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Procedures for the extraction and isolation of bacterial capsular polysaccharides for use as vaccines or linked to proteins as conjugate vaccines
Procedures for the extraction and isolation of bacterial capsular polysaccharides for use as vaccines or linked to proteins as conjugate vaccines
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机译:提取和分离用作疫苗或与蛋白质结合的结合荚膜疫苗的细菌荚膜多糖的程序
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摘要
A procedure to isolate large quantities of capsular polysaccharides (CPS) from culture supernatants as well as bacterial cells of gram-negative and gram-positive bacteria using base extraction is described. The procedure is simple, rapid, reproducible and applicable to a variety of bacterial species. The method also yields novel CPS characterized by their lack of covalent attachment to extraneous peptidoglycan. Vaccines and methods of immunization against bacterial infection using the CPS obtained by the process of the invention are also disclosed. A group B streptococcal capsular polysaccharide produced comprises type Ia CPS having K av approximately of the range 0.010 - 0.005 M w approximately of the range 318 - 311 (kg/mol) M w /M n approximately of the range 1.35 -1.31; type Ib CPS having K av approximately of the range 0.191 - 0.150 M w approximately of the range 218 - 170 (kg/mol) M w /M n approximately of the range 1.61 - 1.20; type II CPS having K av approximately of the range 0.152 - 0.115 M w approximately of the range 289-246 (kg/mol) M w /M n approximately of the range 1.46; type III CPS having K av approximately of the range 0.343 - 0.268 M w approximately of the range 108 - 104 (kg/mol) M w /M n approximately of the range 1.24 - 1.22; type V CPS having K av approximately of the range 0.257 - 0.156 M w approximately of the range 179 - 92 (kg/mol) M w /M n approximately of the range 1.28 - 1.15.
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机译:描述了使用碱基提取从培养上清液以及革兰氏阴性和革兰氏阳性细菌的细菌细胞中分离大量荚膜多糖(CPS)的方法。该过程简单,快速,可重现,适用于多种细菌。该方法还产生了新颖的CPS,其特征在于它们与外源肽聚糖缺乏共价连接。还公开了使用通过本发明的方法获得的CPS的针对细菌感染的疫苗和免疫方法。产生的B组链球菌荚膜多糖包括Ia类型的CPS,其K av大约在0.010-0.005的范围内M w大约为318-311(kg / mol)M w / M n大约在1.35 -1.31的范围内;Ib CPS类型,其K av约为0.191-0.150M w大约为218-170(kg / mol)M w / M n大约在1.61-1.20范围内;II型CPS,K av约为0.152-0.115M w约为289-246(kg / mol)M w / M n大约在1.46的范围内;K av大约在0.343-0.268范围内的III型CPSM w大约在108-104(kg / mol)的范围内M w / M n大约为1.24-1.22;Kav大约在0.257-0.156范围内的V型CPSM w大约为179-92(kg / mol)M w / M n大约为1.28-1.15。
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