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

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.