CN112194728A - Chimeric antigen receptor for endotoxin removal and uses thereof - Google Patents

Chimeric antigen receptor for endotoxin removal and uses thereof Download PDF

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CN112194728A
CN112194728A CN202011000494.5A CN202011000494A CN112194728A CN 112194728 A CN112194728 A CN 112194728A CN 202011000494 A CN202011000494 A CN 202011000494A CN 112194728 A CN112194728 A CN 112194728A
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CN112194728B (en
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胡适
傅文燕
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Fengchao Medical Technology Shanghai Co ltd
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Abstract

The present invention provides a chimeric antigen receptor polypeptide for endotoxin clearance having (a) an extracellular domain comprising one or more endotoxin-specific binding regions; (b) a transmembrane domain; (c) at least one copy of an intracellular signaling domain of a phagocyte-specific phagocytic signal receptor. Wherein the endotoxin-specific binding region comprises: (1) an extracellular domain of a pattern recognition receptor or a functional variant or fragment thereof; (2) an endotoxin-specific antibody or a functional fragment thereof. Further, the invention also provides polynucleotides encoding the polypeptides, genetic vectors, recombinant cells carrying chimeric antigen receptors for endotoxin clearance, and compositions. The chimeric polypeptide aiming at the endotoxin provided by the invention has the advantages that the extracellular domain can specifically recognize the endotoxin, the intracellular signal domain can activate and mediate cell phagocytosis to clear the endotoxin, and the process does not mediate the release of cytokines and does not mediate tissue damage.

Description

Chimeric antigen receptor for endotoxin removal and uses thereof
Technical Field
The invention relates to a design scheme and application of a Chimeric Antigen Receptor (CAR) for endotoxin removal, wherein the CAR specifically binds pathogenic bacteria endotoxin, and simultaneously activates phagocyte-specific phagocytic signals in cells of the CAR to mediate phagocytic recombinant cells to phagocytize and remove the endotoxin.
Background
Endotoxin refers to the toxin factor released after rupture of gram-negative bacteria and is the product of the cell wall of gram-negative bacteria. It is not released from pathogenic microorganisms without rupture of the bacteria, but only after death and lysis of the bacteria or artificial destruction of the bacterial cells, and is called endotoxin. Endotoxin is toxic to a host, when a patient suffers infection, massive cell death and destruction are caused due to the action of antibiotics, immune cells and the like, massive endotoxin enters blood, immune cells are continuously activated, and immune system disorder and various pathophysiological disorders of the patient are caused.
Disclosure of Invention
The present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a chimeric antigen receptor for endotoxin removal and use thereof.
In a first aspect of the invention, a chimeric polypeptide is disclosed, comprising: (a) an extracellular domain comprising one or more endotoxin-specific binding regions; (b) a transmembrane domain; (c) the phagocytic cell-specific phagocytes at least one copy of an intracellular signaling domain of a signaling receptor.
In some cases, the chimeric polypeptide further comprises a signal peptide derived from a signal peptide of the transmembrane domain, which can be genetically engineered to be linked upstream of the extracellular domain, e.g., the N-terminus of the polypeptide chain.
In the extracellular domain, the endotoxin-specific binding domain comprises: (1) an extracellular domain of a pattern recognition receptor or a functional variant or fragment thereof; (2) an endotoxin-specific antibody or a functional fragment thereof. The antibody or functional fragment thereof includes an antigen binding fragment (Fab), a single chain variable fragment (scFv), a nanobody, a VH domain, a VL domain, a single domain antibody (sdAb), a VNAR domain and a VHH domain, a bispecific antibody, a diabody (diabody), or a functional fragment of the above.
The term "pattern recognition receptor" is an immunological concept, and a Pattern Recognition Receptor (PRR) is a type of molecular pattern (PAMP) that is mainly expressed on the surface of innate immune cells, is non-clonally distributed, and can recognize one or more pathogen-associated molecules. Is representative of immune receptors in innate immunity, encoded by a limited number of germline genes, which are evolutionarily well conserved, and also suggests that such receptors are of great importance to the survival of organisms. Its recognition and interaction with PAMPs is critical for the initiation of innate immune responses. Any pattern recognition receptor is suitable for use in the structural scheme of the chimeric polypeptides of the invention.
In certain preferred embodiments of the invention, the pattern recognition receptor comprises TLR1(Gene ID:7096), TLR2(Gene ID:7097), TLR3(Gene ID:7098), TLR4(Gene ID:7099), TLR5(Gene ID:7100), TLR6(Gene ID:10333), TLR7(Gene ID:51284), TLR8(Gene ID:51311), TLR9(Gene ID:54106), TLR10(Gene ID:81793), Dectin-1(Gene ID:64581), Dectin-2(Gene ID:93978), Mincle (Gene ID:26253), CLEC2(Gene ID:51266), CLEC5A (Gene ID:23601), CLEC12A (Gene ID:160364), DCIR (Gene ID: 505085), CLSF 8(Gene ID: 58643), and LBP 2329 (Gene ID: 9239), and LBID: LBP 856).
In certain preferred embodiments of the invention, the endotoxin-specific antibody comprises WN1222-5, S25-39, Ab-52, S55-3, S1-15 or S25-23.
In the transmembrane domain, the transmembrane domain is derived from the stem (stalk) and/or transmembrane domain of CD8, TYRO3, AXL, MERK, ADGRB1, ADGRB3, HAVCR1, TIMD4, Stablin-1, Stablin-2, Integrin subbunit α v, Integrin subbunit β 3, Integrin subbunit β 5, SCARF1, AGER, CD300LF, CD36, LRP1, MRC1, or TREM 2. In some embodiments, the transmembrane domain may be linked downstream of the extracellular domain and upstream of the intracellular signaling domain by genetic engineering methods.
Signal peptides include those derived from CD8, TYRO3, AXL, MERK, ADGRB1, ADGRB3, HAVCR1, TIMD4, Stablin-1, Stablin-2, Integrinsubbunit α v, Integrinsubbunit β 3, Integrinsubbunit β 5, SCARF1, AGER, CD300LF, CD36, LRP1, MRC1, and TREM 2.
Among intracellular domains, the intracellular signaling domain of a phagocyte-specific phagocytic signal receptor refers to a phagocytic signal receptor specific to phagocytes, such as monocytes, which naturally mediates phagocytic events of phagocytes. In some embodiments, the phagocyte-specific phagocytic signal receptor comprises a TAM family receptor tyrosine kinases (TAM family of receptor tyrosine kinases), a Phosphatidylserine receptor family (phospholipidase receptors), a low-density lipoprotein receptor-related protein family (low-density lipoprotein receptor-related proteins), a mannose receptor family (mannose receptors), a scavenger receptor family (scavenger receptors), a TREM receptor family (triggered on myoelectric cells). Preferably, TYRO3 (Uniport: Q06418), AXL (Uniport: P30530), MERRTK (Uniport: Q12866), ADGRB1 (Uniport: O14514), ADGRB3 (Uniport: O60242), HAVCR1 (Uniport: Q96D42), TIMD4 (Uniport: Q96H15), Stablin-1 (Uniport: Q9NY15), Stablin-2 (Uniport: Q8WWQ8), Integrin subBunt α v (Uniport: P06756), Integrin subBunt β 3 (Uniport: P05106), Integrin subBunt β 5 (Uniport: P18084), RF1 (Uniport: Q62), AGER (Uniport: Q0609), CD300 (Uniport: 82300), AXL (Uniport: P3058), ZC 0719, ZC 369611, Uniport: MRT 369611, Uniport: CD 898, and ZC 369611 (Uniport: CD 8911).
The intracellular signaling domain of phagocyte-specific phagocytic signal receptors does not contain immunoreceptor tyrosine-based activation motifs (ITAMs). Most preferred are TYRO3 (Uniport: Q06418), AXL (Uniport: P30530), and MERRTK (Uniport: Q12866), which belong to the TAM receptor family and do not contain the ITAM motif.
In some embodiments of the present disclosure, preferred structures of the chimeric polypeptide include: (a) a signal peptide; (b) an extracellular domain comprising one or more endotoxin-specific binding regions; (c) a transmembrane domain; (d) at least one copy of an intracellular signaling domain from a phagocyte-specific phagocytic signal receptor. Preferably, the chimeric polypeptide comprises: (a) a signal peptide from CD 8; (b) an extracellular domain of a pattern recognition receptor; (c) a transmembrane domain from CD 8; (d) intracellular signaling domains from TYRO3, AXL, merk.
In certain preferred embodiments of the invention, the amino acid sequence of the chimeric polypeptide is as shown in SEQ ID Nos. 1-8.
Accordingly, in a second aspect of the invention, there is provided a polynucleotide encoding the chimeric polypeptide described above, a gene vector carrying the polynucleotide, and a recombinant cell comprising such a gene vector.
The expression vector provided by the invention comprises the following elements which can be connected by a genetic engineering method: a transcription promoter, a DNA region encoding a suitable signal peptide, a DNA region encoding the chimeric polypeptide described above, and a transcription terminator. Vectors are well known to those skilled in the art, such as viral vectors [ Morstutl, et al. cell,2016,164(4): 780-.
In some embodiments, the recombinant cell is a prokaryotic cell or a eukaryotic cell. In some embodiments, the recombinant cell is a human cell. In some embodiments, the recombinant cell is an immune cell. In some embodiments, the recombinant cell is a phagocytic cell. In some embodiments, the phagocytic function cells include macrophages, such as dendritic cells, mast cells, monocytes, neutrophils, microglia, and astrocytes. In some embodiments, the phagocytic cell is a bone marrow-derived macrophage (BMDM) or a bone marrow-derived dendritic cell (BMDC).
In treating a subject having the disease, an effective amount of recombinant phagocytic cells that have been modified to express a chimeric polypeptide having specific affinity for endotoxin are administered to the subject. Preferably, the phagocytic cells are derived from the same individual to whom the recombinant cells are administered.
When constructing the gene vector and the recombinant cell, it is preferable that the recombinant cell simultaneously expresses at least two chimeric polypeptides having different endotoxin-specific binding regions. Experiments prove that compared with a recombinant cell carrying only one chimeric polypeptide, when the recombinant cell simultaneously expresses at least two chimeric polypeptides with different endotoxin-specific binding regions, the phagocytosis and the elimination of the endotoxin by the recombinant cell can be obviously improved. The combined use of recombinant cells carrying a single chimeric polypeptide does not have the technical effect described above.
In a third aspect of the invention, there is also provided a method for genetically modifying a cell. The method comprises contacting (a) a chimeric polypeptide as described herein; and/or (b) introduction of an isolated, synthetic or recombinant polynucleotide molecule as described herein into a cell. In some embodiments, the cell is a phagocytic cell, preferably including macrophages, dendritic cells, mast cells, monocytes, neutrophils, microglia and astrocytes.
In a fourth aspect, the invention also relates to a cell culture comprising at least one recombinant cell as disclosed herein and a culture medium.
In the fifth aspect of the invention, a pharmaceutical composition is also provided. The composition comprises the chimeric polypeptide, the polynucleotide, the vector, the recombinant cell and at least one pharmaceutically acceptable carrier. In addition, the pharmaceutical composition may further comprise a diluent or excipient for medical or pharmaceutical use.
In a sixth aspect of the invention, the use of the chimeric antigen receptor for endotoxin removal as described above in the preparation of a reagent, a kit or a therapeutic drug for diagnosing or treating a disease associated with excessive endotoxin accumulation is provided.
The use comprises the following steps: 1) direct binding to endotoxin, mitigating the hyperactivation of innate immune signals by endotoxin, as described in example 2; 2) inducing cells to phagocytose directly to clear endotoxin as described in examples 2-5; 3) compared with current universal CARs, immune cells carrying the CARs do not significantly release pro-and or anti-inflammatory cytokines, inhibit endogenous Toll-like receptor signaling pathways after phagocytosis of endotoxins. As described in examples 4-5. 4) Reducing the over-expression and release of organ inflammation mediators, relieving organ inflammation injury, enhancing the stress resistance of organs, resisting organ injury, reducing the content of endotoxin in blood, and the like, as described in example 5.
