文献类型：期刊文献

中文题名：表达重组蛋白的CHO-GS细胞的无血清培养

英文题名：Serum-free culture of CHO-GS cells for the production of recombinant protein

作者：张芳[1];张立[1];易小萍[1];孙祥明[1];张元兴[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海,200237

年份：2005

卷号：15

期号：7

起止页码：73

中文期刊名：高技术通讯

外文期刊名：Chinese High Technology Letters

收录：CSTPCD;;EI(收录号：2005339305149);Scopus;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

基金：国家高技术研究发展计划(863计划)

语种：中文

中文关键词：重组蛋白;HADAMARD;DMEM/F12;无血清培养基;谷氨酰胺;蛋白表达;细胞生长;细胞密度;统计方法;低分子量;添加浓度;外源蛋白;分批培养;CHO;多聚糖;葡聚糖;牛血清;GS;硫酸;转瓶

摘要：采用DOEHLERT和HADAMARD统计方法建立了一个适于重组CHO-GS细胞生长和外源蛋白表达的无谷氨酰胺无血清培养基SFMB.对多种硫酸多聚糖进行了考察,选取低分子量硫酸葡聚糖,添加浓度为25 mg/L,可以避免细胞结团.用SFMB在方瓶中培养重组CHO-GS细胞,细胞生长优于含10%小牛血清的无谷氨酰胺DMEM/F12培养,最大细胞密度提高了40.7%,外源蛋白的表达水平也得到提高.在100 ml转瓶中分批培养,最大细胞密度和蛋白表达分别达到1.5×106 cells/ml和0.48 mg/L.
A new glutamine-free and serum-free medium SFMB was formulated by using DOEHLERT and HADAMARD statistical methods for the culture of recombinant Chinese hamster ovary cells, transfected with the glutamine synthetase gene, and the expression of recombinant protein. Several kinds of sulfated polysaccharides were investigated to prevent cells from aggregating in serum-free cell culture. This was achieved by adding 25 mg/L dextran sulfate of low molecule weight (5000 Da). In comparison with the culture in glutamine-free DMEM/F12 with 10% new calf serum, the maximum cell density was increased by 40.7% in flask suspension culture in SFMB, and the production of protein was also improved. In the batch culture of rCHO-GS with SFMB in a 100 ml spinner bottle with agitation, cell aggregation did not happened, and cell density and protein production were 1.5 × 106 cells/ml and 0.48 mg/L, respectively.