丁酸钠联合Feed促进293F细胞表达PD-1嵌合抗体

Enhancing effect of sodium butyrate and Feed on PD-1 chimeric antibody expression in 293F cells

目的:探讨丁酸钠和CHO CD Efficient Feed(以下简称Feed)对293F细胞表达程序性死亡受体-1(PD-1嵌合抗体产量的影响。方法:采用无血清培养基,通过哺乳动物293F细胞瞬时表达抗人PD-1嵌合抗体;在细胞对数生长期添加不同浓度丁酸钠(0,0. 5,1,2,4 mmol/L)处理293F细胞,连续培养7 d,分别在第1,3,5天添加Feed。同时,每24 h对培养细胞进行取样,自动细胞计数仪检测细胞总量和活性,流式细胞术检测细胞周期。结果:丁酸钠联合Feed可有效提高293F细胞表达PD-1嵌合抗体,最高表达量为57.4 mg/L,是对照组产量的近8倍;丁酸钠可抑制细胞过度增殖,并阻滞细胞周期G_1期,同时添加Feed可增加抗体产量。结论:丁酸钠控制293F细胞适度增殖,Feed则有利于提高细胞活力,两者作为细胞培养的有效补充剂可显著提高PD-1嵌合抗体的产量。

Objective: To analyse the effect of sodium butyrate ( NaBu) and CHO CD Efficient Feed ( Feed) 2 supplements on the antibody production of programmed death-1( PD-1) in 293F cells. Methods: Transient transfection of 293F cells for PD-1 chimeric antibody expression in serum-free culture medium were treated with different concentrations of NaBu( 0,0. 5,1,2,4 mmol /L). During the 7-day process,5% Feed was added on day 1,3,and 5,respectively. Transfected 293F cells were tested for cell growth and viability every 24 hours. Flow cytometry was used for DNA content /cell cycle analysis. Results: NaBu combined with Feed could effectively increase the expression of PD-1 chimeric antibody in 293F cells,with the highest expression of 57. 4 mg /L,nearly 8 times the yield of the control group. NaBu inhibited cell proliferation and induced G1 phase cell cycle arrest,meanwhile Feed was effective to improve antibody production. Conclusion: NaBu regulated cell growth while Feed was conducive to improving cell viability;NaBu combined with Feed could effectively increase the expression of PD-1 chimericantibody in 293F cells.