[go: up one dir, main page]

CN111848801B - anti-CD 19 antibody and preparation method and application thereof - Google Patents

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

Info

Publication number
CN111848801B
CN111848801B CN202010870418.3A CN202010870418A CN111848801B CN 111848801 B CN111848801 B CN 111848801B CN 202010870418 A CN202010870418 A CN 202010870418A CN 111848801 B CN111848801 B CN 111848801B
Authority
CN
China
Prior art keywords
ser
gly
thr
leu
gln
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010870418.3A
Other languages
Chinese (zh)
Other versions
CN111848801A (en
Inventor
牟男
张云
马泽龙
袁纪军
曹跃琼
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Genbase Biotechnology Co Ltd
Original Assignee
Shanghai Genbase Biotechnology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Genbase Biotechnology Co Ltd filed Critical Shanghai Genbase Biotechnology Co Ltd
Publication of CN111848801A publication Critical patent/CN111848801A/en
Application granted granted Critical
Publication of CN111848801B publication Critical patent/CN111848801B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • 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
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • 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
    • A61K2039/5156—Animal cells expressing foreign proteins
    • 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
    • A61K2039/5158—Antigen-pulsed cells, e.g. T-cells
    • C—CHEMISTRY; METALLURGY
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • 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]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • 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)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Cell Biology (AREA)
  • Hematology (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Oncology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to the technical field of biology, in particular to an anti-CD 19 antibody and 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. The inventor conducts affinity maturation screening on FMC63 scFv by using a phage display technology, so as to obtain a high-affinity single-chain antibody for CD19.

