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Method for rapid purification of bacterial capsular polysaccharide

A technology of bacterial capsular polysaccharide and capsular polysaccharide, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem of high C-polysaccharide content

Active Publication Date: 2012-09-12
KANVAX BIOPHARM
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  • Abstract
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Problems solved by technology

However, due to the use of sodium deoxycholate lysate for purification, the C-polysaccharide contained in the cell wall of some serotypes is released. Due to the similar chemical properties of C-polysaccharide and capsular polysaccharide, it is not easy to be purified in the subsequent purification. Removed during the process, resulting in too high C-polysaccharide content in the purified polysaccharide

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  • Method for rapid purification of bacterial capsular polysaccharide

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Embodiment 1

[0057] Purify the capsular polysaccharide from the fermentation broth of pneumococcal serotype 19F, the specific method steps are as follows:

[0058] Step 1, fermentation of capsular polysaccharide bacteria: ferment specific capsular polysaccharide bacteria in a fermenter equipped with bacterial culture medium;

[0059] (1), main seed and working seed preparation

[0060] Pneumococcal serotype 19F was obtained from the American Type Culture Collection, inoculated the strains in the freeze-dried seed tube into 5ml of yeast-acid hydrolyzed casein broth, and cultured at 36°C±2°C for 12-24 hours , when the bacteria grow to OD 600 When the reading is 0.6-2.0, inoculate the bacterial solution into 150ml of fresh yeast-acid hydrolyzed casein culture solution, and cultivate at 36°C±2°C for 5-20 hours to the exponential growth phase, stop the cultivation, and pack Freeze-dried and stored at 2-8°C for main seeds.

[0061] Inoculate the strains of the freeze-dried tube of the main se...

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Abstract

The invention discloses a method for rapid purification of bacterial capsular polysaccharide. The method can fast remove pollutants comprising proteins and nucleic acid from a specific bacterial fermentation broth and retains purified capsular polysaccharide. The method comprises the following steps of fermentation of capsular polysaccharide-containing bacteria, acid adjustment of a pH value of a fermentation broth, precipitation of thalli and impurities of the fermentation broth, centrifugation, microfiltration, ultrafiltration condensation and filter wash so that a crude bacterial capsular polysaccharide solution is obtained. A basic principle of the method comprises that through acid adjustment, a pH value of a fermentation broth is in a range of 3 to 5 so that thalli having whole shapes and other impurities are removed and capsular polysaccharide is retained in the fermentation broth and thus purification of capsular polysaccharide is realized. The method can be used for purification of capsular polysaccharide of pneumococcocci, haemophilus influenzae type b, epidemic meningococcocci and typhoid bacillus.

Description

Technical field: [0001] The invention relates to the field of biopharmaceuticals, in particular to a method capable of removing pollutants from bacterial fermentation broth and purifying capsular polysaccharides Background technique: [0002] Pneumococcus, Haemophilus influenzae type b, meningococcus and typhoid bacillus are important pathogenic bacteria that threaten human life. The development of vaccines to prevent infection by these germs has been the goal of the medical community for many years. Since the 1960s, it has been found through experiments that these bacteria have a common morphological structural feature, that is, the surface of the cell membrane is covered with a capsule, which is formed by polysaccharides wrapped on the surface of each cell membrane, so it is called capsule. polysaccharides. In the human body, the capsule plays a decisive role in the pathogenicity of these bacteria because, by preventing antibodies from attaching to the bacterial membrane...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C08B37/00C12R1/46C12R1/21C12R1/01
Inventor 李建平
Owner KANVAX BIOPHARM
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