title: Study on the preparation of recombinant human HSP70 and its presenting-antigen function.
abstract: 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 fo 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.
authors.affiliation: Department of Medical Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030.
authors.forename: G
authors.initials: G
authors.lastname: Zhang
authors.forename: Z
authors.initials: Z
authors.lastname: Feng
authors.forename: R
authors.initials: R
authors.lastname: Yang
authors.forename: D
authors.initials: D
authors.lastname: Li
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000276:immunology*; D002286:Carcinoma, Ehrlich Tumor / Q000276:immunology / Q000473:pathology; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins