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目的:研究乳腺癌细胞中癌基因蛋白Her-2的分离纯化方法。方法:采用裂解液裂解、超声破碎、高速离心、ConA Sepharose亲和层析结合ADP agarose亲和层析方法,从热休克处理后的乳腺癌细胞株中分离纯化出癌基因蛋白Her-2,通过SDS-PAGE电泳及免疫印迹法检测进行蛋白分子质量及性质鉴定。结果:检测结果证明纯化后获得的蛋白复合物中包含相对分子质量为185 KD的蛋白,且能与抗Her-2特异单抗结合。结论:采用本方法可以获得相对纯度较高的Her-2蛋白。
Objective: To study the isolation and purification of oncoprotein Her-2 in breast cancer cells. Methods: The oncoprotein Her-2 was isolated and purified from heat-shock-treated breast cancer cell lines by lysate disruption, sonication, high-speed centrifugation, ConA Sepharose affinity chromatography and ADP agarose affinity chromatography. SDS-PAGE electrophoresis and Western blot detection of protein molecular weight and characterization. Results: The results showed that the protein complex obtained after purification contained a protein with a relative molecular mass of 185 KD, and could bind to anti-Her-2 specific mAb. Conclusion: Her-2 protein with higher relative purity can be obtained by this method.