Description

anti-CD 19 antibody and preparation method and application thereof
The application is a divisional application of an anti-CD 19 antibody, a preparation method and application thereof, and the application date of the original application is as follows: 2017.9.6, application No.: 201710793590.1.
Technical Field
The invention relates to the technical field of biology, in particular to an anti-CD 19 antibody and a preparation method and application thereof.
Background
The CD19 antigen (B lymphocyte antigen CD19, CD 19) is a protein specifically expressed on the surface of human B cells. CD19 is widely expressed on the surface of B cells in various developmental stages and plays an important role, and the CD19 is used as a co-receptor (co-receptor) of a B Cell Receptor (BCR), so that the signal threshold required by antigen-mediated dependent activation of the B cell receptor (antigen-dependent stimulation) can be reduced; b cell receptor activation relies on intracellular domain phosphorylation of CD19 followed by recruitment of Src kinase and PI3K kinase, thereby activating B cells completely. Fully mature B cells, called plasma cells, lose CD19 antigen expression after full maturation.
Hematological tumors associated with B cell malignant proliferation include: b-cell acute lymphoblastic leukemia (B-ALL), B-cell 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 high levels of B-cell malignant proliferation associated with peripheral blood, bone marrow, and the formation of systemic B-cell solid tumors that severely interfere with the patient's blood circulation system. Today, treatment for 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 antibody), and bone marrow transplantation (bone marrow transplantation); in clinical application, the life cycle of patients can be prolonged remarkably by small molecular drugs and antibody drugs, time is won for bone marrow transplantation, but some patients have drug resistance recurrence, and the drugs in clinical use to date have difficult curative effect on the patients with drug resistance recurrence.
A chimeric antigen receptor T cell therapy (CART) belongs to a novel treatment mode, which is characterized in that a patient T cell is used as a base to be expanded in vitro and is subjected to genetic engineering modification, the CART cell can identify a tumor cell specific antigen (such as CD 19), and can specifically lyse the tumor cell after being activated to achieve the purpose of killing tumors, and the CART has the characteristics of short treatment period, quicker curative effect, high complete response rate, capacity of striving for enough time for the patient to wait for bone marrow transplantation and even complete cure. The chimeric antigen receptor is composed of a single-chain antibody (scFv) -hinge region (hinge region) -transmembrane region (transmembrane domain) -costimulatory domain-essential stimulatory domain (addressing domain), wherein a single-chain antibody with high affinity for an antigen is the most important component of a Chimeric Antigen Receptor (CAR), and the affinity of the scFv for the antigen determines whether the CART cell can activate and kill tumor cells, so that the screening of affinity maturation for antigen-specific scFv is particularly important.
FMC63-mIgG2a was a murine antibody raised against CD19 by immunization of animals for the last century. FMC63 scFv has been successfully applied to anti-CD 19 CAR constructs such as Norvatis CTL019 and Juno Therapeutics JCAR015. Clinical trials of CTL019 and JCAR015 in B-cell acute lymphoblastic leukemia gave better results with a complete remission (complete remission) patient rate of >70%.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention aims to provide an anti-CD 19 antibody, a method for preparing the same, and a use thereof, which are used to solve the problems of the prior art.
To achieve the above and other related objects, the present invention provides, in one aspect, an anti-CD 19 antibody, the 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 comprises CDR-H1 having an amino acid sequence shown in SEQ ID No. 1;
<2> the complementarity determining region of the heavy chain variable region comprises CDR-H2 having an amino acid sequence shown in SEQ ID No. 2;
<3> the complementarity determining region of the heavy chain variable region comprises CDR-H3 having an amino acid sequence shown in SEQ ID No. 3;
<4> the complementarity determining region of the light chain variable region comprising CDR-L1 having an 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 shown in SEQ ID No.5, SEQ ID No.6 or SEQ ID No. 7;
<6> the complementarity determining region of the light chain variable region includes CDR-L3 having an amino acid sequence shown in SEQ ID No. 8.
GWSLEDYGVS(SEQ ID No.1)
VIWGSETTYYNSALKS(SEQ ID No.2)
HRYYGGSFDMDY(SEQ ID No.3)
RASQDISKYLN(SEQ ID No.4)
QTSRLAW(SEQ ID No.5)
QTDRKQS(SEQ ID No.6)
HTSRLHS(SEQ ID No.7)
QQGNTLPYT(SEQ ID No.8)
A CDR (complementary determining region) generally refers to a region of an antibody that can sterically complement an antigenic determinant. The variability in antibodies is typically not evenly distributed throughout the variable region of the antibody, and both the heavy and light chain variable regions of a monoclonal antibody typically have 3 hypervariable regions (HVRs) which are generally complementary in spatial structure to antigenic determinants, so the hypervariable regions are also referred to as Complementarity Determining Regions (CDRs), i.e., the heavy chain variable region typically includes three complementarity determining regions, i.e., HCDR1, HCDR2 and HCDR3, and the light chain variable region typically includes 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 include CDR-H1 having an amino acid sequence shown in SEQ ID No.1, CDR-H2 having an amino acid sequence shown in SEQ ID No.2, and CDR-H3 having an amino acid sequence shown in SEQ ID No. 3.
In certain embodiments of the invention, the complementarity determining region of the light chain variable region of the anti-CD 19 antibody includes CDR-L1 having an amino acid sequence shown in SEQ ID No.4, CDR-L2 having an amino acid sequence shown in SEQ ID No.5, SEQ ID No.6, or SEQ ID No.7, and CDR-L3 having an amino acid sequence shown in SEQ ID No. 8.
In some embodiments of the invention, the complementarity determining region of the heavy chain variable region comprises CDR-H1 having an amino acid sequence shown in SEQ ID No.1, CDR-H2 having an amino acid sequence shown in SEQ ID No.2, and CDR-H3 having an amino acid sequence shown in SEQ ID No.3, and the complementarity determining region of the light chain variable region comprises CDR-L1 having an amino acid sequence shown in SEQ ID No.4, CDR-L2 having an amino acid sequence shown in SEQ ID No.5, SEQ ID No.6, or SEQ ID No.7, and CDR-L3 having an amino acid sequence shown in SEQ ID No. 8.
In certain embodiments of the invention, the anti-CD 19 antibody is a monoclonal antibody. A monoclonal antibody generally refers to a population of antibodies in which the antibodies included are substantially identical (except for a few naturally occurring mutations that may be present). Monoclonal antibodies are generally directed against specific determinants on an antigen.
In certain embodiments of the invention, the anti-CD 19 antibody is a single chain antibody (scFv). Single chain antibodies may generally be V comprising antibodies H (variable region of heavy chain) and V L Polypeptide chain of (light chain variable region). Generally, single chain antibodies may also include a linking peptide (like)r), the linker peptide is usually located at V H And V L Such that the scFv forms the desired structure capable of binding to the antigen. For example, the anti-CD 19 antibody can 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 the N segment to the C end H Linker peptide and V L The anti-CD 19 single-chain antibody can also comprise V from the N segment to the C end in sequence L Linker peptide and V H . The linker peptide may be any of a variety of linker peptides suitable for use in the art for forming scFv, for example, the linker peptide may be G4S3 linker, which may be selected or designed as described in Michel Sadelain et, science relative Medicine,2013; jianne et c, science relative Medicine,2015.
In certain embodiments of the invention, the anti-CD 19 antibody is derived from the CD19 specific monoclonal antibody FMC63 (VH: Y14283.1, VL: Y14284.1), the nucleotide sequence of which is shown in SEQ ID No. 9and the amino acid sequence of which is shown in SEQ ID No. 10.
Figure BDA0002650914510000031
Figure BDA0002650914510000041
Figure BDA0002650914510000042
(SEQ ID No.10, wherein the bold underlined portion is a linker peptide preceded by a heavy chain variable region and followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
In certain embodiments of the present invention, the heavy chain variable region and the light chain variable region may further comprise a framework region, and the framework region may be located between the complementarity determining regions or at both ends of the complementarity determining regions. In a specific embodiment of the present invention, the sequence of the framework region is identical to the sequence of the framework region of FMC63, or is a sequence of the framework region of FMC63 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 sequence of the framework region of FMC63, and the sequence of the framework region may have 80%, 85%, 90%, 93%, 95%, 97%, or 99% or more homology to the sequence of the framework region 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 set forth in one of SEQ ID Nos. 79-105; or
