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为建立金黄色葡萄球菌5型荚膜多糖(CP5)纯化与偶联技术方法,获得具有免疫原性的CP5偶联抗原,采用高压破碎菌体的方法释放荚膜多糖,酶处理去除核酸和蛋白质,通过离子交换层析进一步纯化,得到纯化的CP5,并对其纯度和反应原性进行检测;采用己二酸二酰肼(ADH)间桥法制备CP5-牛血清白蛋白(CP5-BSA)偶联抗原,加佐剂免疫小鼠,进行抗体水平测定。结果表明,纯化的CP5纯度为675.1g/kg,经用免疫琼脂双扩散试验检测,纯化多糖仅与同型菌株抗血清发生沉淀反应,偶联抗原CP5-BSA可以刺激小鼠产生CP5免疫应答。此研究结果为奶牛乳房炎金黄色葡萄球菌亚单位疫苗的研制提供了试验依据。
In order to establish a method for purification and conjugation of Staphylococcus aureus type 5 capsular polysaccharide (CP5), an immunogenic CP5 conjugate antigen was obtained. The capsular polysaccharide was released by high-pressure breaking of the bacterial cell. Enzyme treatment was used to remove nucleic acid and protein , Purified further by ion-exchange chromatography to obtain purified CP5, and its purity and reactivity were tested; CP5-bovine serum albumin (CP5-BSA) was prepared by ADH inter- Coupling antigens, plus adjuvants to immunize mice for antibody level determination. The results showed that purified CP5 had a purity of 675.1 g / kg. The immunoprecipitation assay showed that the purified polysaccharide was precipitated only with the antiserum of the same strain. CP5-BSA could stimulate CP5 immune response in mice. The results of this study provide a test basis for the development of dairy mastitis Staphylococcus aureus subunit vaccine.