Diseases associated with excessive endotoxin accumulation include endotoxemia, Systemic Inflammatory Response Syndrome (SIRS) or sepsis (e.g., from viral, bacterial, fungal or parasitic infection), autoimmune disease, surgery, cytotoxic chemotherapy, bone marrow manipulation, major tissue injury or trauma, mesenteric hypoperfusion, gut mucosal injury, malaria, gastrointestinal inflammatory disease, intestinal infection, uterine infection, influenza, acute pneumonia such as acute respiratory distress syndrome or acute lung injury, pulmonary embolism, pancreatitis, autoimmune and collagen vascular diseases, blood transfusion related diseases, burns, smoke or inhaled lung injury, graft versus host disease, ischemia or infarction, reperfusion injury, hemorrhage, anaphylaxis, drug overdose, radiation injury, or chemical injury. In some embodiments, the inflammatory mediator is produced by a disease caused by a pathogen, toxin, or agent of biological warfare, such as viral hemorrhagic fever, aequorin, hantavirus cardiopulmonary syndrome (hantavirus), cholera toxin, botulinum toxin, grass toxin, qotherm [ bornella burnetii ], plaque-specific typhoid (Rickettsia prowaszekii), or psittacosis [ Chlamydia psittaci (Chlamydia psittaci) ].
In a seventh aspect the invention relates to a method of treating an individual in need of endotoxin removal. The method comprises administering to the individual an effective amount of phagocytic cells that have been modified to express the pro-and polypeptide having specific affinity for an virus. In some embodiments, the cell is a phagocytic cell comprising a macrophage, dendritic cell, mast cell, monocyte, neutrophil, microglial cell, and astrocyte. In some embodiments, the phagocytic cell is bmdm (bone marrow derived macrophage) or bmdc (bone marrow derived dendritic cell). In some embodiments, the phagocytic cells are derived from the same individual in need of endotoxin removal therapy.
Compared with the prior art, the invention has the following technical effects:
the invention provides a chimeric polypeptide aiming at endotoxin, wherein an extracellular domain of the chimeric polypeptide can specifically recognize endotoxin, an intracellular signaling domain of the chimeric polypeptide can activate mediated cell phagocytosis to remove the endotoxin, and the process does not mediate the release of cytokines, does not mediate antigen presentation and does not mediate tissue damage. Namely, the chimeric polypeptide of the present invention achieves the outstanding technical effects that: the chimeric antigen receptor of the invention is capable of specifically mediating binding to endotoxins resulting from bacterial lysis, rather than directly to the pathogen itself; simultaneously, the intracellular signal structural domain of the polypeptide is activated to mediate cell phagocytosis to clear endotoxin; and this process does not mediate the release of cytokines.
In addition, the chimeric antigen receptor-mediated macrophage endotoxin phagocytosis disclosed by the invention does not remarkably generate proinflammatory cytokine release and can directly inhibit endogenous Toll-like signals. Therefore, the technical effect of the chimeric antigen receptor is obviously different from that of the existing CAR technology, and the chimeric antigen receptor has wide application prospect.
Detailed Description
The following examples and experimental examples further illustrate the present invention and should not be construed as limiting the present invention. The examples do not include detailed descriptions of conventional methods such as a commonly used antibody engineering method, those for constructing vectors and plastron, a method of inserting a gene encoding a protein into such vectors and plastron or a method of introducing a plasmid into a host cell, a synthetic cell, a device, a construction method of a gene circuit, and the like. Such methods are well known to those having ordinary skill in the art and are described in a number of publications, including Dong Shi Wei, Wang Yan Shu edition, Beijing medical university Press, 2002; sambrook, J., Fritsch, E.F. and Maniais, T. (1989) Molecular Cloning A Laboratory Manual,2nd edition, Cold spring Harbor Laboratory Press; phase Display: A Laboratory Manual, Cold spring Harbor Laboratory Press.
Example 1 preparation of chimeric polypeptide against endotoxin, recombinant cell
Chimeric polypeptides were designed according to the following structure: (a) a signal peptide from CD 8; (b) an extracellular segment of a pattern recognition receptor or an extracellular single chain antibody variable fragment (scFv) having a specific affinity for an endotoxin; (c) a transmembrane domain from CD 8; (d) intracellular domains from FCER1G, CD3, MEGF10, TYRO3, MERKT, AXL. And a chimeric polypeptide without an intracellular region was used as a control, and a Myc tag was added between the hinge and transmembrane domains of CD8 to facilitate detection. Specific structural information of the chimeric polypeptides is shown in table 1:
TABLE 1 chimeric polypeptide Structure
Serial number Name of Source of extracellular domain Intracellular domains Sequence of
1. TLR4-CARMERTK TLR4 MERTK SEQ ID NO.1
2. TLR4-CARTYPO3 TLR4 TYPO3 SEQ ID NO.2
3. TLR4-CARAXL TLR4 AXL SEQ ID NO.3
4. TLR4-CARζ TLR4 CD3ζ
5. TLR4-CARε TLR4 FCER1G
6. TLR4-CAR△ TLR4 basic linker NHRNRRR
7. TLR1-CARMERTK TLR1 MERTK SEQ ID NO.4
8. TLR1-CARζ TLR1 CD3ζ
9. TLR2-CARMERTK TLR2 MERTK SEQ ID NO.5
10. TLR2-CARζ TLR2 CD3ζ
11. TLR6-CARMERTK TLR6 MERTK
12. TLR6-CARζ TLR6 CD3ζ
13. CD14-CARMERTK CD14 MERTK SEQ ID NO.6
14. CD14-CARζ CD14 CD3ζ
15. MD2-CARMERTK MD2 MERTK
16. MD2-CARζ MD2 CD3ζ
17. LBP-CARMERTK LBP MERTK SEQ ID NO.7
18. LBP-CARζ LBP CD3ζ
19. WN1-CARMERTK WN1222-5 MERTK SEQ ID NO.8
20. WN1-CARζ WN1222-5 CD3ζ
Note: CAR Δ is a control chimeric polypeptide that does not contain an intracellular signaling structure
Polynucleotides encoding the chimeric polypeptides were prepared according to the chimeric polypeptide structure described in table 1 using a whole-gene synthesis approach, adding a Myc tag between the extracellular and transmembrane domains to facilitate detection. The nucleotide encoding the chimeric polypeptide is then integrated into the THP-1 cells using a lentiviral expression system. The synthesis of genes based on the structure of polypeptide chains and the expression of artificial polypeptides on the cell surface using a lentiviral system are common and conventional techniques in this field, and the methods are described in the patent literature (Klichinsky M, et al. Nature Biotechnology,2020: 1-7.). Methods for isolation and gene overexpression of primary macrophages are the same as those in the non-patent literature (Klichinsky M, et al. Nature Biotechnology,2020:1-7.), and synthetic receptor cells are finally obtained.
According to the construction mode of the artificial synthetic receptor, the expression of the extracellular domain and the intracellular domain of the receptor are detected by flow cytometry, and the experimental technique for examining the engineered cell surface receptor by flow cytometry is a general technique in the art, and the results are shown in table 2 with reference to non-patent documents [ Fu W, et al. nature communications,2019,10(1):1-12 ].
TABLE 2 expression of chimeric polypeptides
Figure BDA0002694127060000071
Figure BDA0002694127060000081
These results show that the chimeric polypeptides and recombinant cells were successfully prepared and the receptor expression rate was high.
Example 2 binding of chimeric Polypeptides to E.coli endotoxin
And detecting the binding condition of each cell to the endotoxin of the escherichia coli by using a biotin-streptavidin fluorescence system. This method is a routine test in the art, and is described in non-patent literature [ Wangxiangjun, et al, Chongqing medicine, 1999,028(004): 243-. The results are shown in Table 3:
TABLE 3 endotoxin binding
Figure BDA0002694127060000082
Figure BDA0002694127060000091
As can be seen from Table 3, the chimeric polypeptide had a significant ability to enhance endotoxin binding by macrophages.
Example 3 binding phagocytic Clearance of chimeric Polypeptides to endotoxin
Biotin-labeled endotoxins were used to define the antigenic phagocytosis of chimeric polypeptides and recombinant cells, respectively, according to literature procedures [ Gach J S, et al. Loss cytometry endotoxin phagocytosed by cells was detected using streptavidin containing a fluorescent label, and the results are shown in table 4:
TABLE 4 phagocytosis of endotoxin
Figure BDA0002694127060000092
Figure BDA0002694127060000101
These results show that the chimeric polypeptides described herein mediate phagocytosis of endotoxin by phagocytic cells. And unexpectedly shows enhanced phagocytosis when recombinant cells simultaneously express two chimeric polypeptides having different endotoxin-specific binding regions, which is not the case when recombinant cells expressing a single chimeric polypeptide are used in combination.
Further, using the same experimental method, the level of biotin-labeled endotoxin in the culture broth after 24 hours was evaluated to evaluate the antigen-removing ability of the recombinant cells, and the signal intensity was compared between each treatment group and the control group, and the results are shown in Table 5.
TABLE 5 Effect of treatment groups on endotoxin removal
Figure BDA0002694127060000102
Figure BDA0002694127060000111
These results show that the chimeric polypeptides described herein mediate the clearance of the endotoxin by phagocytes. And unexpectedly shows enhanced endotoxin-clearing when recombinant cells simultaneously express two chimeric polypeptides having different endotoxin-specific binding domains, which is not the case when recombinant cells expressing a single chimeric polypeptide are used in combination.
Example 4 detection of cytokine release and downstream signaling pathway following endotoxin phagocytosis by recombinant cells
The results of examining the cytokines in the cell culture broth after endotoxin phagocytosis in example 3 are shown in tables 6 to 8.
TABLE 6 secretory expression of IL-6 by recombinant cells after phagocytosis of endotoxin
Figure BDA0002694127060000112
Figure BDA0002694127060000121
TABLE 7 secretory expression of IL-8 by recombinant cells after phagocytosis of endotoxin
Figure BDA0002694127060000122
Figure BDA0002694127060000131
TABLE 8 secretory expression of TNF- α by recombinant cells after phagocytosis of endotoxin
Figure BDA0002694127060000132
Figure BDA0002694127060000141
These results show that the chimeric polypeptides and recombinant cells described herein do not release pro-inflammatory cytokines and do not mediate pro-inflammatory responses after phagocytosis of endotoxin. The level of NF- κ B activation downstream of the Toll-like receptor in recombinant cells was further examined as shown in Table 9:
TABLE 9 NF- κ B-activated secretory expression after recombinant cells phagocytose endotoxin
Figure BDA0002694127060000142
Figure BDA0002694127060000151
The results show that chimeric polypeptides and recombinant cells described herein do not significantly activate levels of NF-. kappa.B downstream of Toll-like receptors after phagocytosis of in ovo endotoxin as compared to cells not carrying the chimeric polypeptide. The chimeric polypeptides and recombinant cells described herein are shown to inhibit endogenous model Toll-like receptor signaling.