b) An amino acid sequence which has more than 80 percent of homology with the amino acid sequence shown in one of SEQ ID No.79-105 and has the amino acid sequence function defined by a).
Specifically, the amino acid sequence in b) specifically refers to: an amino acid sequence shown as one of SEQ ID Nos. 79-105, which 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 Nos. 79-105. The amino acid sequence in b) may have more than 80%, 85%, 90%, 93%, 95%, 97%, or 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. 106-132; or
d) An amino acid sequence which has more than 80 percent of homology with the amino acid sequence shown in one of SEQ ID No.106-132 and has the amino acid sequence function defined by 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 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 more than 80%, 85%, 90%, 93%, 95%, 97%, or 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 the 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 by inserting the isolated polynucleotide into a multiple cloning site of an expression vector. The expression vector of the present invention is generally referred to various commercially available expression vectors well known in the art, and may be, for example, a bacterial plasmid, a bacteriophage, a yeast plasmid, a plant cell virus, a mammalian cell virus such as adenovirus, retrovirus, or other vectors.
In certain embodiments of the invention, the expression vector is selected from the group consisting of the GV401 expression vector (GV 401 is a commercially available vector from the supplier Ji Kai gene).
In another aspect, the invention provides an antibody expression system comprising the construct or the polynucleotide integrated into the genome from an exogenous source. Any cell suitable for expression of an expression vector may be used as a host cell, for example, 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 a combination of one or more of a T cell, an NK cell.
In another aspect, the present invention provides a method for preparing the anti-CD 19 antibody, comprising the steps of: culturing said antibody expression system under conditions suitable for expression of said antibody, thereby expressing said antibody, and purifying and isolating said antibody.
The host cells used in the present invention are available in the prior art, and can be obtained directly from commercial sources, and the culture medium used in the culture can be various conventional media, and those skilled in the art can select an appropriate medium according to experience and culture the appropriate medium under conditions suitable for the growth of the host cells. After the host cells have been grown to an appropriate cell density, the selected promoter is induced by suitable means (e.g., temperature shift or chemical induction) and the cells are cultured for an additional period of time. The recombinant polypeptide in the above method may be expressed intracellularly or on the cell membrane, or secreted extracellularly. If necessary, the recombinant protein can be isolated and purified by various separation methods using its physical, chemical and other properties. These 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 (such as salt precipitation), centrifugation, cell lysis by osmosis, sonication, ultracentrifugation, molecular sieve chromatography (gel filtration), adsorption chromatography, ion exchange chromatography, high Performance Liquid Chromatography (HPLC), and other various liquid chromatography techniques and combinations thereof.
In another aspect, the invention provides the use of the anti-CD 19 antibody in the preparation or screening of a therapeutic agent, or in the preparation of a diagnostic agent.
The therapeutic agent may be one that targets the CD-19 antigen, binds to or acts on the CD-19 antigen, and thereby treats and/or prevents the indication.
In certain embodiments of the invention, the therapeutic agent may be a tumor therapeutic agent. The tumor treatment drug can be a drug which takes a CD-19 antigen functionally expressed on the surface of a tumor cell as a target, and combines or acts on the CD-19 antigen so as to treat and/or prevent tumors. The tumor may be acute lymphoid leukemia, chronic lymphoid 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) cell therapeutic agent.
The chimeric antigen receptor cell therapeutic agents typically include chimeric antigen receptor cells, which may be chimeric antigen receptor T cells, chimeric antigen receptor NK cells, and the like, which typically include T lymphocytes, which also include a chimeric antigen receptor. The chimeric antigen receptor NK cells typically include NK cells, which also include a chimeric antigen receptor. 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 acting on the cells expressing the CD19 antigen may be killing the cells expressing the CD19 antigen, or the like.
Therefore, the diagnostic drugs specifically refer to reagents for diagnosis that act on a target CD19 antigen and use 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, wherein the extracellular domain comprises 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 α (NM _ 001145873), CD28 (NM _ 006139), DAP10 (NM _ 014266), and the like.
In certain embodiments of the invention, the endodomains can include co-stimulatory domains and/or signaling domains, e.g., the endodomains can include 4-1BB (NM _ 001561), CD28 (NM _ 006139), OX40 (NM _ 003327), ICOS (NM _ 012092), CD3zeta (NM _ 198053), DAP10 (NM _ 014266), and the like.
In some embodiments of the invention, the polypeptide comprises the anti-CD 19 single-chain antibody, the transmembrane domain and the endodomain sequentially from N-terminus to C-terminus. In some embodiments of the invention, the polypeptide comprises, in order from N-terminus to C-terminus, the anti-CD 19 single chain antibody, CD 8a transmembrane region, 4-1BB co-stimulatory domain, CD3zeta signaling domain. In a specific embodiment of the invention, the polypeptide comprises the anti-CD 19 single-chain antibody, CD28 transmembrane region, CD28 costimulatory domain, CD3zeta signaling domain in sequence from N-terminus to C-terminus. In another specific embodiment of the invention, said polypeptide comprises, in order from N-terminus to C-terminus, said anti-CD 19 single chain antibody, CD8 α transmembrane region, OX40 co-stimulatory domain, CD3zeta signaling domain. In another embodiment of the invention, the polypeptide comprises, in order from N-terminus to C-terminus, the anti-CD 19 single chain antibody, CD8 α transmembrane region, ICOS costimulatory domain, CD3zeta signaling domain. In another specific embodiment of the invention, the polypeptide comprises the anti-CD 19 single-chain antibody, the CD8 alpha transmembrane region, the 4-1BB co-stimulatory domain, the CD28 co-stimulatory domain, and the CD3zeta sequentially from N-terminus to C-terminus. In another specific embodiment of the invention, said polypeptide comprises, in order from N-terminus to C-terminus, said anti-CD 19 single chain antibody, CD28 transmembrane region, CD28 costimulatory domain, OX40 costimulatory domain, CD3zeta signaling domain.
In another aspect of the invention, the T lymphocyte comprises the polypeptide bound to a membrane.
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 cells can express the anti-CD 19 antibody on the surface of T lymphocytes, which can direct the T lymphocytes to act on cells expressing the CD19 antigen (e.g., tumor cells), which can be killing cells expressing the CD19 antigen, or the like.
In another aspect of the invention, the NK cell comprises a membrane-bound polypeptide.
In certain embodiments of the invention, the polypeptide is a chimeric antigen receptor.
The NK cells can typically express the polypeptide, which can typically bind to a CD19 antigen, more particularly can bind to a CD19 antigen by comprising an extracellular domain of the anti-CD 19 antibody, and when the polypeptide binds to the antigen, the NK cells can 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 NK cells can express the anti-CD 19 antibody on the surface of NK cells, such that the action of the NK cells on cells expressing the CD19 antigen (e.g., tumor cells) can be directed, such that the action can be killing of cells expressing the CD19 antigen, or the like.
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.
Therefore, the diagnostic kit can generally aim at the CD19 antigen which is the target of action, and the CD19 antigen is taken as a biomarker for diagnosis. The diagnostic kit may further comprise a label for the anti-CD 19 antibody, which may be generally used to label the anti-CD 19 antibody, and the types of labels that may be selected include, but are not limited to, combinations of one or more of fluorescent labels, radioactive labels, enzyme labels, chemiluminescent labels, and the like. Depending on the detection principle of the kit, the kit may also typically comprise one or more reagents required for the detection. In addition, the kit can also comprise the following components according to needs: containers, controls (negative or positive controls), buffers, adjuvants, etc., which can be selected by one skilled in the art as appropriate.
