2, and HCDR3, and the light chain variable regions typically comprise three complementarity determining regions, i.e., LCDR1, LCDR2, and LCDR3.
In certain embodiments of the invention, the complementarity determining regions of the heavy chain variable region of the anti-CD 19 antibody comprise CDR-H1 of the amino acid sequence shown in SEQ ID No.1, CDR-H2 of the amino acid sequence shown in SEQ ID No.2 and CDR-H3 of the amino acid sequence shown in SEQ ID No. 3.
In certain embodiments of the invention, the complementarity determining regions of the light chain variable region of the anti-CD 19 antibody comprise CDR-L1 having the amino acid sequence shown in SEQ ID No.4, CDR-L2 having the amino acid sequence shown in SEQ ID No.5, SEQ ID No.6 or SEQ ID No.7 and CDR-L3 having the amino acid sequence shown in SEQ ID No. 8.
In certain embodiments of the invention, the complementarity determining region of the heavy chain variable region comprises CDR-H1 having the amino acid sequence shown in SEQ ID No.1, CDR-H2 having the amino acid sequence shown in SEQ ID No.2 and CDR-H3 having the amino acid sequence shown in SEQ ID No.3, and the complementarity determining region of the light chain variable region comprises CDR-L1 having the amino acid sequence shown in SEQ ID No.4, CDR-L2 having the amino acid sequence shown in SEQ ID No.5, SEQ ID No.6 or SEQ ID No.7 and CDR-L3 having the amino acid sequence shown in SEQ ID No. 8.
In certain embodiments of the invention, the anti-CD 19 antibody is a monoclonal antibody. Monoclonal antibodies generally refer to a population of antibodies that are substantially identical (except for a few naturally occurring mutations that may be present). Monoclonal antibodies are typically directed against specific determinants on the antigen.
In certain embodiments of the invention, the anti-CD 19 antibody is a single chain Fv (scFv). The single chain antibody may typically be V comprising the antibody H (heavy chain variable region) and V L A polypeptide chain (light chain variable region). In general, single chain antibodies may also include a linker peptide (linker), which is typically located at V H And V L To allow the scFv to form the desired structure for binding to the antigen. For example, the anti-CD 19 antibody may include V H And V L ，V H And V L Can be provided with a connecting peptide, and the single-chain anti-CD 19 antibody can sequentially comprise V from N segment to C end H Linker peptide and V L The anti-CD 19 single strandThe antibody may also include V from N-segment to C-terminal L Linker peptide and V H . The linker peptide may be any of a variety of linker peptides suitable in the art for forming scFv, for example, the linker peptide may be a G4S3 linker, the selection or design of which may be found in references Michel Sadelain etc, science Translational Medicine,2013; carl h.june etc, science Translational Medicine,2015.
In certain embodiments of the invention, the anti-CD 19 antibody is derived from the monoclonal antibody FMC63 (VH: Y14283.1, VL: Y14284.1) specific for CD19, having the nucleotide sequence set forth in SEQ ID No.9 and the amino acid sequence set forth in SEQ ID No. 10.
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.9)
Wherein the underlined parts in bold are the connecting peptides, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order
In certain embodiments of the invention, the heavy and light chain variable regions may further comprise framework regions, which may be located between or at either end of the complementarity determining regions. In a specific embodiment of the present invention, the sequence of the framework region is identical to the framework region sequence of FMC63, or is a framework region sequence obtained by substituting, deleting or adding one or more (specifically, 1 to 50, 1 to 30, 1 to 20, 1 to 10, 1 to 5, or 1 to 3) amino acids to the framework region sequence of FMC63, and the framework region sequence may have homology of 80%, 85%, 90%, 93%, 95%, 97%, or 99% or more to the framework region sequence of FMC 63.
In certain embodiments of the invention, the amino acid sequence of the heavy chain variable region of the anti-CD 19 antibody comprises:
a) An amino acid sequence as shown in one of SEQ ID Nos. 79 to 105; or (b)
b) An amino acid sequence having more than 80% homology with the amino acid sequence shown in one of SEQ ID Nos. 79 to 105 and having the function of the amino acid sequence defined in a).
Specifically, the amino acid sequence in b) specifically refers to: the amino acid sequence shown as one of SEQ ID No.79-105 is obtained by substituting, deleting or adding one or more (specifically, 1-50, 1-30, 1-20, 1-10, 1-5 or 1-3) amino acids, or adding one or more (specifically, 1-50, 1-30, 1-20, 1-10, 1-5 or 1-3) amino acids at the N-terminal and/or C-terminal, and has the amino acid sequence function shown as one of SEQ ID No. 79-105. The amino acid sequence in b) may have 80%, 85%, 90%, 93%, 95%, 97%, or more than 99% homology with one of SEQ ID Nos. 79 to 105.
In certain embodiments of the invention, the amino acid sequence of the light chain variable region of the anti-CD 19 antibody comprises:
c) An amino acid sequence as set forth in one of SEQ ID Nos. 106 to 132; or (b)
d) An amino acid sequence having more than 80% homology with the amino acid sequence shown in one of SEQ ID Nos. 106 to 132 and having the function of the amino acid sequence defined in c).
Specifically, the amino acid sequence in d) specifically refers to: the amino acid sequence shown in one of SEQ ID Nos. 106 to 132 is obtained by substituting, deleting or adding one or more (specifically, 1 to 50, 1 to 30, 1 to 20, 1 to 10, 1 to 5, or 1 to 3) amino acids, or is obtained by adding one or more (specifically, 1 to 50, 1 to 30, 1 to 20, 1 to 10, 1 to 5, or 1 to 3) amino acids at the N-terminal and/or C-terminal and has the amino acid sequence function shown in one of SEQ ID Nos. 106 to 132. The amino acid sequence in b) may have 80%, 85%, 90%, 93%, 95%, 97%, or more than 99% homology with one of SEQ ID Nos. 106 to 132.
In another aspect, the invention provides an isolated polynucleotide encoding the heavy chain variable region and/or the light chain variable region or full length amino acids of the anti-CD 19 antibody.
In another aspect, the invention provides a construct comprising the isolated polynucleotide.
In certain embodiments of the invention, the construct is constructed from the isolated polynucleotide inserted into a multiple cloning site of an expression vector. Expression vectors in the present invention generally refer to a variety of commercially available expression vectors well known in the art, and may be, for example, bacterial plasmids, phage, yeast plasmids, plant cell viruses, mammalian cell viruses such as adenoviruses, retroviruses, or other vectors.
In certain embodiments of the invention, the expression vector is selected from the group consisting of GV401 expression vectors (GV 401 is a commercially available vector, supplier Ji Kai gene).
In another aspect the invention provides an expression system for an antibody, said expression system comprising said construct or said polynucleotide having an exogenous integrated into the genome. Any cell suitable for expression of an expression vector may be used as a host cell, e.g., the host cell may be a prokaryotic cell, such as a bacterial cell; or lower eukaryotic cells, such as yeast cells; or higher eukaryotic cells, such as mammalian cells.
In certain embodiments of the invention, the host cell is selected from one or more of T cells, NK cells.
In another aspect, the invention provides a method for preparing said anti-CD 19 antibody, comprising the steps of: culturing the antibody expression system under conditions suitable for expression of the antibody, thereby expressing the antibody, and purifying and isolating the antibody.
The host cells used in the present invention are all of the prior art, and are commercially available, and the medium used in the culture is also various conventional media, and a person skilled in the art can empirically select a suitable medium to culture under conditions suitable for the growth of the host cells. After the host cells have grown to the appropriate cell density, the selected promoters are induced by suitable means (e.g., temperature switching or chemical induction) and the cells are cultured for an additional period of time. The recombinant polypeptide in the above method may be expressed in a cell, or on a cell membrane, or secreted outside the cell. If desired, the recombinant proteins can be isolated and purified by various separation methods using their physical, chemical and other properties. Such methods are well known to those skilled in the art. Examples of such methods include, but are not limited to: conventional renaturation treatment, treatment with a protein precipitant (salting-out method), centrifugation, osmotic sterilization, super-treatment, super-centrifugation, molecular sieve chromatography (gel filtration), adsorption chromatography, ion exchange chromatography, high Performance Liquid Chromatography (HPLC), and other various liquid chromatography techniques and combinations of these methods.
In another aspect, the invention provides the use of the anti-CD 19 antibody in the preparation or screening of therapeutic agents, or in the preparation of diagnostic agents.
The therapeutic agent may be an agent that targets the CD-19 antigen, binds to or acts on the CD-19 antigen, thereby treating and/or preventing an indication.
In certain embodiments of the invention, the therapeutic agent may be a tumor therapeutic agent. The tumor therapeutic drug can be a drug which takes CD-19 antigen functionally expressed on the surface of tumor cells as a target and binds or acts on the CD-19 antigen so as to treat and/or prevent tumors. The tumor may be acute lymphoblastic leukemia, chronic lymphoblastic leukemia, B-lymphoma, or other tumors associated with malignant proliferation of B cells.
In certain embodiments of the invention, the therapeutic agent is a chimeric antigen receptor (CAR, chimeric antigen receptor) cell therapeutic agent.
