弓形虫主要表膜P30抗原的纯化与鉴定

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。

Isolation and identification of the major membrane protein P30 of Toxoplasma gondii

Objective: To isolate and characterize the major membrane protein P30 of Toxoplasma gondii Methods:The monoclonal antibody against P30 antigen was collected from mouse ascitic fluid by injecting the previously prepared E3 hybridoma cells to adult BALB/c mice It was purified by saturated ammonium precipitation and chromatography on DEAE Trisacryl, and then linked to cyanogen bromide activated sepharose 4B The affinity immunoabsorbent was used to purify P30 protein from the large amount of sonicated antigen of tachyzoites The relatively purified protein P30 was identified by SDS PAGE Results:The result showed that about 8 1 mg P30 protein was obtained from the study Conclusion: The McAb affinity immunoabsorbent is a useful method to isolate the P30 antigen