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丁酸钠联合Feed促进293F细胞表达PD-1嵌合抗体 预览

Enhancing effect of sodium butyrate and Feed on PD -1 chimeric antibody expression in 293F cells
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摘要 目的:探讨丁酸钠和CHO CD EfficientFeed(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(0,0.5,1,2,4mmol/L)处理293F细胞,连续培养7d,分别在第1,3,5天添加Feed。同时,每24h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57.4mg/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G1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.
作者 黄子逸 陈晨 彭雪楠 申春苹 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)
出处 《江苏大学学报:医学版》 CAS 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).
关键词 丁酸钠 FEED 细胞周期 抗体产量 sodium butyrate Feed cell cycle antibody expression
作者简介 黄子逸(1984-),女,江苏苏州人,博士,助理研究员;通讯作者:王雪峰,教授,E-mail:wangxuefeng@suda.edu.cn;通讯作者:张学光,教授,E-mail:xueuguangzh@126.com.
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