Blood group A epitope mimic peptide and application thereof

An anti-tumor and DNA vaccine technology, applied in the field of biochemistry, can solve the problems of high cost, difficult to modify, and the properties of polysaccharide components are not stable enough.

Inactive Publication Date: 2012-07-04
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the synthesis technology of blood group A sugar antigen is complicated, the purification is difficult and the cost is high; in addition, the properties of polysaccharide components are not stable enough, and it is not easy to modify and other operations

Method used

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  • Blood group A epitope mimic peptide and application thereof
  • Blood group A epitope mimic peptide and application thereof
  • Blood group A epitope mimic peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] Discovery and Screening of Example 1A Blood Group Antigen Epitope Mimetic Peptides

[0021] The A blood group antigen epitope mimetic peptide of the present invention is initially screened out from a phage random 12 peptide library, and the specific method is as follows:

[0022] 1. The titer determination of the phage random 12-peptide library ensures that the amount of phage input is about 1.5×10 11 pfu.

[0023] 2. Affinity panning method of phage random peptide library

[0024] (1) Coat 100 mg / L, 100 μl / well anti-blood group A antigen monoclonal antibody on the ELISA plate, shake gently in a wet box, and incubate overnight at 4°C. Take 10 μl of the peptide library stock solution and dilute it to 100 μl with TBS, add it to the wells of the ELISA plate, 100 μl / well, shake slowly at room temperature for 1 hour. Plates were washed 10 times with 0.1% TBST. Add 100 μl of 0.2M glycine-hydrochloric acid (pH2.2), dilute and amplify the eluate with LB medium, and measure ...

example 2

[0031] Example 2 Study on P / F-pIRES, P / F-M-pIRES Recombinant Plasmids and Their Antitumor Effects

[0032] 1. Construction of P / F-pIRES and P / F-M-pIRES recombinant plasmids

[0033] ①. Construction of P / F-pIRES recombinant plasmid

[0034] Using the sequence of SEQ NO.2 as a template, and using dATP, dGTP, dCTP and dTTP as raw materials, the transmembrane region of the mimetic peptide and FAS gene was formed in an ABI 394 DNA / RNA synthesizer through deprotection group activation, ligation, closure, oxidation and other steps And the P / F fusion gene fused with the intracellular segment, the upstream of the fusion gene contains the Xho I restriction site, and the downstream contains the Mlu I restriction site. The P / Fas fusion gene and pIRES plasmid were double digested with Xho I and Mlu I. The digestion reaction system was 40 μL, including: 8 μL 10×Tango Buffer, 2 μL Xho I, 2 μL Mlu I, 10 μL DNA and 18 μL sterile water. The reaction system was vortexed and mixed, then placed...

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Abstract

The invention provides a blood group A epitope mimic peptide. The amino acid sequence of the peptide is Tyr-Val-Asp-Ser-Lys-Ala-Phe-Arg-Ala-Val-Val-Arg. The blood group A epitope mimic peptide can mimic carbohydrate epitope of blood group A and is used for preparing anti-tumor DNA vaccines. The invention also provides the anti-tumor DNA vaccines which are constructed by inserting the genes codingthe peptide according to the claim 1 and the fusion protein with intracellular domain of Fas into the multiple clone sites of the pIRES plasmids. After being injected into part of the tumor, the DNA vaccines can be absorbed by the tumor cells and express the mimic peptide and the fusion protein of the anti-tumor factors so that the mimic peptide is bonded with the complement and the naturally existing antibody to give play to CDC and ADCC to kill the tumor cells.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to short peptides, especially antigenic epitope mimic peptides. Background technique [0002] Human beings have been committed to the research of tumor immunity for more than 100 years, forming a complete theoretical system and establishing a series of immunotherapy including non-specific immunotherapy, tumor vaccine, adoptive cellular immunotherapy, cytokine immunotherapy, etc. The basic idea of ​​tumor immunotherapy is to mobilize the anti-tumor immune response ability of the host's immune system through related technical methods to eliminate the formed tumor cells or inhibit their further development. Most immunotherapy methods can enhance the immune function of the patient's body, but the anti-tumor effect is much different from expectations. The main reason is that the immune function status of tumor patients cannot directly reflect the anti-tumor immune effect of the body. Even if...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C12N15/62A61K48/00A61P35/00
Inventor 张积仁岑东芝邹建军李许锋罗敏
Owner SOUTHERN MEDICAL UNIVERSITY
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