Example 5 use of recombinant cells in a small animal model of endotoxin
According to the method disclosed by the patent (CN111018999A), a small animal model of cecal ligature perforation sepsis is prepared by using BALB/c and is moldedThe recombinant cells described in the invention were subsequently subjected to a therapeutic study in animal models, 10 mice per group, and the treatment group used the mouse monocyte cell line RAW264.7 carrying the chimeric polypeptide described in the invention. The treatment groups were administered every 6 hours for 2 times on the day of molding, each dose being 1X 106The control group was administered a mouse monocyte cell line (UTD) not carrying recombinant polypeptide, and the blank model group was given only to the cell culture medium. Mice survival was observed and recorded every 12 hours until 7 days post-surgery. Blood samples were taken from the mice 48 hours after the operation and the levels of each marker in each group of plasma were measured using an ELISA kit. The results are shown in tables 10 to 14:
table 10 mouse survival (%)
Figure BDA0002694127060000152
Figure BDA0002694127060000161
TABLE 11 TNF α expression levels
Figure BDA0002694127060000162
TABLE 12 serum SOD Activity of each group
Figure BDA0002694127060000163
Figure BDA0002694127060000171
TABLE 13 serum MDA content (. mu.mol/L) Activity of each group
Figure BDA0002694127060000172
TABLE 14 relative endotoxin content in blood of each group
Figure BDA0002694127060000181
The results prove that the chimeric polypeptide and the recombinant cell have the effects of reducing acute inflammatory exudation, relieving in-vivo inflammatory factor release, relieving tissue injury, enhancing SOD activity, reducing serum MDA content and reducing blood endotoxin level, and can be used for endotoxemia caused by acute tissue injury of viruses. And when the recombinant cell simultaneously expresses two chimeric polypeptides with different endotoxin-specific binding regions, the technical effect is unexpectedly enhanced, but the combined application of the recombinant cells expressing the single chimeric polypeptide does not have the technical effect.
Sequence listing
<110> Feng tide pharmaceutical technology (Shanghai) Co Ltd
<120> chimeric antigen receptor for endotoxin removal and use thereof
<130> specification of claims
<160> 8
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1171
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Glu Ser Trp Glu Pro Cys Val Glu Val Val Pro
20 25 30
Asn Ile Thr Tyr Gln Cys Met Glu Leu Asn Phe Tyr Lys Ile Pro Asp
35 40 45
Asn Leu Pro Phe Ser Thr Lys Asn Leu Asp Leu Ser Phe Asn Pro Leu
50 55 60
Arg His Leu Gly Ser Tyr Ser Phe Phe Ser Phe Pro Glu Leu Gln Val
65 70 75 80
Leu Asp Leu Ser Arg Cys Glu Ile Gln Thr Ile Glu Asp Gly Ala Tyr
85 90 95
Gln Ser Leu Ser His Leu Ser Thr Leu Ile Leu Thr Gly Asn Pro Ile
100 105 110
Gln Ser Leu Ala Leu Gly Ala Phe Ser Gly Leu Ser Ser Leu Gln Lys
115 120 125
Leu Val Ala Val Glu Thr Asn Leu Ala Ser Leu Glu Asn Phe Pro Ile
130 135 140
Gly His Leu Lys Thr Leu Lys Glu Leu Asn Val Ala His Asn Leu Ile
145 150 155 160
Gln Ser Phe Lys Leu Pro Glu Tyr Phe Ser Asn Leu Thr Asn Leu Glu
165 170 175
His Leu Asp Leu Ser Ser Asn Lys Ile Gln Ser Ile Tyr Cys Thr Asp
180 185 190
Leu Arg Val Leu His Gln Met Pro Leu Leu Asn Leu Ser Leu Asp Leu
195 200 205
Ser Leu Asn Pro Met Asn Phe Ile Gln Pro Gly Ala Phe Lys Glu Ile
210 215 220
Arg Leu His Lys Leu Thr Leu Arg Asn Asn Phe Asp Ser Leu Asn Val
225 230 235 240
Met Lys Thr Cys Ile Gln Gly Leu Ala Gly Leu Glu Val His Arg Leu
245 250 255
Val Leu Gly Glu Phe Arg Asn Glu Gly Asn Leu Glu Lys Phe Asp Lys
260 265 270
Ser Ala Leu Glu Gly Leu Cys Asn Leu Thr Ile Glu Glu Phe Arg Leu
275 280 285
Ala Tyr Leu Asp Tyr Tyr Leu Asp Asp Ile Ile Asp Leu Phe Asn Cys
290 295 300
Leu Thr Asn Val Ser Ser Phe Ser Leu Val Ser Val Thr Ile Glu Arg
305 310 315 320
Val Lys Asp Phe Ser Tyr Asn Phe Gly Trp Gln His Leu Glu Leu Val
325 330 335
Asn Cys Lys Phe Gly Gln Phe Pro Thr Leu Lys Leu Lys Ser Leu Lys
340 345 350
Arg Leu Thr Phe Thr Ser Asn Lys Gly Gly Asn Ala Phe Ser Glu Val
355 360 365
Asp Leu Pro Ser Leu Glu Phe Leu Asp Leu Ser Arg Asn Gly Leu Ser
370 375 380
Phe Lys Gly Cys Cys Ser Gln Ser Asp Phe Gly Thr Thr Ser Leu Lys
385 390 395 400
Tyr Leu Asp Leu Ser Phe Asn Gly Val Ile Thr Met Ser Ser Asn Phe
405 410 415
Leu Gly Leu Glu Gln Leu Glu His Leu Asp Phe Gln His Ser Asn Leu
420 425 430
Lys Gln Met Ser Glu Phe Ser Val Phe Leu Ser Leu Arg Asn Leu Ile
435 440 445
Tyr Leu Asp Ile Ser His Thr His Thr Arg Val Ala Phe Asn Gly Ile
450 455 460
Phe Asn Gly Leu Ser Ser Leu Glu Val Leu Lys Met Ala Gly Asn Ser
465 470 475 480
Phe Gln Glu Asn Phe Leu Pro Asp Ile Phe Thr Glu Leu Arg Asn Leu
485 490 495
Thr Phe Leu Asp Leu Ser Gln Cys Gln Leu Glu Gln Leu Ser Pro Thr
500 505 510
Ala Phe Asn Ser Leu Ser Ser Leu Gln Val Leu Asn Met Ser His Asn
515 520 525
Asn Phe Phe Ser Leu Asp Thr Phe Pro Tyr Lys Cys Leu Asn Ser Leu
530 535 540
Gln Val Leu Asp Tyr Ser Leu Asn His Ile Met Thr Ser Lys Lys Gln
545 550 555 560
Glu Leu Gln His Phe Pro Ser Ser Leu Ala Phe Leu Asn Leu Thr Gln
565 570 575
Asn Asp Phe Ala Cys Thr Cys Glu His Gln Ser Phe Leu Gln Trp Ile
580 585 590
Lys Asp Gln Arg Gln Leu Leu Val Glu Val Glu Arg Met Glu Cys Ala
595 600 605
Thr Pro Ser Asp Lys Gln Gly Met Pro Val Leu Ser Leu Asn Ile Thr
610 615 620
Cys Gln Met Asn Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
625 630 635 640
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
645 650 655
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
660 665 670
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
675 680 685
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Val Gln Glu Thr
690 695 700
Lys Phe Gly Asn Ala Phe Thr Glu Glu Asp Ser Glu Leu Val Val Asn
705 710 715 720
Tyr Ile Ala Lys Lys Ser Phe Cys Arg Arg Ala Ile Glu Leu Thr Leu
725 730 735
His Ser Leu Gly Val Ser Glu Glu Leu Gln Asn Lys Leu Glu Asp Val
740 745 750
Val Ile Asp Arg Asn Leu Leu Ile Leu Gly Lys Ile Leu Gly Glu Gly
755 760 765
Glu Phe Gly Ser Val Met Glu Gly Asn Leu Lys Gln Glu Asp Gly Thr
770 775 780
Ser Leu Lys Val Ala Val Lys Thr Met Lys Leu Asp Asn Ser Ser Gln
785 790 795 800
Arg Glu Ile Glu Glu Phe Leu Ser Glu Ala Ala Cys Met Lys Asp Phe
805 810 815
Ser His Pro Asn Val Ile Arg Leu Leu Gly Val Cys Ile Glu Met Ser
820 825 830
Ser Gln Gly Ile Pro Lys Pro Met Val Ile Leu Pro Phe Met Lys Tyr
835 840 845
Gly Asp Leu His Thr Tyr Leu Leu Tyr Ser Arg Leu Glu Thr Gly Pro
850 855 860
Lys His Ile Pro Leu Gln Thr Leu Leu Lys Phe Met Val Asp Ile Ala
865 870 875 880
Leu Gly Met Glu Tyr Leu Ser Asn Arg Asn Phe Leu His Arg Asp Leu
885 890 895
Ala Ala Arg Asn Cys Met Leu Arg Asp Asp Met Thr Val Cys Val Ala
900 905 910
Asp Phe Gly Leu Ser Lys Lys Ile Tyr Ser Gly Asp Tyr Tyr Arg Gln
915 920 925
Gly Arg Ile Ala Lys Met Pro Val Lys Trp Ile Ala Ile Glu Ser Leu
930 935 940
Ala Asp Arg Val Tyr Thr Ser Lys Ser Asp Val Trp Ala Phe Gly Val
945 950 955 960
Thr Met Trp Glu Ile Ala Thr Arg Gly Met Thr Pro Tyr Pro Gly Val
965 970 975
Gln Asn His Glu Met Tyr Asp Tyr Leu Leu His Gly His Arg Leu Lys
980 985 990
Gln Pro Glu Asp Cys Leu Asp Glu Leu Tyr Glu Ile Met Tyr Ser Cys
995 1000 1005
Trp Arg Thr Asp Pro Leu Asp Arg Pro Thr Phe Ser Val Leu Arg Leu
1010 1015 1020
Gln Leu Glu Lys Leu Leu Glu Ser Leu Pro Asp Val Arg Asn Gln Ala
1025 1030 1035 1040
Asp Val Ile Tyr Val Asn Thr Gln Leu Leu Glu Ser Ser Glu Gly Leu
1045 1050 1055
Ala Gln Gly Ser Thr Leu Ala Pro Leu Asp Leu Asn Ile Asp Pro Asp
1060 1065 1070
Ser Ile Ile Ala Ser Cys Thr Pro Arg Ala Ala Ile Ser Val Val Thr
1075 1080 1085
Ala Glu Val His Asp Ser Lys Pro His Glu Gly Arg Tyr Ile Leu Asn
1090 1095 1100
Gly Gly Ser Glu Glu Trp Glu Asp Leu Thr Ser Ala Pro Ser Ala Ala
1105 1110 1115 1120
Val Thr Ala Glu Lys Asn Ser Val Leu Pro Gly Glu Arg Leu Val Arg
1125 1130 1135
Asn Gly Val Ser Trp Ser His Ser Ser Met Leu Pro Leu Gly Ser Ser
1140 1145 1150
Leu Pro Asp Glu Leu Leu Phe Ala Asp Asp Ser Ser Glu Gly Ser Glu