The inventor conducts affinity maturation screening on FMC63 scFv by using phage display technology, so as to obtain high-affinity single-chain antibodies to CD19, wherein the single-chain antibodies can competitively block the combination of wild-type FMC63 mIgG2a to CD19, and thus the antigen binding sites of the single-chain antibodies are consistent with that of FMC63 mIgG2 a. Furthermore, the present inventors further engineered high affinity single chain antibodies into chimeric antigen receptors, for example, using T cells and NK cells expressing anti-CD 19 chimeric antigen receptors for CD 19-expressing hematologic cancer (B-ALL, B-CLL, B-Lymphoma, etc.) therapy, thereby verifying that the mutated chimeric antigen receptors can improve the killing ability against tumor cells.
Drawings
FIGS. 1 to 5 are graphs showing the results of the experiment in example 3 of the present invention.
FIGS. 6 to 10 are graphs showing the results of the experiment in example 4 of the present invention.
FIGS. 11 to 14 are graphs showing the results of the experiment in example 6 of the present invention.
FIG. 15 shows a schematic diagram of the GV400 vector structure.
FIG. 16 shows a schematic diagram of the vector structure of GV 401.
FIGS. 17 to 19 are graphs showing the results of the experiment in 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
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. 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, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts. These techniques are well described in the literature, and may be found in particular in the study of the MOLECULAR CLONING, sambrook et al: 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; (iii) METHODS IN ENZYMOLOGY, vol.304, chromatin (P.M.Wassarman and A.P.Wolffe, eds.), academic Press, san Diego,1999; and METHODS IN MOLECULAR BIOLOGY, vol.119, chromatography Protocols (P.B.Becker, ed.) Humana Press, totowa,1999, etc.
Example 1
Construction of CDR1, CDR2 and CDR3 mutant libraries of the heavy chain (H) and light chain (L) of FMC63 scFv:
the selection of the CDR regions of the heavy and light chains of FMC63 was based on the amino acid numbering of the variable domains (template monoclonal antibody FMC63, VH: Y14283.1, VL: Y14284.1), kabat numbering (Kabat number scheme. Bioinf. Org. Uk).
The pCAN-FMC63 scFv plasmid (constructed by inserting a template sequence into the multiple cloning site of pCANTAB 5E plasmid (purchased from GE) as a template, wherein the template sequence is VH: Y14283.1 and VL: Y14284.1) was used for introducing mutation by random primer PCRThe results are shown in Table 2. The obtained CDR1, CDR2, CDR 3mutation library PCR products of heavy chain (H) and light chain (L) of FMC63 scFv were named H1, H2, H3, L1, L2 and L3, respectively. After the PCR product and pCANTAB 5E were digested and recovered with Sfi I and Not I, they were ligated with T4 DNA ligase at 16 ℃ overnight. Electrically transferring the connecting product to TG1 competent cells, resuspending a 2xYT culture medium, recovering the cells at 37 ℃ for 1 hour, taking bacterial liquid for gradient dilution, and counting plates to obtain at least 10 library volumes of all mutation libraries 8 All the other bacterial solutions were spread on 2XYT (GA) plates (2% glucose, 100ug/ml penicillin). From the mutation library, 20 single clones were randomly picked for sequencing, and the diversity was 100%.
TABLE 1
Figure BDA0002650914510000101
Example 2
Panning of phage antibody library:
20nM CD19-his-biotin antigen and phage antibody library were added and incubated for 2h at room temperature, and the mixture was transferred to streptavidin magnetic beads and incubated for 15min at room temperature. PBST-PBS washed away the unbound phage, and then pancreatin was added at 37 ℃ for 30min to elute the bound phage. Infecting 4ml of TG1 thallus with the phage eluted by pancreatin digestion, standing at 37 deg.C for 30min, taking part of the bacteria liquid to dilute in gradient for plate counting, and coating the rest bacteria liquid on 2xYT (GA) plate for the next round of packaging. The packaged phage can be used for the next round of panning, 4 rounds of panning enrichment are carried out, the antigen concentration is reduced by 10-fold dilution gradient of each round of panning, and the PBST-PBS washing times are increased one by one (the CD19-his-biotin antigen concentrations of four rounds of panning are 20nM, 2nM, 0.2nM and 0.02nM respectively, and the PBST-PBS washing times are 7, 10, 15 and 20 times respectively).
Example 3
Screening and identification of high affinity scFv: after four rounds of panning, randomly picking single clone, inducing with IPTG, picking supernatant to carry out ELISA, after ELISA primary screening, picking clone with positive signal at least 2 times larger than negative signal, sending the clone to sequencing, analyzing sequencing result, and extracting clone corresponding to CDR area with more enrichment. The ELISA preliminary screening comprises the following specific steps:
colonies were picked and cloned in 4ml 2xYT medium, cultured overnight at 37 ℃ 200rpm, 1:100 is transferred into 500ml 2xYT culture medium and cultured at 37 ℃ and 200rpm to logarithmic phase; adding M13K07 helper phage (MOI = 20), incubating at 37 ℃ for 30 minutes at rest followed by 30 minutes at 37 ℃ and 200 rpm; centrifuging to remove supernatant, and resuspending the thallus into 500ml 2xYT/Kan/Amp resistant culture medium for overnight culture; the supernatant after overnight incubation was precipitated with 1/5 volume of PEG/NaCl solution for 1 hour and centrifuged, and the precipitate was resuspended in 30% glycerol-containing PBS solution and stored at 4 ℃. A268 was measured after the phage solution was diluted 2-10 fold, and the specific results are shown in Table 2 (in Table 2, clone numbers represent the numbers of picked colonies).
Avidin was coated onto 96-well plates, biotin-labeled CD19 antigen was bound to 96-well plates, and after 1 hour incubation after adding gradient diluted phage (original concentration of phage a268 before dilution see table 2), detection of range-bound CD19 with Anti-range antibody (Anti M13 range antibody, purchased from GE corporation), read at OD450nm, with results as shown in table 3: the smaller the EC50 (A268) (EC 50 indicates the concentration of phage corresponding to 50% saturation signal, and EC50 (A268) in Table 3 indicates that the absorbance of A268 corresponding to EC50 is proportional to the concentration of phage), the larger the dilution factor of the corresponding clone required for reaching the same saturation signal, i.e., the stronger the binding capacity of the corresponding clone (A268 is measured by Nanodrop (Thermofisher)); positive control FMC63WT phage, negative control anti-CD 19 single chain antibody expressing phage, using the same method as for other clones). Specific results are shown in FIGS. 1-5.
TABLE 2
Figure BDA0002650914510000111
Figure BDA0002650914510000121
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 screen:
TG1 clones with more repeated CDR mutations were plated, monoclonal antibodies were picked, and the purified phage was purified after IPTG induction, after concentration determination (A268 =5.4 diluted by 3-fold concentration gradient), the dilution gradient competed with 3ug/ml FMC63 mIgG2a (well plate: avidin was coated to 96 well plate, biotin-labeled CD19 antigen was bound to 96 well), detection was performed using anti-mouse Fc secondary antibody, substrate (TMB) was added followed by incubation for 5mins,1M sulfuric acid termination reaction and OD450nm reading, specific results are shown in Table 4, FIGS. 6-10. The different clones were compared for EC50 and Emax (A268 by Nanodrop assay (thermolither); the positive control was FMC63WT phage and the negative control was phage expressing anti-non-CD 19 single chain antibody, using the same method as for the other clones). The smaller the EC50 (A268) (EC 50 indicates the concentration of phage corresponding to the maximum inhibition rate Emax 50%, and EC50 (A268) in Table 4 indicates the absorbance of A268 corresponding to EC 50) is, the stronger the ability of the single-chain antibody of the corresponding clone to compete for FMC63 mIgG2a to 50% binding is; a larger Emax (maximum inhibition, the percentage of all cloned phage that compete for wild-type antibody binding at an absorbance of 5.4) indicates a stronger competition for the same A268 concentration of single-chain antibody of the corresponding clone.
TABLE 4
Clone number EC50(A268) Emax (maximum inhibition)
3H4 1.93 60％
3C2 1.87 45％
3C11 1.25 66％
3H9 1.75 26％
4E10 >5.4 10.6％
3F12 >5.4 9.5％
5A12 2.97 27％
8C4 1.58 45％
5G6 1.97 51％
8C7 1.92 70％
8B6 0.83 58％
8C10 1.46 56％
5A1 0.96 37％
8B11 2.04 59％
8D12 2.02 32％
8A1 1.71 48％
6D5 1.81 46％
8C1 2.36 48％
8F12 >5.4 6.7％
Positive control (FMC 63 WT) >5.4 20％
Negative control >5.4 19％
Based on the above results, 3H4,3C2,3C11,4E10,5A12,8C4,8B6 was selected for combinatorial mutation testing (HCDR 2-rich sequences were not abundant, so 4E10 was still selected even though it had poor competitive activity).
