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Group-A neisseria meningitides capsular polysaccharide conjugated vaccine and preparation method

A Neisseria and capsular polysaccharide technology, which is applied in the direction of antibacterial drugs, pharmaceutical formulas, and medical preparations of non-active ingredients, etc., can solve the problems of inapplicable vaccines, reduce vaccine immunogenicity, etc., and achieve low production costs , Improve quality stability, simple process

Inactive Publication Date: 2013-04-03
CANSINO BIOLOGICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Too high and too low combination ratio will greatly reduce the immunogenicity of the vaccine, making it unsuitable for vaccine use

Method used

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  • Group-A neisseria meningitides capsular polysaccharide conjugated vaccine and preparation method
  • Group-A neisseria meningitides capsular polysaccharide conjugated vaccine and preparation method
  • Group-A neisseria meningitides capsular polysaccharide conjugated vaccine and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Production of serogroup A meningococcal polysaccharide:

[0032]The meningitis capsular polysaccharide production strain (Neisseria meningitides) of group A was purchased from the China Medical Culture Collection and Management Center (strain number: CMCC (B) 29201). Neisseria meningococcus group A was fermented separately in a 50L fermenter. The obtained 30L fermentation broth was centrifuged to obtain supernatant and placed in a stainless steel tank, and concentrated to 3L by ultrafiltration using 100KD membrane bags, and adding 300ml of hexadecyltrimethyl bromide with a mass concentration of 10% to the concentrated solution Aqueous ammonium solution, stirred evenly, and then kept at 4°C for 2 hours. The quiescent solution was centrifuged to obtain a precipitate, to which was added sterile 1 M aqueous sodium chloride solution to 1 L, and dissolution of the precipitate was effected at 40°C. Add anhydrous ethanol to the lysate to a final volume concentration of 25%, s...

Embodiment 2

[0035] Preparation of carrier protein

[0036] The production strain Corynebacterium diphtheriae was purchased by the American Culture Collection Center (ATCC), and the strain number is 39255. The lyophilized diphtheria CRM197 protein-producing seeds were inoculated into test tubes containing culture medium for 16 hours. Transfer an aliquot of the culture to a 0.5 L shake flask containing growth medium and incubate the flask on a rotary shaker at 34.5-36.5°C for 8 hours. liter flask and incubate the flask on a rotary shaker at 34.5-36.5°C for 18 hours. A fermentor containing 30 L of growth medium was inoculated with this 4 liter shake flask culture. The fermenters were incubated at 30-36.5°C, pH 7.4 for 28 hours. The fermenter contents were filtered through a centrifuge and depth filter into a collector.

[0037] The obtained 30L fermentation broth was concentrated to 2L with a 30KD membrane package, and pH 7.5, 500mM phosphate buffer (sodium salt) was added to the concent...

Embodiment 3

[0041] A preparation method for a group A Neisseria meningitidis capsular polysaccharide conjugate vaccine, comprising the following steps: (1) loading 2.5 g of purified group A meningococcal capsular polysaccharide powder into a reactor, adding pure water to Meningococcal capsular polysaccharide was dissolved at a temperature of 4°C when the concentration of meningoencephalic capsular polysaccharide was 4 g / L. Then, the reaction tank was heated and heated, and the meningeal capsular polysaccharide was diluted with pure water to make the concentration of 1.25 g / L for the subsequent reaction. The meningococcal capsular polysaccharide solution was heated to the set temperature according to Table 1, and 30% hydrogen peroxide was added to make the hydrogen peroxide mass concentration shown in Table 1; the temperature and reaction time were shown in Table 1, and the heater was turned off and passed through The solution was rapidly cooled to room temperature by ice-water bath circul...

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Abstract

The invention discloses a group-A neisseria meningitides capsular polysaccharide conjugated vaccine and a preparation method. The preparation method comprises the following steps of: dissolving purified group-A neisseria meningitides capsular polysaccharide in water, heating, and adding hydrogen peroxide to react; performing ultrafiltration and removing impurities, and concentrating to obtain depolymerized meningococcal liquid; adding ADH (antidiuretic hormone) and EDAC (carbodiimide) to react, adding sodium cyanoborohydride to react, and performing ultrafiltration and removing impurities to obtain derived group-A meningococcal capsular polysaccharides; dissolving the derived group-A meningococcal capsular polysaccharides and vector protein CRM197 in buffer liquid, regulating pH, and adding 1-(3-dimethylaminopropyl)-3-ethylcarbodimide hydrochloride to react. The vaccine with molecular weight of over 200KD has stronger immunogenicity. The method is simple in process flow, high in yield and low in production cost, and the polysaccharide-protein ratio is relatively stable and can be controlled by human being, so that the quality stability of combined products can be greatly improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Neisseria meningitidis capsular polysaccharide conjugated vaccine and a preparation method. Background technique [0002] Vaccines based on meningococcal polysaccharides have been described, which elicit an immune response against capsular polysaccharides. These antibodies can cause complement-mediated bacteriolysis against serotype-specific meningococci. Meningococcal polysaccharide vaccines have been shown to be effective in children and adults, but have limited efficacy in infants and young children. Subsequent administrations of this polysaccharide produced weak or no booster responses in young populations. The period of protection of the meningococcal polysaccharide vaccine does not last very long and is estimated to be 3 to 5 years in adults and children older than 4 years. The protection period is less than 3 years for children aged 1 to 4 years. [0003] Pol...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61K39/095A61K47/48A61P31/04
Inventor 朱涛宇学锋毛慧华邵忠琦刘正
Owner CANSINO BIOLOGICS INC
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