1155 1160 1165
Val Leu Met
1170
<210> 2
<211> 1138
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Glu Ser Trp Glu Pro Cys Val Glu Val Val Pro
20 25 30
Asn Ile Thr Tyr Gln Cys Met Glu Leu Asn Phe Tyr Lys Ile Pro Asp
35 40 45
Asn Leu Pro Phe Ser Thr Lys Asn Leu Asp Leu Ser Phe Asn Pro Leu
50 55 60
Arg His Leu Gly Ser Tyr Ser Phe Phe Ser Phe Pro Glu Leu Gln Val
65 70 75 80
Leu Asp Leu Ser Arg Cys Glu Ile Gln Thr Ile Glu Asp Gly Ala Tyr
85 90 95
Gln Ser Leu Ser His Leu Ser Thr Leu Ile Leu Thr Gly Asn Pro Ile
100 105 110
Gln Ser Leu Ala Leu Gly Ala Phe Ser Gly Leu Ser Ser Leu Gln Lys
115 120 125
Leu Val Ala Val Glu Thr Asn Leu Ala Ser Leu Glu Asn Phe Pro Ile
130 135 140
Gly His Leu Lys Thr Leu Lys Glu Leu Asn Val Ala His Asn Leu Ile
145 150 155 160
Gln Ser Phe Lys Leu Pro Glu Tyr Phe Ser Asn Leu Thr Asn Leu Glu
165 170 175
His Leu Asp Leu Ser Ser Asn Lys Ile Gln Ser Ile Tyr Cys Thr Asp
180 185 190
Leu Arg Val Leu His Gln Met Pro Leu Leu Asn Leu Ser Leu Asp Leu
195 200 205
Ser Leu Asn Pro Met Asn Phe Ile Gln Pro Gly Ala Phe Lys Glu Ile
210 215 220
Arg Leu His Lys Leu Thr Leu Arg Asn Asn Phe Asp Ser Leu Asn Val
225 230 235 240
Met Lys Thr Cys Ile Gln Gly Leu Ala Gly Leu Glu Val His Arg Leu
245 250 255
Val Leu Gly Glu Phe Arg Asn Glu Gly Asn Leu Glu Lys Phe Asp Lys
260 265 270
Ser Ala Leu Glu Gly Leu Cys Asn Leu Thr Ile Glu Glu Phe Arg Leu
275 280 285
Ala Tyr Leu Asp Tyr Tyr Leu Asp Asp Ile Ile Asp Leu Phe Asn Cys
290 295 300
Leu Thr Asn Val Ser Ser Phe Ser Leu Val Ser Val Thr Ile Glu Arg
305 310 315 320
Val Lys Asp Phe Ser Tyr Asn Phe Gly Trp Gln His Leu Glu Leu Val
325 330 335
Asn Cys Lys Phe Gly Gln Phe Pro Thr Leu Lys Leu Lys Ser Leu Lys
340 345 350
Arg Leu Thr Phe Thr Ser Asn Lys Gly Gly Asn Ala Phe Ser Glu Val
355 360 365
Asp Leu Pro Ser Leu Glu Phe Leu Asp Leu Ser Arg Asn Gly Leu Ser
370 375 380
Phe Lys Gly Cys Cys Ser Gln Ser Asp Phe Gly Thr Thr Ser Leu Lys
385 390 395 400
Tyr Leu Asp Leu Ser Phe Asn Gly Val Ile Thr Met Ser Ser Asn Phe
405 410 415
Leu Gly Leu Glu Gln Leu Glu His Leu Asp Phe Gln His Ser Asn Leu
420 425 430
Lys Gln Met Ser Glu Phe Ser Val Phe Leu Ser Leu Arg Asn Leu Ile
435 440 445
Tyr Leu Asp Ile Ser His Thr His Thr Arg Val Ala Phe Asn Gly Ile
450 455 460
Phe Asn Gly Leu Ser Ser Leu Glu Val Leu Lys Met Ala Gly Asn Ser
465 470 475 480
Phe Gln Glu Asn Phe Leu Pro Asp Ile Phe Thr Glu Leu Arg Asn Leu
485 490 495
Thr Phe Leu Asp Leu Ser Gln Cys Gln Leu Glu Gln Leu Ser Pro Thr
500 505 510
Ala Phe Asn Ser Leu Ser Ser Leu Gln Val Leu Asn Met Ser His Asn
515 520 525
Asn Phe Phe Ser Leu Asp Thr Phe Pro Tyr Lys Cys Leu Asn Ser Leu
530 535 540
Gln Val Leu Asp Tyr Ser Leu Asn His Ile Met Thr Ser Lys Lys Gln
545 550 555 560
Glu Leu Gln His Phe Pro Ser Ser Leu Ala Phe Leu Asn Leu Thr Gln
565 570 575
Asn Asp Phe Ala Cys Thr Cys Glu His Gln Ser Phe Leu Gln Trp Ile
580 585 590
Lys Asp Gln Arg Gln Leu Leu Val Glu Val Glu Arg Met Glu Cys Ala
595 600 605
Thr Pro Ser Asp Lys Gln Gly Met Pro Val Leu Ser Leu Asn Ile Thr
610 615 620
Cys Gln Met Asn Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
625 630 635 640
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
645 650 655
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
660 665 670
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
675 680 685
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Leu Arg Lys Arg Arg Lys
690 695 700
Glu Thr Arg Phe Gly Gln Ala Phe Asp Ser Val Met Ala Arg Gly Glu
705 710 715 720
Pro Ala Val His Phe Arg Ala Ala Arg Ser Phe Asn Arg Glu Arg Pro
725 730 735
Glu Arg Ile Glu Ala Thr Leu Asp Ser Leu Gly Ile Ser Asp Glu Leu
740 745 750
Lys Glu Lys Leu Glu Asp Val Leu Ile Pro Glu Gln Gln Phe Thr Leu
755 760 765
Gly Arg Met Leu Gly Lys Gly Glu Phe Gly Ser Val Arg Glu Ala Gln
770 775 780
Leu Lys Gln Glu Asp Gly Ser Phe Val Lys Val Ala Val Lys Met Leu
785 790 795 800
Lys Ala Asp Ile Ile Ala Ser Ser Asp Ile Glu Glu Phe Leu Arg Glu
805 810 815
Ala Ala Cys Met Lys Glu Phe Asp His Pro His Val Ala Lys Leu Val
820 825 830
Gly Val Ser Leu Arg Ser Arg Ala Lys Gly Arg Leu Pro Ile Pro Met
835 840 845
Val Ile Leu Pro Phe Met Lys His Gly Asp Leu His Ala Phe Leu Leu
850 855 860
Ala Ser Arg Ile Gly Glu Asn Pro Phe Asn Leu Pro Leu Gln Thr Leu
865 870 875 880
Ile Arg Phe Met Val Asp Ile Ala Cys Gly Met Glu Tyr Leu Ser Ser
885 890 895
Arg Asn Phe Ile His Arg Asp Leu Ala Ala Arg Asn Cys Met Leu Ala
900 905 910
Glu Asp Met Thr Val Cys Val Ala Asp Phe Gly Leu Ser Arg Lys Ile
915 920 925
Tyr Ser Gly Asp Tyr Tyr Arg Gln Gly Cys Ala Ser Lys Leu Pro Val
930 935 940
Lys Trp Leu Ala Leu Glu Ser Leu Ala Asp Asn Leu Tyr Thr Val Gln
945 950 955 960
Ser Asp Val Trp Ala Phe Gly Val Thr Met Trp Glu Ile Met Thr Arg
965 970 975
Gly Gln Thr Pro Tyr Ala Gly Ile Glu Asn Ala Glu Ile Tyr Asn Tyr
980 985 990
Leu Ile Gly Gly Asn Arg Leu Lys Gln Pro Pro Glu Cys Met Glu Asp
995 1000 1005
Val Tyr Asp Leu Met Tyr Gln Cys Trp Ser Ala Asp Pro Lys Gln Arg
1010 1015 1020
Pro Ser Phe Thr Cys Leu Arg Met Glu Leu Glu Asn Ile Leu Gly Gln
1025 1030 1035 1040
Leu Ser Val Leu Ser Ala Ser Gln Asp Pro Leu Tyr Ile Asn Ile Glu
1045 1050 1055
Arg Ala Glu Glu Pro Thr Ala Gly Gly Ser Leu Glu Leu Pro Gly Arg
1060 1065 1070
Asp Gln Pro Tyr Ser Gly Ala Gly Asp Gly Ser Gly Met Gly Ala Val
1075 1080 1085
Gly Gly Thr Pro Ser Asp Cys Arg Tyr Ile Leu Thr Pro Gly Gly Leu
1090 1095 1100
Ala Glu Gln Pro Gly Gln Ala Glu His Gln Pro Glu Ser Pro Leu Asn
1105 1110 1115 1120
Glu Thr Gln Arg Leu Leu Leu Leu Gln Gln Gly Leu Leu Pro His Ser
1125 1130 1135
Ser Cys
<210> 3
<211> 1120
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 3
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Glu Ser Trp Glu Pro Cys Val Glu Val Val Pro
20 25 30
Asn Ile Thr Tyr Gln Cys Met Glu Leu Asn Phe Tyr Lys Ile Pro Asp
35 40 45
Asn Leu Pro Phe Ser Thr Lys Asn Leu Asp Leu Ser Phe Asn Pro Leu
50 55 60
Arg His Leu Gly Ser Tyr Ser Phe Phe Ser Phe Pro Glu Leu Gln Val
65 70 75 80
Leu Asp Leu Ser Arg Cys Glu Ile Gln Thr Ile Glu Asp Gly Ala Tyr
85 90 95
Gln Ser Leu Ser His Leu Ser Thr Leu Ile Leu Thr Gly Asn Pro Ile
100 105 110
Gln Ser Leu Ala Leu Gly Ala Phe Ser Gly Leu Ser Ser Leu Gln Lys
115 120 125
Leu Val Ala Val Glu Thr Asn Leu Ala Ser Leu Glu Asn Phe Pro Ile
130 135 140
Gly His Leu Lys Thr Leu Lys Glu Leu Asn Val Ala His Asn Leu Ile
145 150 155 160
Gln Ser Phe Lys Leu Pro Glu Tyr Phe Ser Asn Leu Thr Asn Leu Glu
165 170 175
His Leu Asp Leu Ser Ser Asn Lys Ile Gln Ser Ile Tyr Cys Thr Asp
180 185 190
Leu Arg Val Leu His Gln Met Pro Leu Leu Asn Leu Ser Leu Asp Leu
195 200 205
Ser Leu Asn Pro Met Asn Phe Ile Gln Pro Gly Ala Phe Lys Glu Ile
210 215 220
Arg Leu His Lys Leu Thr Leu Arg Asn Asn Phe Asp Ser Leu Asn Val
225 230 235 240
Met Lys Thr Cys Ile Gln Gly Leu Ala Gly Leu Glu Val His Arg Leu
245 250 255
Val Leu Gly Glu Phe Arg Asn Glu Gly Asn Leu Glu Lys Phe Asp Lys
260 265 270
Ser Ala Leu Glu Gly Leu Cys Asn Leu Thr Ile Glu Glu Phe Arg Leu
275 280 285
Ala Tyr Leu Asp Tyr Tyr Leu Asp Asp Ile Ile Asp Leu Phe Asn Cys
290 295 300
Leu Thr Asn Val Ser Ser Phe Ser Leu Val Ser Val Thr Ile Glu Arg
305 310 315 320
Val Lys Asp Phe Ser Tyr Asn Phe Gly Trp Gln His Leu Glu Leu Val
325 330 335
Asn Cys Lys Phe Gly Gln Phe Pro Thr Leu Lys Leu Lys Ser Leu Lys
340 345 350
Arg Leu Thr Phe Thr Ser Asn Lys Gly Gly Asn Ala Phe Ser Glu Val
355 360 365
Asp Leu Pro Ser Leu Glu Phe Leu Asp Leu Ser Arg Asn Gly Leu Ser
370 375 380
Phe Lys Gly Cys Cys Ser Gln Ser Asp Phe Gly Thr Thr Ser Leu Lys
385 390 395 400
Tyr Leu Asp Leu Ser Phe Asn Gly Val Ile Thr Met Ser Ser Asn Phe
405 410 415
Leu Gly Leu Glu Gln Leu Glu His Leu Asp Phe Gln His Ser Asn Leu
420 425 430
Lys Gln Met Ser Glu Phe Ser Val Phe Leu Ser Leu Arg Asn Leu Ile
435 440 445
Tyr Leu Asp Ile Ser His Thr His Thr Arg Val Ala Phe Asn Gly Ile
450 455 460
Phe Asn Gly Leu Ser Ser Leu Glu Val Leu Lys Met Ala Gly Asn Ser
465 470 475 480
Phe Gln Glu Asn Phe Leu Pro Asp Ile Phe Thr Glu Leu Arg Asn Leu
485 490 495
Thr Phe Leu Asp Leu Ser Gln Cys Gln Leu Glu Gln Leu Ser Pro Thr
500 505 510
