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Poliomyelitis inactivated vaccine and its preparing method

A technology for polio and inactivated vaccine, applied in the field of inactivated polio vaccine, can solve the problems of inconvenient large-scale production, increased vaccine production cost, and the threat of laboratory leakage

Inactive Publication Date: 2005-08-03
BEIJING INST OF BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, the production of IPV using wild-type viruses needs to be carried out in a biosafety level 3 laboratory. Through the airflow control in the operation room and strict disinfection of the discharged substances, the virus is prevented from leaking into the environment or infecting the operators. Such production not only Increase the cost of vaccine production, and it is not convenient for large-scale production
In addition, as the goal of global polio eradication is getting closer, the opportunities for humans to be naturally infected with poliovirus are decreasing, and the possible laboratory leakage caused by the use of wild-type virus strains to produce vaccines (W-IPV) is harmful to humans. The threat appears to be growing

Method used

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preparation example Construction

[0017] In the present invention, the inactivated polio vaccine is carried out with an attenuated strain as the virus species. In one embodiment, the present invention provides a method for preparing an inactivated polio vaccine, including the following steps:

[0018] a, cultivating cells from cell lines;

[0019] b. When the cells grow into a dense monolayer, inoculate the attenuated polio strain virus;

[0020] c, the virus multiplies in the cell;

[0021] d. Collect the virus suspension produced by the cells;

[0022] e. Remove cell residues through clarification and filtration;

[0023] f, Concentrate the virus suspension by ultrafiltration;

[0024] g. Purify the virus liquid through a gel filtration chromatography step and an ion exchange chromatography step;

[0025] h, the virus is inactivated by an inactivating agent;

[0026] i. Prepare an inactivated vaccine.

[0027] In the present invention, the cell line used can be any cell conventionally used to propagate attenuate...

Embodiment 1

[0051] Example 1 Vaccine production process flow

[0052] Cells: An African green monkey kidney cell line (Vero cell line) was used. The cell was isolated from an African green monkey kidney in 1963 by a Japanese scholar. The 112th generation was sent to the American Type Culture Collection (ATCC). After identification, the cells were non-tumorigenic, free of bacteria, mold and mycoplasma contamination, and no virus contamination, and were suitable for the production of biological products.

[0053] In December 1988, the Beijing Institute of Biological Products obtained the 117th generation Vero cell (numbered CCL81) two square flask from ATCC in the United States, which was immediately frozen as the original seed in liquid nitrogen. A master cell bank was prepared from a primary seed bank by passage and expansion, and a working cell bank was prepared by passage and expansion from a master seed bank. The verification of the master cell bank and the working cell bank are in complia...

Embodiment 2

[0066] Example 2 Vaccine purity analysis

[0067] Because the cell matrix of the vaccine is passaged cells, the purity of the vaccine directly affects the safety of the vaccine. The WHO's "Regulations on Passaging Cells as Cellular Substrates for Biological Products" requires that the residual cell DNA in the vaccine must be less than 100pg / dose. Therefore, the residual Vero cell DNA in the vaccine of the present invention is detected by nucleic acid hybridization. The method proceeds as follows:

[0068] Culture collection about 10 7 Vero cells, according to the method provided by "Molecular Cloning" to extract Vero cell chromosomal DNA. The DNA was reacted with proteinase K and SDS at 50°C for 3 hours, extracted three times with a phenol / form / isoamyl alcohol (25:24:1) mixture, and then the DNA was precipitated with absolute ethanol. The purified DNA is used as a standard for DNA determination, and a part of it is labeled with photosensitive biotin to prepare probes. Use the same...

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Abstract

The present invention relates to inactivated poliomyelitis vaccine and its preparation process, and is especially inactivated poliomyelitis vaccine prepared with attenuated poliomyelitis virus as initial strain. The present invention also relates to the method of preventing the testee from suffering from poliomyelitis.

Description

Technical field [0001] The invention relates to an inactivated polio vaccine and a preparation method thereof. Specifically, the inactivated polio vaccine of the present invention is prepared using attenuated polio virus as the virus species. The present invention also relates to a method of preventing polio. Background technique [0002] Poliomyelitis is an acute infectious disease caused by the polio virus, which spreads widely and is extremely harmful. In the late 1950s, inactivated polio vaccines and oral live vaccines (IPV and OPV) came out one after another. The use of IPV and OPV effectively reduces the incidence of polio. Since the WHO put forward the goal of eradicating polio in 1988, the number of reported cases worldwide has dropped from 350,000 cases in 1988 to less than 700 cases in 2003, and the number of cases has been reduced from 125 countries in 1988 to six. [0003] my country successfully developed OPV at the end of 1959. The production process is to inoculate...

Claims

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Application Information

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IPC IPC(8): A61K39/02
CPCY02A50/30
Inventor 石慧颖李懿田龙赵敏翁捷张冲陈明支惠庞成华
Owner BEIJING INST OF BIOLOGICAL PROD
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