人舌癌Tca8113细胞热休克蛋白多肽复合物(HSP70-PC)的提取和鉴定  

Purification and identification of HSP70 tumor peptide extracted from heated human Tca8113 cells

作　　者：肖林[1] 梁新华[2] 胡静[2] 肖胤[1] 毛祖彝[2] 

机构地区：[1]涪陵中心医院口腔医院口腔颌面外科,重庆408000 [2]四川大学卫生部口腔生物医学工程重点实验室

出　　处：《实用口腔医学杂志》2010年第5期675-676,共2页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:30300391);四川大学华西口腔医学院青年科学研究基金资助项目(编号:2003年)

摘　　要：采用ADP-Agarose亲和层析结合DEAE离子交换对Tca8113细胞的HSP70-PC进行分离纯化。结果显示在NaCl250～350mmol/L浓度处出现一洗脱峰,收集此峰各组分,经10%SDS-PAGE、在相对分子质量70000处可见清晰单一蛋白条带,电泳后于第5峰发现相对分子质量为70000的蛋白,此蛋白被洗脱的盐浓度为NaCl250～350mmol/L,此即为高纯度的HSP70-PC。HSP70获得率为1g湿重的Tca8113细胞能纯化85μg的HSP70-PC。采取低渗匀浆、超速离心、ConA-Sepharose亲和层析、ADP-Agarose亲和层析及DEAE离子交换的纯化方案,可获得高纯度的HSP70-PC。To study the purification and identification of HSP70 tumor peptide extracted from heated human Tca8113 cells.HSP70 associated Tca8113 cells tumor peptides were purified from tumor cell pellet treated in water bath（43 ℃/30 min）,by ConA-Sepharose column,ADP-Agarose column and DEAE ino-exchange chromatography.The purified fractions were tested by SDS-PAGE,immunoblotted with anti-HSP70 mAb.HSP70 tumor peptide was purified from heated Tca8113 cells,which was a high purity protein with 70 kD molecular weight and integrated with anti-HSP70 mAb.The highly purified HSP70-PC can be obtained successfully by using the affinity chromatography and DEAE ion-exchange chromatography.

关 键 词：舌癌 热休克蛋白 纯化 

分 类 号：R739.8[医药卫生—肿瘤]