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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 establish a method for the purification of human epidermal growth factor receptor 2 (HER2) protein in breast cancer. Methods Homogenate was prepared from fresh breast cancer tissue and crude protein was extracted from breast cancer tissue by 60% saturated ammonium sulfate precipitation method. The crude protein was extracted by DEAE-Sephadex A-50 ion exchange chromatography and twice Sephacryl S-200 molecular sieve The HER2 protein was further purified by chromatography. The purified protein was identified by SDS-PAGE and Western blot. Results After purification, the relative molecular mass of HER2 protein was 185 000; HER2 protein could specifically bind with rabbit anti-human HER2 polyclonal antibody. Conclusion The established purification method has obtained immunologically active HER2 protein, which laid the foundation for the preparation of monoclonal antibody HER2.