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Prepn process of specific fusion vaccine

A fusion vaccine, a specific technology, applied in the direction of medical preparations containing active ingredients, drug combinations, pharmaceutical formulas, etc.

Inactive Publication Date: 2003-05-14
朱远源
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0103] A method for preparing a specific fusion vaccine, comprising the following steps in turn:

[0104] ①Synthesis of target gene or target antigen by PCR method: For organisms belonging to DNA genome (such as bacteria, DN virus, etc.), extract purified DNA molecules as templates, and use PCR amplification to isolate target genes and antigens;

[0105] ② Translocation and recombination of antigens within and / or between genes: use restriction endonucleases to reconnect the digested fragments to complete the recombination.

[0106] The above-mentioned intragenic and / or intergenic translocation and recombination of antigens mainly includes:

[0107] 1) Antigen random translocation recombination method: Since the target gene is known, according to the requirements of recombination, restriction endonuclease can be selected according to the conventional method, and the digested fragments are reconnected to complete the recombination;

[0108] 2) Form:

[0109] (a) Random translo...

Embodiment 2

[0167] Synthesize target gene or target antigen by PCR method:

[0168] For organisms with RNA genomes (such as RNA viruses, etc.), first extract and purify double-stranded RNA molecules, then reverse-transcribe the double-stranded RNA molecules into DNA molecules, and use this as a PCR template to isolate The target gene or antigen, the whole process is called RT-PCR, in which the specific steps of reverse transcription of RNA into DNA molecules include:

[0169] a) The double-stranded RNA molecule and the random primer are mixed in a molecular ratio of 1:50 (or 1:100 or 1:500 or 1:1000 or 1:10000). Random primer sequence:

[0170] 5′-----------------------XXXX-3′

[0171] 5′-----------------------XXXXX-3′

[0172] 5′-----------------------XXXXXX-3′

[0173] 5′-----------------------XXXXXXX-3′

[0174] 5′-----------------------XXXXXXXX-3′

[0175] 5′-----------------------XXXXXXXXX-3′

[0176] 5′-----------------------XXXXXXXXXX-3′

[0177] 5′-----------------------XX...

Embodiment 3

[0197]Synthesize target gene or target antigen with PCR method: as organism is the parasite or mammal, human body etc. that have POLY (A) RNA, the extracting of total RNA can be used Chinese patent application number: 01113523.9 extracting liquid extraction (or Extract by other methods of the prior art), the method for synthesizing cDNA from total RNA can be carried out by the corresponding method of Chinese patent application number: 01113524.7 (or carry out with other methods of the prior art), with this synthetic cDNA as template, Use the PCR amplification method to isolate the target gene or antigen; if it is to prepare a cancer-specific fusion vaccine, and the gene is a rare gene, it is necessary to add a subtractive screening step for tumor-related genes before the above-mentioned PCR amplification, to remove and normal The same genes of the cells are used to screen out the tumor genes to prevent the vaccine from producing an autoimmune response. This subtractive screenin...

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Abstract

The preparation process of specific fusion vaccine includes successively: synthesizing target gene or target antigen via PCR method; translocating and recombining the antigen inside and / or between genes; cloning to expressino vector to express fusion antigen; screening high-efficiency fusion antigen cloning; and preparing specific fusion vaccine. The novel and unique process can prepare vaccine with high immunological effect for preventing invasion of outer pathogen and self mutation cell reproduction and difffusion and can stimulate cytotoxic T lymphocyte to produce effective immunological treatment of outer pathogen and self mutation cell with antigen carried cell as medium.

Description

Technical field: [0001] The invention relates to a preparation method of a vaccine. Background technique: [0002] Vaccine is an ancient term, and the concept of traditional vaccine is limited to the prevention of external pathogens (such as bacteria, viruses, etc.) and avoids being infected by them. Due to the breakthrough progress in tumor immunity in recent years, therapeutic cancer vaccines have brought new hope to cancer patients. Since then, the vaccine has been liberated from its traditional concept, not only for passive prevention, but also for active treatment. With the progress of the concept of vaccines, the material basis of vaccines has also changed: preventive vaccines have developed from using inactivated pathogens as vaccines to synthetic polypeptides and synthetic nucleic acids (DNA and RNA) as vaccines. The principle is that synthetic polypeptide antigens and nucleic acid antigens directly or indirectly stimulate the immune cells of the body to produce im...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K45/00A61P37/02
Inventor 朱远源
Owner 朱远源
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