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目的 应用单克隆抗体免疫亲和层析技术提纯弓形虫主要表膜P30抗原并进行鉴定。方法 将我们已建立的分泌抗表膜P30抗原单克隆抗体的E3杂交瘤细胞注入BALB/c小鼠腹腔 ,收集腹水 ,用饱和硫酸胺沉淀和DEAE—纤维柱层析法提纯单克隆抗体 ,将其偶联至溴化氢活化的琼脂糖 4B上装柱 ,经免疫亲和层析技术 ,从大量的低功率超声粉碎速殖子抗原中分离纯化P30蛋白质 ,并用SDS—PAGE进行鉴定。结果 本次实验约获得了 8 1mg的P30蛋白质 ,且纯度较高。结论 单克隆抗体免疫亲和层析技术可获得一定量的P30抗原。
OBJECTIVE: To purify and identify P30 antigen of the major surface membrane of Toxoplasma gondii by monoclonal antibody immunoaffinity chromatography. Methods E3 hybridoma cells secreting McAb against P30 antigen were prepared and injected into the peritoneal cavity of BALB / c mice. The ascites was collected and the monoclonal antibodies were purified by saturated ammonium sulphate precipitation and DEAE-fiber column chromatography. It was coupled to hydrogen bromide-activated Sepharose 4B column and purified by immunoaffinity chromatography. The P30 protein was isolated and purified from a large number of low-power sonication tachyzoites and identified by SDS-PAGE. Results In this experiment, 8 1 mg of P30 protein was obtained with high purity. Conclusion Monoclonal antibody immunoaffinity chromatography can obtain a certain amount of P30 antigen.