丙戊酸钠和锌离子对CHO工程细胞抗体产量的影响  

Effect of sodium valproate and zinc ion on antibody production in engineered CHO cells

作　　者：宋瑞卿 苏杭 张玲 张路明 马宁宁 SONG Rui-qing;SU Hang;ZHANG Ling;ZHANG Lu-ming;MA Ning-ning(Wuyang College,Shengyang Pharmaceutical University Shengyang 110016,Liaoning Province,China)

机构地区：[1]沈阳药科大学无涯学院,辽宁沈阳110016

出　　处：《中国生物制品学杂志》2021年第7期798-803,共6页Chinese Journal of Biologicals

基　　金：重大新药创制——创新抗体药物新品种研发及其关键创新技术体系建设(2019ZX09732001-019)。

摘　　要：目的考察流加补料培养工艺中丙戊酸钠、锌离子及其浓度对表达单克隆抗体的CHO工程细胞抗体产量的影响。方法通过单因素试验筛选丙戊酸钠提高抗体产量的最佳浓度,以及可促进细胞生长并提高抗体产量的流加物质(氨基酸类、维生素类、金属离子等)及其最佳浓度。利用Minitab 17软件考察丙戊酸钠和锌离子两因子的交互作用,得到其最优水平,并进行验证。结果细胞培养第8天单次添加终浓度为1.5 mmol/L的丙戊酸钠,可使抗体产量提高(81.5±9.7)%;锌离子可促进细胞生长并明显提高抗体产量,细胞培养第3天开始,每24 h添加终浓度为90μmol/L的锌离子,可使抗体产量提高(38.3±4.3)%。丙戊酸钠和锌离子组合使用可使抗体产量分别比其单独使用提高27.0%和63.8%。结论组合使用丙戊酸钠、锌离子对CHO工程细胞抗体产量提高的比例明显优于单独使用,可能是丙戊酸钠促进抗体表达的同时抑制细胞生长,限制其提高抗体产量的潜力,锌离子的加入可减弱丙戊酸钠对细胞生长的影响,组合使用可更高效地表达抗体。Objective To investigate the effect of sodium valproate and zinc ion in fed batch culture process of engineered CHO cells for expression of monoclonal antibody(McAb).Methods By using single factor experiment,the optimal sodium valproate concentration for increasing the antibody production and the additives such as amino acids,vitamins and metal ions as well as their concentrations were screened.The interaction of sodium valproate and zinc ion was evaluated by using MINITAB 17 software,then optimized and further verified.Results A single addition of sodium valproate to a final concentration of 1.5 mmol/L on day 8 after cell culture increased the antibody production by(81.5±9.7)%.Zinc ion promoted the cell growth and increased the antibody production significantly.The addition of zinc ion to a final concentration of 90μmol/L every 24 h starting from day 8 after cell culture increased the antibody production by(38.3±4.3)%.However,compared with application of sodium valproate and zinc ion alone,the combination of the two factors increased the antibody production by 27.0% and 63.8% respectively.Conclusion The combined application of sodium valproate and zinc ion was significantly superior to application alone in increasing the antibody production,of which the possible mechanism was that sodium valproate promoted the antibody expression while inhibited the cell growth and limited the potential in increasing antibody production.However,the addition of zinc ion weakened the adverse effect of sodium valproate on cell growth,and the combination of sodium valproate and zinc ion was beneficial to highly effective expression of antibody.

关 键 词：丙戊酸钠 锌离子 抗体产量 CHO工程细胞株 流加补料培养 

分 类 号：R915[医药卫生—微生物与生化药学]