Example 5
FMC63 scFv mutant Kd/Koff assay:
transferring FMC63 scFv mutants (the clone numbers corresponding to the mutants, and the nucleotide sequences and amino acid sequences of scFv corresponding to the clone numbers are shown below, SEQ ID Nos. 25-51 and SEQ ID Nos. 52-78) to pGCIgGH1 (Ji Kai gene) (the vector sequence is shown as SEQ ID No. 133), constructing hIgG1 Fc fusion protein capable of expressing secretory FMC63 scFv mutants, and expressing the hIgG1 Fc fusion protein by using a eukaryotic expression system; purification was performed using proteinA column.
The CD19 antigen was coupled to AR2G chip (Fortie Bio) through amino group, FMC63 scFv mutant was used as mobile phase, and the Ocet Red96 system was used to determine the corresponding scFv Koff rate, and the test results showed that (Table 5), the above scFv mutants all had good affinity to CD19 antigen.
TABLE 5
Figure BDA0002650914510000131
Figure BDA0002650914510000141
The above results indicate that each clone has a Koff value substantially identical to that of FMC63 WT.
Example 6
Acquisition of high affinity scFv:
referring to ELISA binding, competition and Koff test data in examples 1-5, clones were selected in which the corresponding CDRs were aligned and combined by PCR (clone numbers corresponding to the scFv obtained after combination, and nucleotide and amino acid sequences of the scFv corresponding to the clone numbers are shown below, and CDRs in the combined scFv sequence were selected from SEQ ID Nos. 25-51, SEQ ID Nos. 52-78), and screened by ELISA assay for binding of CD19 to scFv 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
Preparation of E.coli periplasm (PPE):
TG1 clones with more repeated CDR mutations are plated, single clones are picked, thalli are collected after IPTG induction and are resuspended in TES (0.2M Tris-HC1,0.5mM EDTA,0.5M sucrose), 1/5 multiplied by TES is added for mixing, the mixture is placed on ice for 30min, supernatant is obtained by centrifugation at 13000rpm, and the supernatant is stored at 4 ℃ after being filtered by a 0.22 mu M filter membrane. The results of ELISA (Experimental reference example 4) performed by diluting the periplasm in a certain ratio (2,4,8, 16, 32, 64, 128) in a CD 19-coated 96-well plate are shown in FIGS. 11-14. From the experimental results, it is seen that combo5, combo7, combo9, combo10, combo14 bound weakly to CD19-his, combo6, combo8, combo11, combo13 bound strongly to CD19 (higher OD450nm light absorption at low dilution) by PPE dilution in combination ELISA experiments, with combo6 binding the highest amount.
Clone number: 3H4
Figure BDA0002650914510000151
Figure BDA0002650914510000152
Figure BDA0002650914510000153
(SEQ ID No.52, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.79)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.106)
Clone number: 3C2
Figure BDA0002650914510000154
Figure BDA0002650914510000161
Figure BDA0002650914510000162
Figure BDA0002650914510000163
(SEQ ID No.53, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHRYYGGSFDMDYWGQGTSVTVSS(SEQ ID No.80)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.107)
Clone number: 3C11
Figure BDA0002650914510000164
Figure BDA0002650914510000165
Figure BDA0002650914510000166
(SEQ ID No.54, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHWYYGGSFAMDYWGQGTSVTVSS(SEQ ID No.81)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.108)
Clone number: 4E10
Figure BDA0002650914510000167
Figure BDA0002650914510000171
Figure BDA0002650914510000172
Figure BDA0002650914510000173
(SEQ ID No.55, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.82)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.109)
Clone number: 3F12
Figure BDA0002650914510000174
Figure BDA0002650914510000175
Figure BDA0002650914510000176
(SEQ ID No.56, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGLIQGSWHTYYGAALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.83)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.110)
Clone number: 8F12
Figure BDA0002650914510000177
Figure BDA0002650914510000181
Figure BDA0002650914510000182
Figure BDA0002650914510000183
(SEQ ID No.57, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.84)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYQTDRKQSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.111)
Clone number: 3H9
Figure BDA0002650914510000184
Figure BDA0002650914510000185
Figure BDA0002650914510000186
(SEQ ID No.58, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSMAMDYWGQGTSVTVSS(SEQ ID No.85)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.112)
Clone number: 5A12
Figure BDA0002650914510000191
Figure BDA0002650914510000192
Figure BDA0002650914510000193
(SEQ ID No.59, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.86)
DIQMTQTTSSLSASLGDRVTISCRASQPIRRYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.113)
Clone number: 8C4
Figure BDA0002650914510000194
Figure BDA0002650914510000195
Figure BDA0002650914510000196
(SEQ ID No.60, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.87)
DIQMTQTTSSLSASLGDRVTISCRASMDIKKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.114)
Clone number: 5G6
Figure BDA0002650914510000201
Figure BDA0002650914510000202
Figure BDA0002650914510000203
(SEQ ID No.61, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.88)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYQTSRLAWGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.115)
Clone number: 8C7
Figure BDA0002650914510000204
Figure BDA0002650914510000205
Figure BDA0002650914510000206
(SEQ ID No.62, wherein the bold underlined portion is the connecting peptide, the connecting peptide is the heavy chain variable region, the connecting peptide is followed by the light chain variable region, the underlined portion is CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3) EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.89)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKAAPLTFGGGTKLEIT(SEQ ID No.116)
Clone number: 8B6
Figure BDA0002650914510000211
Figure BDA0002650914510000212
Figure BDA0002650914510000213
(SEQ ID No.63, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.90)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.117)
Clone number: 8C10
Figure BDA0002650914510000214
Figure BDA0002650914510000215
Figure BDA0002650914510000221
(SEQ ID No.64, wherein the bold underlined portion is a linker peptide preceded by a heavy chain variable region and followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.91)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKGPLTFGGGTKLEIT(SEQ ID No.118)
Clone number: 5A1
Figure BDA0002650914510000222
Figure BDA0002650914510000223
Figure BDA0002650914510000224
(SEQ ID No.65, wherein the bold underlined is a linker peptide, preceded by a linker peptideThe heavy chain variable region is followed by the linker peptide and the light chain variable region is followed by CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.92)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKRAPLTFGGGTKLEIT(SEQ ID No.119)
Clone number: 8B11
Figure BDA0002650914510000225
Figure BDA0002650914510000231
Figure BDA0002650914510000232
(SEQ ID No.66, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.93)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKRGPLTFGGGTKLEIT(SEQ ID No.120)
Clone number: 8D12
Figure BDA0002650914510000233
Figure BDA0002650914510000234
Figure BDA0002650914510000235
(SEQ ID No.67, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.94)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKTAPLTFGGGTKLEIT(SEQ ID No.121)
Clone number: 8A1
Figure BDA0002650914510000236
Figure BDA0002650914510000241
Figure BDA0002650914510000242
Figure BDA0002650914510000243
(SEQ ID No.68, wherein the bold underlined portion is a linker peptide preceded by a heavy chain variable region and followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.95)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKTGPLTFGGGTKLEIT(SEQ ID No.122)
Clone number: 6D5
Figure BDA0002650914510000244
Figure BDA0002650914510000245
Figure BDA0002650914510000246
(SEQ ID No.69, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.96)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKTLPFTFGGGTKLEIT(SEQ ID No.123)
Clone number: 8C1
Figure BDA0002650914510000247
Figure BDA0002650914510000251
Figure BDA0002650914510000252
Figure BDA0002650914510000253
(SEQ ID No.70, wherein the bold underlined portion is a linker peptide preceded by a heavy chain variable region and followed by a linker peptideThe underlined parts in the light chain variable region are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3 in that order)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.97)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNKAPLTFGGGTKLEIT(SEQ ID No.124)
Clone number: combo-5
Figure BDA0002650914510000254
Figure BDA0002650914510000255
Figure BDA0002650914510000256
(SEQ ID No.71, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHRYYGGSFDMDYWGQGTSVTVSS(SEQ ID No.98)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.125)
Clone number: combo-6
Figure BDA0002650914510000257
Figure BDA0002650914510000261
Figure BDA0002650914510000262
Figure BDA0002650914510000263
