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Method for preparing pneumococcal capsular polysaccharide by viscosity control

A technology for pneumococcus and capsular polysaccharide is applied in the field of preparation of pneumococcal capsular polysaccharide, which can solve the problems of increasing downstream process pressure, excessive dilution, affecting stirring effect, etc., and achieves good quality, improved production efficiency and recovery rate. high effect

Active Publication Date: 2019-02-15
BEIJING ZHIFEI LVZHU BIOPHARM +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In the stage of polysaccharide polymerization by CTAB, only the final concentration of CTAB is controlled, and the polymerization process may not be sufficient due to the difference in sugar concentration.
The solution with too high viscosity has a high concentration of sugar, which may lead to insufficient total amount of CTAB, and affect the stirring effect, make the local reaction concentration too high, and affect the uniformity of the reaction; the dilution factor is too large, increasing the amount of CTAB added, and at the same time increasing pressure on downstream processes

Method used

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  • Method for preparing pneumococcal capsular polysaccharide by viscosity control
  • Method for preparing pneumococcal capsular polysaccharide by viscosity control
  • Method for preparing pneumococcal capsular polysaccharide by viscosity control

Examples

Experimental program
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Effect test

Embodiment 1、1

[0091] The viscosity control of embodiment 1, type 1 pneumococcal polysaccharide solution ultrafiltration

[0092] (a) Type 1 pneumococcus fermentation broth is inactivated, centrifuged, clarified and filtered, and then concentrated until the viscosity reaches 10 cp, and then dialyzed. After dialysis, continue to over-concentrate the solution, wash the membrane bag 2-3 times with buffer solution, collect the concentrated solution, and stir evenly.

[0093] (b) Detect the viscosity of the type 1 polysaccharide solution, dilute the solution viscosity with buffer solution to 10cp for clarification and filtration, continue to dilute with buffer solution to a viscosity of 5cp after filtration, and add CTAB solution under continuous stirring.

[0094] (c) Centrifuge the CTAB polysaccharide solution, collect the precipitate, place in the sodium chloride solution and continue to stir, add the sodium chloride solution until the viscosity increases to 5cp until the viscosity is 3cp; add...

Embodiment 2

[0102] The viscosity control of embodiment 2, type 2 pneumococcal polysaccharide solution ultrafiltration

[0103] (a) After inactivation, centrifugation, clarification and filtration of the type 2 pneumococcal fermentation liquid, it is concentrated first, and when the viscosity reaches 8cp, dialysis is started. After dialysis, continue to over-concentrate the solution, wash the membrane bag 2-3 times with buffer solution, collect the concentrated solution, and stir evenly.

[0104] (b) Detect the viscosity of the type 2 polysaccharide solution, dilute the solution viscosity with buffer solution to 8cp for clarification and filtration, continue to dilute with buffer solution to a viscosity of 3cp after filtration, and add CTAB solution in a state of continuous stirring.

[0105] (c) Centrifuge the CTAB polysaccharide solution, collect the precipitate, place in the sodium chloride solution and continue to stir, add the sodium chloride solution until the viscosity increases to ...

Embodiment 3

[0113] The viscosity control of embodiment 3, type 3 pneumococcal polysaccharide solution ultrafiltration

[0114] (a) Type 3 pneumococcus fermentation broth is inactivated, centrifuged, clarified and filtered, and then concentrated, until the viscosity reaches 10cp, and then dialyzed. After dialysis, continue to over-concentrate the solution, wash the membrane bag 2-3 times with buffer solution, collect the concentrated solution, and stir evenly.

[0115] (b) Detect the viscosity of the type 3 polysaccharide solution, use buffer to dilute the solution viscosity to 10cp for clarification and filtration, use buffer to continue diluting to a viscosity of 5cp after filtration, and add CTAB solution in a state of continuous stirring.

[0116] (c) Centrifuge the CTAB polysaccharide solution, collect the precipitate, place in the sodium chloride solution and continue to stir, add the sodium chloride solution until the viscosity increases to 5cp until the viscosity is 3cp; add the so...

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Abstract

The invention belongs to the field of biological products and relates to a preparation method of pneumococcal capsular polysaccharide. According to the preparation method, pneumococcus inactivated andclarified fermentation liquid as an initial feed liquid is used for purification and preparation by steps as follows: ultrafiltering and concentrating the clarified fermentation liquid, adding CTAB to form a precipitate with polysaccharide, performing centrifugation to collect the precipitate, and performing depolymerization by sodium chloride and chromatography purification to finally obtain pneumococcal capsular polysaccharide; viscosity control comprises steps of ultrafiltering, clarifying and filtering, precipitation by CTAB, depolymerization by sodium chloride and chromatography.

Description

technical field [0001] The invention belongs to the field of biological products and relates to a preparation method of pneumococcal capsular polysaccharide. Background technique [0002] Pneumococci are Gram-positive bacteria with capsules and polysaccharide antigens. Pneumococcus can cause a variety of diseases, such as pneumonia, otitis media, bacteremia, meningitis, etc. The condition is serious and the mortality rate is high. Therefore, the development of preventive vaccines has become an urgent need. At present, there are 23-valent pneumococcal polysaccharide vaccines, 10-valent and 13-valent pneumococcal polysaccharide conjugate vaccines on the market for the prevention of pneumococcal infection, of which 23-valent pneumococcal polysaccharide vaccines are used for adults, and 10-valent and 13-valent pneumococcal polysaccharide vaccines are used for children. Both the polysaccharide vaccine and the polysaccharide conjugate vaccine need to use the capsular polysacchari...

Claims

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Application Information

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IPC IPC(8): C08B37/00
CPCC08B37/0003
Inventor 林彦彬朱卫华杜琳
Owner BEIJING ZHIFEI LVZHU BIOPHARM
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