肺腺癌组织中HSP70多肽复合物的分离及纯化

Separation and purification of heat shock protein 70 peptide complex from human lung adenocarcinoma tissue

目的:探讨肺腺癌组织中HSP70多肽复合物分离及纯化的可行性方案。方法:采用裂解液裂解、超声破碎、ConA-sepharose亲和层析和DEAE阴离子交换层析方法,从肺腺癌组织中分离纯化HSP70蛋白复合物;通过SDS-PAGE电泳、Westernblot检测获得蛋白;运用Bradford法对其进行定量分析。结果:通过该实验方法可以从1g肺癌组织中获取110μg左右的HSP70多肽复合物,其相对分子质量为70000。结论:采用ConA-sepharose亲和层析和DEAE阴离子亲和层析相结合的蛋白纯化法可以获取高纯度的HSP70多肽复合物。

Aim：To explore a method of separating and purifying HSP70 peptide complex from human lung adenocarcinoma.Methods：HSP70 peptide complex was separated and purified from human lung adenocarcinoma tissue by means of cell lysis,ultrasonication,ConA-sepharose column,and DEAE ion-exchange chromatogaraphy.The purified fractions were tested by SDS-PAGE and Western blot.And quantitative analysis was made by Bradford.Results：About 110 μg HSP70 peptide complex was obtained from 1 g lung adenocarcinoma tissue by this experimental method,with relative molecular weight of 70 000.Conclusion：The high purity HSP70 peptide complex could be obtained by ConA-sepharose column and DEAE ion-exchange chromatogaraphy.