(SEQ ID No.72, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHRYYGGSFDMDYWGQGTSVTVSS(SEQ ID No.99)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.126)
Clone number: combo-7
Figure BDA0002650914510000264
Figure BDA0002650914510000265
Figure BDA0002650914510000266
(SEQ ID No.73, wherein the bold underlined portion is a linker peptide preceded by a heavy chain variable region and followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGGIIGSQTTYYLAGLKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHWYYGGSFAMDYWGQGTSVTVSS(SEQ ID No.100)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.127)
Clone number: combo-8
Figure BDA0002650914510000271
Figure BDA0002650914510000272
Figure BDA0002650914510000273
(SEQ ID No.74, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGWSLEDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHWYYGGSFAMDYWGQGTSVTVSS(SEQ ID No.101)
DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT(SEQ ID No.128)
Clone number: combo-9
Figure BDA0002650914510000274
Figure BDA0002650914510000275
Figure BDA0002650914510000276
(SEQ ID No.75, wherein the bold underlined portion is a linker peptide preceded by a heavy chain variable region and followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.102)
DIQMTQTTSSLSASLGDRVTISCRASQPIRRYLNWYQQKPDGTVKLLIYQTSRLAWGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.129)
Clone number: combo-10
Figure BDA0002650914510000281
Figure BDA0002650914510000282
Figure BDA0002650914510000283
(SEQ ID No.76, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.103)
DIQMTQTTSSLSASLGDRVTISCRASQPIRRYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.130)
Clone number: combo-13
Figure BDA0002650914510000284
Figure BDA0002650914510000285
Figure BDA0002650914510000286
(SEQ ID No.77, wherein the bold underlined is a linker peptide, ligationThe heavy chain variable region is in front of the peptide, the light chain variable region is in back of the connecting peptide, and the underlined parts are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 in sequence)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.104)
DIQMTQTTSSLSASLGDRVTISCRASMDIKKYLNWYQQKPDGTVKLLIYQTSRLAWGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.131)
Clone number: combo-14
Figure BDA0002650914510000291
Figure BDA0002650914510000292
Figure BDA0002650914510000293
(SEQ ID No.78, wherein the bold underlined portion is a linker peptide, the linker peptide is preceded by a heavy chain variable region, the linker peptide is followed by a light chain variable region, and the underlined portions are CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, CDR-L3, respectively)
EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQSPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(SEQ ID No.105)
DIQMTQTTSSLSASLGDRVTISCRASMDIKKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGKKAPLTFGGGTKLEIT(SEQ ID No.132)
The vector pGCIgGH1 full sequence:
AgtgGGAATTGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTGTCGTGAGGAAGATCTCTAGAAGCTGGGTACCTTGTGCCCGGGCGCCACCATGGAGTTTGGGCTGAGCTGGCTTTTTCTTGTCGCGATTCTTAAGGGTGTCCAGTGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGAGCGGCCGCTCGAGGCCGGCAAGGCCGGATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTATGGCTGATTATGATCCGGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGATACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGAGGTCGACTCTAGAGGATCGATCCCCGCCCCGGACGAACTAAACCTGACTACGACATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCTTCAGTTGGTTGGTACAACTTGCCAACTGGGCCCTGTTCCACATGTGACACGGGGGGGGACCAAACACAAAGGGGTTCTCTGACTGTAGTTGACATCCTTATAAATGGATGTGCACATTTGCCAACACTGAGTGGCTTTCATCCTGGAGCAGACTTTGCAGTCTGTGGACTGCAACACAACATTGCCTTTATGTGTAACTCTTGGCTGAAGCTCTTACACCAATGCTGGGGGACATGTACCTCCCAGGGGCCCAGGAAGACTACGGGAGGCTACACCAACGTCAATCAGAGGGGCCTGTGTAGCTACCGATAAGCGGACCCTCAAGAGGGCATTAGCAATAGTGTTTATAAGGCCCCCTTGTTAACCCTAAACGGGTAGCATATGCTTCCCGGGTAGTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGCTATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATATCTCCCACCCCATGAGCTGTCACGGTTTTATTTACATGGGGTCAGGATTCCACGAGGGTAGTGAACCATTTTAGTCACAAGGGCAGTGGCTGAAGATCAAGGAGCGGGCAGTGAACTCTCCTGAATCTTCGCCTGCTTCTTCATTCTCCTTCGTTTAGCTAATAGAATAACTGCTGAGTTGTGAACAGTAAGGTGTATGTGAGGTGCTCGAAAACAAGGTTTCAGGTGACGCCCCCAGAATAAAATTTGGACGGGGGGTTCAGTGGTGGCATTGTGCTATGACACCAATATAACCCTCACAAACCCCTTGGGCAATAAATACTAGTGTAGGAATGAAACATTCTGAATATCTTTAACAATAGAAATCCATGGGGTGGGGACAAGCCGTAAAGACTGGATGTCCATCTCACACGAATTTATGGCTATGGGCAACACATAATCCTAGTGCAATATGATACTGGGGTTATTAAGATGTGTCCCAGGCAGGGACCAAGACAGGTGAACCATGTTGTTACACTCTATTTGTAACAAGGGGAAAGAGAGTGGACGCCGACAGCAGCGGACTCCACTGGTTGTCTCTAACACCCCCGAAAATTAAACGGGGCTCCACGCCAATGGGGCCCATAAACAAAGACAAGTGGCCACTCTTTTTTTTGAAATTGTGGAGTGGGGGCACGCGTCAGCCCCCACACGCCGCCCTGCGGTTTTGGACTGTAAAATAAGGGTGTAATAACTTGGCTGATTGTAACCCCGCTAACCACTGCGGTCAAACCACTTGCCCACAAAACCACTAATGGCACCCCGGGGAATACCTGCATAAGTAGGTGGGCGGGCCAAGATAGGGGCGCGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTTGGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCATGCATATACAGTCAGCATATGATACCCAGTAGTAGAGTGGGAGTGCTATCCTTTGCATATGCCGCCACCTCCCAAGGGGGCGTGAATTTTCGCTGCTTGTCCTTTTCCTGCTGCTTATCGATGATAAGCTGTCAAACATGAGAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGAAGCTGTCCCTGATGGTCGTCATCTACCTGCCTGGACAGCATGGCCTGCAACGCGGGCATCCCGATGCCGCCGGAAGCGAGAAGAATCATAATGGGGAAGGCCATCCAGCCTCGCGTCGCGAACGCCAGCAAGACGTAGCCCAGCGCGTCGGCCCCGAGATGCGCCGCGTGCGGCTGCTGGAGATGGCGGACGCGATGGATATGTTCTGCCAAGGGTTGGTTTGCGCATTCACAGTTCTCCGCAAGAATTGATTGGCTCCAATTCTTGGAGTGGTGAATCCGTTAGCGAGGTGCCGCCCTGCTTCATCCCCGTGGCCCGTTGCTCGCGTTTGCTGGCGGTGTCACTGGCCCCGTGGGTTAGGGACGGGGTCCCCCATGGGGAATGGTTTATGGTTCGTGGGGGTTATTATTTTGGGCGTTGCGTGGGGTCAGGTCCACGACTGGACTGAGCAGACAGACCCATGGTTTTTGGATGGCCTGGGCATGGACCGCATGTACTGGCGCGACACGAACACCGGGCGTCTGTGGCTGCCAAACACCCCCGACCCCCAAAAACCACCGCGCGGATTTCTGGCGTGCCAAGCTAGTCGACCAATTCTCATGTTTGACAGCTTATCATCGCAGATCCGGGCAACGTTGTTGCCATTGCTGCAGGCGCAGAACTGGTAGGTATGGAAGATCT(SEQ ID No.133)
example 7
Engineering high affinity scFv to CAR:
construction of the scFv-BBz vector:
the FMC63-BBz chimeric antigen receptor is mutated according to CDR mutated amino acids by referring to a scFv sequence shown in SEQ ID No.25-132, specifically, the scFv sequence is connected with a CD8 alpha-4-1 BB-CD3zeta by a PCR method to obtain the mutated FMC63 Mutant-BBz chimeric antigen receptor, the scFv-BBz is inserted into a GV400 vector (between BamHI and EcoRI, ji Kai gene, a vector schematic diagram is shown in FIG. 15) between an EF-1 promoter and a WPRE structure by a standard molecular biology method, and the following clones are selected: 3F12,5A12,5G6,8C4,8F12,8D12 and Combo6.
Constructing scFv-BBz-P2A-EGFP vector:
the scFv-BBz mutant obtained after mutation was inserted between the EF-1 promoter and the P2A structure (ATNFSLLKQAGDVEENPGP, SEQ ID No. 134) in the GV401 vector (between BamHI and EcoRI, ji Kai gene, the vector schematic diagram is shown in FIG. 16) by standard molecular biology methods.
Example 8
FMC63 Mutant-BBz in vitro validation
Packaging FMC63 Mutant-BBz obtained by construction of each scFv sequence into Lentivirus (Lentivirus), and infecting human primary PBMC; infected T cells are expanded and cultured in vitro, and whether the function of the FMC63mutation for killing the tumor cells can be enhanced or not is evaluated through cytokine release and killing experiments, and specific results are as follows:
and (3) detecting infection efficiency:
after the GV401 shuttle vector is packaged into lentivirus, the human T cells are infected, scFv-BBz-2A-EGFP is transcribed into mRNA, the mRNA is translated by ribosome, and the two independent protein chains are formed by the breakage between glycine and auxiliary amino acid at the tail end of a 2A sequence (ATNFSLLKQAGDVEENPGP, SEQ ID No. 135), so that the expression mole numbers of scFv-BBz and EGFP protein are kept consistent, and the expression efficiency of EGFP can be detected by detecting an FITC channel through a flow cytometer, which represents the infection efficiency of the lentivirus.
Cytokine release assay:
human PBMCs were activated with CD3 and CD28 antibodies (OKT 3 clone and 15E8 clone, miltenyi Biotec) for 24 hours, after packaging FMC63 Mutant-BBz obtained from each scFv sequence construction into Lentivirus (Lentivirus) the PBMCs were infected, infected to express anti-CD 19 chimeric antigen receptor, after PBMCs continued to be cultured for 8-10 days, T cells were collected, and T cells and Raji cells (expressing CD 19) were cultured in the ratio 1:1 Mixed culture in 2% serum RPMI1640 medium for 16 hours the mixed culture supernatants were assayed for cytokine release using the BD Cytometric bead array kit. Experiments show that PBMC obtained by infection (FMC 63 Mutant-BBz obtained by construction of each scFv sequence is packaged into a lentivirus infection to obtain, and expresses FMC63 Mutant-BBz chimeric antigen receptor) has strong binding capacity with Raji cells and can release cytokines compared with FMC63 WT-BBz. The infection efficiency of each clone was as follows:
TABLE 7
Cloning The 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％
The cytokine release fold changes for each clone were as follows:
TABLE 8
Cloning Multiple of IL2 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
Watch 10
Cloning Multiple of IL2 IFN gamma multiple
FMC63 WT 123 109
5G6 87 107
8C4 73 98
Cytokine release experimental results show that 8D12 and 3F12 clones did not significantly release cytokines upon tumor cell stimulation, suggesting that 8D12 and 3F12 clonal mutations are detrimental to T cell binding to CD19 expressing tumor cells.
