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HPV (human papillomavirus) antigen epitope as well as identification method and application thereof

A technology of HPV16E7 and antigen, which is applied in the fields of cancer antigen components, chemical instruments and methods, and vertebrate antigen components, etc. It can solve the problems that HPV patients cannot be cured, the development and clinical application of HPV therapeutic vaccines are urgent, and the infection cannot be cleared.

Active Publication Date: 2022-05-13
SHENZHEN GINO BIOTECHNOLOGY CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, preventive vaccines focus on prevention and cannot clear existing infections. They cannot cure HPV-infected patients. The research and development and clinical application of HPV therapeutic vaccines have become urgent

Method used

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  • HPV (human papillomavirus) antigen epitope as well as identification method and application thereof
  • HPV (human papillomavirus) antigen epitope as well as identification method and application thereof
  • HPV (human papillomavirus) antigen epitope as well as identification method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0073] According to some specific embodiments of the present invention, the HPV antigen epitope includes at least one of the amino acid sequences shown in SEQ ID NO: 1-7.

[0074] In yet another aspect, the present invention provides a mutant, which has at least one mutation site in addition to the anchor site, compared to the aforementioned isolated peptide (HPV E6, SEQ ID NO: 1). According to a specific embodiment of the present invention, when the obtained HPV antigenic epitope is an octapeptide, the 2nd and 8th amino acids of the obtained HPV antigenic epitope are the anchor sites of HLA, and the amino acids at other sites in the epitope are After substitution, for example, the E6 HPV epitope in this application is mutated at the first, third, fifth, sixth, and seventh positions, and it can still have the same or related immunogenicity and Its potential therapeutic effect is that it can be presented by HLA-I molecules, recognized by CTL cells or TIL cells, and then present...

Embodiment 1

[0159] Example 1 Identification of HPV antigenic epitopes

[0160] In this example, the immunoaffinity method is used to obtain the immune polypeptide, and the specific operation process is as follows: figure 2 As shown, the specific operation steps are as follows:

[0161] 1.1 Acquisition of HLA protein complexes (major histocompatibility complex (MHC) class I complex molecules)

[0162] The specific experimental operation of this experiment is as follows:

[0163] 1) Take 1mL protein A Sepharose CL-4B (GE healthcare) 50% resin into Poly-PrepChromatography Columns, flow through ultrapure water and PBS to wash, and flow through 4mg anti-HLA-class A, B, C antibodies ( W6 / 32, ATCC HB-95);

[0164] 2) After flowing through PBS, flow through cross-linking equilibrium solution (200mM triethanolamine);

[0165] 3) Flow through the cross-linking reaction solution, retain 1mL of the cross-linking reaction solution (50mM DMP), close the lower channel, and let it stand at room temp...

Embodiment 2

[0194] Example 2 Evaluation of HPV-specific tumor antigen peptide immunogenicity by enzyme-linked immunospot assay (ELISPOT)

[0195] In the experiment, T2 cells were loaded with the newly identified 7 kinds of HPV16 type-specific mixed antigen polypeptides described in Example 1, which were presented to cytotoxic T cell CTL through the antigen presentation function of T2 cells, and TAP (antigen presentation and transport) of T2 cells Molecular defects activate immune responses upon presentation of experimental antigens by payloads of exogenous polypeptides. The experiment set up positive control group, negative control group and test product group. After the living cells of T2 (ATCC: CRL-1992) in the logarithmic growth phase were collected after cell culture treatment, they were diluted with medium and resuspended to 5e 5 cells / mL for later use. Take 1mL T2 cell group from each group, add phytohemagglutinin PHA (Sigma, product number L8902-25MG) to the positive control grou...

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Abstract

The invention provides an HPV (human papillomavirus) antigen epitope as well as an identification method and application thereof. The HPV antigen epitope comprises 1) 75-83 amino acids of HPV E7 protein; 2) 75-82 amino acids of the HPV E7 protein; 3) 77-86 amino acids of the HPV E7 protein; 4) 77-90 amino acids of the HPV E7 protein; 5) 79-88 amino acids of the HPV E7 protein; and 6) 4-11 amino acids of the HPV E7 protein. An HPV antigen epitope is obtained by utilizing an identification method of the screened and optimized HPV antigen epitope, the obtained HPV antigen epitope is applied, various dominant mutants are further obtained, drugs and vaccines prepared from the HPV antigen epitope or the mutants can effectively treat various cancers, and the HPV antigen epitope or the mutants are higher in safety and smaller in side effect.

Description

technical field [0001] The present invention relates to the field of biotechnology, specifically, the present invention relates to HPV antigenic epitopes and identification methods and applications thereof, more specifically, the present invention relates to a method for identifying HPV antigenic epitopes, isolated peptides, mutants, and expression vectors , antigen-presenting cells, immune effector cells, use of reagents in the preparation of kits, kits, use of the isolated peptide or expression vector or antigen-presenting cells or immune cells in the preparation of medicines, medicines, vaccines and diagnostic systems. Background technique [0002] Human papillomavirus (HPV) infection is the cause of almost all invasive cervical cancers and some anogenital malignancies and oral cancers. Taking cervical cancer as an example, the persistent infection of human papillomavirus is the main pathogenic factor leading to cervical cancer. It has developed into the fourth most commo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/025C12N15/85C12N5/10G01N33/574A61K39/00A61P35/00
CPCC07K14/005C12N15/85C12N5/0639C12N5/0635C12N5/0645G01N33/574A61K39/0011A61P35/00C12N2510/02A61K2039/5154A61K2039/5156
Inventor 李波周璀张攀
Owner SHENZHEN GINO BIOTECHNOLOGY CO LTD
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