The chimeric antigen receptor cell therapeutic drug generally includes a chimeric antigen receptor cell, which may be a chimeric antigen receptor T cell, a chimeric antigen receptor NK cell, or the like, which generally includes a T lymphocyte, which also includes a chimeric antigen receptor. The chimeric antigen receptor NK cells generally include NK cells, which also include chimeric antigen receptors. The chimeric antigen receptor includes a transmembrane domain, an intracellular domain, and an extracellular domain. In certain embodiments of the invention, the extracellular domain comprises the anti-CD 19 antibody, i.e., the chimeric antigen receptor cell can express the anti-CD 19 antibody on the cell surface, thereby directing the cell to act on cells expressing the CD19 antigen (e.g., tumor cells). The action on the cells expressing the CD19 antigen may be killing the cells expressing the CD19 antigen, or the like.
Therefore, the diagnosis medicine specifically refers to a reagent for diagnosing the action target CD19 antigen by taking the CD19 antigen as a biomarker.
In another aspect, the invention provides an isolated polypeptide comprising a transmembrane domain, an intracellular domain, and an extracellular domain comprising the anti-CD 19 antibody.
In certain embodiments of the invention, the polypeptide is a chimeric antigen receptor.
In some embodiments of the invention, the transmembrane domain may include CD8 a (NM-001145873), CD28 (NM-006139), DAP10 (NM-014266), and the like.
In certain embodiments of the invention, the intracellular domain may comprise a co-stimulatory domain and/or a signaling domain, e.g., the intracellular domain may comprise 4-1BB (NM_001561), CD28 (NM_006139), OX40 (NM_ 003327), ICOS (NM_ 012092), CD3zeta (NM_ 198053), DAP10 (NM_ 014266), and the like.
In certain embodiments of the invention, the polypeptide comprises the anti-CD 19 single chain antibody, a transmembrane domain, and an intracellular domain in order from the N-terminus to the C-terminus. In some embodiments of the invention, the polypeptide comprises the anti-CD 19 single chain antibody, a CD 8. Alpha. Transmembrane region, a 4-1BB costimulatory domain, and a CD3zeta signaling domain in that order from the N-terminus to the C-terminus. In a specific embodiment of the present invention, the polypeptide comprises the anti-CD 19 single chain antibody, CD28 transmembrane region, CD28 co-stimulatory domain, CD3zeta signaling domain, in order from N-terminus to C-terminus. In another embodiment of the present invention, the polypeptide comprises the anti-CD 19 single chain antibody, a CD8 a transmembrane region, an OX40 co-stimulatory domain, a CD3zeta signaling domain, in order from the N-terminus to the C-terminus. In another embodiment of the invention, the polypeptide comprises the anti-CD 19 single chain antibody, a CD8 a transmembrane region, an ICOS costimulatory domain, a CD3zeta signaling domain in order from the N-terminus to the C-terminus. In another embodiment of the present invention, the polypeptide comprises the anti-CD 19 single chain antibody, CD 8. Alpha. Transmembrane region, 4-1BB co-stimulatory domain, CD28 co-stimulatory domain, CD3zeta, in order from N-terminus to C-terminus. In another embodiment of the present invention, the polypeptide comprises the anti-CD 19 single chain antibody, CD28 transmembrane region, CD28 co-stimulatory domain, OX40 co-stimulatory domain, CD3zeta signaling domain, in order from N-terminus to C-terminus.
In another aspect of the invention, a T lymphocyte comprises the polypeptide membrane bound thereto.
In certain embodiments of the invention, the polypeptide is a chimeric antigen receptor.
The T lymphocytes may typically express the polypeptide, which may typically bind to a CD19 antigen, more particularly may bind to a CD19 antigen via an extracellular domain comprising the anti-CD 19 antibody, and when the polypeptide binds to the CD19 antigen, the T lymphocytes may typically be activated and/or stimulated to proliferate. In certain embodiments of the invention, the extracellular domain comprises the anti-CD 19 antibody, i.e., the chimeric antigen receptor T cell can express the anti-CD 19 antibody on the surface of a T lymphocyte, thereby directing the T lymphocyte to act on cells expressing the CD19 antigen (e.g., tumor cells), which may be killing cells expressing the CD19 antigen, etc.
In another aspect of the invention, an NK cell contains the polypeptide membrane bound.
In certain embodiments of the invention, the polypeptide is a chimeric antigen receptor.
The NK cells may generally express the polypeptide, which may generally bind to the CD19 antigen, more particularly may bind to the CD19 antigen via an extracellular domain comprising the anti-CD 19 antibody, and when the polypeptide binds to the antigen, the NK cells may generally be activated and/or stimulated to proliferate. In certain embodiments of the invention, the extracellular domain comprises the anti-CD 19 antibody, i.e., the chimeric antigen receptor NK cells can express the anti-CD 19 antibody on the surface of NK cells, thereby allowing for the induction of an effect by NK cells on cells expressing the CD19 antigen (e.g., tumor cells), which can be killing of cells expressing the CD19 antigen, etc.
In another aspect, the invention provides a diagnostic kit comprising a diagnostically effective dose of said anti-CD 19 antibody or immunoconjugate thereof. An effective amount generally refers to an amount that provides a diagnostic benefit.
The diagnostic kit can generally diagnose the action target CD19 antigen with the CD19 antigen as a biomarker. The diagnostic kit may also include a label for the anti-CD 19 antibody, which may generally be used to label the anti-CD 19 antibody, optionally in a variety of labels including, but not limited to, fluorescent labels, radioactive labels, enzyme-labeled labels, chemiluminescent labels, and the like. The kit may also typically comprise one or more reagents required for detection, depending on the detection principle of the kit. In addition, the kit may further include, as required: containers, controls (negative or positive controls), buffers, adjuvants, etc., which can be selected by the skilled artisan according to the particular circumstances.
The present inventors performed affinity maturation screening on FMC63 scFv using phage display technology, resulting in high affinity single chain antibodies to CD19 that were able to competitively block the binding of wild type FMC63 mIgG2a to CD19, suggesting that the antigen binding sites of these single chain antibodies are identical to FMC63 mIgG2 a. Furthermore, the present inventors have further engineered high affinity single chain antibodies into chimeric antigen receptors, for example, using T cells, NK cells expressing anti-CD 19 chimeric antigen receptor for the treatment of hematological cancers (B-ALL, B-CLL, B-Lymphoma, etc.) expressing CD19, thereby verifying that the mutated chimeric antigen receptor is capable of improving the killing ability against tumor cells.
Drawings
FIGS. 1-5 are schematic illustrations of experimental results of example 3 of the present invention.
FIGS. 6-10 are schematic diagrams showing experimental results of example 4 of the present invention.
FIGS. 11-14 are schematic illustrations of experimental results of example 6 of the present invention.
Fig. 15 shows a schematic view of GV400 carrier structure.
Fig. 16 shows a schematic view of GV401 carrier structure.
FIGS. 17-19 are schematic diagrams showing experimental results of example 8 of the present invention.
FIG. 20 is a schematic diagram showing the experimental results of example 8 of the present invention.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention.
Before the embodiments of the invention are explained in further detail, it is to be understood that the invention is not limited in its scope to the particular embodiments described below; it is also to be understood that the terminology used in the examples of the invention is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the invention; in the description and claims of the invention, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
Where numerical ranges are provided in the examples, it is understood that unless otherwise stated herein, both endpoints of each numerical range and any number between the two endpoints are significant both in the numerical range. Unless defined otherwise, 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. In addition to the specific methods, devices, materials used in the embodiments, any methods, devices, and materials of the prior art similar or equivalent to those described in the embodiments of the present invention may be used to practice the present invention according to the knowledge of one skilled in the art and the description of the present invention.
Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed in the present invention employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA techniques, and related arts. These techniques are well described in the prior art literature and see, in particular, sambrook et al MOLECULAR CLONING: a LABORATORY MANUAL, second edition, cold Spring Harbor Laboratory Press,1989and Third edition,2001; ausubel et al, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, john Wiley & Sons, new York,1987and periodic updates; the series METHODS IN ENZYMOLOGY, academic Press, san Diego; wolffe, CHROMATIN STRUCTURE AND FUNCTION, third edition, academic Press, san Diego,1998; METHODS IN ENZYMOLOGY, vol.304, chromatin (p.m. wassman and a.p. wolffe, eds.), academic Press, san Diego,1999; and METHODS IN MOLECULAR BIOLOGY, vol.119, chromatin Protocols (p.b. becker, ed.) Humana Press, totowa,1999, etc.
Example 1
Construction of CDR1, CDR2, CDR3 mutant libraries of heavy chain (H) and light chain (L) of FMC63 scFv:
the selection of the FMC63 heavy and light chain CDR regions is based on amino acid counting of the variable regions (template monoclonal antibody FMC63, VH: Y14283.1, VL: Y14284.1), kabat counting (Kabat number scheme.Bioinf. Org. Uk).
The pCAN-FMC63 scFv plasmid (constructed from a multiple cloning site with the template sequence inserted into the pCANTAB 5E plasmid (purchased from GE) and the template sequence VH: Y14283.1, VL: Y14284.1) was used as template to introduce mutations by PCR using random primers as shown in Table 1. The obtained CDR1, CDR2, CDR3 mutant library PCR products of the heavy chain (H) and light chain (L) of FMC63 scFv were designated as H1, H2, H3, L1, L2 and L3, respectively. The PCR product was digested with Sfi I and Not I, recovered, and then ligated overnight at 16℃with T4 DNA ligase. Electrotransformation of the connection product into TG1 competent cells, resuspension of the 2XYT culture medium, resuscitating at 37 ℃ for 1h, and performing plate counting by taking bacterial liquid gradient dilution to obtain at least 10 mutant libraries 8 All the remaining bacterial solutions were plated on 2XYT (GA) plates (glucose 2%, penicillin 100 ug/ml). And randomly picking 20 monoclone from the mutation library to send sequence, wherein the diversity is 100%.
TABLE 1
Example 2
Panning of phage antibody library:
20nM of CD19-his-biotin antigen was added and incubated with phage antibody library at room temperature for 2h, and the mixture was transferred to streptavidin beads for 15min at room temperature. The unbound phage was washed off with PBST-PBS, and pancreatin was added thereto for 30min at 37℃to elute the bound phage. Infecting 4ml of logarithmic phase TG1 thalli with phage eluted by pancreatin digestion, standing at 37 ℃ for 30min, taking part of bacterial liquid for gradient dilution for plate counting, and coating all other bacterial liquid on a 2XYT (GA) plate for the next round of packaging. The packaged phage were used for the next round of panning, with 4 rounds of panning enrichment, each round of panning with a 10-fold dilution gradient decreasing antigen concentration, and increasing PBST-PBS wash times from round to round (20 nM, 2nM, 0.2nM and 0.02nM for CD19 his-biotin antigen concentration, and 7, 10, 15, 20 times for PBST-PBS wash, respectively).
Example 3
Screening and identification of high affinity scFv: after four rounds of panning, single clone is randomly selected, supernatant is selected for ELISA after IPTG induction, after ELISA preliminary screening, clone with positive signal at least 2 times larger than negative signal is selected for sequencing, sequencing result is analyzed, and clone corresponding to CDR region with more enrichment is extracted. The ELISA primary screening comprises the following specific steps:
Colonies were picked and cloned in 4ml 2XYT medium, cultured overnight at 37℃at 200rpm, at 1:100 is transferred into 500ml of 2XYT culture medium, and cultured to logarithmic phase at 37 ℃ and 200 rpm; M13K07 helper phage (moi=20) was added, incubated at 37 ℃ for 30 min at 200rpm after incubation at 37 ℃ for 30 min; the supernatant was removed by centrifugation and the cells were resuspended in 500ml of 2XYT/Kan/Amp resistant medium and cultured overnight; the supernatant from overnight incubation was precipitated with 1/5 volume of PEG/NaCl solution, allowed to stand for 1 hour, centrifuged, and the precipitate was resuspended in 30% glycerol in PBS and stored at 4 ℃. The phage solution was diluted 2-10 times and A268 was assayed, and the specific results are shown in Table 2 (in Table 2, clone numbers represent the numbers of picked colony clones).
Avidin was coated onto 96-well plates, biotin-labeled CD19 antigen was bound to 96-well plates, incubated for 1 hour after addition of gradient diluted phage (original concentration of phage before dilution a268, see table 2), and the phage bound CD19 was detected with Anti-phage antibody (Anti M13 phage antibody, purchased by GE corporation), and OD450nm was read, with the results shown in table 3: the smaller the EC50 (a 268) (EC 50 indicates phage concentration corresponding to 50% saturation signal, EC50 (a 268) in table 3 indicates that absorbance of a268 corresponding to EC50 is proportional to phage concentration), the greater the dilution of the corresponding clone required to reach the same saturation signal, i.e., the greater the corresponding clone binding capacity (a 268 measured by Nanodrop (thermofsher)); the positive control was FMC63WT phage, the negative control was phage expressing anti-non-CD 19 single chain antibodies, and the method of use was the same as the other clones). The specific results are shown in FIGS. 1-5.
TABLE 2
Cloning A268
3H4 22.36
3C2 13.04
3C11 11.48
4E10 29.2
3F12 8.88
3H9 8.36
5A12 6.00
8C4 11.16
5G6 9.64
5A1 17.4
6D5 17.36
8A1 13.00
8B6 10.20
8B11 12.84
8C1 14.60
8C7 12.52
8C10 11.08
8D12 53.24
8F12 26.0
Positive control (FMC 63 WT) 71.44
Negative control 77.76
TABLE 3 Table 3
Clone number EC50(A268)
3H4 0.60
3C2 0.03
3C11 0.11
4E10 0.46
3F12 0.24
3H9 0.10
5A12 0.21
8C4 0.27
5G6 0.13
8C7 0.03
8B6 0.02
8C10 0.03
5A1 0.04
8B11 0.01
8D12 0.27
8A1 0.06
6D5 0.11
8C1 0.07
8F12 0.12
Positive control (FMC 63 WT) 7.14
Negative control 19.44
Example 4
FMC63 scFv affinity competition ELISA screening:
the CDR mutations were repeated more than in TG1 clone plates, monoclonal was picked, IPTG induced and the purified phase was purified, after concentration measurement (A268=5.4 diluted in a 3-fold concentration gradient), the dilution gradient was competitive with 3ug/ml FMC63 mIgG2a (well plate: avidin was coated onto 96 well plate, biotin-labeled CD19 antigen was bound to 96 well plate), anti-mouse Fc secondary antibody was used for detection, 5mins was incubated after addition of substrate (TMB), 1M sulfuric acid was used to terminate the reaction and OD450nm was read, and specific results are shown in Table 4, FIGS. 6-10. The EC50 and Emax (A268 determined by Nanodrop (thermoshier)) of the different clones were compared, the positive control was FMC63WT phage, the negative control was phage expressing anti-non-CD 19 single chain antibody, and the method of use was the same as the other clones. The smaller the EC50 (a 268) (EC 50 indicates the phage concentration corresponding to 50% of the maximum inhibition Emax, and EC50 (a 268 indicates the absorbance of a268 corresponding to EC 50) in table 4) represents the stronger the single chain antibody of the corresponding clone competes for FMC63 mIgG2a to 50% binding; the larger Emax (maximum inhibition, the percentage of all cloned phages that were able to compete for the binding capacity of the wild-type antibody at an absorbance of 5.4) represents the more competitive a single chain antibody of the corresponding clone at the same A268 concentration.
TABLE 4 Table 4
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Based on the above results, 3H4,3C2,3C11,4E10,5A12,8C4,8B6 was selected for the combinatorial mutation test (HCDR 2 enriched sequence was not so much, so even 4E10 was poorly competitive, but still selected).
Example 5
FMC63 scFv mutant Kd/Koff assay:
the FMC63 scFv mutant (the clone numbers corresponding to the mutants and the nucleotide sequence and amino acid sequence of scFv corresponding to the clone numbers are shown in the following, SEQ ID No.25-51 and SEQ ID No. 52-78) is transferred to pGCIgGH1 (Ji Kai gene) (the vector sequence is shown as SEQ ID No. 133), so that a secretion type FMC63 scFv mutant hIgG1 Fc fusion protein which can be expressed is constructed, and the eukaryotic expression system is utilized for expression; purification was performed using protein A column.
The CD19 antigen was coupled to an AR2G chip (Fortie Bio) via amino groups, FMC63 scFv mutants as mobile phase, and the corresponding scFv Koff rates were determined using the Octet Red96 system, and the test results showed (table 5) that all of the scFv mutants had good affinity for CD19 antigen.
TABLE 5
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The above results indicate that each clone possesses a Koff value that is substantially identical to FMC63 WT.
Example 6
Obtaining high affinity scFv:
referring to ELISA binding, competition and Koff test data in examples 1-5, clones were selected for alignment and combination of corresponding CDRs by PCR, and the results of the combination were as follows (clone numbers corresponding to scFv obtained after the combination, and nucleotide sequences and amino acid sequences of scFv corresponding to clone numbers are shown below, and CDRs of the combined scFv sequences were selected from SEQ ID Nos. 25-51 and SEQ ID Nos. 52-78), and screening was performed by ELISA experiments for binding scFv to CD19 contained in E.coli periplasm (PPE). FMC63 mutant library combinations were as follows:
TABLE 6
Heavy chain Combos CDR-H1 CDR-H2 CDR-H3
Combo5 3H4 4E10 3C2
Combo6 3H4 FMC63 WT 3C2
Combo7 3H4 4E10 3C11
Combo8 3H4 FMC63 WT 3C11
Light chain Combos CDR-L1 CDR-L2 CDR-L3
Combo9 5A12 5G6 8B6
Combo10 5A12 FMC63 WT 8B6
Combo13 8C4 5G6 8B6
Combo14 8C4 FMC63 WT 8B6
E.coli periplasm (PPE) preparation:
the TG1 clone with more repeated CDR mutation is coated, monoclonal is selected, the strain is collected after IPTG induction and resuspended in TES (0.2M Tris-HC1,0.5mM EDTA,0.5M sucrose), 1/5 XTES is added to be mixed uniformly, the mixture is placed on ice for 30min, the supernatant is obtained by centrifugation at 13000rpm, and the supernatant is filtered by a 0.22 mu M filter membrane and stored at 4 ℃. The peripheral materials were diluted in a certain ratio (2, 4,8, 16, 32, 64, 128) and added to a CD 19-coated 96-well plate, and ELISA (experimental method reference example 4) was performed, and the results are shown in FIGS. 11 to 14. As can be seen from the experimental results, combo5, combo7, combo9, combo10, combo14 bound poorly to CD19-his, combo6, combo8, combo11, combo13 bound strongly to CD19 (with higher OD450nm light absorption at low dilution) by PPE dilution binding ELISA, with combo6 binding highest.