The 5a12 clone released higher levels of cytokines IL2 and IFN γ at infection efficiencies substantially consistent with FMC63WT (26.6% vs 26.9%).
Combo6 releases IL2 levels significantly higher than FMC63WT at lower infection efficiency.
Mutations in 5a12 (CDRL 1), 3H4 (CDRH 1), 3C2 (CDRH 3) are suggested to favor increased cytokine release levels for CD19 binding.
Tumor killing experiment:
the T cells obtained from the above preparation (T cells were collected after PBMC were further cultured for 8-10 days) and Raji cells were cultured according to a certain E: t ratio, for example 30:1,10: 1,3:1,1:1 proportion of the culture medium was mixed and cultured in 2% serum RPMI1640 medium for 4 hours, and the culture supernatant was mixed with LDH substrate (CytoTox 96 Non-Radioactive Cytoaxity Assay Kit, promega) 1:1 volume mixed and incubated at room temperature for 30 minutes before reading the 490nm light absorption. Experimental results show that PBMC obtained by infection (obtained by packaging FMC63 Mutant-BBz obtained by construction of each scFv sequence into lentivirus infection) has strong tumor killing capacity compared with FMC63 WT-BBz.
Cell killing experimental results (fig. 17-19) indicate that 3F12 clone has no significant ability to kill tumor cells, suggesting that 3F12 mutation is not favorable for binding CD19. Compared with FMC63WT, the 8F12 and 5A12 clones have no significant difference in killing capacity to tumor cells. Compared with FMC63WT, the Combo6 clone has a significant improvement in tumor cell killing. The summarized experimental results are shown in fig. 20, and compared with FMC63WT, the Combo6 clone has significantly improved tumor cell killing under the condition that the ratio of E to T is 10 and 3, which indicates that the Combo6 combinatorial mutation is beneficial to T cell recognition of tumor cells.
Example 9
FMC63 Mutant-BBz in vitro validation
After FMC63 Mutant-BBz obtained from each scFv sequence construction was packaged as Lentivirus (Lentivirus), NK92 cells were infected (culture conditions: RPMI1640+20% FBS +200IU/ml hIL 2); the infected NK92 cells are amplified and cultured in vitro, and whether the function of the FMC63mutation for killing tumor cells can be enhanced by the NK92 cells is evaluated through cytokine release and killing experiments, and the specific results are as follows:
cytokine release assay:
NK92 cell line was cultured in RPMI1640+20% FBS +200IU/ml hIL 2; packaging FMC63 Mutant-BBz into lentivirus, infecting NK92 cells, and performing amplification and culture in vitro; NK92 cells and Raji cells (expressing CD 19) were mixed in a ratio of 1:1 Mixed culture in 2% serum RPMI1640 medium for 16 hours, and the mixed culture supernatant using BD Cytometric bead array kit determination of cytokine release. Experiments show that NK92 obtained by infection (FMC 63 Mutant-BBz obtained by construction of each scFv sequence is packaged into lentivirus infection to obtain the expression of FMC63 Mutant-BBz chimeric antigen receptor) has stronger binding capacity with Raji cells and can release cytokines compared with FMC63 WT-BBz.
Tumor killing experiment:
the NK92 cells and Raji cells obtained by the preparation are mixed according to a certain E: t ratio, for example 30:1,10: 1,3:1,1:1 ratio mixed culture in 2% serum RPMI1640 medium for 4 hours, and mixing the culture supernatant with LDH substrate (CytoTox 96 Non-Radioactive Cytoxicity Assay Kit, promega) 1:1 volume mixed and incubated at room temperature for 30 minutes before reading the 490nm light absorption. Experimental results show that NK92 cells obtained by infection (obtained by packaging FMC63 Mutant-BBz obtained by construction of each scFv sequence into lentivirus infection) have strong tumor killing capacity compared with FMC63 WT-BBz.
In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Sequence listing
<110> GbI Biotechnology Co., ltd
<120> anti-CD 19 antibody, preparation method and use thereof
<150> 2016108048313
<151> 2016-09-06
<150> 2016108335750
<151> 2016-09-20
<160> 135
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Gly Trp Ser Leu Glu Asp Tyr Gly Val Ser
1 5 10
<210> 2
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<400> 3
His Arg Tyr Tyr Gly Gly Ser Phe Asp Met Asp Tyr
1 5 10
<210> 4
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Arg Ala Ser Gln Asp Ile Ser Lys Tyr Leu Asn
1 5 10
<210> 5
<211> 7
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 5
Gln Thr Ser Arg Leu Ala Trp
1 5
<210> 6
<211> 7
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 6
Gln Thr Asp Arg Lys Gln Ser
1 5
<210> 7
<211> 7
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 7
His Thr Ser Arg Leu His Ser
1 5
<210> 8
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 8
Gln Gln Gly Asn Thr Leu Pro Tyr Thr
1 5
<210> 9
<211> 726
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<400> 11
tcacatgcac tgtctcaggg gtctcattac ccgactatgg tgtaagctgg attcgccagc 60
ctccacg 67
<210> 12
<211> 79
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 12
agggtctgga gtggctggga gtaatatggg gtagtgaaac cacatactat aattcagctc 60
tcaaatccag actgaccat 79
<210> 13
<211> 72
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 13
ccatttacta ctgtgccaaa cattattact acggtggtag ctatgctatg gactactggg 60
gccaaggaac ct 72
<210> 14
<211> 72
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 14
acagagtcac catcagttgc agggcaagtc aggacattag taaatattta aattggtatc 60
agcagaaacc ag 72
<210> 15
<211> 61
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 15
ctgttaaact cctgatctac catacatcaa gattacactc aggagtccca tcaaggttca 60
g 61
<210> 16
<211> 64
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 16
ttgccactta cttttgccaa cagggtaata cgcttccgta cacgttcgga ggggggacca 60
agct 64
<210> 17
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 17
ccctcatagt tagcgtaacg 20
<210> 18
<211> 24
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 18
agcggataac aatttcacac agga 24
<210> 19
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 19
ccctgagaca gtgcatgtga 20
<210> 20
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 20
tcccagccac tccagaccct 20
<210> 21
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 21
tttggcacag tagtaaatgg 20
<210> 22
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 22
gcaactgatg gtgactctgt 20
<210> 23
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 23
gtagatcagg agtttaacag 20
<210> 24
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 24
ttggcaaaag taagtggcaa 20
<210> 25
<211> 726
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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 Sequence (Artificial Sequence)
<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, the complementarity determining region of the heavy chain variable region of the anti-CD 19 antibody 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; the complementarity determining region of the light chain variable region of the anti-CD 19 antibody comprises CDR-L1 with an amino acid sequence shown in SEQ ID No.4, CDR-L2 with an amino acid sequence shown in SEQ ID No. 7and CDR-L3 with an amino acid sequence shown in SEQ ID No. 8.
2. The anti-CD 19 antibody of 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 comprises the amino acid sequence shown as SEQ ID No. 99;
and/or, the amino acid sequence of the light chain variable region of the anti-CD 19 antibody comprises the amino acid sequence shown as SEQ ID No. 126;
and/or the amino acid sequence of the anti-CD 19 antibody is shown as SEQ ID NO. 72.
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 antibody expression system comprising the construct or genome of claim 4 having integrated therein an exogenous polynucleotide according to claim 3.
6. A method of producing 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 therapeutic agent.
8. An isolated polypeptide comprising a transmembrane domain, an intracellular domain and an extracellular domain, the extracellular domain comprising the anti-CD 19 antibody of any one of claims 1-2.
9. The polypeptide of claim 8, wherein said polypeptide is a chimeric antigen receptor;
and/or, the transmembrane domain comprises CD 8a, CD28, or DAP10;
and/or, the endodomain comprises 4-1BB, CD28, OX40, ICOS, CD3zeta, or 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 in sequence.
10. A cell comprising a membrane-bound polypeptide according to any one of claims 8-9, said cell being a T lymphocyte and/or an NK cell;
and/or, the polypeptide is a chimeric antigen receptor;
and/or, when said polypeptide binds to a CD19 antigen, said T lymphocytes and/or NK cells can be activated and/or stimulated to proliferate;
and/or, the T lymphocytes and/or NK cells express the anti-CD 19 antibody on the surface.
11. A diagnostic kit comprising a diagnostically effective amount of an anti-CD 19 antibody or immunoconjugate according to any one of claims 1-2.
CN202010870418.3A 2016-09-06 2017-09-06 anti-CD 19 antibody and preparation method and application thereof Active CN111848801B (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
CN201610804831 2016-09-06
CN2016108048313 2016-09-06
CN2016108335750 2016-09-20
CN201610833575 2016-09-20
CN201710793590.1A CN107793479B (en) 2016-09-06 2017-09-06 An anti-CD19 antibody and its preparation method and use

