Role of iron and sodium citrate in animal protein‐free CHO cell culture medium on cell growth and monoclonal antibody production

柠檬酸钠 单克隆抗体 生物化学 细胞培养 化学 抗体 生物 医学 遗传学 病理 免疫学
作者
Yunling Bai,Changjian Wu,Jia Zhao,Yan‐Hui Liu,Wei Ding,Wai Lam W. Ling
出处
期刊:Biotechnology Progress [Wiley]
卷期号:27 (1): 209-219 被引量:49
标识
DOI:10.1002/btpr.513
摘要

Chemically defined iron compounds were investigated for the development of animal protein-free cell culture media to support growth of CHO cells and production of monoclonal antibodies (mAb). Using a multivessel approach of 96-well plates, shake flasks, and bioreactors, we identified iron and its chemical partner citrate as critical components for maintenance of continuous cell growth and mAb production. The optimized iron concentration range was determined to be 0.1-0.5 mM and that for citrate 0.125-1 mM. This complete formulation is able to maintain cell growth to similar levels as those supplemented with iron compounds alone; however, mAb productivity was enhanced by 30-40% when citrate was present. The addition of sodium citrate (SC) did not affect product quality as determined by size exclusion chromatography, ion exchange chromatography, reversed phase and normal phase-HPLC. No significant changes in glucose and lactate profiles, amino acid utilization, or mAb heavy and light chain expression ratios were observed. Cellular ATP level was ∼30% higher when SC was included suggesting that SC may have a role in enhancing cellular energy content. When cell lysates were analyzed by LC-MS to assess the overall cellular protein profile, we identified that in the SC-containing sample, proteins involved in ribosome formation and protein folding were upregulated, and those functions in protein degradation were downregulated. Taken together, this data demonstrated that iron and citrate combination significantly enhanced mAb production without altering product quality and suggested these compounds had a role in upregulating the protein synthetic machinery to promote protein production.
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