Clone number: 3H4
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGTGGTCATTAGAGGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.25)
(SEQ ID No.52, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.79)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.106)
Clone number: 3C2
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATCGGTACTACGGTGGTAGCTTTGATATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.26)
(SEQ ID No.53, wherein the bold underlined section is a connecting peptide, even The heavy chain variable region is before the peptide connection, the light chain variable region is after the peptide connection, and the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 in sequence
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHRYYGGSFDMDYWGQGTSVTVSS(SEQ ID No.80)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.107)
Clone number: 3C11
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTGGTACTACGGTGGTAGCTTTGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.27)
(SEQ ID No.54, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHWYYGGSFAMDYWGQGTSVTVSS(SEQ ID No.81)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.108)
Clone number: 4E10
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGGGATAATTGGTAGTCAGACCACATACTATCTGGCTGGGCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.28)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSGGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT (SEQ ID No.55, wherein the underlined parts in bold are the connecting peptides, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.82)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.109)
Clone number: 3F12
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGACTGATACAGGGTAGTTGGCATACATACTATGGTGCGGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.29)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGLIQGSWHTYYGAALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSGGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT (SEQ ID No.56, wherein the underlined parts in bold are the connecting peptides, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGLIQGSWHTYYGAALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.83)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.110)
Clone number: 8F12
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCAGACAGATAGAAAGCAGTCGGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.30)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSGGGGSGGGGSGGGGSDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYQTDRKQSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT (SEQ ID No.57, wherein the underlined parts in bold are the connecting peptides, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.84)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYQTDRKQSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.111)
Clone number: 3H9
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTATTACGGTGGTAGCATGGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.31)
(SEQ ID No.58, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSMAMDYWGQGTSVTVSS(SEQ ID No.85)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.112)
Clone number: 5A12
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGCCTATTCGGAGGTATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.32)
(SEQ ID No.59, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.86)
DIQMTQTTSSLSASLGDRVTISCRASQPIRRYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.113)
Clone number: 8C4
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTATGGACATTAAGAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.33)
(SEQ ID No.60, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.87)
DIQMTQTTSSLSASLGDRVTISCRASMDIKKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.114)
Clone number: 5G6
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCAGACATCAAGATTAGCCTGGGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.34)
(SEQ ID No.61, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.88)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYQTSRLAWGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.115)
Clone number: 8C7
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGGCGGCTCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.35)
(SEQ ID No.62, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.89)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKAAPLTFGGGTKLEIT(SEQ ID No.116)
Clone number: 8B6
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCATCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGGAAGAAGGCGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.36)
(SEQ ID No.63, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.90)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.117)
Clone number: 8C10
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGAAGGGGCCGCTTACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.37)
(SEQ ID No.64, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIK DNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.91)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDY SLTISNLEQEDIATYFCQQGKKGPLTFGGGTKLEIT(SEQ ID No.118)
Clone number: 5A1
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGAGGGCGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.38)
(SEQ ID No.65, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.92)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKRAPLTFGGGTKLEIT(SEQ ID No.119)
Clone number: 8B11
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCGTCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGAGGGGGCCGTTGACTTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.39)
(SEQ ID No.66, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.93)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKRGPLTFGGGTKLEIT(SEQ ID No.120)
Clone number: 8D12
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGACGGCTCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.40)
(SEQ ID No.67, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the CDR-H1, the CDR in this order -H2、CDR-H3、CDR-L1、CDR-L2、CDR-L3)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.94)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKTAPLTFGGGTKLEIT(SEQ ID No.121)
Clone number: 8A1
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGACGGGGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.41)
(SEQ ID No.68, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.95)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKTGPLTFGGGTKLEIT(SEQ ID No.122)
Clone number: 6D5
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAAGACGCTTCCGTTTACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.42)
(SEQ ID No.69, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.96)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKTLPFTFGGGTKLEIT(SEQ ID No.123)
Clone number: 8C1
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATAAGGCGCCGCTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.43)
(SEQ ID No.70, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.97)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNKAPLTFGGGTKLEIT(SEQ ID No.124)
Clone number: combo-5
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGTGGTCATTAGAGGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGGGATAATTGGTAGTCAGACCACATACTATCTGGCTGGGCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATCGGTACTACGGTGGTAGCTTTGATATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.44)
(SEQ ID No.71, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHRYYGGSFDMDYWGQGTSVTVSS(SEQ ID No.98)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.125)
Clone number: combo-6
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGTGGTCATTAGAGGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATCGGTACTACGGTGGTAGCTTTGATATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.45)
(SEQ ID No.72, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHRYYGGSFDMDYWGQGTSVTVSS(SEQ ID No.99)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.126)
Clone number: combo-7
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGTGGTCATTAGAGGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGGGATAATTGGTAGTCAGACCACATACTATCTGGCTGGGCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTGGTACTACGGTGGTAGCTTTGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.46)
(SEQ ID No.73, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHWYYGGSFAMDYWGQGTSVTVSS(SEQ ID No.100)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.127)
Clone number: combo-8
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGTGGTCATTAGAGGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTGGTACTACGGTGGTAGCTTTGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.47)
(SEQ ID No.74, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHWYYGGSFAMDYWGQGTSVTVSS(SEQ ID No.101)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.128)
Clone number: combo-9
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGCCTATTCGGAGGTATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCAGACATCAAGATTAGCCTGGGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGGAAGAAGGCGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.48)
(SEQ ID No.75, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.102)
DIQMTQTTSSLSASLGDRVTISCRASQPIRRYLNWYQQKPDGTVKLLIYQTSRLAWGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.129)
Clone number: combo-10
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGCCTATTCGGAGGTATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGGAAGAAGGCGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.49)
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(SEQ ID No.76, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.103)
DIQMTQTTSSLSASLGDRVTISCRASQPIRRYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.130)
Clone number: combo-13
