重组人HSP70的制备及其提呈抗原作用的研究  

Study on the Preparation of Recombinant Human Heat Shock Protein 70 and Its Presenting-Antigen Function

作　　者：杨瑞[1] 冯作化[1] 张桂梅[1] 李东[1] 

机构地区：[1]华中科技大学同济医学院医学分子生物学研究室,武汉430030

出　　处：《药物生物技术》2001年第5期244-247,共4页Pharmaceutical Biotechnology

基　　金：国家自然科学基金 ( 39970 32 2 );教育部跨世纪人才培养计划基金资助项目

摘　　要：研究重组人热休克蛋白 70 (HSP70 )的制备及其提呈抗原的作用。采用不含葡萄糖的M9ZB培养基及终浓度为 0 .0 2mmol/LIPTG和 5mmol/L乳糖诱导带有人HSP70基因表达质粒的工程菌 ,表达效率达到 6 0 %,经DEAE阴离子交换层析柱可获得纯度 >90 %的HSP70。该纯化产物可于体外结合肿瘤抗原肽。利用含 5 %甘油的非变性PAGE分析可以快速鉴定HSP70与肿瘤抗原肽的结合。HSP70 -肿瘤抗原肽复合物在体外可以激活淋巴细胞的特异性杀肿瘤功能。The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in M9ZB medium without glucose and induced with IPTG at a final concentration of 0.02 mmol/L and lactose at an final concentration of 5 mmol/L, the engineered bacteria carrying expression vector of human HSP70 gene could express at a efficiency of 60%. After the purification with DEAE ion exchange chromatography, the purity of HSP70 was higher than 90%. The purified product could bind with tumor antigen peptide in vitro , and the binding can be identified by non denaturating PAGE containing 5% glycerol. HSP70 peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells. The results indicated that the recombinant human HSP70 can be used in tumor therapy as a factor presenting antigen.

关 键 词：重组 热休克蛋白70 抗原肽 肿瘤 

分 类 号：R394.8[医药卫生—医学遗传学] R392－33[医药卫生—基础医学]