Protein-free fed-batch culture of non-GS NS0 cell lines for production of recombinant antibodies
Protein-free fed-batch culture of non-GS NS0 cell lines for production of recombinant antibodies
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DOI：
10.1002/bit.21060
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发表时间：
2007-02-01
影响因子：
3.8
通讯作者：
Saner, Paul W.
中科院分区：
文献类型：
--
作者：
Burky, John E.;Wesson, Mark C.;Saner, Paul W.
本文介绍了一种基于重组NS0衍生细胞系补料分批培养的新型抗体生产平台。针对5种非谷氨酰胺合成酶（GS）NS0细胞系，使用富集且优化的无蛋白、无胆固醇且化学成分确定的基础培养基和补料培养基，开发了一种标准化的补料分批培养工艺。该工艺可重复进行，并且从2升规模到100升生物反应器规模都能可靠地放大，实现了每天每升>120毫克的体积生产率。当细胞进入稳定期或死亡期时，所有5种细胞系的补料分批培养都表现出显著的乳酸消耗。相对于先前开发的补料分批工艺（FBP），乳酸的峰值浓度和最终浓度都较低。考虑到补料分批培养基础培养基具有非传统的高初始葡萄糖浓度（15克/升），且总体葡萄糖消耗量超过17克/升，如此低的乳酸产生量和高的乳酸消耗率是出乎意料的。通过进一步的培养基优化，该工艺平台的潜力得到了进一步证明，对于5种细胞生产系中的一种，在为期13天的补料分批工艺中，最终抗体浓度达到2.64 ± 0.19克/升，体积生产率为每天每升>200毫克。使用这种标准化的无蛋白、无胆固醇的NS0补料分批工艺平台，能够在治疗性抗体的生产中保证研发时间的一致性和成本效益。
Presented is a novel antibody production platform based on the fed-batch culture of recombinant NS0-derived cell lines. A standardized fed-batch culture process was developed for five non-GS NS0 cell lines using enriched and optimized protein-free, cholesterol-free, and chemically defined basal and feed media. The process performed reproducibly and scaled faithfully from the 2-L to the 100-L bioreactor scale achieving a volumetric productivity of > 120 mg/L per day. Fed-batch cultures for all five cell lines exhibited significant lactate consumption when the cells entered the stationary or death phase. Peak and final lactate concentrations were low relative to a previously developed fed-batch process (FBP). Such low lactate production and high lactate consumption rates were unanticipated considering the fed-batch culture basal medium has an unconventionally high initial glucose concentration of 15 g/L, and an overall glucose consumption in excess of 17 g/L. The potential of this process platform was further demonstrated through additional media optimization, which has resulted in a final antibody concentration of 2.64 +/- 0.19 g/L and volumetric productivity of > 200 mg/L per day in a 13-day FBP for one of the five cell production lines. Use of this standardized protein-free, cholesterol-free NS0 FBP platform enables consistency in development time and cost effectiveness for manufacturing of therapeutic antibodies.