Purification of capsular polysaccharide from Streptococcus pneumoniae serotype 23F by a procedure suitable for scale-up
Purification of capsular polysaccharide from Streptococcus pneumoniae serotype 23F by a procedure suitable for scale-up
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DOI：
10.1042/ba20020075
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发表时间：
2003-06-01
影响因子：
2.8
通讯作者：
Tanizaki, MM
中科院分区：
文献类型：
--
作者：
Gonçalves, VMM;Takagi, M;Tanizaki, MM
肺炎链球菌是一种致病性荚膜细菌，可引起肺炎、菌血症和脑膜炎。结合载体蛋白的荚膜多糖已被广泛用作疫苗抗原。血清型23F是全球流行的肺炎球菌之一。建立了一种简便、高效、易于规模化提纯23F型荚膜多糖的方法。该方法使用0.22 mm膜切向微滤获得的培养液，在30 kDa螺旋膜上切向超滤浓缩肉汤微滤液，分馏乙醇沉淀（28-60%），核酸酶和蛋白酶处理，在30 kDa盒式膜上浓缩/滤过。最终多糖回收率为89%。最终蛋白和核苷酸污染分别为1.5% （w/w）和0.3% （w/w）。最终得到的纯多糖符合世界卫生组织的要求，成品中未发现残留的蛋白酶。
Streptococcus pneumoniae is a pathogenic encapsulated bacterium, which causes pneumonia, bacteraemia and meningitis. Capsular polysaccharide conjugated to a carrier protein has been widely used as a vaccine antigen. Serotype 23F is one of the prevalent worldwide pneumococci. A simple and efficient method for capsular polysaccharide serotype 23F purification that can easily be scaled-up was developed. This method consisted of using culture broth obtained by tangential microfiltration through a 0.22 mum membrane, broth microfiltrate concentration by tangential ultrafiltration in a 30 kDa spiral membrane, fractional ethanol precipitation (28-60%), nuclease and proteinase treatment, and concentration/diafiltration in a 30 kDa cassette membrane. The final polysaccharide recovery was 89%. The final protein and nucleotide contamination was 1.5% (w/w) and 0.3% (w/w) respectively. The final pure polysaccharide meets the requirements of the World Health Organization and residual proteinase was not found in the final product.