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目的 单克隆抗体免疫亲和层析技术提纯弓形虫主要表膜 P30抗原 SDS- PAGE进行鉴定。 方法 用已建立的能分泌抗弓形虫 P30抗原的 E3杂交瘤细胞腹腔注入 BAL B/ c小鼠自腹水中收集单克隆抗体 (Mc Ab) ,经饱和硫酸胺沉淀后 DEAE-纤维柱层析纯化 ,偶联至溴化氢活化的琼脂糖 4B载体上装柱 ,将大量的低频超声粉碎速殖子抗原反复过柱洗脱以收集纯化的 P30蛋白质并通过 SDS- PAGE进行鉴定。 结果 经单克隆抗体免疫亲合层析约获得了 8.1mg较纯的 P30蛋白质 ,在 SDS- PAGE上呈现单一条带。 结论 单克隆抗体免疫亲合层析技术为一较好的提纯 P30抗原的方法
Objective Monoclonal antibody immunoaffinity chromatography purification of Toxoplasma gondii P30 surface antigen was identified by SDS-PAGE. Methods BALB / c mice were injected intraperitoneally with E3 hybridoma cells secreting anti-toxoplasma P30 antigen. The monoclonal antibodies (McAb) were collected from ascites fluid and purified by DEAE-fiber column chromatography after saturated with ammonium sulfate precipitation. Coupled to a hydrogen bromide-activated Sepharose 4B support, a column was loaded and a number of low frequency sonication tachyzoites were repeatedly column eluted to collect the purified P30 protein and identified by SDS-PAGE. Results The monoclonal antibody immunoaffinity chromatography obtained about 8.1 mg of pure P30 protein, showing a single band on SDS-PAGE. Conclusion Monoclonal antibody immunoaffinity chromatography is a better method to purify P30 antigen