乳癌组织中人类表皮生长因子受体2蛋白的纯化及鉴定  

Purification and identification of human epidermal growth factor receptor 2 in breast cancer tissue

作　　者：施雨露[1] 孙成艳 张贯石[1] 聂彩霞 潘润泽[4] 付勤[5] 刘永茂[1] 

机构地区：[1]吉林大学基础医学院实验教学中心,吉林长春130021 [2]长春迪瑞医疗科技股份有限公司,吉林长春130012 [3]沈阳二〇四医院,辽宁沈阳110043 [4]吉林大学第一医院放射线科,吉林长春130021 [5]吉林省肿瘤医院,吉林长春100018

出　　处：《中国生物制品学杂志》2015年第5期536-539,共4页Chinese Journal of Biologicals

基　　金：吉林省自然科学基金(20105071)

摘　　要：目的建立乳腺癌组织中人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)蛋白的纯化方法。方法采用新鲜的乳腺癌组织制备匀浆液,经60%饱和硫酸铵盐析沉淀法进行乳腺癌组织蛋白的粗提;再通过DEAE-Sephadex A-50离子交换层析和两次Sephacryl S-200分子筛层析进一步纯化HER2蛋白;纯化蛋白进行SDS-PAGE和Western blot鉴定。结果纯化后HER2蛋白相对分子质量为185 000;HER2蛋白可与兔抗人HER2多克隆抗体发生特异性结合。结论建立的纯化方法获得了具有免疫学活性的HER2蛋白,为HER2单克隆抗体的制备奠定了基础。Objective To develop a method for purification of human epidermal growth factor receptor 2 （HER2） from breast cancer tissue. Methods Fresh breast cancer tissue was prepared into homogenate, from which protein was crudely extracted by salting out with 60% saturated ammonium sulfate and further purified by DEAE-Sephadex A-50 ion exchange chromatography and 2 times of Sephacryl S-200 molecular sieve chromatography, then identified by SDS-PAGE and Western blot. Results The purified HER2 protein, with a relative molecular mass of 185 000, which showed specific binding to rabbit anti-human HER2 polyclonal antibody. Conclusion HER2 protein with immune activity was obtained by the developed purification methods, which laid a foundation of preparation of monoclonal antibody against HER2.

关 键 词：乳腺癌 人类表皮生长因子受体2蛋白 纯化 

分 类 号：R373.24[医药卫生—病原生物学] R392-33[医药卫生—基础医学]