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTATGGACATTAAGAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCAGACATCAAGATTAGCCTGGGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGGAAGAAGGCGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.50)
(SEQ ID No.77, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.104)
DIQMTQTTSSLSASLGDRVTISCRASMDIKKYLNWYQQKPDGTVKLLIYQTSRLAWGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.131)
Clone number: combo-14
GAGGTGAAACTGCAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAAGCTGGATTCGCCAGTCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAATTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGCAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGGGCCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTATGGACATTAAGAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGGAAGAAGGCGCCGTTGACGTTCGGAGGGGGGACCAAGCTGGAGATCACA(SEQ ID No.51)
(SEQ ID No.78, wherein the underlined parts in bold are the connecting peptide, the heavy chain variable region before the connecting peptide, the light chain variable region after the connecting peptide, the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in this order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.105)
DIQMTQTTSSLSASLGDRVTISCRASMDIKKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.132)
Vector pGCIgGH1 full sequence:
AgtgGGAATTGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTGTCGTGAGGAAGATCTCTAGAAGCTGGGTACCTTGTGCCCGGGCGCCACCATGGAGTTTGGGCTGAGCTGGCTTTTTCTTGTCGCGATTCTTAAGGGTGTCCAGTGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGAGCGGCCGCTCGAGGCCGGCAAGGCCGGATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTATGGCTGATTATGATCCGGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGATACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGAGGTCGACTCTAGAGGATCGATCCCCGCCCCGGACGAACTAAACCTGACTACGACATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCTTCAGTTGGTTGGTACAACTTGCCAACTGGGCCCTGTTCCACATGTGACACGGGGGGGGACCAAACACAAAGGGGTTCTCTGACTGTAGTTGACATCCTTATAAATGGATGTGCACATTTGCCAACACTGAGTGGCTTTCATCCTGGAGCAGACTTTGCAGTCTGTGGACTGCAACACAACATTGCCTTTATGTGTAACTCTTGGCTGAAGCTCTTACACCAATGCTGGGGGACATGTACCTCCCAGGGGCCCAGGAAGACTACGGGAGGCTACACCAACGTCAATCAGAGGGGCCTGTGTAGCTACCGATAAGCGGACCCTCAAGAGGGCATTAGCAATAGTGTTTATAAGGCCCCCTTGTTAACCCTAAACGGGTAGCATATGCTTCCCGGGTAGTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGCTATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATATCTCCCACCCCATGAGCTGTCACGGTTTTATTTACATGGGGTCAGGATTCCACGAGGGTAGTGAACCATTTTAGTCACAAGGGCAGTGGCTGAAGATCAAGGAGCGGGCAGTGAACTCTCCTGAATCTTCGCCTGCTTCTTCATTCTCCTTCGTTTAGCTAATAGAATAACTGCTGAGTTGTGAACAGTAAGGTGTATGTGAGGTGCTCGAAAACAAGGTTTCAGGTGACGCCCCCAGAATAAAATTTGGACGGGGGGTTCAGTGGTGGCATTGTGCTATGACACCAATATAACCCTCACAAACCCCTTGGGCAATAAATACTAGTGTAGGAATGAAACATTCTGAATATCTTTAACAATAGAAATCCATGGGGTGGGGACAAGCCGTAAAGACTGGATGTCCATCTCACACGAATTTATGGCTATGGGCAACACATAATCCTAGTGCAATATGATACTGGGGTTATTAAGATGTGTCCCAGGCAGGGACCAAGACAGGTGAACCATGTTGTTACACTCTATTTGTAACAAGGGGAAAGAGAGTGGACGCCGACAGCAGCGGACTCCACTGGTTGTCTCTAACACCCCCGAAAATTAAACGGGGCTCCACGCCAATGGGGCCCATAAACAAAGACAAGTGGCCACTCTTTTTTTTGAAATTGTGGAGTGGGGGCACGCGTCAGCCCCCACACGCCGCCCTGCGGTTTTGGACTGTAAAATAAGGGTGTAATAACTTGGCTGATTGTAACCCCGCTAACCACTGCGGTCAAACCACTTGCCCACAAAACCACTAATGGCACCCCGGGGAATACCTGCATAAGTAGGTGGGCGGGCCAAGATAGGGGCGCGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTTGGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCATGCATATACAGTCAGCATATGATACCCAGTAGTAGAGTGGGAGTGCTATCCTTTGCATATGCCGCCACCTCCCAAGGGGGCGTGAATTTTCGCTGCTTGTCCTTTTCCTGCTGCTTATCGATGATAAGCTGTCAAACATGAGAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGAAGCTGTCCCTGATGGTCGTCATCTACCTGCCTGGACAGCATGGCCTGCAACGCGGGCATCCCGATGCCGCCGGAAGCGAGAAGAATCATAATGGGGAAGGCCATCCAGCCTCGCGTCGCGAACGCCAGCAAGACGTAGCCCAGCGCGTCGGCCCCGAGATGCGCCGCGTGCGGCTGCTGGAGATGGCGGACGCGATGGATATGTTCTGCCAAGGGTTGGTTTGCGCATTCACAGTTCTCCGCAAGAATTGATTGGCTCCAATTCTTGGAGTGGTGAATCCGTTAGCGAGGTGCCGCCCTGCTTCATCCCCGTGGCCCGTTGCTCGCGTTTGCTGGCGGTGTCACTGGCCCCGTGGGTTAGGGACGGGGTCCCCCATGGGGAATGGTTTATGGTTCGTGGGGGTTATTATTTTGGGCGTTGCGTGGGGTCAGGTCCACGACTGGACTGAGCAGACAGACCCATGGTTTTTGGATGGCCTGGGCATGGACCGCATGTACTGGCGCGACACGAACACCGGGCGTCTGTGGCTGCCAAACACCCCCGACCCCCAAAAACCACCGCGCGGATTTCTGGCGTGCCAAGCTAGTCGACCAATTCTCATGTTTGACAGCTTATCATCGCAGATCCGGGCAACGTTGTTGCCATTGCTGCAGGCGCAGAACTGGTAGGTATGGAAGATCT(SEQ ID No.133)
example 7
High affinity scFv was engineered to CAR:
construction of scFv-BBz vector:
referring to the scFv sequence shown in SEQ ID No.25-132, the chimeric antigen receptor of FMC63-BBz is mutated according to amino acids mutated in CDRs, specifically, the scFv sequence is connected with CD8 alpha-4-1 BB-CD3zeta by a PCR method to obtain a mutated chimeric antigen receptor of FMC 63-Mutant-BBz series, scFv-BBz is inserted into GV400 vector (between BamHI and EcoRI, ji Kai genes, vector schematic diagram is shown in FIG. 15) by a standard molecular biology method, and cloning is selected as follows: 3F12,5A12,5G6,8C4,8F12,8D12 and Combo6.
Construction of scFv-BBz-P2A-EGFP vector:
the scFv-BBz mutant obtained after the mutation was inserted between EF-1 promoter and P2A structure (ATNFSLLKQAGDVEENPGP, SEQ ID No. 134) in GV401 vector (between BamHI and EcoRI, ji Kai gene, vector schematic shown in FIG. 16) by standard molecular biology method.
Example 8
FMC 63-Mutant-BBz in vitro verification
After FMC 63-BBz obtained by constructing each scFv sequence is packaged into lentiviruses (Lentivirus), primary PBMC of human beings are infected; the infected T cells are expanded and cultured in vitro, and whether the FMC63 antigen can enhance the function of T cells in killing tumor cells is evaluated through cytokine release and killing experiments, and the specific results are as follows:
infection efficiency detection:
after the GV401 shuttle vector is packaged into lentiviruses, the human T cells are infected, scFv-BBz-2A-EGFP is transcribed into mRNA, and is translated through a ribosome, and two independent protein chains are formed by cleavage between glycine and auxiliary amino acid at the tail end of a 2A sequence (ATNFSLLKQAGDVEENPGP, SEQ ID No. 135), so that the expression mole number of scFv-BBz and the EGFP protein are kept consistent, and the expression efficiency of the EGFP can be detected by detecting a FITC channel through a flow cytometer to represent the infection efficiency of the lentiviruses.
Cytokine release assay:
human PBMC were stimulated with CD3 and CD28 antibodies (OKT 3 clone and 15E8 clone, miltenyi Biotec) for 24 hours, FMC 63-Mutant-BBz obtained by construction of the respective scFv sequences was packaged into lentiviruses (Lentivirus) and then the PBMC were infected, the anti-CD 19 chimeric antigen receptor was expressed, and after the PBMC continued to be cultured for 8-10 days, T cells were collected and T cells and Raji cells (expressing CD 19) were cultured in accordance with the ratio 1:1 Mixed culture in 2% serum RPMI1640 medium for 16 hours the mixed culture supernatants were assayed for cytokine release using BD Cytometric bead array kit. Experiments show that PBMC obtained by infection (FMC 63 Mutant-BBz obtained by constructing scFv sequences is packaged into lentivirus for infection, and the chimeric antigen receptor of FMC63 Mutant-BBz is expressed) have stronger binding capacity to Raji cells and can release cytokines compared with FMC63 WT-BBz. The infection efficiency of each clone was as follows:
TABLE 7
Cloning Infection efficiency%
FMC63 WT 26.9％
3F12 62.8％
5A12 26.6％
5G6 58.5％
8C4 55.9％
8F12 55.0％
8D12 65.7％
Combo6 15.6％
Fold change in cytokine release for each clone was as follows:
TABLE 8
Cloning IL2 multiple IFN gamma multiple
FMC63 WT 106 126
Combo6 146 158
TABLE 9
Cloning IL2 multiple IFN gamma multiple
FMC63 WT 46 16
5A12 47 41
8D12 3 7
3F12 1 1
8F12 32 15
Table 10
Cloning IL2 multiple IFN gamma multiple
FMC63 WT 123 109
5G6 87 107
8C4 73 98
The cytokine release experimental results show that the 8D12 and 3F12 clones have no significant cytokine release for tumor cell stimulation, suggesting that the 8D12 and 3F12 clone mutations are unfavorable for T cell binding to CD19 expressing tumor cells.
The 5a12 clone released higher levels of cytokines IL2 and ifnγ at an infection efficiency (26.6% vs 26.9%) substantially consistent with FMC63 WT.
Combo6 released significantly higher levels of IL2 than FMC63 WT at lower infection efficiency.
It was suggested that 5a12 (CDRL 1), 3H4 (CDRH 1), 3C2 (CDRH 3) mutations favoured increased cytokine release levels for CD19 binding.
Tumor killing experiments:
t cells (PBMC were collected after further culturing for 8-10 days) and Raji cells obtained as described above were cultured according to a certain E: t ratio, e.g. 30:1,10: 1,3:1,1:1 ratio in RPMI1640 medium with 2% serum for 4 hours, the culture supernatant was mixed with LDH substrate (CytoTox 96 Non-Radioactive Cytotoxicity Assay Kit, promega) 1:1 volume of the mixture, after incubation at room temperature for 30 minutes, 490nm light absorption was read. Experimental results show that PBMC obtained by infection (FMC 63 Mutant-BBz obtained by constructing scFv sequences is packaged into lentivirus infection) have strong tumor killing capability compared with FMC63 WT-BBz.
Cell killing experiments (FIGS. 17-19) showed that the 3F12 clone did not have significant killing ability against tumor cells, suggesting that the 3F12 mutation was detrimental to binding to CD19.8F12, 5A12 clones showed no significant difference in tumor cell killing ability compared to FMC63 WT. Compared with FMC63 WT, the Combo6 clone has significantly improved tumor cell killing. The assembled experimental results are shown in FIG. 20, and compared with FMC63 WT, the Combo6 clone has obviously improved killing on tumor cells under the conditions that E: T is 10 and 3 proportion, which suggests that Combo6 combined mutation is beneficial to T cell recognition on tumor cells.
