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

作　　者：黄子逸[1,2,3] 陈晨 彭雪楠 申春苹[2,3,4] 朱一蓓 王雪峰 张学光 HUANG Zi-yi;CHEN Chen;PENG Xue-nan;SHEN Chun-ping;ZHU Yi-bei;WANG Xue-feng;ZHANG Xue-guang(Jiangsu Institute of Clinical Immunology,The First Affiliated Hospital of Soochow University,Suzhou Jiangsu 215006;Jiangsu Key Laboratory of Clinical Immunology,Suzhou Jiangsu 215006;Jiangsu Key Laboratory of Gastrointestinal Tumor Immunology,Suzhou Jiangsu 215006;Department of Biochemistry and Molecular Biology,School of Biology and Basic Medical Sciences,Soochow University,Suzhou Jiangsu 215123;Department of Immunology,School of Biology and Basic Medical Sciences,Soochow University,Suzhou Jiangsu 215123,China)

机构地区：[1]苏州大学附属第一医院江苏省临床免疫研究所,江苏苏州215006 [2]江苏省临床免疫学重点实验室,江苏苏州215006 [3]江苏省胃肠道肿瘤免疫重点实验室,江苏苏州215006 [4]苏州大学基础医学与生物科学学院生物化学与分子生物学系,江苏苏州215123 [5]苏州大学基础医学与生物科学学院免疫学系,江苏苏州215123

出　　处：《江苏大学学报（医学版）》2019年第2期93-97,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(31700778;31570889;91642103;81874110;81502454);江苏省"六大人才高峰"高层次人才计划资助项目(2017-SWYY-065);江苏省自然科学基金资助项目(BK20181174)

摘　　要：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.

关 键 词：丁酸钠 FEED 细胞周期 抗体产量 

分 类 号：R393[医药卫生—基础医学]