Purification method of streptococcus pneumoniae capsular polysaccharide

Abstract

The invention relates to a purification method of streptococcus pneumoniae capsular polysaccharide. The purification method comprises the following steps: subjecting streptococcus pneumoniae lysate to ultrafiltration and condensation, adding sodium deoxycholate, adjusting the pH to 4-5, carrying out a centrifugal treatment, collecting the supernate, adding nuclease into the supernate, carrying out reactions for 3-8 hours to obtain acidified lysate, adding sodium deoxycholate into the acidified lysate, subjecting acidified lysate to ultrafiltration, performing acidification and centrifugation, collecting the supernate, and finally subjecting the supernate to ultrafiltration and condensation to obtain a purified polysaccharide solution. In the provided purification method, phenol is not used; after ethanol drying, the obtained purified streptococcus pneumoniae capsular polysaccharide is the cell lysate after one nuclease treatment, the product is more environment-friendly and safer, the process time is short, the production cost is low, and massive production can be achieved.

Description

technical field [0001] The field of the present invention relates to a purification method of capsular polysaccharide of Streptococcus pneumoniae, especially a simplified purification method. Background technique [0002] Streptococcus pneumoniae is the primary pathogen of respiratory tract infection, not only the most common pathogen causing community-acquired pneumonia in children, but also the main pathogen causing fatal pneumonia infection in the elderly. At present, more than 90 serotypes have been found, of which about 30 serotype strains are pathogenic to humans. Capsular polysaccharide is the main virulence factor of Streptococcus pneumoniae. The research results show that the capsular polysaccharide vaccine of Streptococcus pneumoniae has good immunogenicity and safety. [0003] The general process of preparing the Streptococcus pneumoniae capsular polysaccharide vaccine is as follows: the bacteria are grown in a fermenter, and at the end of the fermentation, sodiu...

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Problems solved by technology

US20100129881 and NormaSuarez et al. (NORMASUA′REZ, LAURAFRANCO, et al., FRAGUAS APPLIEDANDENVIRONMENTALMICROBIOLOGY, 2001, p.969–971Vol.67, No.2), introduced the method of column chromatography in the purification process of Streptococcus pneumoniae capsular polysaccharide, However, this method has high requirements for operators, takes a long time for sample processing, and the operation and maintenance costs of equipment are high, so it is difficult to meet the needs of large-scale production

CA1206905 proposed enzymatic hydrolysis method to purify capsular polysaccharide of Streptococcus pneumoniae, VivianeMaimoni (Viviane Maimoni etal., Simple and efficient method of bacterial polysaccharides purification for vaccines production using hydrolytic enzymes and tangential flow ultrafiltration, Communicating Current Research and Educational Topics and Trends in Applied Microbiology, 2007, 1(13):450–457; Viviane Maimoni etal., PurificationofCapsularpolysaccharides form streptococcus spneumoniaeserotype23Fbyaproceduresuitableforscale‐up, Biotechnol.Appl.Biochem., 2003,37(3):283‐287) and others, made some improvements, but these methods all need to use a variety of nucleases, proteases, more steps , the cost is high, and ethanol graded precipitation treatment is still required, which has potential safety hazards

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