Example 9
FMC 63-Mutant-BBz in vitro verification
After packaging FMC 63-Mutant-BBz obtained by constructing each scFv sequence into Lentivirus (Lentivirus), NK92 cells are infected (culture conditions: RPMI1640+20% FBS+200IU/ml hIL 2); the NK92 cells after infection are amplified and cultured in vitro, and whether the FMC63 mutation can enhance the function of NK92 cells in killing tumor cells is evaluated through cytokine release and killing experiments, and the specific results are as follows:
cytokine release assay:
NK92 cell lines were cultured in RPMI1640+20% FBS+200IU/ml hIL 2; packaging FMC 63-Mutant-BBz into lentivirus, then infecting NK92 cells, and carrying out in vitro amplification and culture; NK92 cells and Raji cells (expressing CD 19) were mixed in proportion 1:1 Mixed culture in RPMI1640 medium with 2% serum for 16 hours, the release of cytokines was measured by BD Cytometric bead array kit from the mixed culture supernatant. Experiments show that NK92 obtained by infection (FMC 63 Mutant-BBz obtained by constructing scFv sequences is packaged into lentivirus to be obtained by infection, and the FMC63 Mutant-BBz chimeric antigen receptor is expressed) has stronger binding capacity to Raji cells and can release cytokines compared with FMC63 WT-BBz.
Tumor killing experiments:
NK92 cells and Raji cells obtained by the preparation are prepared according to a certain E: t ratio, e.g. 30:1,10: 1,3:1,1:1 ratio in RPMI1640 medium with 2% serum for 4 hours, the culture supernatant was mixed with LDH substrate (CytoTox 96 Non-Radioactive Cytotoxicity Assay Kit, promega) 1:1 volume of the mixture, after incubation at room temperature for 30 minutes, 490nm light absorption was read. Experimental results show that NK92 cells obtained by infection (FMC 63 Mutant-BBz obtained by constructing scFv sequences is packaged into lentivirus infection) have stronger tumor killing capacity compared with FMC63 WT-BBz.
In summary, the present invention effectively overcomes the disadvantages of the prior art and has high industrial utility value.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Sequence listing
<110> Shanghai Ji-times Biotechnology Co., ltd
<120> an anti-CD 19 antibody, and preparation method and use thereof
<160> 135
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> Artificial
<400> 1
Gly Trp Ser Leu Glu Asp Tyr Gly Val Ser
1 5 10
<210> 2
<211> 16
<212> PRT
<213> Artificial
<400> 2
Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys Ser
1 5 10 15
<210> 3
<211> 12
<212> PRT
<213> Artificial
<220>
<221> UNSURE
<222> (2)..(2)
<223> The 'Xaa' at location 2 stands for Arg, Tyr, or Trp.
<220>
<221> UNSURE
<222> (8)..(8)
<223> The 'Xaa' at location 8 stands for Phe, or Met.
<220>
<221> UNSURE
<222> (9)..(9)
<223> The 'Xaa' at location 9 stands for Asp, Glu, or Ala.
<400> 3
His Xaa Tyr Tyr Gly Gly Ser Xaa Xaa Met Asp Tyr
1 5 10
<210> 4
<211> 11
<212> PRT
<213> Artificial
<220>
<221> UNSURE
<222> (4)..(4)
<223> The 'Xaa' at location 4 stands for Gln, or Met.
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Asp, or Pro.
<220>
<221> UNSURE
<222> (7)..(7)
<223> The 'Xaa' at location 7 stands for Arg, or Lys.
<220>
<221> UNSURE
<222> (8)..(8)
<223> The 'Xaa' at location 8 stands for Arg, or Lys.
<400> 4
Arg Ala Ser Xaa Xaa Ile Xaa Xaa Tyr Leu Asn
1 5 10
<210> 5
<211> 7
<212> PRT
<213> Artificial
<400> 5
Gln Thr Ser Arg Leu Ala Trp
1 5
<210> 6
<211> 7
<212> PRT
<213> Artificial
<400> 6
Gln Thr Asp Arg Lys Gln Ser
1 5
<210> 7
<211> 7
<212> PRT
<213> Artificial
<400> 7
His Thr Ser Arg Leu His Ser
1 5
<210> 8
<211> 9
<212> PRT
<213> Artificial
<220>
<221> UNSURE
<222> (4)..(4)
<223> The 'Xaa' at location 4 stands for Asn, or Lys.
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Thr, Ala, Lys, or Arg.
<220>
<221> UNSURE
<222> (6)..(6)
<223> The 'Xaa' at location 6 stands for Leu, Ala, or Gly.
<220>
<221> UNSURE
<222> (8)..(8)
<223> The 'Xaa' at location 8 stands for Phe, or Tyr.
<400> 8
Gln Gln Gly Xaa Xaa Xaa Pro Xaa Thr
1 5
<210> 9
<211> 726
<212> DNA
<213> Artificial
<400> 9
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggcggtg gctcgggcgg tggtgggtcg ggtggcggcg gatctgacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 10
<211> 242
<212> PRT
<213> Artificial
<400> 10
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 11
<211> 67
<212> DNA
<213> Artificial
<400> 11
tcacatgcac tgtctcaggg gtctcattac ccgactatgg tgtaagctgg attcgccagc 60
ctccacg 67
<210> 12
<211> 79
<212> DNA
<213> Artificial
<400> 12
agggtctgga gtggctggga gtaatatggg gtagtgaaac cacatactat aattcagctc 60
tcaaatccag actgaccat 79
<210> 13
<211> 72
<212> DNA
<213> Artificial
<400> 13
ccatttacta ctgtgccaaa cattattact acggtggtag ctatgctatg gactactggg 60
gccaaggaac ct 72
<210> 14
<211> 72
<212> DNA
<213> Artificial
<400> 14
acagagtcac catcagttgc agggcaagtc aggacattag taaatattta aattggtatc 60
agcagaaacc ag 72
<210> 15
<211> 61
<212> DNA
<213> Artificial
<400> 15
ctgttaaact cctgatctac catacatcaa gattacactc aggagtccca tcaaggttca 60
g 61
<210> 16
<211> 64
<212> DNA
<213> Artificial
<400> 16
ttgccactta cttttgccaa cagggtaata cgcttccgta cacgttcgga ggggggacca 60
agct 64
<210> 17
<211> 20
<212> DNA
<213> Artificial
<400> 17
ccctcatagt tagcgtaacg 20
<210> 18
<211> 24
<212> DNA
<213> Artificial
<400> 18
agcggataac aatttcacac agga 24
<210> 19
<211> 20
<212> DNA
<213> Artificial
<400> 19
ccctgagaca gtgcatgtga 20
<210> 20
<211> 20
<212> DNA
<213> Artificial
<400> 20
tcccagccac tccagaccct 20
<210> 21
<211> 20
<212> DNA
<213> Artificial
<400> 21
tttggcacag tagtaaatgg 20
<210> 22
<211> 20
<212> DNA
<213> Artificial
<400> 22
gcaactgatg gtgactctgt 20
<210> 23
<211> 20
<212> DNA
<213> Artificial
<400> 23
gtagatcagg agtttaacag 20
<210> 24
<211> 20
<212> DNA
<213> Artificial
<400> 24
ttggcaaaag taagtggcaa 20
<210> 25
<211> 726
<212> DNA
<213> Artificial
<400> 25
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcagggtg gtcattagag gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 26
<211> 726
<212> DNA
<213> Artificial
<400> 26
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acatcggtac 300
tacggtggta gctttgatat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 27
<211> 726
<212> DNA
<213> Artificial
<400> 27
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattggtac 300
tacggtggta gctttgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 28
<211> 726
<212> DNA
<213> Artificial
<400> 28
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggaggg ataattggta gtcagaccac atactatctg 180
gctgggctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 29
<211> 726
<212> DNA
<213> Artificial
<400> 29
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggactg atacagggta gttggcatac atactatggt 180
gcggctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 30
<211> 726
<212> DNA
<213> Artificial
<400> 30
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accagacaga tagaaagcag tcgggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 31
<211> 726
<212> DNA
<213> Artificial
<400> 31
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattat 300
tacggtggta gcatggctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 32
<211> 726
<212> DNA
<213> Artificial
<400> 32
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcagccta ttcggaggta tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 33
<211> 726
<212> DNA
<213> Artificial
<400> 33
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtatggaca ttaagaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 34
<211> 726
<212> DNA
<213> Artificial
<400> 34
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accagacatc aagattagcc tggggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 35
<211> 726
<212> DNA
<213> Artificial
<400> 35
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa ggcggctccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 36
<211> 726
<212> DNA
<213> Artificial
<400> 36
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc catcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacaggggaa gaaggcgccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 37
<211> 726
<212> DNA
<213> Artificial
<400> 37
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa gaaggggccg cttacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 38
<211> 726
<212> DNA
<213> Artificial
<400> 38
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa gagggcgccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 39
<211> 726
<212> DNA
<213> Artificial
<400> 39
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc cgtcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa gagggggccg ttgactttcg gaggggggac caagctggag 720
atcaca 726
<210> 40
<211> 726
<212> DNA
<213> Artificial
<400> 40
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa gacggctccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 41
<211> 726
<212> DNA
<213> Artificial
<400> 41
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa gacggggccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 42
<211> 726
<212> DNA
<213> Artificial
<400> 42
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa gacgcttccg tttacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 43
<211> 726
<212> DNA
<213> Artificial
<400> 43
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa taaggcgccg ctgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 44
<211> 726
<212> DNA
<213> Artificial
<400> 44
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcagggtg gtcattagag gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggaggg ataattggta gtcagaccac atactatctg 180
gctgggctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acatcggtac 300
tacggtggta gctttgatat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 45
<211> 726
<212> DNA
<213> Artificial
<400> 45
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcagggtg gtcattagag gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acatcggtac 300
tacggtggta gctttgatat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 46
<211> 726
<212> DNA
<213> Artificial
<400> 46
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcagggtg gtcattagag gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggaggg ataattggta gtcagaccac atactatctg 180
gctgggctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattggtac 300
tacggtggta gctttgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 47
<211> 726
<212> DNA
<213> Artificial
<400> 47
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcagggtg gtcattagag gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattggtac 300
tacggtggta gctttgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcaggaca ttagtaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacagggtaa tacgcttccg tacacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 48
<211> 726
<212> DNA
<213> Artificial
<400> 48
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcagccta ttcggaggta tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accagacatc aagattagcc tggggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacaggggaa gaaggcgccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 49
<211> 726
<212> DNA
<213> Artificial
<400> 49
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagcct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtcagccta ttcggaggta tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacaggggaa gaaggcgccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 50
<211> 726
<212> DNA
<213> Artificial
<400> 50
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtatggaca ttaagaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accagacatc aagattagcc tggggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacaggggaa gaaggcgccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 51
<211> 726
<212> DNA
<213> Artificial
<400> 51
gaggtgaaac tgcaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccgtc 60
acatgcactg tctcaggggt ctcattaccc gactatggtg taagctggat tcgccagtct 120
ccacgaaagg gtctggagtg gctgggagta atatggggta gtgaaaccac atactataat 180
tcagctctca aatccagact gaccatcatc aaggacaact ccaagagcca agttttctta 240
aaaatgaaca gtctgcaaac tgatgacaca gccatttact actgtgccaa acattattac 300
tacggtggta gctatgctat ggactactgg ggccaaggaa cctcagtcac cgtctcctca 360
ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggacat ccagatgaca 420
cagactacat cctccctgtc tgcctctctg ggagacagag tcaccatcag ttgcagggca 480
agtatggaca ttaagaaata tttaaattgg tatcagcaga aaccagatgg aactgttaaa 540
ctcctgatct accatacatc aagattacac tcaggagtcc catcaaggtt cagtggcagt 600
gggtctggaa cagattattc tctcaccatt agcaacctgg agcaagaaga tattgccact 660
tacttttgcc aacaggggaa gaaggcgccg ttgacgttcg gaggggggac caagctggag 720
