工业微生物2017，Vol.47Issue(1)：43-51,9.DOI:10.3969/j.issn.1001-6678.2017.01.007
运用氨基酸分析法和DOE法优化抗PD-1单克隆抗体生产用培养基
Optimiztion of anti-programmed death-1 monoclonal antibody medium using amino acid analysis and design of experiment
摘要
Abstract
The purpose of this study was to optimize anti-PD-1 monoclonal antibody basic and feed medium by using amino acid analysis and design of experiment.The better basic and feed medium was obtained according to the cell growth and antibody expression through single factor screening a variety of basic and feed medium from different vendors.The kinds of key amino acids affecting cell culture and antibody expression through analysis amino acid consumption of better basic and feed medium were determined using amino acid analysis method.The anti-PD-1 monoclonal antibody basic and feed medium were optimized according to cell growth and antibody expression through adding different concentrations of amino acids using design of experiment analysis software.The ultimately determined optimal basic medium formula were as follows:Hycell CHO Medium,1.04 mmol/L L-asparagine,0.76 mmol/L L-glutamine.The optimal feed medium formula were the following:OPM CHOCD Feed 1,38.7 mmol/L L-histidine,75.0 mmol/L L-tyrosine,64.0 mmol/L L-serine,49.2 mmol/L L-glutamine,18.7 mmol/L L-cysteine.Through the 3 L test validation,the optimized medium peak viable cell density (PVCD)increased by 62.7％ than that of the unoptimized and the anti-PD-1 monoclonal antibody expression increased by 71.5％ than that of unoptimized,respectively.The biological activity of anti-PD-1 monoclonal antibody in optimized medium was not significantly reduced if compared with that of the unoptimized medium.关键词
抗PD-1单克隆抗体/培养基优化/DOE分析/氨基酸分析Key words
anti-PD-1 monoclonal antibody/optimizing medium/design of experiment analysis/amino acid analysis引用本文复制引用
高玲梅,张慧娟,李若竹,于玉根..运用氨基酸分析法和DOE法优化抗PD-1单克隆抗体生产用培养基[J].工业微生物,2017,47(1):43-51,9.基金项目
2014年度深圳市科技创新委员会技术创新计划战略新兴产业发展专项资金资助项目,项目编号:JSGG20141117163744687. （）
