High cellª²density fermentation of engineered E.coli and antitumor immunity of recombinant human MAGE3/HSP70 in vivo

    HU Peiª²Zhen£¬LU Fan£¬WANG Xiaoª²Gong£¬LI Xia£¬SI Shaoª²Yan£¬HUANG Yang£¬MA Bin£¬ZHANG Xiuª²Min£¬SUI Yanª²Fang

    Department of Pathology, Xijing Hospital, Department of Molecular Biology and Biochemistry£¬ School of Basic Medicine, Fourth Military Medical University£¬Xi¡¯an 710033, China

    ¡¾Abstract¡¿ AIM: To realize high cellª²density fermentation of MAGE3/HSP70 engineered E.coli£¬and to evaluate the antitumor immunity of MAGE3/HSP70 fusion protein in vivo. METHODS: High cellª²density fermentation of MAGE3/HSP70 enginered E. coli was carried out with constant soluble oxygen and continuous cultivation in selfª²control fermentor. The mouse was immunized by purified MAGE3/HSP70 fusion protein in E.coli, and the antitumor immunity was detected by ELISPOT and LDH release assay. RESULTS: High cellª²density fermentation of MAGE3/HSP70 engineered E.coli was successful, and the expression of MAGE3/HSP70 in fermented bacterial was the same as the flask culture. The results of ELISPOT and LDH release assay showed that MAGE3/HSP70 generated tumor specific cytotoxic T lymphocyte (CTL) to damage tumor cel1. CONCLUSION: High cellª²density fermentation of MAGE3/HSP70 engineered E.coli was realized and MAGE3/HSP70 fusion protein can generate specific CTL to kill tumor cel1. ......