atcaca 726
<210> 52
<211> 242
<212> PRT
<213> Artificial
<400> 52
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 53
<211> 242
<212> PRT
<213> Artificial
<400> 53
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 54
<211> 242
<212> PRT
<213> Artificial
<400> 54
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Trp Tyr Tyr Gly Gly Ser Phe Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 55
<211> 242
<212> PRT
<213> Artificial
<400> 55
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Gly Ile Ile Gly Ser Gln Thr Thr Tyr Tyr Leu Ala Gly Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 56
<211> 242
<212> PRT
<213> Artificial
<400> 56
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Leu Ile Gln Gly Ser Trp His Thr Tyr Tyr Gly Ala Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 57
<211> 242
<212> PRT
<213> Artificial
<400> 57
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr Gln Thr Asp Arg Lys Gln Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 58
<211> 242
<212> PRT
<213> Artificial
<400> 58
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Met Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 59
<211> 242
<212> PRT
<213> Artificial
<400> 59
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Pro Ile Arg Arg Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 60
<211> 242
<212> PRT
<213> Artificial
<400> 60
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Met Asp Ile Lys Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 61
<211> 242
<212> PRT
<213> Artificial
<400> 61
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr Gln Thr Ser Arg Leu Ala Trp Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 62
<211> 242
<212> PRT
<213> Artificial
<400> 62
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Ala Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 63
<211> 242
<212> PRT
<213> Artificial
<400> 63
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Lys Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 64
<211> 242
<212> PRT
<213> Artificial
<400> 64
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Lys Gly Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 65
<211> 242
<212> PRT
<213> Artificial
<400> 65
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Arg Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 66
<211> 242
<212> PRT
<213> Artificial
<400> 66
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Arg Gly Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 67
<211> 242
<212> PRT
<213> Artificial
<400> 67
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Thr Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 68
<211> 242
<212> PRT
<213> Artificial
<400> 68
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Thr Gly Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 69
<211> 242
<212> PRT
<213> Artificial
<400> 69
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Thr Leu Pro Phe Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 70
<211> 242
<212> PRT
<213> Artificial
<400> 70
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Lys Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 71
<211> 242
<212> PRT
<213> Artificial
<400> 71
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Gly Ile Ile Gly Ser Gln Thr Thr Tyr Tyr Leu Ala Gly Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 72
<211> 242
<212> PRT
<213> Artificial
<400> 72
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 73
<211> 242
<212> PRT
<213> Artificial
<400> 73
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Gly Ile Ile Gly Ser Gln Thr Thr Tyr Tyr Leu Ala Gly Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Trp Tyr Tyr Gly Gly Ser Phe Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 74
<211> 242
<212> PRT
<213> Artificial
<400> 74
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Trp Tyr Tyr Gly Gly Ser Phe Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 75
<211> 242
<212> PRT
<213> Artificial
<400> 75
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Pro Ile Arg Arg Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr Gln Thr Ser Arg Leu Ala Trp Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Lys Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 76
<211> 242
<212> PRT
<213> Artificial
<400> 76
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Gln Pro Ile Arg Arg Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Lys Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 77
<211> 242
<212> PRT
<213> Artificial
<400> 77
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Met Asp Ile Lys Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr Gln Thr Ser Arg Leu Ala Trp Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Lys Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 78
<211> 242
<212> PRT
<213> Artificial
<400> 78
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
115 120 125
Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Thr Thr Ser
130 135 140
Ser Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala
145 150 155 160
Ser Met Asp Ile Lys Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp
165 170 175
Gly Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly
180 185 190
Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu
195 200 205
Thr Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln
210 215 220
Gln Gly Lys Lys Ala Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Thr
<210> 79
<211> 120
<212> PRT
<213> Artificial
<400> 79
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 80
<211> 120
<212> PRT
<213> Artificial
<400> 80
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 81
<211> 120
<212> PRT
<213> Artificial
<400> 81
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Trp Tyr Tyr Gly Gly Ser Phe Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 82
<211> 120
<212> PRT
<213> Artificial
<400> 82
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Gly Ile Ile Gly Ser Gln Thr Thr Tyr Tyr Leu Ala Gly Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 83
<211> 120
<212> PRT
<213> Artificial
<400> 83
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Leu Ile Gln Gly Ser Trp His Thr Tyr Tyr Gly Ala Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 84
<211> 120
<212> PRT
<213> Artificial
<400> 84
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 85
<211> 120
<212> PRT
<213> Artificial
<400> 85
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Met Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 86
<211> 120
<212> PRT
<213> Artificial
<400> 86
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 87
<211> 120
<212> PRT
<213> Artificial
<400> 87
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 88
<211> 120
<212> PRT
<213> Artificial
<400> 88
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 89
<211> 120
<212> PRT
<213> Artificial
<400> 89
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 90
<211> 120
<212> PRT
<213> Artificial
<400> 90
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 91
<211> 120
<212> PRT
<213> Artificial
<400> 91
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 92
<211> 120
<212> PRT
<213> Artificial
<400> 92
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 93
<211> 120
<212> PRT
<213> Artificial
<400> 93
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 94
<211> 120
<212> PRT
<213> Artificial
<400> 94
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 95
<211> 120
<212> PRT
<213> Artificial
<400> 95
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 96
<211> 120
<212> PRT
<213> Artificial
<400> 96
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 97
<211> 120
<212> PRT
<213> Artificial
<400> 97
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 98
<211> 120
<212> PRT
<213> Artificial
<400> 98
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Gly Ile Ile Gly Ser Gln Thr Thr Tyr Tyr Leu Ala Gly Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 99
<211> 120
<212> PRT
<213> Artificial
<400> 99
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 100
<211> 120
<212> PRT
<213> Artificial
<400> 100
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Gly Ile Ile Gly Ser Gln Thr Thr Tyr Tyr Leu Ala Gly Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Trp Tyr Tyr Gly Gly Ser Phe Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 101
<211> 120
<212> PRT
<213> Artificial
<400> 101
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Trp Ser Leu Glu Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Trp Tyr Tyr Gly Gly Ser Phe Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 102
<211> 120
<212> PRT
<213> Artificial
<400> 102
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 103
<211> 120
<212> PRT
<213> Artificial
<400> 103
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 104
<211> 120
<212> PRT
<213> Artificial
<400> 104
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 105
<211> 120
<212> PRT
<213> Artificial
<400> 105
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln
1 5 10 15
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr
20 25 30
Gly Val Ser Trp Ile Arg Gln Ser Pro Arg Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys
50 55 60
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu
65 70 75 80
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Ser Val Thr Val Ser Ser
115 120
<210> 106
<211> 107
<212> PRT
<213> Artificial
<400> 106
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 107
<211> 107
<212> PRT
<213> Artificial
<400> 107
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 108
<211> 107
<212> PRT
<213> Artificial
<400> 108
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 109
<211> 107
<212> PRT
<213> Artificial
<400> 109
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 110
<211> 107
<212> PRT
<213> Artificial
<400> 110
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 111
<211> 107
<212> PRT
<213> Artificial
<400> 111
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr Gln Thr Asp Arg Lys Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 112
<211> 107
<212> PRT
<213> Artificial
<400> 112
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 113
<211> 107
<212> PRT
<213> Artificial
<400> 113
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Pro Ile Arg Arg Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 114
<211> 107
<212> PRT
<213> Artificial
<400> 114
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Met Asp Ile Lys Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 115
<211> 107
<212> PRT
<213> Artificial
<400> 115
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr Gln Thr Ser Arg Leu Ala Trp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 116
<211> 107
<212> PRT
<213> Artificial
<400> 116
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Ala Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 117
<211> 107
<212> PRT
<213> Artificial
<400> 117
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Lys Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 118
<211> 107
<212> PRT
<213> Artificial
<400> 118
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Lys Gly Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 119
<211> 107
<212> PRT
<213> Artificial
<400> 119
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Arg Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 120
<211> 107
<212> PRT
<213> Artificial
<400> 120
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Arg Gly Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 121
<211> 107
<212> PRT
<213> Artificial
<400> 121
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Thr Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 122
<211> 107
<212> PRT
<213> Artificial
<400> 122
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Thr Gly Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 123
<211> 107
<212> PRT
<213> Artificial
<400> 123
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Thr Leu Pro Phe
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 124
<211> 107
<212> PRT
<213> Artificial
<400> 124
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Lys Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 125
<211> 107
<212> PRT
<213> Artificial
<400> 125
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 126
<211> 107
<212> PRT
<213> Artificial
<400> 126
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 127
<211> 107
<212> PRT
<213> Artificial
<400> 127
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 128
<211> 107
<212> PRT
<213> Artificial
<400> 128
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 129
