Group ABC meningococcus combined vaccine and preparing method thereof

A meningococcal and combined vaccine technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, vaccines, etc., can solve the problem of no bactericidal power, and achieve the effect of reducing the number of vaccinations and reducing the pain of vaccination

Pending Publication Date: 2016-12-14
BEIJING ZHIFEI LVZHU BIOPHARM +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Antibodies produced by immunization with recombinant fHBP can cause complement-mediated bactericidal effects against the same subfami...

Method used

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  • Group ABC meningococcus combined vaccine and preparing method thereof
  • Group ABC meningococcus combined vaccine and preparing method thereof
  • Group ABC meningococcus combined vaccine and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1A

[0066] The preparation of embodiment 1A group, C group polysaccharide:

[0067] After fermentation of group A and group C meningococci, the supernatant was collected by centrifugation in a disc centrifuge, and cetyltrimethylammonium bromide with a final concentration of 0.15% was added to precipitate the polysaccharide, and the polysaccharide precipitate was collected by centrifugation, and the final concentration was The precipitate was dissolved with 0.5 mol / L calcium chloride, and the nucleic acid was removed by ethanol precipitation with a final concentration of 25%, the supernatant was collected by centrifugation, and the polysaccharide was precipitated with cold ethanol with a final concentration of 75%. After dissolving the polysaccharide and treating it with 1-3% sodium deoxycholate, use GE Capto adhere and CaptoDEAE two kinds of gel medium chromatography in series to collect the flow-through peak, and filter the flow-through peak with a 30KD membrane bag ultrafiltratio...

Embodiment 2

[0068] The preparation of embodiment 2 carrier protein TT:

[0069]The fermentation culture was centrifuged in a disc centrifuge to collect the supernatant, after detoxification by 0.5% formaldehyde, 30KD ultrafiltration was used to remove impurities, 30% ammonium sulfate was used to precipitate TT, and Sephacryl S-300HR gel filtration chromatography was used to collect TT protein The monomer peak was concentrated by 30KD ultrafiltration to prepare TT stock solution. The detoxification process needs to be checked for detoxification, and the original solution needs to be checked for specific toxicity and toxicity reversal.

Embodiment 3

[0070] Preparation of embodiment 3 polysaccharide-protein conjugate (taking cyanogen bromide activation method as example):

[0071] 3.1 Polysaccharide activation:

[0072] Dissolve the polysaccharide to 5-15 mg / ml, adjust the pH value to about 10.8, add 1 / 10 (g / g) cyanogen bromide to the polysaccharide solution, and activate the polysaccharide for 8 minutes at room temperature in an alkaline environment with a pH value of 10.8.

[0073] 3.2 Polysaccharide derivation:

[0074] Add 0.5mol / L adipic dihydrazide at a ratio of 1:1 (volume ratio to activated polysaccharide), and react at room temperature for 50-70min. Cyanogen bromide and adipic dihydrazide were removed by ultrafiltration with a 30KD membrane bag to obtain polysaccharide derivatives.

[0075] 3.3 Polysaccharide protein binding:

[0076] Mix and stir the polysaccharide derivative and the carrier protein TT at a ratio of 1:1 (g / g), add carbodiimide at a ratio of carbodiimide: polysaccharide = 1:10, room temperature...

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Abstract

The invention provides a group ABC meningococcus combined vaccine and a preparing method thereof. The group ABC meningococcus combined vaccine is prepared from a group A and group C meningococcus polysaccharide-protein conjugate, and recombinant protein of a human H factor binding protein (fHBP) subgroup A and a human H factor binding protein (fHBP) subgroup B of group B meningococcus. The invention further provides a preparing method of the recombinant fHBP-A protein and fHBP-B protein. The combined vaccine is used for immunity of children 2 or more years old, and used for preventing invasive diseases such as cerebrospinal meningitis, bacteremia, pneumonia and pericarditis caused by group A or group B or group C meningococcus, and providing a better and wider protection effect on meningococcus.

Description

technical field [0001] The present invention relates to the preparation of a vaccine, in particular to an ABC group meningococcal combined vaccine and a preparation method thereof. A combined vaccine preparation prepared by mixing polysaccharide protein conjugates, group B meningococcal human factor H binding protein (fHBP) A and B subfamily two recombinant proteins and a preparation method thereof. Background technique [0002] 1 Classification and prevalence of epidemic cerebrospinal meningitis: [0003] Meningococcal meningitis (referred to as meningococcal meningitis) is a respiratory infectious disease mainly caused by meningitis and bacteremia caused by Neisseria meningitides (NM). At present, infants and young children are the main cases, showing endemic, outbreak, and sporadic forms. The invasive diseases caused by NM include meningitis, bacteremia, and pneumonia, etc., and the case fatality rate is about 12% to 20%. Among the survivors, 11% % ~ 19% left sequelae. ...

Claims

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Application Information

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IPC IPC(8): A61K39/116A61P31/04C07K14/22C12N15/70C12N15/31
CPCA61K39/095A61K2039/545A61K2039/55505A61K2039/70C07K14/22C12N15/70C12N2800/101C12N2800/22
Inventor 苏桂民杜琳朱卫华冀颖
Owner BEIJING ZHIFEI LVZHU BIOPHARM
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