摘要
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 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
出处
《泰山医学院学报》
CAS
2000年第4期256-266,共11页
Journal of Taishan Medical College
基金
山东省科委
山东省计生委科研资助!( 961165 3 18)
关键词
弓形虫
表膜
P30抗原
纯化
鉴定
制备
toxoplasma gondii
P30 antigen
mcAb
affinity immuoabsorbant