<211> 107
<212> PRT
<213> Artificial
<400> 129
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Pro Ile Arg Arg Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr Gln Thr Ser Arg Leu Ala Trp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Lys Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 130
<211> 107
<212> PRT
<213> Artificial
<400> 130
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Pro Ile Arg Arg Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Lys Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 131
<211> 107
<212> PRT
<213> Artificial
<400> 131
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Met Asp Ile Lys Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr Gln Thr Ser Arg Leu Ala Trp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Lys Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 132
<211> 107
<212> PRT
<213> Artificial
<400> 132
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Met Asp Ile Lys Lys Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Lys Lys Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr
100 105
<210> 133
<211> 7252
<212> DNA
<213> Artificial
<400> 133
agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga 60
gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa 120
ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta 180
tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca 240
ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt 300
gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg 360
aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt 420
tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg 480
tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct 540
ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt 600
ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg 660
gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc 720
tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg 780
gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc 840
aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag 900
ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag 960
gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt 1020
ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca 1080
cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa 1140
gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaagatctct 1200
agaagctggg taccttgtgc ccgggcgcca ccatggagtt tgggctgagc tggctttttc 1260
ttgtcgcgat tcttaagggt gtccagtgcg acaaaactca cacatgccca ccgtgcccag 1320
cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc 1380
tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc 1440
ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc 1500
cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc 1560
aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc 1620
ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc 1680
tgcccccatc ccgggatgag ctgaccaaga accaggtcag cctgacctgc ctggtcaaag 1740
gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaact 1800
acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac agcaagctca 1860
ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcatgagg 1920
ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa tgagcggccg 1980
ctcgaggccg gcaaggccgg atccagacat gataagatac attgatgagt ttggacaaac 2040
cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 2100
atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 2160
gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg 2220
tggtatggct gattatgatc cggctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc 2280
tgacacatgc agctcccgga tacggtcaca gcttgtctgt aagcggatgc cgggagcaga 2340
caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgaggtcg 2400
actctagagg atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc 2460
ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc 2520
cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg 2580
acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag 2640
actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc 2700
tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct 2760
acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca 2820
ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg 2880
ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga 2940
agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac 3000
cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca 3060
ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct 3120
tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag 3180
taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt 3240
ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct 3300
tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc 3360
atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg 3420
gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg 3480
accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac 3540
gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc 3600
acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt 3660
gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt 3720
aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac 3780
cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc 3840
gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt 3900
tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta 3960
gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat 4020
aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat 4080
atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat 4140
atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat 4200
atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat 4260
atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc 4320
tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc 4380
tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc 4440
tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc 4500
tatcctaatc tgtatccggg tagcatatgc tatcctcatg catatacagt cagcatatga 4560
tacccagtag tagagtggga gtgctatcct ttgcatatgc cgccacctcc caagggggcg 4620
tgaattttcg ctgcttgtcc ttttcctgct gcttatcgat gataagctgt caaacatgag 4680
aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 4740
ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 4800
tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 4860
atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 4920
attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 4980
gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 5040
agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 5100
aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt 5160
cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 5220
cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 5280
actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 5340
cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 5400
ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa 5460
ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 5520
gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 5580
gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 5640
ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 5700
cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 5760
caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 5820
taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 5880
cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 5940
cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 6000
gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 6060
aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 6120
cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 6180
tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 6240
acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 6300
ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 6360
ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 6420
tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 6480
tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 6540
ctggcctttt gctggccttt tgctcacatg aagctgtccc tgatggtcgt catctacctg 6600
cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat 6660
aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc 6720
ggccccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc 6780
caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga 6840
gtggtgaatc cgttagcgag gtgccgccct gcttcatccc cgtggcccgt tgctcgcgtt 6900
tgctggcggt gtcactggcc ccgtgggtta gggacggggt cccccatggg gaatggttta 6960
tggttcgtgg gggttattat tttgggcgtt gcgtggggtc aggtccacga ctggactgag 7020
cagacagacc catggttttt ggatggcctg ggcatggacc gcatgtactg gcgcgacacg 7080
aacaccgggc gtctgtggct gccaaacacc cccgaccccc aaaaaccacc gcgcggattt 7140
ctggcgtgcc aagctagtcg accaattctc atgtttgaca gcttatcatc gcagatccgg 7200
gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat ct 7252
<210> 134
<211> 19
<212> PRT
<213> Artificial
<400> 134
Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn
1 5 10 15
Pro Gly Pro
<210> 135
<211> 19
<212> PRT
<213> Artificial sequence (Artificial Sequence)
<400> 135
Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn
1 5 10 15
Pro Gly Pro

Claims (11)

1. An anti-CD 19 antibody comprising a heavy chain variable region and a light chain variable region, the anti-CD 19 antibody having the following technical characteristics:
<1> complementarity determining regions of the heavy chain variable region include CDR-H1 having the amino acid sequence: GVSLPDYGVS;
<2> complementarity determining regions of the heavy chain variable region include CDR-H2, which has the amino acid sequence: VIWGSETTYYNSALKS;
<3> complementarity determining regions of the heavy chain variable region include CDR-H3, which has the amino acid sequence: HYYYGGSYAMDY;
<4> complementarity determining regions of the light chain variable region include CDR-L1 having the amino acid sequence: RASQPIRRYLN;
<5> complementarity determining regions of the light chain variable region include CDR-L2, which has the amino acid sequence: HTSRLHS;
<6> complementarity determining regions of the light chain variable region include CDR-L3, which has the amino acid sequence: QQGNTLPYT.
2. An anti-CD 19 antibody according to claim 1, wherein the anti-CD 19 antibody is a monoclonal antibody;
and/or, the anti-CD 19 antibody is a single chain antibody;
and/or, the anti-CD 19 antibody is derived from the monoclonal antibody FMC63 specific for CD 19;
And/or the amino acid sequence of the heavy chain variable region of the anti-CD 19 antibody is shown as the amino acid sequence of SEQ ID No. 86;
and/or the amino acid sequence of the light chain variable region of the anti-CD 19 antibody is shown as the amino acid sequence of SEQ ID No. 113.
3. An isolated polynucleotide encoding the anti-CD 19 antibody of any one of claims 1-2.
4. A construct comprising the isolated polynucleotide of claim 3.
5. An expression system for an antibody comprising the construct or the polynucleotide of claim 3 integrated with an exogenous source in the genome of claim 4.
6. The method of preparing an anti-CD 19 antibody according to any one of claims 1-2, comprising the steps of: culturing the antibody expression system of claim 5 under conditions suitable for expression of said antibody, thereby expressing said antibody, and purifying and isolating said antibody.
7. Use of an anti-CD 19 antibody according to any one of claims 1-2 for the preparation or screening of a medicament for the treatment of a tumor.
8. An isolated polypeptide comprising a transmembrane domain, an intracellular domain and an extracellular domain comprising the anti-CD 19 antibody of any one of claims 1-2.
9. The polypeptide of claim 8, wherein the polypeptide is a chimeric antigen receptor;
and/or, the transmembrane domain comprises CD8 a, CD28, DAP10;
and/or, the intracellular domain comprises 4-1BB, CD28, OX40, ICOS, CD3zeta, DAP10;
and/or the polypeptide comprises the anti-CD 19 antibody, a transmembrane domain and an intracellular domain from the N end to the C end.
10. A cell comprising a membrane-bound polypeptide according to any one of claims 8-9, which is a T lymphocyte and/or an NK cell;
and/or the polypeptide is a chimeric antigen receptor;
and/or, when the polypeptide binds to CD19 antigen, the T lymphocytes and/or NK cells can be activated and/or stimulated to proliferate;
and/or, the T lymphocytes and/or NK cells surface express the anti-CD 19 antibody.
11. A diagnostic kit comprising a diagnostically effective amount of an anti-CD 19 antibody or immunoconjugate thereof of any one of claims 1-2.
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