卵巢癌单抗183B2相应抗原的纯化及其特性分析

Purification and characterization of the antigen recognized by monoclonal antibody 183B2 against ovarian carcinoma

目的 纯化卵巢癌单克隆抗体 1 83B2相应抗原 ,并鉴定该抗原的理化特性。方法 将STRE AMLINErProteinA纯化的单抗 1 83B2与CNBr活化的sepharose 4B相偶联 ,制备亲和层析柱 ,再将用饱和硫酸铵粗提的病人腹水上柱纯化得到 1 83B2相应抗原。提纯抗原经高碘酸钠、胰蛋白酶、链酶蛋白酶、神经氨酸酶、去糖脂混合液及加热处理后进行间接ELISA检测。结果 经亲和层析纯化得到活性好的 1 83B2相应抗原。经高碘酸钠、神经氨酸酶、去糖脂混合液处理后抗原活性不变 ,而胰蛋白酶、链酶蛋白酶、及加热处理后抗原活性消失。结论 单抗 1 83B2相应的抗原决定簇位于肽链上 。

Objective To purify and study the physical and chemical characteristics of MAb183B2 corresponding antigen of ovarian carcinomas. Methods The antigen was purified from ascitic fluid by ammonium sulfate precipitation followed by affinity chromatography on CNBr activated-sepharose 4B column pre bound with monoclonal antibody 183B2, which was purified by STREAMLINE rProtein A column. The activity of the purified antigen was determined after treatment of NaIO4, trypsin, pronase, neuraminidase, deglycolipid mixture or heating. Results The activity of the antigen was still positive after treated by NaIO4, neuraminidase or deglycolipid mixture, while became negative after treatment with trypsin, pronase or heating. Conclusions The epitope of 183B2-Antigen is at its peptide core and this antigen might be novel in ovarian carcinomas.