目的:探讨肺腺癌细胞中热休克蛋白70多肽复合物分离及纯化的方法。方法:采用裂解液裂解、超声破碎、ConA—Sepharose亲和层析、ADP-agarose亲和层析结合DEAE阴离子交换层析方法,从体外培养的热休克(43℃)处理后的人肺腺癌细胞株A549中分离纯化HSP70多肽复合物,通过SDS—PAGE电泳、Western-blot检测获得蛋白。结果:获得蛋白的相对分子质量约为70000,能与抗HSP70特异单抗结合。结论:采用ConA-Sepharose亲和层析、ADP—agarose亲和层析结合DEAE阴离子交换层析从肺腺癌细胞株A549中可获得高纯度的HSP70多肽复合物,为进一步研究以HSP70多肽复合物为基础的肺癌免疫治疗提供了依据。
Aim : To research a method of separating and purifying HSP70-associate tumor peptides from human lung a cell( A549 ). Methods : HSP70-associate tumor peptides were separated and purified from tumor cells treated with water bath(43℃ ) by ConA-Sepharose column, ADP-agarose column, and DEAE ion-exchange chromatography. The purified fractions were tested by SDS-PAGE and Western blot. Results: The final purified fractions had a high purity and specificity. Conclusion: HSP70-associate tumor peptides could be purified by ConA-Sepharose column, ADP-agarose affinity chromatography and DEAE ion-exchange chromatography, which provides reference for immunity treatment of lung cancer.