论文部分内容阅读
The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in glucose free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol/L and 5 mmol/L respectively, the engineered bacteria carrying expression vector of human HSP70 gene expressed rHSP70 at an efficiency of 60 %. After the purification with DEAE ion exchange chromatography, HSP70 with a purity of higher than 90 % was obtained. The purified product could bind tumor antigen peptide in vitro , and the binding was identified by native PAGE containing 5 % glycerol. HSP70 peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells, suggesting that the recombinant human HSP70 could be used as an antigen presenting reagent in tumor therapy.
The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in glucose free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol / L and 5 mmol / L respectively, the engineered bacteria carrying expression vector of human HSP70 gene expressed rHSP70 at an efficiency of 60%. After purification with DEAE ion exchange chromatography, HSP70 with a purity of higher than 90% was obtained. The purified product could bind tumor antigen peptide in vitro, and the binding was identified by native PAGE containing 5% glycerol. HSP70 peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells, suggesting that the recombinant human HSP70 could be used as an antigen presenting reagent in tumor therapy.