Method for purifying A-group and C-group meningococcus polysaccharides

A meningococcal and purification method technology, applied in the field of purification of group A group C meningococcal polysaccharide, can solve problems such as water body and air pollution, environmental hazards of phenol, etc., and achieve the effects of preventing damage, avoiding large-scale use and simple operation.

Inactive Publication Date: 2016-11-16
CHENGDU OLYMVAX BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, phenol has serious harm to the environment

Method used

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  • Method for purifying A-group and C-group meningococcus polysaccharides
  • Method for purifying A-group and C-group meningococcus polysaccharides

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Purification of Group A Meningococcal Polysaccharide

[0018] Dissolve group A meningococcal jaggery in buffer (20mM Tris-HCl, 0.5% sodium deoxycholate, pH 8.0) at a concentration of 5-10mg / ml. After the dissolved sample was clarified and filtered through a 0.45 μm filter membrane, it was loaded on a DEAE-Sepharose Fast Flow chromatography column pre-equilibrated with buffer solution (20mM Tris-HCl, 0.5% sodium deoxycholate, pH 8.0), and the unadsorbed The flow-through peak was then loaded on a CaptoAdhere chromatographic column pre-equilibrated with buffer (20mM Tris-HCl, pH 8.0), eluted with 20mM Tris-HCl, pH 8.0 buffer as the mobile phase, and the polysaccharide-containing target peak. Then the collected polysaccharide solution is desalted with water for injection as the ultrafiltrate with an ultrafiltration membrane bag with a molecular weight cutoff of 300KD. The solution after desalination is the meningococcal polysaccharide solution of group A and group C, and ...

Embodiment 2

[0021] Preparation of Group C Meningococcal Polysaccharide

[0022] Dissolve group C meningococcal jaggery in buffer (20mM Tris-HCl, 0.5% sodium deoxycholate, pH 8.0) at a concentration of 6-8mg / ml. The dissolved sample was clarified and filtered through a 0.45 μm filter membrane, and loaded onto a DEAE-Sepharose Fast Flow column pre-equilibrated with buffer (20mM Tris-HCl, 0.5% sodium deoxycholate, pH 8.0). Adsorbed flow-through peaks were then loaded on a Capto Adhere chromatographic column pre-equilibrated with buffer (20mM Tris-HCl, pH 8.0), eluted with 20mM Tris-HCl, pH 8.0 buffer as the mobile phase, and collected Target peaks containing polysaccharides. Then the collected polysaccharide solution is desalted with water for injection as the ultrafiltrate with an ultrafiltration membrane bag with a molecular weight cutoff of 300KD. The solution after desalination is the meningococcal polysaccharide solution of group A and group C, and the quality index is tested by sampl...

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Abstract

The invention discloses a method for purifying A-group and C-group meningococcus polysaccharides. The method comprises the following steps: (1) carrying out fermentation culture on meningococcus, and carrying out sterilizing treatment, so as to obtain a sterilized culture solution; (2) preparing meningococcus capsular polysaccharides; (3) carrying out chromatographic eluting on a crude prepared polysaccharide solution, collecting polysaccharide containing target peaks, carrying out desalting on the collected polysaccharide solution by using an ultrafiltration membrane pack in a manner of taking water for injection as ultrafiltrate, so as to obtain a desalted solution, i.e., an A-group and C-group meningococcus polysaccharide solution, wherein a buffer solution A is prepared from 20mM Tris-HCl and 0.5% sodium deoxycholate and has the pH of 8.0, and a buffer solution B is prepared from 20mM Tris-HCl and has the pH of 8.0. According to the method, chromatography is adopted creatively, so that the large-amount consumption of phenol in the traditional purification methods is avoided, and the environmental disruption caused by the phenol is effectively avoided on the basis of guaranteeing the quality of purifying; the polysaccharide recovery ratio is close to that of the traditional phenol extraction methods, and the operation is simple, so that the method is applicable to large-scale production.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for purifying polysaccharides of group A and group C meningococci. Background technique [0002] Epidemic meningitis (referred to as meningitis, the same below) is caused by Neisseria meningococcus ( Neisseria meningitidis ) caused by acute respiratory infectious diseases. For more than one hundred years, it has been popular or sporadic all over the world. It can cause sepsis and meningitis after infection with pathogenic bacteria. The susceptible population is mainly children, and the case fatality rate of fulminant type is the highest, which can reach 40% to 60%. The incidence rate in all continents of the world today is 1 / 100,000 to 10 / 100,000, and the total case fatality rate is 5% to 15%. Up to 20% of meningitis patients will have neurological sequelae, including mental impairment and deafness. Serological classification according to the type of capsular polysaccharide...

Claims

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Application Information

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IPC IPC(8): C08B37/00
CPCC08B37/0003
Inventor 吴强陈元芬王宇
Owner CHENGDU OLYMVAX BIOPHARM
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