Ala Phe Asn Ser Leu Ser Ser Leu Gln Val Leu Asn Met Ser His Asn
515 520 525
Asn Phe Phe Ser Leu Asp Thr Phe Pro Tyr Lys Cys Leu Asn Ser Leu
530 535 540
Gln Val Leu Asp Tyr Ser Leu Asn His Ile Met Thr Ser Lys Lys Gln
545 550 555 560
Glu Leu Gln His Phe Pro Ser Ser Leu Ala Phe Leu Asn Leu Thr Gln
565 570 575
Asn Asp Phe Ala Cys Thr Cys Glu His Gln Ser Phe Leu Gln Trp Ile
580 585 590
Lys Asp Gln Arg Gln Leu Leu Val Glu Val Glu Arg Met Glu Cys Ala
595 600 605
Thr Pro Ser Asp Lys Gln Gly Met Pro Val Leu Ser Leu Asn Ile Thr
610 615 620
Cys Gln Met Asn Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro
625 630 635 640
Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys
645 650 655
Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala
660 665 670
Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu
675 680 685
Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys His Arg Arg Lys Lys Glu
690 695 700
Thr Arg Tyr Gly Glu Val Phe Glu Pro Thr Val Glu Arg Gly Glu Leu
705 710 715 720
Val Val Arg Tyr Arg Val Arg Lys Ser Tyr Ser Arg Arg Thr Thr Glu
725 730 735
Ala Thr Leu Asn Ser Leu Gly Ile Ser Glu Glu Leu Lys Glu Lys Leu
740 745 750
Arg Asp Val Met Val Asp Arg His Lys Val Ala Leu Gly Lys Thr Leu
755 760 765
Gly Glu Gly Glu Phe Gly Ala Val Met Glu Gly Gln Leu Asn Gln Asp
770 775 780
Asp Ser Ile Leu Lys Val Ala Val Lys Thr Met Lys Ile Ala Ile Cys
785 790 795 800
Thr Arg Ser Glu Leu Glu Asp Phe Leu Ser Glu Ala Val Cys Met Lys
805 810 815
Glu Phe Asp His Pro Asn Val Met Arg Leu Ile Gly Val Cys Phe Gln
820 825 830
Gly Ser Glu Arg Glu Ser Phe Pro Ala Pro Val Val Ile Leu Pro Phe
835 840 845
Met Lys His Gly Asp Leu His Ser Phe Leu Leu Tyr Ser Arg Leu Gly
850 855 860
Asp Gln Pro Val Tyr Leu Pro Thr Gln Met Leu Val Lys Phe Met Ala
865 870 875 880
Asp Ile Ala Ser Gly Met Glu Tyr Leu Ser Thr Lys Arg Phe Ile His
885 890 895
Arg Asp Leu Ala Ala Arg Asn Cys Met Leu Asn Glu Asn Met Ser Val
900 905 910
Cys Val Ala Asp Phe Gly Leu Ser Lys Lys Ile Tyr Asn Gly Asp Tyr
915 920 925
Tyr Arg Gln Gly Arg Ile Ala Lys Met Pro Val Lys Trp Ile Ala Ile
930 935 940
Glu Ser Leu Ala Asp Arg Val Tyr Thr Ser Lys Ser Asp Val Trp Ser
945 950 955 960
Phe Gly Val Thr Met Trp Glu Ile Ala Thr Arg Gly Gln Thr Pro Tyr
965 970 975
Pro Gly Val Glu Asn Ser Glu Ile Tyr Asp Tyr Leu Arg Gln Gly Asn
980 985 990
Arg Leu Lys Gln Pro Ala Asp Cys Leu Asp Gly Leu Tyr Ala Leu Met
995 1000 1005
Ser Arg Cys Trp Glu Leu Asn Pro Gln Asp Arg Pro Ser Phe Thr Glu
1010 1015 1020
Leu Arg Glu Asp Leu Glu Asn Thr Leu Lys Ala Leu Pro Pro Ala Gln
1025 1030 1035 1040
Glu Pro Asp Glu Ile Leu Tyr Val Asn Met Asp Glu Gly Gly Gly Tyr
1045 1050 1055
Pro Glu Pro Pro Gly Ala Ala Gly Gly Ala Asp Pro Pro Thr Gln Pro
1060 1065 1070
Asp Pro Lys Asp Ser Cys Ser Cys Leu Thr Ala Ala Glu Val His Pro
1075 1080 1085
Ala Gly Arg Tyr Val Leu Cys Pro Ser Thr Thr Pro Ser Pro Ala Gln
1090 1095 1100
Pro Ala Asp Arg Gly Ser Pro Ala Ala Pro Gly Gln Glu Asp Gly Ala
1105 1110 1115 1120
<210> 4
<211> 1119
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Ser Glu Phe Leu Val Asp Arg Ser Lys Asn Gly
20 25 30
Leu Ile His Val Pro Lys Asp Leu Ser Gln Lys Thr Thr Ile Leu Asn
35 40 45
Ile Ser Gln Asn Tyr Ile Ser Glu Leu Trp Thr Ser Asp Ile Leu Ser
50 55 60
Leu Ser Lys Leu Arg Ile Leu Ile Ile Ser His Asn Arg Ile Gln Tyr
65 70 75 80
Leu Asp Ile Ser Val Phe Lys Phe Asn Gln Glu Leu Glu Tyr Leu Asp
85 90 95
Leu Ser His Asn Lys Leu Val Lys Ile Ser Cys His Pro Thr Val Asn
100 105 110
Leu Lys His Leu Asp Leu Ser Phe Asn Ala Phe Asp Ala Leu Pro Ile
115 120 125
Cys Lys Glu Phe Gly Asn Met Ser Gln Leu Lys Phe Leu Gly Leu Ser
130 135 140
Thr Thr His Leu Glu Lys Ser Ser Val Leu Pro Ile Ala His Leu Asn
145 150 155 160
Ile Ser Lys Val Leu Leu Val Leu Gly Glu Thr Tyr Gly Glu Lys Glu
165 170 175
Asp Pro Glu Gly Leu Gln Asp Phe Asn Thr Glu Ser Leu His Ile Val
180 185 190
Phe Pro Thr Asn Lys Glu Phe His Phe Ile Leu Asp Val Ser Val Lys
195 200 205
Thr Val Ala Asn Leu Glu Leu Ser Asn Ile Lys Cys Val Leu Glu Asp
210 215 220
Asn Lys Cys Ser Tyr Phe Leu Ser Ile Leu Ala Lys Leu Gln Thr Asn
225 230 235 240
Pro Lys Leu Ser Asn Leu Thr Leu Asn Asn Ile Glu Thr Thr Trp Asn
245 250 255
Ser Phe Ile Arg Ile Leu Gln Leu Val Trp His Thr Thr Val Trp Tyr
260 265 270
Phe Ser Ile Ser Asn Val Lys Leu Gln Gly Gln Leu Asp Phe Arg Asp
275 280 285
Phe Asp Tyr Ser Gly Thr Ser Leu Lys Ala Leu Ser Ile His Gln Val
290 295 300
Val Ser Asp Val Phe Gly Phe Pro Gln Ser Tyr Ile Tyr Glu Ile Phe
305 310 315 320
Ser Asn Met Asn Ile Lys Asn Phe Thr Val Ser Gly Thr Arg Met Val
325 330 335
His Met Leu Cys Pro Ser Lys Ile Ser Pro Phe Leu His Leu Asp Phe
340 345 350
Ser Asn Asn Leu Leu Thr Asp Thr Val Phe Glu Asn Cys Gly His Leu
355 360 365
Thr Glu Leu Glu Thr Leu Ile Leu Gln Met Asn Gln Leu Lys Glu Leu
370 375 380
Ser Lys Ile Ala Glu Met Thr Thr Gln Met Lys Ser Leu Gln Gln Leu
385 390 395 400
Asp Ile Ser Gln Asn Ser Val Ser Tyr Asp Glu Lys Lys Gly Asp Cys
405 410 415
Ser Trp Thr Lys Ser Leu Leu Ser Leu Asn Met Ser Ser Asn Ile Leu
420 425 430
Thr Asp Thr Ile Phe Arg Cys Leu Pro Pro Arg Ile Lys Val Leu Asp
435 440 445
Leu His Ser Asn Lys Ile Lys Ser Ile Pro Lys Gln Val Val Lys Leu
450 455 460
Glu Ala Leu Gln Glu Leu Asn Val Ala Phe Asn Ser Leu Thr Asp Leu
465 470 475 480
Pro Gly Cys Gly Ser Phe Ser Ser Leu Ser Val Leu Ile Ile Asp His
485 490 495
Asn Ser Val Ser His Pro Ser Ala Asp Phe Phe Gln Ser Cys Gln Lys
500 505 510
Met Arg Ser Ile Lys Ala Gly Asp Asn Pro Phe Gln Cys Thr Cys Glu
515 520 525
Leu Gly Glu Phe Val Lys Asn Ile Asp Gln Val Ser Ser Glu Val Leu
530 535 540
Glu Gly Trp Pro Asp Ser Tyr Lys Cys Asp Tyr Pro Glu Ser Tyr Arg
545 550 555 560
Gly Thr Leu Leu Lys Asp Phe His Met Ser Glu Leu Ser Cys Asn Ile
565 570 575
Thr Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile
580 585 590
Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala
595 600 605
Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr
610 615 620
Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu
625 630 635 640
Val Ile Thr Leu Tyr Cys Lys Arg Val Gln Glu Thr Lys Phe Gly Asn
645 650 655
Ala Phe Thr Glu Glu Asp Ser Glu Leu Val Val Asn Tyr Ile Ala Lys
660 665 670
Lys Ser Phe Cys Arg Arg Ala Ile Glu Leu Thr Leu His Ser Leu Gly
675 680 685
Val Ser Glu Glu Leu Gln Asn Lys Leu Glu Asp Val Val Ile Asp Arg
690 695 700
Asn Leu Leu Ile Leu Gly Lys Ile Leu Gly Glu Gly Glu Phe Gly Ser
705 710 715 720
Val Met Glu Gly Asn Leu Lys Gln Glu Asp Gly Thr Ser Leu Lys Val
725 730 735
Ala Val Lys Thr Met Lys Leu Asp Asn Ser Ser Gln Arg Glu Ile Glu
740 745 750
Glu Phe Leu Ser Glu Ala Ala Cys Met Lys Asp Phe Ser His Pro Asn
755 760 765
Val Ile Arg Leu Leu Gly Val Cys Ile Glu Met Ser Ser Gln Gly Ile
770 775 780
Pro Lys Pro Met Val Ile Leu Pro Phe Met Lys Tyr Gly Asp Leu His
785 790 795 800
Thr Tyr Leu Leu Tyr Ser Arg Leu Glu Thr Gly Pro Lys His Ile Pro
805 810 815
Leu Gln Thr Leu Leu Lys Phe Met Val Asp Ile Ala Leu Gly Met Glu
820 825 830
Tyr Leu Ser Asn Arg Asn Phe Leu His Arg Asp Leu Ala Ala Arg Asn
835 840 845
Cys Met Leu Arg Asp Asp Met Thr Val Cys Val Ala Asp Phe Gly Leu
850 855 860
Ser Lys Lys Ile Tyr Ser Gly Asp Tyr Tyr Arg Gln Gly Arg Ile Ala
865 870 875 880
Lys Met Pro Val Lys Trp Ile Ala Ile Glu Ser Leu Ala Asp Arg Val
885 890 895
Tyr Thr Ser Lys Ser Asp Val Trp Ala Phe Gly Val Thr Met Trp Glu
900 905 910
Ile Ala Thr Arg Gly Met Thr Pro Tyr Pro Gly Val Gln Asn His Glu
915 920 925
Met Tyr Asp Tyr Leu Leu His Gly His Arg Leu Lys Gln Pro Glu Asp
930 935 940
Cys Leu Asp Glu Leu Tyr Glu Ile Met Tyr Ser Cys Trp Arg Thr Asp
945 950 955 960
Pro Leu Asp Arg Pro Thr Phe Ser Val Leu Arg Leu Gln Leu Glu Lys
965 970 975
Leu Leu Glu Ser Leu Pro Asp Val Arg Asn Gln Ala Asp Val Ile Tyr
980 985 990
Val Asn Thr Gln Leu Leu Glu Ser Ser Glu Gly Leu Ala Gln Gly Ser
995 1000 1005
Thr Leu Ala Pro Leu Asp Leu Asn Ile Asp Pro Asp Ser Ile Ile Ala
1010 1015 1020
Ser Cys Thr Pro Arg Ala Ala Ile Ser Val Val Thr Ala Glu Val His
1025 1030 1035 1040
Asp Ser Lys Pro His Glu Gly Arg Tyr Ile Leu Asn Gly Gly Ser Glu
1045 1050 1055
Glu Trp Glu Asp Leu Thr Ser Ala Pro Ser Ala Ala Val Thr Ala Glu
1060 1065 1070
Lys Asn Ser Val Leu Pro Gly Glu Arg Leu Val Arg Asn Gly Val Ser
1075 1080 1085
Trp Ser His Ser Ser Met Leu Pro Leu Gly Ser Ser Leu Pro Asp Glu
1090 1095 1100
Leu Leu Phe Ala Asp Asp Ser Ser Glu Gly Ser Glu Val Leu Met
1105 1110 1115
<210> 5
<211> 1131
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 5
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Glu Ser Ser Asn Gln Ala Ser Leu Ser Cys Asp
20 25 30
Arg Asn Gly Ile Cys Lys Gly Ser Ser Gly Ser Leu Asn Ser Ile Pro
35 40 45
Ser Gly Leu Thr Glu Ala Val Lys Ser Leu Asp Leu Ser Asn Asn Arg
50 55 60
Ile Thr Tyr Ile Ser Asn Ser Asp Leu Gln Arg Cys Val Asn Leu Gln
65 70 75 80
Ala Leu Val Leu Thr Ser Asn Gly Ile Asn Thr Ile Glu Glu Asp Ser
85 90 95
Phe Ser Ser Leu Gly Ser Leu Glu His Leu Asp Leu Ser Tyr Asn Tyr
100 105 110
Leu Ser Asn Leu Ser Ser Ser Trp Phe Lys Pro Leu Ser Ser Leu Thr
115 120 125
Phe Leu Asn Leu Leu Gly Asn Pro Tyr Lys Thr Leu Gly Glu Thr Ser
130 135 140
Leu Phe Ser His Leu Thr Lys Leu Gln Ile Leu Arg Val Gly Asn Met
145 150 155 160
Asp Thr Phe Thr Lys Ile Gln Arg Lys Asp Phe Ala Gly Leu Thr Phe
165 170 175
Leu Glu Glu Leu Glu Ile Asp Ala Ser Asp Leu Gln Ser Tyr Glu Pro
180 185 190
Lys Ser Leu Lys Ser Ile Gln Asn Val Ser His Leu Ile Leu His Met
195 200 205
Lys Gln His Ile Leu Leu Leu Glu Ile Phe Val Asp Val Thr Ser Ser
210 215 220
Val Glu Cys Leu Glu Leu Arg Asp Thr Asp Leu Asp Thr Phe His Phe
225 230 235 240
Ser Glu Leu Ser Thr Gly Glu Thr Asn Ser Leu Ile Lys Lys Phe Thr
245 250 255
Phe Arg Asn Val Lys Ile Thr Asp Glu Ser Leu Phe Gln Val Met Lys
260 265 270
Leu Leu Asn Gln Ile Ser Gly Leu Leu Glu Leu Glu Phe Asp Asp Cys
275 280 285
Thr Leu Asn Gly Val Gly Asn Phe Arg Ala Ser Asp Asn Asp Arg Val
290 295 300
Ile Asp Pro Gly Lys Val Glu Thr Leu Thr Ile Arg Arg Leu His Ile
305 310 315 320
Pro Arg Phe Tyr Leu Phe Tyr Asp Leu Ser Thr Leu Tyr Ser Leu Thr
325 330 335
Glu Arg Val Lys Arg Ile Thr Val Glu Asn Ser Lys Val Phe Leu Val
340 345 350
Pro Cys Leu Leu Ser Gln His Leu Lys Ser Leu Glu Tyr Leu Asp Leu
355 360 365
Ser Glu Asn Leu Met Val Glu Glu Tyr Leu Lys Asn Ser Ala Cys Glu
370 375 380
Asp Ala Trp Pro Ser Leu Gln Thr Leu Ile Leu Arg Gln Asn His Leu
385 390 395 400
Ala Ser Leu Glu Lys Thr Gly Glu Thr Leu Leu Thr Leu Lys Asn Leu
405 410 415
Thr Asn Ile Asp Ile Ser Lys Asn Ser Phe His Ser Met Pro Glu Thr
420 425 430
Cys Gln Trp Pro Glu Lys Met Lys Tyr Leu Asn Leu Ser Ser Thr Arg
435 440 445
Ile His Ser Val Thr Gly Cys Ile Pro Lys Thr Leu Glu Ile Leu Asp
450 455 460
Val Ser Asn Asn Asn Leu Asn Leu Phe Ser Leu Asn Leu Pro Gln Leu
465 470 475 480
Lys Glu Leu Tyr Ile Ser Arg Asn Lys Leu Met Thr Leu Pro Asp Ala
485 490 495
Ser Leu Leu Pro Met Leu Leu Val Leu Lys Ile Ser Arg Asn Ala Ile
500 505 510
Thr Thr Phe Ser Lys Glu Gln Leu Asp Ser Phe His Thr Leu Lys Thr
515 520 525
Leu Glu Ala Gly Gly Asn Asn Phe Ile Cys Ser Cys Glu Phe Leu Ser
530 535 540
Phe Thr Gln Glu Gln Gln Ala Leu Ala Lys Val Leu Ile Asp Trp Pro
545 550 555 560
Ala Asn Tyr Leu Cys Asp Ser Pro Ser His Val Arg Gly Gln Gln Val
565 570 575
Gln Asp Val Arg Leu Ser Val Ser Glu Cys His Arg Thr Thr Thr Thr
580 585 590
Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro
595 600 605
Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val
610 615 620
His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro
625 630 635 640
Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu
645 650 655
Tyr Cys Lys Arg Val Gln Glu Thr Lys Phe Gly Asn Ala Phe Thr Glu
660 665 670
Glu Asp Ser Glu Leu Val Val Asn Tyr Ile Ala Lys Lys Ser Phe Cys
675 680 685
Arg Arg Ala Ile Glu Leu Thr Leu His Ser Leu Gly Val Ser Glu Glu
690 695 700
Leu Gln Asn Lys Leu Glu Asp Val Val Ile Asp Arg Asn Leu Leu Ile
705 710 715 720
Leu Gly Lys Ile Leu Gly Glu Gly Glu Phe Gly Ser Val Met Glu Gly
725 730 735
Asn Leu Lys Gln Glu Asp Gly Thr Ser Leu Lys Val Ala Val Lys Thr
740 745 750
Met Lys Leu Asp Asn Ser Ser Gln Arg Glu Ile Glu Glu Phe Leu Ser
755 760 765
Glu Ala Ala Cys Met Lys Asp Phe Ser His Pro Asn Val Ile Arg Leu
770 775 780
Leu Gly Val Cys Ile Glu Met Ser Ser Gln Gly Ile Pro Lys Pro Met
785 790 795 800
Val Ile Leu Pro Phe Met Lys Tyr Gly Asp Leu His Thr Tyr Leu Leu
805 810 815
Tyr Ser Arg Leu Glu Thr Gly Pro Lys His Ile Pro Leu Gln Thr Leu
820 825 830
Leu Lys Phe Met Val Asp Ile Ala Leu Gly Met Glu Tyr Leu Ser Asn
835 840 845
Arg Asn Phe Leu His Arg Asp Leu Ala Ala Arg Asn Cys Met Leu Arg
850 855 860
Asp Asp Met Thr Val Cys Val Ala Asp Phe Gly Leu Ser Lys Lys Ile
865 870 875 880
Tyr Ser Gly Asp Tyr Tyr Arg Gln Gly Arg Ile Ala Lys Met Pro Val
885 890 895
Lys Trp Ile Ala Ile Glu Ser Leu Ala Asp Arg Val Tyr Thr Ser Lys
900 905 910
Ser Asp Val Trp Ala Phe Gly Val Thr Met Trp Glu Ile Ala Thr Arg
915 920 925
Gly Met Thr Pro Tyr Pro Gly Val Gln Asn His Glu Met Tyr Asp Tyr
930 935 940
Leu Leu His Gly His Arg Leu Lys Gln Pro Glu Asp Cys Leu Asp Glu
945 950 955 960
Leu Tyr Glu Ile Met Tyr Ser Cys Trp Arg Thr Asp Pro Leu Asp Arg
965 970 975
Pro Thr Phe Ser Val Leu Arg Leu Gln Leu Glu Lys Leu Leu Glu Ser
980 985 990
Leu Pro Asp Val Arg Asn Gln Ala Asp Val Ile Tyr Val Asn Thr Gln
995 1000 1005
Leu Leu Glu Ser Ser Glu Gly Leu Ala Gln Gly Ser Thr Leu Ala Pro
1010 1015 1020
Leu Asp Leu Asn Ile Asp Pro Asp Ser Ile Ile Ala Ser Cys Thr Pro
1025 1030 1035 1040
Arg Ala Ala Ile Ser Val Val Thr Ala Glu Val His Asp Ser Lys Pro
1045 1050 1055
His Glu Gly Arg Tyr Ile Leu Asn Gly Gly Ser Glu Glu Trp Glu Asp
1060 1065 1070
Leu Thr Ser Ala Pro Ser Ala Ala Val Thr Ala Glu Lys Asn Ser Val
1075 1080 1085
Leu Pro Gly Glu Arg Leu Val Arg Asn Gly Val Ser Trp Ser His Ser
1090 1095 1100
Ser Met Leu Pro Leu Gly Ser Ser Leu Pro Asp Glu Leu Leu Phe Ala
1105 1110 1115 1120
Asp Asp Ser Ser Glu Gly Ser Glu Val Leu Met
1125 1130
<210> 6
<211> 889
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 6
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Thr Thr Pro Glu Pro Cys Glu Leu Asp Asp Glu
20 25 30
Asp Phe Arg Cys Val Cys Asn Phe Ser Glu Pro Gln Pro Asp Trp Ser
35 40 45
Glu Ala Phe Gln Cys Val Ser Ala Val Glu Val Glu Ile His Ala Gly
50 55 60
Gly Leu Asn Leu Glu Pro Phe Leu Lys Arg Val Asp Ala Asp Ala Asp
65 70 75 80
Pro Arg Gln Tyr Ala Asp Thr Val Lys Ala Leu Arg Val Arg Arg Leu
85 90 95
Thr Val Gly Ala Ala Gln Val Pro Ala Gln Leu Leu Val Gly Ala Leu
100 105 110
Arg Val Leu Ala Tyr Ser Arg Leu Lys Glu Leu Thr Leu Glu Asp Leu
115 120 125
Lys Ile Thr Gly Thr Met Pro Pro Leu Pro Leu Glu Ala Thr Gly Leu
130 135 140
Ala Leu Ser Ser Leu Arg Leu Arg Asn Val Ser Trp Ala Thr Gly Arg
145 150 155 160
Ser Trp Leu Ala Glu Leu Gln Gln Trp Leu Lys Pro Gly Leu Lys Val
165 170 175
Leu Ser Ile Ala Gln Ala His Ser Pro Ala Phe Ser Cys Glu Gln Val
180 185 190
Arg Ala Phe Pro Ala Leu Thr Ser Leu Asp Leu Ser Asp Asn Pro Gly
195 200 205
Leu Gly Glu Arg Gly Leu Met Ala Ala Leu Cys Pro His Lys Phe Pro
210 215 220
Ala Ile Gln Asn Leu Ala Leu Arg Asn Thr Gly Met Glu Thr Pro Thr
225 230 235 240
Gly Val Cys Ala Ala Leu Ala Ala Ala Gly Val Gln Pro His Ser Leu
245 250 255
Asp Leu Ser His Asn Ser Leu Arg Ala Thr Val Asn Pro Ser Ala Pro
260 265 270
Arg Cys Met Trp Ser Ser Ala Leu Asn Ser Leu Asn Leu Ser Phe Ala
275 280 285
Gly Leu Glu Gln Val Pro Lys Gly Leu Pro Ala Lys Leu Arg Val Leu
290 295 300
Asp Leu Ser Cys Asn Arg Leu Asn Arg Ala Pro Gln Pro Asp Glu Leu
305 310 315 320
Pro Glu Val Asp Asn Leu Thr Leu Asp Gly Asn Pro Phe Leu Val Pro
325 330 335
Gly Thr Ala Leu Pro His Glu Gly Ser Met Asn Thr Thr Thr Pro Ala
340 345 350
Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser
355 360 365
Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr
370 375 380
Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala
385 390 395 400
Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys
405 410 415
Lys Arg Val Gln Glu Thr Lys Phe Gly Asn Ala Phe Thr Glu Glu Asp
420 425 430
Ser Glu Leu Val Val Asn Tyr Ile Ala Lys Lys Ser Phe Cys Arg Arg
435 440 445
Ala Ile Glu Leu Thr Leu His Ser Leu Gly Val Ser Glu Glu Leu Gln
