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CTL (cytotoxic lymphocyte) epitope polypeptide of vascular endothelial growth factors and application of CTL epitope polypeptide

A growth factor receptor and vascular endothelium technology, which is applied in the field of CTL epitope polypeptides of vascular endothelial growth factor receptors, can solve the problems of limited production of heat shock protein gp96 protein, affecting the binding of exogenous polypeptides, and difficult industrial application, etc. Enhanced immunological function, stable expression system, strong binding ability

Inactive Publication Date: 2016-08-31
HEHO XIAMEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Due to the limited source of tissue, the heat shock protein gp96 protein extracted from tissue is limited in production, which is difficult for industrial application, and the gp96 protein extracted from tissue binds to the antigenic polypeptide in the tissue cells, affecting its combination with exogenous polypeptide, thus affecting the efficacy of the vaccine
In addition, constrained by ethical and moral factors, gp96 extracted from tissues is difficult to promote directly as a drug

Method used

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  • CTL (cytotoxic lymphocyte) epitope polypeptide of vascular endothelial growth factors and application of CTL epitope polypeptide
  • CTL (cytotoxic lymphocyte) epitope polypeptide of vascular endothelial growth factors and application of CTL epitope polypeptide
  • CTL (cytotoxic lymphocyte) epitope polypeptide of vascular endothelial growth factors and application of CTL epitope polypeptide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1, preparation of nonapeptide derived from VEGFR

[0075] The derived nonapeptide is a VEGFR211-219 polypeptide fragment, its amino acid sequence is shown in SEQ ID NO.3, and the amino acid sequence from the nitrogen terminal to the carbon terminal is: Ser Ile Met Tyr Ile Val Val Val Val.

[0076] The VEGFR211-219 polypeptide fragment was synthesized by Gill Biochemical (Shanghai) Co., Ltd.

[0077] Control polypeptide: hepatitis B virus HBc82-90 peptide is derived from hepatitis B virus core protein, non-tumor antigen, its amino acid sequence is shown in SEQ ID NO.4, and the amino acid sequence from the nitrogen terminal to the carbon terminal direction is: Arg Glu Leu Val Val Ser Tyr Val Asn.

Embodiment 2

[0078] Example 2. Experimental verification of the binding force between VEGFR211-219 polypeptide fragments and HLA-A2 molecules

[0079] The binding force of VEGFR211-219 polypeptide fragments to HLA-A2 molecules was detected by T2 cell binding assay and in vitro refolding assay respectively.

[0080] 1. T2 cell binding assay to detect the binding ability of peptides to HLA-A2 molecules on the cell surface

[0081] T2 cells express HLA-A0201 (HLA-A0201 can also be called HLA-A * 0201, which is a branch of HLA-A2 molecules), but its TAP gene is defective, so the cells cannot process and present endogenous polypeptides, so the HLA-A0201 molecules on the cell surface are in an unstable and low-expression state. When there are exogenous antigenic polypeptides that can be bound by HLA-A0201 molecules, these polypeptides bind to HLA-A0201 molecules on the surface of T2 cells, which will greatly increase the stability of HLA-A0201 on the cell surface and greatly increase the expres...

Embodiment 3

[0109] Example 3, Insect Baculovirus Expression System Expressing Heat Shock Protein gp96

[0110] In the embodiment,

[0111] FastBac TM 1 Empty plasmid was purchased from Protin Biotechnology (Beijing) Co., Ltd., product number BioVector1058 19-1.

[0112] DH10Bac TM E.coli (abbreviated as DH10Bac) was purchased from Beijing Yuanpinghao Biotechnology Co., Ltd., product number CL108-01.

[0113] HiTrap-Q Sepharose: The chromatographic column is HiTrap Q HP, purchased from GE Company, product number 17-1153-01, and the inner diameter and column length of the chromatographic column are 0.7×2.5 cm.

[0114] Superdex 200 10 / 300 GL: The chromatographic column is Superdex 200 10 / 300 GL, purchased from GE Company, product number 17-5175-01, and the inner diameter and column length of the chromatographic column are 1.0×3.0 cm.

[0115] 1. Build pFastBac TM 1-gp96 plasmid

[0116] 1. Design and synthesis of gp96 primers: using the human gp96 gene sequence in GenBank as a temp...

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Abstract

The invention relates to a CTL (cytotoxic lymphocyte) epitope polypeptide of vascular endothelial growth factors and application of the CTL epitope polypeptide. Amino acid sequences of the CTL epitope polypeptide are shown as SEQ ID NO.3; a derivative sequence or a plurality of derivative sequences of amino acid is substituted, deleted, displaced, inserted and / or added to the CTL epitope polypeptide on the basis of the amino acid sequences show as the SEQ ID NO.3. The invention further provides vaccine. The vaccine comprises the CTL epitope polypeptide of the vascular endothelial growth factors and an adjuvant. The adjuvant is gp96 protein. The CTL epitope polypeptide, the application and the vaccine have the advantages that the adjuvant and the CTL epitope polypeptide of the vascular endothelial growth factors are matched with each other, accordingly, the immunity can be improved, tumor treatment effects can be obviously improved, and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a CTL epitope polypeptide of a vascular endothelial growth factor receptor and an application thereof. Background technique [0002] Lung cancer is one of the malignant tumors with the fastest-growing morbidity and mortality in China and the world, and the greatest threat to the health and life of the population. The incidence of lung cancer ranks first among malignant tumors in my country. The incidence of lung cancer among long-term smokers is as high as 2 / 1000. With the increasing urban smog, the incidence of lung cancer has and will continue to show rapid growth. Common treatments for lung cancer include surgery, chemotherapy, radiation therapy, targeted therapy, immunotherapy, and combinations of different treatments. In recent years, even though the treatment methods of lung cancer have been continuously innovated and improved, the treatment effect is still unsatis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/71C07K14/47C07K1/18C07K1/16C12N15/12C12N15/866A61K38/17A61K39/00A61K39/39A61P35/00
CPCC07K14/71A61K38/00A61K39/00A61K39/39A61K2039/55516C07K14/47C12N15/86C12N2710/14043C12N2800/105
Inventor 孟颂东王刚李鑫
Owner HEHO XIAMEN BIOTECH CO LTD
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