一种适用于多种沙门氏菌血清型的 O 抗原可扩展纯化方法。
A scalable method for O-antigen purification applied to various Salmonella serovars.
机构信息
Novartis Vaccines Institute for Global Health, 53100 Siena, Italy.
出版信息
Anal Biochem. 2013 Mar 1;434(1):136-45. doi: 10.1016/j.ab.2012.10.038. Epub 2012 Nov 7.
Abstract
The surface lipopolysaccharide of gram-negative bacteria is both a virulence factor and a B cell antigen. Antibodies against O-antigen of lipopolysaccharide may confer protection against infection, and O-antigen conjugates have been designed against multiple pathogens. Here, we describe a simplified methodology for extraction and purification of the O-antigen core portion of Salmonella lipopolysaccharide, suitable for large-scale production. Lipopolysaccharide extraction and delipidation are performed by acetic acid hydrolysis of whole bacterial culture and can take place directly in a bioreactor, without previous isolation and inactivation of bacteria. Further O-antigen core purification consists of rapid filtration and precipitation steps, without using enzymes or hazardous chemicals. The process was successfully applied to various Salmonella enterica serovars (Paratyphi A, Typhimurium, and Enteritidis), obtaining good yields of high-quality material, suitable for conjugate vaccine preparations.
摘要
革兰氏阴性菌的表面脂多糖既是一种毒力因子，也是 B 细胞抗原。针对脂多糖 O 抗原的抗体可能提供针对感染的保护，并且已经针对多种病原体设计了 O 抗原缀合物。在这里，我们描述了一种从沙门氏菌脂多糖中提取和纯化 O 抗原核心部分的简化方法，适用于大规模生产。通过整个细菌培养物的乙酸水解进行脂多糖提取和去脂，并且可以直接在生物反应器中进行，而无需事先分离和灭活细菌。进一步的 O 抗原核心纯化包括快速过滤和沉淀步骤，而不使用酶或危险化学品。该工艺已成功应用于各种沙门氏菌血清型（副伤寒 A、伤寒和肠炎），获得了高质量的高产量物质，适用于结合疫苗的制备。
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