Multi-component antigen acellular pertussis vaccine and preparation method thereof

A pertussis, cell-free technology, applied in the direction of antibacterial drugs, bacterial antigen components, drug combinations, etc., can solve problems such as the inability to screen out the best aP vaccine, and the unresolved adverse reactions of wP vaccines.

Active Publication Date: 2009-08-19
复星安特金(成都)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For decades, the immune effect of wP vaccine has been fully confirmed, but the problem of adverse reactions after wP vaccination has not been resolved
It can be seen that it is still impossible to screen out the best aP vaccine according to the amount of antigenic components, the content of each antigenic component, and the preparation ratio between the antigenic components from the existing aP vaccines at present (Zhang Yanling, Vaccinology, Science Press, 2004 834 pages)

Method used

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  • Multi-component antigen acellular pertussis vaccine and preparation method thereof
  • Multi-component antigen acellular pertussis vaccine and preparation method thereof
  • Multi-component antigen acellular pertussis vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Subculture of Bordetella pertussis strain and culture in fermenter

[0083] The strain of Bordetella pertussis used in the present invention is purchased from the Medical Bacteria Collection (CMCC), and its strain number is CMCC 58003, which is a phase I CS strain of pertussis. The full name of the Medical Bacteria Collection Center is the China Medical Microbiology Collection Management Center, and the Bordetella pertussis strains are preserved in the center's affiliated unit: National Institute for the Control of Pharmaceutical and Biological Products (NICPBP), Beijing. The agency produces, sells and sells Bordetella pertussis strains that are available for purchase by the public. Other suitable strains of Bordetella pertussis can also be used if desired, and the strains given here are merely illustrative of the invention.

[0084] Open the freeze-dried strain of Bordetella pertussis, inoculate it on Bao-ginger medium or other suitable medium, and culture it at 35-37°C...

Embodiment 2

[0086] Preparation of multicomponent cellular structural antigens of Bordetella pertussis

[0087] Add sodium deoxycholate phosphate buffer solution with a final concentration of 0.05-0.20% to the harvested bacterial cell sediment, shake at 2-8°C for 2-4 hours, then stir at 2-8°C for 18-24 hours to lyse cell. Add ammonium sulfate to a final concentration of 15-25%, place at 2-8°C for 5-7 days, centrifuge at 3,000-6,000 rpm for 30 minutes, and collect the precipitate. Add concentrated salt extract to the precipitate, stir at 2-8°C for 5-7 days, centrifuge at 9,000-18,000 rpm for 30-60 minutes, and collect the supernatant. Add ammonium sulfate to a final concentration of 25-35%, place at 2-8°C for 5-7 days, centrifuge at 4,000-9,000 rpm for 30-60 minutes, and collect the precipitate. Add concentrated salt extract to the precipitate, stir at 2-8°C for 3-5 days, then dialyze for 5-7 days, centrifuge at 10,000-30,000 rpm for 1-3 hours, and collect the supernatant. The supernatan...

Embodiment 3

[0089] Preparation of multicomponent cell-secreted antigen of Bordetella pertussis

[0090] Add ammonium sulfate to the harvested culture supernatant to a final concentration of 15-25%, place it at 2-8°C for 5-7 days, centrifuge at 3,000-6,000 rpm for 30 minutes, and collect the precipitate. Add concentrated salt extract to the precipitate, stir at 2-8°C for 5-7 days, centrifuge at 9,000-18,000 rpm for 30-60 minutes, and collect the supernatant. Add ammonium sulfate to a final concentration of 25-35%, place at 2-8°C for 5-7 days, centrifuge at 4,000-9,000 rpm for 30-60 minutes, and collect the precipitate. Add concentrated salt extract to the precipitate, stir at 2-8°C for 3-5 days, then dialyze for 5-7 days, centrifuge at 10,000-30,000 rpm for 1-3 hours, and collect the supernatant. The supernatant is purified by 10-30% sucrose density gradient centrifugation, centrifuged at 20,000-35,000 rpm for 16-24 hours, and the original antigen solution is collected. Add glutaraldehyd...

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Abstract

The invention relates to a multi-component antigen cell-free pertussis vaccine for preventing Bordetella pertussis infection and a preparation method thereof. The cell-free pertussis vaccine comprises multi-component cell structure antigen which is obtained by cracking, separating, extracting and purifying thalli cells of Bordetella pertussis liquid cultures, and can further comprise multi-component cell secretion-type antigen which is obtained by separating, extracting and purifying the supernatant part of the Bordetella pertussis liquid cultures. The combined vaccine is suitable for infants, children, teenagers and adults for preventing the Bordetella pertussis infection.

Description

technical field [0001] The invention relates to a pertussis vaccine, in particular to an acellular pertussis vaccine containing multi-component antigens. The invention also relates to a preparation method of the acellular pertussis vaccine. Background technique [0002] Whooping cough disease is an important cause of infant and child mortality worldwide and remains a public health concern even in countries with high vaccination coverage. According to the latest estimate of WHO, there were about 17.6 million whooping cough cases in the world in 2003, 90% of which occurred in developing countries, and about 279,000 people died of it (WHO. Weekly Epidemiological Record. No. 4, 2005, pp. 31-39). [0003] Bordetella pertussis, the causative agent of whooping cough, is a tiny, Gram-negative coccus with complex nutritional requirements that can specifically attach to the mucous membrane layer of the human respiratory tract. Bordetella genus can undergo phenotypic changes due to c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/10A61K39/08A61K39/05A61P31/04A61P11/00
Inventor 薛平
Owner 复星安特金(成都)生物制药有限公司
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