450 455 460
Asn Lys Leu Glu Asp Val Val Ile Asp Arg Asn Leu Leu Ile Leu Gly
465 470 475 480
Lys Ile Leu Gly Glu Gly Glu Phe Gly Ser Val Met Glu Gly Asn Leu
485 490 495
Lys Gln Glu Asp Gly Thr Ser Leu Lys Val Ala Val Lys Thr Met Lys
500 505 510
Leu Asp Asn Ser Ser Gln Arg Glu Ile Glu Glu Phe Leu Ser Glu Ala
515 520 525
Ala Cys Met Lys Asp Phe Ser His Pro Asn Val Ile Arg Leu Leu Gly
530 535 540
Val Cys Ile Glu Met Ser Ser Gln Gly Ile Pro Lys Pro Met Val Ile
545 550 555 560
Leu Pro Phe Met Lys Tyr Gly Asp Leu His Thr Tyr Leu Leu Tyr Ser
565 570 575
Arg Leu Glu Thr Gly Pro Lys His Ile Pro Leu Gln Thr Leu Leu Lys
580 585 590
Phe Met Val Asp Ile Ala Leu Gly Met Glu Tyr Leu Ser Asn Arg Asn
595 600 605
Phe Leu His Arg Asp Leu Ala Ala Arg Asn Cys Met Leu Arg Asp Asp
610 615 620
Met Thr Val Cys Val Ala Asp Phe Gly Leu Ser Lys Lys Ile Tyr Ser
625 630 635 640
Gly Asp Tyr Tyr Arg Gln Gly Arg Ile Ala Lys Met Pro Val Lys Trp
645 650 655
Ile Ala Ile Glu Ser Leu Ala Asp Arg Val Tyr Thr Ser Lys Ser Asp
660 665 670
Val Trp Ala Phe Gly Val Thr Met Trp Glu Ile Ala Thr Arg Gly Met
675 680 685
Thr Pro Tyr Pro Gly Val Gln Asn His Glu Met Tyr Asp Tyr Leu Leu
690 695 700
His Gly His Arg Leu Lys Gln Pro Glu Asp Cys Leu Asp Glu Leu Tyr
705 710 715 720
Glu Ile Met Tyr Ser Cys Trp Arg Thr Asp Pro Leu Asp Arg Pro Thr
725 730 735
Phe Ser Val Leu Arg Leu Gln Leu Glu Lys Leu Leu Glu Ser Leu Pro
740 745 750
Asp Val Arg Asn Gln Ala Asp Val Ile Tyr Val Asn Thr Gln Leu Leu
755 760 765
Glu Ser Ser Glu Gly Leu Ala Gln Gly Ser Thr Leu Ala Pro Leu Asp
770 775 780
Leu Asn Ile Asp Pro Asp Ser Ile Ile Ala Ser Cys Thr Pro Arg Ala
785 790 795 800
Ala Ile Ser Val Val Thr Ala Glu Val His Asp Ser Lys Pro His Glu
805 810 815
Gly Arg Tyr Ile Leu Asn Gly Gly Ser Glu Glu Trp Glu Asp Leu Thr
820 825 830
Ser Ala Pro Ser Ala Ala Val Thr Ala Glu Lys Asn Ser Val Leu Pro
835 840 845
Gly Glu Arg Leu Val Arg Asn Gly Val Ser Trp Ser His Ser Ser Met
850 855 860
Leu Pro Leu Gly Ser Ser Leu Pro Asp Glu Leu Leu Phe Ala Asp Asp
865 870 875 880
Ser Ser Glu Gly Ser Glu Val Leu Met
885
<210> 7
<211> 1019
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 7
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Ala Asn Pro Gly Leu Val Ala Arg Ile Thr Asp
20 25 30
Lys Gly Leu Gln Tyr Ala Ala Gln Glu Gly Leu Leu Ala Leu Gln Ser
35 40 45
Glu Leu Leu Arg Ile Thr Leu Pro Asp Phe Thr Gly Asp Leu Arg Ile
50 55 60
Pro His Val Gly Arg Gly Arg Tyr Glu Phe His Ser Leu Asn Ile His
65 70 75 80
Ser Cys Glu Leu Leu His Ser Ala Leu Arg Pro Val Pro Gly Gln Gly
85 90 95
Leu Ser Leu Ser Ile Ser Asp Ser Ser Ile Arg Val Gln Gly Arg Trp
100 105 110
Lys Val Arg Lys Ser Phe Phe Lys Leu Gln Gly Ser Phe Asp Val Ser
115 120 125
Val Lys Gly Ile Ser Ile Ser Val Asn Leu Leu Leu Gly Ser Glu Ser
130 135 140
Ser Gly Arg Pro Thr Val Thr Ala Ser Ser Cys Ser Ser Asp Ile Ala
145 150 155 160
Asp Val Glu Val Asp Met Ser Gly Asp Leu Gly Trp Leu Leu Asn Leu
165 170 175
Phe His Asn Gln Ile Glu Ser Lys Phe Gln Lys Val Leu Glu Ser Arg
180 185 190
Ile Cys Glu Met Ile Gln Lys Ser Val Ser Ser Asp Leu Gln Pro Tyr
195 200 205
Leu Gln Thr Leu Pro Val Thr Thr Glu Ile Asp Ser Phe Ala Asp Ile
210 215 220
Asp Tyr Ser Leu Val Glu Ala Pro Arg Ala Thr Ala Gln Met Leu Glu
225 230 235 240
Val Met Phe Lys Gly Glu Ile Phe His Arg Asn His Arg Ser Pro Val
245 250 255
Thr Leu Leu Ala Ala Val Met Ser Leu Pro Glu Glu His Asn Lys Met
260 265 270
Val Tyr Phe Ala Ile Ser Asp Tyr Val Phe Asn Thr Ala Ser Leu Val
275 280 285
Tyr His Glu Glu Gly Tyr Leu Asn Phe Ser Ile Thr Asp Asp Met Ile
290 295 300
Pro Pro Asp Ser Asn Ile Arg Leu Thr Thr Lys Ser Phe Arg Pro Phe
305 310 315 320
Val Pro Arg Leu Ala Arg Leu Tyr Pro Asn Met Asn Leu Glu Leu Gln
325 330 335
Gly Ser Val Pro Ser Ala Pro Leu Leu Asn Phe Ser Pro Gly Asn Leu
340 345 350
Ser Val Asp Pro Tyr Met Glu Ile Asp Ala Phe Val Leu Leu Pro Ser
355 360 365
Ser Ser Lys Glu Pro Val Phe Arg Leu Ser Val Ala Thr Asn Val Ser
370 375 380
Ala Thr Leu Thr Phe Asn Thr Ser Lys Ile Thr Gly Phe Leu Lys Pro
385 390 395 400
Gly Lys Val Lys Val Glu Leu Lys Glu Ser Lys Val Gly Leu Phe Asn
405 410 415
Ala Glu Leu Leu Glu Ala Leu Leu Asn Tyr Tyr Ile Leu Asn Thr Phe
420 425 430
Tyr Pro Lys Phe Asn Asp Lys Leu Ala Glu Gly Phe Pro Leu Pro Leu
435 440 445
Leu Lys Arg Val Gln Leu Tyr Asp Leu Gly Leu Gln Ile His Lys Asp
450 455 460
Phe Leu Phe Leu Gly Ala Asn Val Gln Tyr Met Arg Val Thr Thr Thr
465 470 475 480
Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro
485 490 495
Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val
500 505 510
His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro
515 520 525
Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu
530 535 540
Tyr Cys Lys Arg Val Gln Glu Thr Lys Phe Gly Asn Ala Phe Thr Glu
545 550 555 560
Glu Asp Ser Glu Leu Val Val Asn Tyr Ile Ala Lys Lys Ser Phe Cys
565 570 575
Arg Arg Ala Ile Glu Leu Thr Leu His Ser Leu Gly Val Ser Glu Glu
580 585 590
Leu Gln Asn Lys Leu Glu Asp Val Val Ile Asp Arg Asn Leu Leu Ile
595 600 605
Leu Gly Lys Ile Leu Gly Glu Gly Glu Phe Gly Ser Val Met Glu Gly
610 615 620
Asn Leu Lys Gln Glu Asp Gly Thr Ser Leu Lys Val Ala Val Lys Thr
625 630 635 640
Met Lys Leu Asp Asn Ser Ser Gln Arg Glu Ile Glu Glu Phe Leu Ser
645 650 655
Glu Ala Ala Cys Met Lys Asp Phe Ser His Pro Asn Val Ile Arg Leu
660 665 670
Leu Gly Val Cys Ile Glu Met Ser Ser Gln Gly Ile Pro Lys Pro Met
675 680 685
Val Ile Leu Pro Phe Met Lys Tyr Gly Asp Leu His Thr Tyr Leu Leu
690 695 700
Tyr Ser Arg Leu Glu Thr Gly Pro Lys His Ile Pro Leu Gln Thr Leu
705 710 715 720
Leu Lys Phe Met Val Asp Ile Ala Leu Gly Met Glu Tyr Leu Ser Asn
725 730 735
Arg Asn Phe Leu His Arg Asp Leu Ala Ala Arg Asn Cys Met Leu Arg
740 745 750
Asp Asp Met Thr Val Cys Val Ala Asp Phe Gly Leu Ser Lys Lys Ile
755 760 765
Tyr Ser Gly Asp Tyr Tyr Arg Gln Gly Arg Ile Ala Lys Met Pro Val
770 775 780
Lys Trp Ile Ala Ile Glu Ser Leu Ala Asp Arg Val Tyr Thr Ser Lys
785 790 795 800
Ser Asp Val Trp Ala Phe Gly Val Thr Met Trp Glu Ile Ala Thr Arg
805 810 815
Gly Met Thr Pro Tyr Pro Gly Val Gln Asn His Glu Met Tyr Asp Tyr
820 825 830
Leu Leu His Gly His Arg Leu Lys Gln Pro Glu Asp Cys Leu Asp Glu
835 840 845
Leu Tyr Glu Ile Met Tyr Ser Cys Trp Arg Thr Asp Pro Leu Asp Arg
850 855 860
Pro Thr Phe Ser Val Leu Arg Leu Gln Leu Glu Lys Leu Leu Glu Ser
865 870 875 880
Leu Pro Asp Val Arg Asn Gln Ala Asp Val Ile Tyr Val Asn Thr Gln
885 890 895
Leu Leu Glu Ser Ser Glu Gly Leu Ala Gln Gly Ser Thr Leu Ala Pro
900 905 910
Leu Asp Leu Asn Ile Asp Pro Asp Ser Ile Ile Ala Ser Cys Thr Pro
915 920 925
Arg Ala Ala Ile Ser Val Val Thr Ala Glu Val His Asp Ser Lys Pro
930 935 940
His Glu Gly Arg Tyr Ile Leu Asn Gly Gly Ser Glu Glu Trp Glu Asp
945 950 955 960
Leu Thr Ser Ala Pro Ser Ala Ala Val Thr Ala Glu Lys Asn Ser Val
965 970 975
Leu Pro Gly Glu Arg Leu Val Arg Asn Gly Val Ser Trp Ser His Ser
980 985 990
Ser Met Leu Pro Leu Gly Ser Ser Leu Pro Asp Glu Leu Leu Phe Ala
995 1000 1005
Asp Asp Ser Ser Glu Gly Ser Glu Val Leu Met
1010 1015
<210> 8
<211> 805
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 8
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu
1 5 10 15
His Ala Ala Arg Pro Asp Ile Gln Met Asn Gln Ser Pro Ser Ser Leu
20 25 30
Ser Ala Ser Leu Gly Asp Thr Ile Ser Ile Thr Cys Arg Ala Ser Gln
35 40 45
Asn Ile Asn Ile Trp Leu Ser Trp Tyr Gln Gln Lys Pro Gly Asn Val
50 55 60
Pro Lys Leu Leu Ile Tyr Lys Ala Ser Asn Leu His Thr Gly Val Pro
65 70 75 80
Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ile Ile
85 90 95
Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Gly
100 105 110
Gln Ser Tyr Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
115 120 125
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu
130 135 140
Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser
145 150 155 160
Leu Ser Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Tyr
165 170 175
Met Thr Trp Val Arg Gln Ala Pro Gly Lys Ala Pro Glu Trp Leu Ala
180 185 190
Leu Ile Arg Asn Lys Arg Asn Gly Asp Thr Ala Glu Tyr Ser Ala Ser
195 200 205
Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Tyr Ser Arg Ser Ile Leu
210 215 220
His Leu Gln Met Asn Ala Leu Arg Thr Glu Asp Ser Ala Thr Tyr Tyr
225 230 235 240
Cys Val Arg Gln Gly Arg Gly Tyr Thr Leu Asp Tyr Trp Gly Gln Gly
245 250 255
Thr Ser Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro
260 265 270
Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu
275 280 285
Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp
290 295 300
Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly
305 310 315 320
Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Val Gln
325 330 335
Glu Thr Lys Phe Gly Asn Ala Phe Thr Glu Glu Asp Ser Glu Leu Val
340 345 350
Val Asn Tyr Ile Ala Lys Lys Ser Phe Cys Arg Arg Ala Ile Glu Leu
355 360 365
Thr Leu His Ser Leu Gly Val Ser Glu Glu Leu Gln Asn Lys Leu Glu
370 375 380
Asp Val Val Ile Asp Arg Asn Leu Leu Ile Leu Gly Lys Ile Leu Gly
385 390 395 400
Glu Gly Glu Phe Gly Ser Val Met Glu Gly Asn Leu Lys Gln Glu Asp
405 410 415
Gly Thr Ser Leu Lys Val Ala Val Lys Thr Met Lys Leu Asp Asn Ser
420 425 430
Ser Gln Arg Glu Ile Glu Glu Phe Leu Ser Glu Ala Ala Cys Met Lys
435 440 445
Asp Phe Ser His Pro Asn Val Ile Arg Leu Leu Gly Val Cys Ile Glu
450 455 460
Met Ser Ser Gln Gly Ile Pro Lys Pro Met Val Ile Leu Pro Phe Met
465 470 475 480
Lys Tyr Gly Asp Leu His Thr Tyr Leu Leu Tyr Ser Arg Leu Glu Thr
485 490 495
Gly Pro Lys His Ile Pro Leu Gln Thr Leu Leu Lys Phe Met Val Asp
500 505 510
Ile Ala Leu Gly Met Glu Tyr Leu Ser Asn Arg Asn Phe Leu His Arg
515 520 525
Asp Leu Ala Ala Arg Asn Cys Met Leu Arg Asp Asp Met Thr Val Cys
530 535 540
Val Ala Asp Phe Gly Leu Ser Lys Lys Ile Tyr Ser Gly Asp Tyr Tyr
545 550 555 560
Arg Gln Gly Arg Ile Ala Lys Met Pro Val Lys Trp Ile Ala Ile Glu
565 570 575
Ser Leu Ala Asp Arg Val Tyr Thr Ser Lys Ser Asp Val Trp Ala Phe
580 585 590
Gly Val Thr Met Trp Glu Ile Ala Thr Arg Gly Met Thr Pro Tyr Pro
595 600 605
Gly Val Gln Asn His Glu Met Tyr Asp Tyr Leu Leu His Gly His Arg
610 615 620
Leu Lys Gln Pro Glu Asp Cys Leu Asp Glu Leu Tyr Glu Ile Met Tyr
625 630 635 640
Ser Cys Trp Arg Thr Asp Pro Leu Asp Arg Pro Thr Phe Ser Val Leu
645 650 655
Arg Leu Gln Leu Glu Lys Leu Leu Glu Ser Leu Pro Asp Val Arg Asn
660 665 670
Gln Ala Asp Val Ile Tyr Val Asn Thr Gln Leu Leu Glu Ser Ser Glu
675 680 685
Gly Leu Ala Gln Gly Ser Thr Leu Ala Pro Leu Asp Leu Asn Ile Asp
690 695 700
Pro Asp Ser Ile Ile Ala Ser Cys Thr Pro Arg Ala Ala Ile Ser Val
705 710 715 720
Val Thr Ala Glu Val His Asp Ser Lys Pro His Glu Gly Arg Tyr Ile
725 730 735
Leu Asn Gly Gly Ser Glu Glu Trp Glu Asp Leu Thr Ser Ala Pro Ser
740 745 750
Ala Ala Val Thr Ala Glu Lys Asn Ser Val Leu Pro Gly Glu Arg Leu
755 760 765
Val Arg Asn Gly Val Ser Trp Ser His Ser Ser Met Leu Pro Leu Gly
770 775 780
Ser Ser Leu Pro Asp Glu Leu Leu Phe Ala Asp Asp Ser Ser Glu Gly
785 790 795 800
Ser Glu Val Leu Met
805

Claims (10)

1. A chimeric antigen receptor for endotoxin clearance, comprising: (a) an extracellular domain comprising one or more endotoxin-specific binding regions; (b) a transmembrane domain; (c) at least one copy of an intracellular signaling domain of a phagocyte-specific phagocytic signal receptor,
wherein the endotoxin-specific binding domain comprises: (1) an extracellular domain of a pattern recognition receptor or a functional variant or fragment thereof; (2) an antibody specific for endotoxin or a functional fragment thereof,
the phagocyte-specific phagocytic signal receptor includes: a TAM family receptor tyrosine kinase, a phosphatidylserine receptor family, a low density lipoprotein receptor-related protein family, a mannose receptor family, a scavenger receptor family, or a TREM receptor family.
2. The chimeric antigen receptor for endotoxin clearance of claim 1, wherein:
wherein the pattern recognition receptor comprises TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, Dectin-1, Dectin-2, Mincle, CLEC2, CLEC5A, CLEC12A, DCIR, CLECSF8, CD14, MD-2 or LBP,
the endotoxin-specific antibody comprises WN1222-5, S25-39, Ab-52, S55-3, S1-15 or S25-23.
3. The chimeric antigen receptor for endotoxin clearance of claim 2, wherein:
wherein the transmembrane domain comprises a stem or transmembrane domain derived from CD8, TYRO3, AXL, MERK, ADGRB1, ADGRB3, HAVCR1, TIMD4, Stablin-1, Stablin-2, Integrin subbunit α v, Integrin subbunit β 3, Integrin subbunit β 5, SCARF1, AGER, CD300LF, CD36, LRP1, MRC1 or TREM 2;
the intracellular signaling domain of the phagocyte-specific phagocytic signal receptor does not contain an immunoreceptor tyrosine-based activation motif.
4. The chimeric antigen receptor for endotoxin clearance of claim 3, wherein:
wherein the intracellular signaling domain of the phagocyte-specific phagocytic signal receptor comprises FCER1G, CD3, MEGF10, TYRO3, MERKT, or AXL.
5. The chimeric antigen receptor for endotoxin clearance of claim 1, wherein:
wherein the structure of the chimeric antigen receptor is selected from any one of the following cases:
(1) the extracellular domain is derived from TLR4, the intracellular signal domain is derived from MERRTK, and the amino acid sequence is shown as SEQ ID NO. 1;
(2) the extracellular domain is derived from TLR4, the intracellular signal domain is derived from TYRO3, and the amino acid sequence is shown in SEQ ID NO. 2;
(3) the extracellular domain is derived from TLR4, the intracellular signal domain is derived from AXL, and the amino acid sequence is shown in SEQ ID NO. 3;
(4) the extracellular domain is derived from TLR1, the intracellular signal domain is derived from MERRTK, and the amino acid sequence is shown as SEQ ID NO. 4;
(5) the extracellular domain is derived from TLR2, the intracellular signal domain is derived from MERRTK, and the amino acid sequence is shown as SEQ ID NO. 5;
(6) the extracellular domain is derived from CD14, the intracellular signal domain is derived from MERRTK, and the amino acid sequence is shown as SEQ ID NO. 6;
(7) the extracellular domain is derived from LBP, the intracellular signal domain is derived from MERRTK, and the amino acid sequence is shown as SEQ ID NO. 7;
(8) the extracellular domain is derived from WN1222-5, the intracellular signal domain is derived from MERTK, and the amino acid sequence is shown in SEQ ID NO. 8.
6. A polynucleotide encoding the chimeric antigen receptor for endotoxin removal as claimed in any one of claims 1 to 5, a gene vector carrying the polynucleotide, and a recombinant cell comprising the gene vector.
7. The polynucleotide of the chimeric antigen receptor for endotoxin clearance of claim 6, the gene vector carrying the polynucleotide and the recombinant cell comprising the gene vector, wherein:
wherein the gene vector carries at least two polynucleotides aiming at different epitopes of the same endotoxin.
8. The polynucleotide of the chimeric antigen receptor for endotoxin clearance of claim 7, the gene vector carrying the polynucleotide and the recombinant cell comprising the gene vector, wherein:
wherein, the gene vector carries two polynucleotides aiming at different epitopes of the same endotoxin, and the combination is TLR4-CARMERTKAnd CD14-CARMERTKOr TLR4-CARMERTKAnd LBP-CARMERTK，
The two polynucleotides are linked by nucleotides encoding a 2A peptide.
9. Use of the chimeric antigen receptor for endotoxin removal, polynucleotide encoding the same, vector, recombinant cell or composition according to any one of claims 1 to 5 for the preparation of an agent, a kit or a therapeutic drug for diagnosing or treating diseases associated with excessive endotoxin accumulation.
10. A pharmaceutical composition comprising an active ingredient which is the chimeric antigen receptor for endotoxin removal of any one of claims 1 to 5 or the polynucleotide of any one of claims 6 to 8, a gene vector carrying the polynucleotide or a recombinant cell comprising the gene vector, and a pharmaceutically acceptable diluent or excipient.
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