丁酸钠提高重组CHO细胞表达量的研究  

SODIUM BUTYRATE ENHANCE THE EXPRESSION OF RECOMBINANT CHO CELLS

作　　者：顾琪[1] 俞吉安[2] 张月丽[1] 施泓[1] 汪家愉[1] 

机构地区：[1]华东医院上海市老年医学研究所,上海市200040 [2]上海交通大学生命科学技术学院,200030

出　　处：《老年医学与保健》2003年第3期153-154,170,共3页Geriatrics & Health Care

摘　　要：目的提高CHO细胞表达外源目标蛋白产量。方法用不同浓度的丁酸钠处理重组CHO细胞,观察细胞数量,细胞形态,表达率,表达量的变化。结果当丁酸钠的浓度为1mM,2mM,5mM时,重组细胞的表达量有不同程度的提高。表达率与丁酸钠的浓度有关,也与丁酸钠加入的时间有关。传代24小时加药的细胞,经2mM,5mM丁酸钠处理后,表达量虽比对照组高,但细胞生长情况不好;传代48小时加药的细胞,经5mM丁酸钠处理后,表达量可提高5倍左右,但细胞数大大减少。当细胞长满瓶后(第7天)加入丁酸钠5mM,加药次日的表达量为对照组(不加药组)的9.5倍。细胞数也有所减少。上述实验表明,由于丁酸钠对细胞生长有抑制作用,表达量虽提高,但细胞数减少,不能达到增产的目的。因此我们设计了用正常培液与丁酸钠交替使用的方法,获得了良好的效果。5mM丁酸钠与正常培液交替处理,在加药后7天内所得的表达物产量为对照组的2.1倍。提高了重组CHO细胞表达目标蛋白的产量。结论丁酸钠的合理使用可提高目标蛋白的产量。Objective To enhance the protein product of the recombinant CHO cells.Methods Different concen-trations of Sodium Butyrate(NaBu)were added to the culture medium.Their influence to cell number,cell morphology ,ex-pression rate and expression amount s were observed.Results When the concentrations were 1mM,2m M,and 5mM,the expression rate of the recombinant cells increased at different degrees.The expression rate related to the c oncentration of NaBu,and also to the time when we put the NaB u into the medium.When put the NaBu at24h after passage,the concentratio ns were2mM and 5mM,although the expression rates were higher than control,the c ell growth went down.When put the NaB u at 48h after passage,the concentration wa s 5mM,the expression rate raised 5ti mes .But the number of cells decrease d.When put the NaBu at the 7th day after passage ,the concentration was 5mM,the expre ssion rate reached to 9.5times comparing to the control.But the cell number also dec reased.These experimental data showed that the addition of NaBu could en hance the expression rate,but the number of ce lls decreased.It was because of the e ffects of cell inhibition by the NaBu.So we designed an alternate inducing methods,whic h used medium with and without NaBu alternately.When use medium with and w ithout NaBu,its concentration was 5mM,the total amount of the protein product within 7days were 2.1times comparing to the control.Conclusions Use NaBu in a suitable way can enhance the product of the target protein of t he recombinant CHO cells.[

关 键 词：丁酸类 CH0细胞 重组蛋白质类 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]