BRIAN, BAHLER,TSU-SHUN, LEE,ARNOLD JASON, LOTVIN,EDWARD MARK, RUPPEN,FINK SUE PAMELA, CHARBONNEAU
申请号：
TR201820018
公开号：
TR201820018T4
申请日:
2006.03.31
申请国别(地区):
TR
年份:
2019
代理人:
摘要：
Disclosed is a process for reducing the protein content and preserving the capsular polysaccharide content in a complex cellular Streptococcus pneumoniae lysate broth prior to purification, the process comprising the steps of: (a) growing a selected S. pneumoniae serotype in a soy-based medium, which includes: (i) inoculating a first container containing the soy-based medium with seed stock of the selected serotype, and incubating the first container until growth requirements are met, (ii) inoculating a second container containing the soy-based medium with the culture from step (i) while maintaining a stable pH and temperature in the second container, and (b) lysing with a detergent the bacterial cells produced in step (a), thereby producing a lysate containing soluble proteins, cell debris, nucleic acids and polysaccharide； (c) agitating the cellular lysate for a time sufficient to assure complete lysis and polysaccharide release； (d) lowering the pH of the cellular lysate to less than 5.5 to precipitate out the detergent and most of the soluble proteins； (e) holding the solution and precipitate formed in step (d) without agitation for a time sufficient to allow settling of the precipitate； and (f) processing the solution and precipitate by centrifugation and/or filtration, whereby the capsular polysaccharide in solution is preserved and the soluble protein is effectively reduced.Saflaştırma öncesinde bir kompleks hücresel O Streptococcus pneumoniae lisatı besiyeri içerisindeki protein içeriğinin azaltılmasına ve kapsüler polisakarid içeriğinin korunmasına yönelik bir proses tarif edilir. Hücre lizizi sonrasında pH düşürmeden faydalanılması, protein spesifikasyonunu tutarlı şekilde karşılayan saflaştırılmış bir polisakarid ve saflaştırma prosesi sırasında daha yüksek polisakarid geri kazanım verimleri ile sonuçlanmıştır.