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN201710793590.1A Division CN107793479B (en) 2016-09-06 2017-09-06 An anti-CD19 antibody and its preparation method and use

Publications (2)

Publication Number Publication Date
CN111848801A CN111848801A (en) 2020-10-30
CN111848801B true CN111848801B (en) 2022-10-11

Family

ID=61531889

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202010870418.3A Active CN111848801B (en) 2016-09-06 2017-09-06 anti-CD 19 antibody and preparation method and application thereof
CN201710793590.1A Active CN107793479B (en) 2016-09-06 2017-09-06 An anti-CD19 antibody and its preparation method and use

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN201710793590.1A Active CN107793479B (en) 2016-09-06 2017-09-06 An anti-CD19 antibody and its preparation method and use

Country Status (1)

Country Link
CN (2) CN111848801B (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110003338B (en) * 2019-04-16 2021-04-23 北京免疫方舟医药科技有限公司 Anti-OX40 antibody and its application
CN115867581A (en) * 2020-03-25 2023-03-28 北京艺妙神州医药科技有限公司 Chimeric antigen receptors containing protective peptides and uses thereof
CN115515984A (en) * 2020-05-08 2022-12-23 亘喜生物科技（上海）有限公司 A kind of anti-CD19 antibody antibody and its preparation and application
CN111718422A (en) * 2020-05-26 2020-09-29 广州重磅生物科技有限公司 Chimeric antigen receptor targeting CD19 and application thereof
CN111848799B (en) * 2020-07-31 2023-09-12 杭州皓阳生物技术有限公司 Specific detection antibody and application thereof in preparation of detection kit
US20230406922A1 (en) * 2020-11-20 2023-12-21 Simcere Zaiming Pharmaceutical Co., Ltd. Humanized cd19 antibody and use thereof
CN112375146B (en) * 2021-01-07 2021-04-13 北京百普赛斯生物科技股份有限公司 Anti-idiotypic antibody for detecting Anti-CD19 CAR expression level and application thereof
CN112394179B (en) * 2021-01-18 2021-04-09 北京百普赛斯生物科技股份有限公司 FMC63 CAR-T cell immunogenicity ELISA detection kit
TW202448956A (en) 2023-06-09 2024-12-16 大陸商上海藥明巨諾生物醫藥研發有限公司 Fusion protein and medical use thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102209556A (en) * 2008-11-07 2011-10-05 埃姆诺医药有限公司 Modified anti-CD19 antibody
WO2015188135A1 (en) * 2014-06-06 2015-12-10 The California Institute For Biomedical Research Constant region antibody fusion proteins and compositions thereof
CN105330750A (en) * 2015-11-20 2016-02-17 上海细胞治疗研究院 Molecular brake for rapidly stopping killing effect of CAR-T (T cell engineered with chimeric antigen receptors) and application of molecular brake
CN105837689A (en) * 2015-01-13 2016-08-10 博生吉医药科技(苏州)有限公司 Anti-CD19 monoclonal antibody and preparation method thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090220501A1 (en) * 2006-01-30 2009-09-03 Fey Georg H Anti-CD19 Antibody, Immunotoxin and Treatment Method
TWI805109B (en) * 2014-08-28 2023-06-11 美商奇諾治療有限公司 Antibodies and chimeric antigen receptors specific for cd19

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102209556A (en) * 2008-11-07 2011-10-05 埃姆诺医药有限公司 Modified anti-CD19 antibody
WO2015188135A1 (en) * 2014-06-06 2015-12-10 The California Institute For Biomedical Research Constant region antibody fusion proteins and compositions thereof
CN105837689A (en) * 2015-01-13 2016-08-10 博生吉医药科技(苏州)有限公司 Anti-CD19 monoclonal antibody and preparation method thereof
CN105330750A (en) * 2015-11-20 2016-02-17 上海细胞治疗研究院 Molecular brake for rapidly stopping killing effect of CAR-T (T cell engineered with chimeric antigen receptors) and application of molecular brake

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"CD19 与B细胞恶性肿瘤的免疫治疗";刘亚丹 等;《中国肿瘤生物治疗杂志》;20150228;第22卷(第1期);第99-103页 *
"Successful Construction and Massive Expression of a Novel Anti-CD19 Human-Mouse Chimeric Antibody Hm2E8b";Diying Shen et al.;《MONOCLONAL ANTIBODIES IN IMMUNODIAGNOSIS AND IMMUNOTHERAPY》;20141231;第33卷(第4期);第215-220页 *
"T cells expressing CD19/CD20 bi-specific chimeric antigen receptors prevent antigen escape by malignant B cells";Eugenia Zah et al.;《Cancer Immunol Res》;20160408;第4卷(第6期);第498-508页 *

Also Published As

Publication number Publication date
CN107793479B (en) 2024-02-06
CN111848801A (en) 2020-10-30
CN107793479A (en) 2018-03-13

Similar Documents

Publication Publication Date Title
CN107793480B (en) anti-CD 19 antibody and preparation method and application thereof
CN111848801B (en) anti-CD 19 antibody and preparation method and application thereof
CA2436671C (en) Rationally designed antibodies
AU2018289428B2 (en) Methods and compositions for chimeric antigen receptor targeting cancer cells
US20040253242A1 (en) Rationally designed antibodies
KR102502293B1 (en) Plasma kallikrein binding proteins
RU2727451C2 (en) T-cells modified with a chimeric antigen receptor targeting cs1
US6261558B1 (en) Synthetic human neutralizing monoclonal antibodies to human immunodeficiency virus
JP2024150445A (en) Claudin 6 antibodies and methods for treating cancer
KR20070034487A (en) Determination of Anti-Il-13 Antibody, Anti-Il-13 Antibody and Complex Containing the Same
KR102102239B1 (en) Method for treating breast cancer
WO1995034664A1 (en) Method of detecting ligand interactions
KR101970428B1 (en) Recombinant viral Viral serine Vaccine
KR101654890B1 (en) Repebody Against Immunoglobulin G and Uses Thereof
KR101572219B1 (en) Method for Improving Repebody Containing Repeat Module
KR20220107233A (en) Method of obtaining CAR-NK cells
HK1248245A1 (en) Anti-cd19 antibody, preparation method and usage thereof
HK1248245B (en) Anti-cd19 antibody, preparation method and usage thereof
CN110857320A (en) Antibodies capable of binding to norovirus, composite materials, detection devices, and methods using the same
CN110857321B (en) Antibodies capable of binding norovirus, composite materials using same, detection devices and methods
RU2745452C2 (en) Only-hccd r 3 libraries with reduced combinatorial redundancy and optimized loop length distribution
CN114621929B (en) Antitumor dendritic cell, preparation method thereof, expression vector and application
CN110461323A (en) Oncolytic virus propagation method and antitumor agent
HK40069346A (en) Cs1 targeted chimeric antigen receptor-modified t cells
HK40031291A (en) Methods of treating triple negative breast cancer and basal-like breast cancer using inhibitors against kidney associated 1 (kaag1) inhibitor

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant