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High cell-density fermentation of engineered E.coli and antitumor immunity of recombinant human MAGE3/HSP70 in vivo
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 3
- DOI:
- 10.3321/j.issn:1000-2790.2007.03.003
- Key Word:
- 黑色素瘤抗原;发酵;大肠杆菌
Abstract： 目的:实现重组人黑色素瘤抗原MAGE3/HSP70融合蛋白工程菌的高密度发酵,并测定MAGE3/HSP70融合蛋白在体内的抗肿瘤免疫效应. 方法:利用自控发酵罐发酵,采用溶氧反馈,分批补料的方法对MAGE3/HSP70融合蛋白工程菌进行了高密度发酵,从菌体中纯化MAGE3/HSP70融合蛋白,并用所纯化的MAGE3/HSP70融合蛋白免疫小鼠,运用酶联免疫斑点法(ELISPOT)和细胞毒性杀伤实验(LDH)检测了融合蛋白激活机体细胞免疫反应的状况. 结果:高密度发酵取得成功,发酵菌中MAGE3/HSP70融合蛋白的表达量与摇瓶中的相当,摸索出发酵水平纯化MAGE3/HSP70融合蛋白工艺,ELISPOT和LDH显示MAGE3/HSP70可以激活机体免疫反应,并产生针对MAGE3的CTL,特异性杀伤表达MAGE3的肿瘤细胞. 结论:MAGE3/HSP70融合蛋白工程菌成功实现高密度发酵,MAGE3/HSP70融合蛋白可以有效激活机体免疫反应,产生针对特异性抗原MAGE3的CTL,免疫学活性良好,为新型抗肿瘤疫苗临床前研究奠定了良好基础.
- [1]马加海,隋延仿,叶菁,等.肿瘤抗原基因MAGE-3的克隆、原核表达与纯化[J].2004,(6).
- [2]JIANG Xiao,Hong-Song Chen.Biological functions of melanoma-associated antigens[J].2004,(13).
- [3]孟洁如,颜真,赵宁,等.导向性人干扰素α2a工程菌的高密度发酵[J].2004,(23).
- [4]Minami K,Yamaguchi Y,Hihara J,etc.Generation of antigen-presenting cells using cultured dendritic cells and amplified autologous tumor mRNA.[J].2005,69(5).
- [5]Anna Maria, Sandén,Ingela, Prytz,Ioannis, Tubulekas,etc.Limiting factors in Escherichia coli fed-batch production of recombinant proteins.[J].2003,81(2).
- [6]De Anda R,Lara AR,Hernandez V,etc.Replacement of the glucose phosphotransferase transport system by galactose permease reduces acetate accumulation and improves process performance of Escherichia coli for recombinant protein production without impairment of growth rate.[J].2006,8(3).
- [7]R, Khalilzadeh,S A, Shojaosadati,N, Maghsoudi,etc.Process development for production of recombinant human interferon-gamma expressed in Escherichia coli.[J].2004,31(2).
- [8]James, Zawada,James, Swartz.Maintaining rapid growth in moderate-density Escherichia coli fermentations.[J].2005,89(4).