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CCR5 autogenous polypeptide vaccine and preparation method thereof

A peptide vaccine, autologous technology, applied in the field of medicine and biology, can solve the problems of B cell immune inability to break

Inactive Publication Date: 2007-07-11
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it is not possible to stimulate specific B-cell clones that are knocked out, it is possible that B-cell anergy can be broken through appropriate immunization strategies

Method used

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  • CCR5 autogenous polypeptide vaccine and preparation method thereof
  • CCR5 autogenous polypeptide vaccine and preparation method thereof
  • CCR5 autogenous polypeptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0100] 1 Construction, expression and purification of CCR5 autologous peptide vaccine PADRE-rCCR5

[0101] 1.1 Construction of rCCR5 gene

[0102] Analyze the structure of CCR5 in the Swiss-Pro protein database, combine the literature, intercept the 4 extracellular N-TERM ECL1, ECL2, ECL3 amino acid sequences of CCR5, and use flexible linker to connect them in series to obtain the amino acid sequence of the analog CCR5 extracellular fragment (named: rCCR5), the four amino acid fragments outside the membrane of the receptor CCR5 and the Linker sequence are as follows:

[0103] N-TERM: MDYQVSSPIYDINYYTSEPCQKINVKQIAAR

(31aa)

ECL1: YAAAQWDFGNTMCQ(14aa)

ECL2: RSQKEGLHYTCSSHFPYSQYQFWKNFQTLK(30aa)

[0104] ECL3: NTFQEFFGLNNCSSSNRLDQAM(22aa)

Linker: GGGGS

[0105] 1.1.1 Synthesis of rCCR5 gene

[0106] According to the gene sequence of human CCR5 published by Gene-bank, the EcoRI restriction site is inserted at the 5 end, and the stop...

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Abstract

The invention relates to a method for producing CCR5 autovaccine, wherein based on the structure of CCR5, it uses linker whose amino acid sequence is GGGGS to connect external four sections of CCR5 cell, to simulate the external structure as rCCR5; inserts T cell assist epitope PADRE into N end of rCCR5; artificially synthesizes PADRE- rCCR5 gene; the target gene is colon into represent carrier of pBV-220 corn; transfers bacillus coli, to represent effect in it; washes with inclusion body; chromatography in gel post; obtaining purified protein; the target protein PADRE-rCCR5 is used as CCR5 polypeptide vaccine antigen, while immunity animal represents that the PADRE-rCCR5 can induce body to generate the antibody with high CCR5 specificity; and the invention uses flow cell check immunity antiserum to test U937 cell of CCR5, which proves that the average combine rate can reach 75. 8%. The invention supports CCR5 autovaccine construction and AIDS vaccine research.

Description

Technical field [0001] The invention belongs to the field of medical biotechnology, and specifically relates to technologies such as gene synthesis, protein vaccine construction, animal immunity experiments, and in vitro detection of induced antiserum, especially CCR5 autologous polypeptide vaccine and a preparation method thereof. Background technique [0002] AIDS (acquired immunodeficiency syndrome abbreviated as AIDS) is a chronic infectious disease with extremely high fatality rate caused by human immunodeficiency virus (human immunodeficiency virus abbreviated as HIV) infection, which is raging around the world. The number of HIV-infected people worldwide has reached 40.3 million, and 25 million have died. In 2005, there were 4.9 million newly infected people with HIV, of which 4.2 million were adults, and 3.1 million died of AIDS. At present, there are no very effective measures for the treatment and prevention of AIDS. Treatment is mainly focused on antiretroviral treatme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/18C12N15/09C12P21/02
Inventor 张英起吴孔田韩苇李萌薛晓畅孟洁如包春杰郝强李维娜王增禄
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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