Tumor genetically-engineered vaccine utilizing mucin-1 and survivin as target sites

A vaccine composition and site technology, which is applied in the fields of tumor DNA vaccines, viral vector vaccines and protein vaccines, can solve the problems of weak immunogenicity of DNA vector vaccines, relatively large potential safety hazards and difficulties in preparation of recombinant poxvirus vaccines in primary vaccination, etc. question

Active Publication Date: 2012-10-17
CHANGCHUN BCHT BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

From the discovery of MUC1 to the present, a lot of vaccine research work has been carried out with it, and some vaccines have entered the stage of human clinical research, but most of them are traditional polypeptide, protein or dendritic cell (DC) vaccines, Most of these vaccines have problems such as poor immunogenicity or difficulty in preparation; the genetic vaccines that have entered the clinic are mainly recombinant poxvirus vaccines. Although no toxicity has been detected, there are still relatively large safety hazards in primary vaccination-boosting of recombinant poxvirus vaccines directly. , and the clinical effect is not very satisfactory; DNA carrier vaccine is still basically in the preclinic...

Method used

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  • Tumor genetically-engineered vaccine utilizing mucin-1 and survivin as target sites
  • Tumor genetically-engineered vaccine utilizing mucin-1 and survivin as target sites
  • Tumor genetically-engineered vaccine utilizing mucin-1 and survivin as target sites

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Experimental program
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Effect test

Embodiment Construction

[0042] One. The preparation method and conditions of the above-mentioned fragment 33M, S8 and CpG motif

[0043] Preparation of 1.33M

[0044] MUC1VNTR is a repeat fragment containing 60 bases, GenBank number is NM_002456 (SEQ ID NO: 1). Two pairs of primers were designed according to the base sequence of a VNTR, and a VNTR (1m) repeat region fragment was synthesized by overlapping extension PCR (SOE PCR), and then digested with restriction endonucleases SalI and XhoI to produce the same sticky end characteristics, sequentially connected to construct 33M gene fragments. The specific method is as follows:

[0045] 1) Overlap extension PCR technology

[0046] PCR reaction conditions are: 95°C for 20s, 55°C for 20s, 72°C for 30s, 30 cycles, using primers P1 (SEQ ID NO: 2) and P2 (SEQ ID NO: 3) to amplify 1 copy of MUC1 without ATG VNTR (abbreviated as m), using P2 and P3 (SEQ ID NO: 4) as primers to amplify 1 copy of MUC1 VNTR (abbreviated as Am) fragment containing ATG.

[...

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Abstract

The invention relates to fields of tumor DNA vaccines, viral vector vaccines and protein vaccines and especially relates to a tumor genetically-engineered vaccine utilizing mucin-1 and survivin as target sites. The tumor genetically-engineered vaccine comprises recombinant DNA vectors of CpVR-MS and CpDV-IL2-MS. The invention also relates to a use of recombinant protein vaccines of S8 and 9M, a DNA vaccine, a viral vaccine, a protein vaccine and an optimized combination of immune methods of the DNA vaccine, the viral vaccine and the protein vaccine in preparation of an anti-tumor vaccine.

Description

technical field [0001] The invention relates to the fields of tumor DNA vaccines, virus vector vaccines and protein vaccines. Specifically, the present invention relates to recombinant DNA vectors CpVR-MS and CpDV-IL2-MS; recombinant protein vaccines S8 and 9M and DNA vaccines, viral vector vaccines, protein vaccines and the optimal combination of immune methods between these three forms of vaccines Use in the preparation of anti-tumor vaccines. Background technique [0002] Survivin is a member of the inhibitor of apoptosis protein (IAP) family, has dual functions of anti-apoptosis and regulation of cell division, and is widely expressed in various embryonic tissues and cancer cells, but not in normal terminally differentiated cells . These characteristics make survivin a new target in the fields of tumorigenesis, development and therapy. [0003] Current research shows that survivin has potential value in tumor gene therapy: its tumor-specific expression is positively c...

Claims

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Application Information

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IPC IPC(8): C12N15/63A61K48/00A61K38/17A61P35/00
Inventor 孔维张海红于湘晖于永慧王玉倩张丽星
Owner CHANGCHUN BCHT BIOTECH
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