Method for rapidly purifying bacteria capsular polysaccharide

A bacterial capsular polysaccharide, capsular polysaccharide technology, applied in the direction of 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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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. Since the chemical characteristics of C-polysaccharide and

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  • Method for rapidly purifying bacteria capsular polysaccharide

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Embodiment 1

[0055] Purify the capsular polysaccharide from the fermentation broth of pneumococcal serotype 19F, the specific method steps are as follows:

[0056] Step 1, fermentation of capsular polysaccharide bacteria: ferment specific capsular polysaccharide bacteria in a fermenter equipped with bacterial culture medium;

[0057] (1), main seed and working seed preparation

[0058] Pneumococcal serotype 19F was obtained from the American Type Culture Collection. The strains in the freeze-dried seed tube were inoculated into 5ml of yeast-acid hydrolyzed casein broth, and cultured at 36°C±2°C for 12-24 hours, when bacteria grow to OD 600 When the reading is 1.0-1.5, inoculate the bacterial solution into 150ml of fresh yeast-acid hydrolyzed casein culture solution, and cultivate at 36°C±2°C for 5-10 hours to the exponential growth phase, stop the cultivation, and pack Freeze-dried and stored at 2-8°C for main seeds.

[0059] Inoculate the strains of the freeze-dried tube of the main se...

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Abstract

The invention discloses a method for rapidly purifying bacteria capsular polysaccharide, which can rapidly remove the pollutants in a specific bacteria broth, protein and nucleic acid are included, the purified capsular polysaccharide can be reserved, capsular polysaccharide-containing bacteria is included, the processes of fermentation, centrifugation and microfiltration are carried out for removing thalline and indissoluble particle substance, acid is added for adjusting the pH value of the solution for deposing the impurities, the solution is subjected to ultrafiltration concentration and filter washing to obtain a crude bacteria capsular polysaccharide solution. A basic principle of the present invention is characterized in that acid is added to regulate the pH value to a range of 3-5, the capsular polysaccharide with impurities is purified in the solution. The capsular polysaccharide can be used for purifying pneumococcus, b-type haemophilus influenza, epidemic meningococcus 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, experiments have found 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 the bacterial cell membrane, so it is called capsular polysaccharide. 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, the c...

Claims

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Application Information

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IPC IPC(8): C12P19/04C08B37/00C12R1/46C12R1/21C12R1/01
Inventor 李建平
Owner KANVAX BIOPHARM
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