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首页> 外文期刊>Biotechnology Progress >Use of a Plackett-Burman Statistical Design to Determine the Effect of Selected Amino Acids on Monoclonal Antibody Production in CHO Cells
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Use of a Plackett-Burman Statistical Design to Determine the Effect of Selected Amino Acids on Monoclonal Antibody Production in CHO Cells

机译：使用Plackett-Burman统计设计来确定所选氨基酸对CHO细胞中单克隆抗体产生的影响

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摘要

Culture media design is central to the optimization of monoclonal antibody (mAb) production. Although general strategies do not currently exist for optimization of culture media, the combined use of statistical design and analysis of experiments and strategies based on simple material balances can facilitate culture media design. In this study, we evaluate the effect of selected amino acids on the growth rate and monoclonal antibody production of a Chinese hamster ovary DG-44 (CHO-DG44) cell line. These amino acids were selected based on their relative mass fraction in the specific mAb produced in this study, their consumption rate during bioreactor experiments, and also through a literature review. A Plackett-Burman statistical design was conducted to minimize the number of experiments needed to obtain statistically relevant information. The effect of this set of amino acids was evaluated during exponential cell culture (considering viable cell concentration and the specific growth rate as main output variables) and during the high cell-density stage (considering mAb final concentration and specific productivity as relevant output variables). For this particular cell line, leucine (Leu) and arginine (Arg) had the highest negative and positive effects on cell viability, respectively; Leu and threonine (Thr) had the highest negative effect on growth rate, and valine (Vol) and Arg demonstrated the highest positive impact on mAb final concentration. Results suggest the pertinence of a two-stage strategy for amino acid supplementation, with a mixture optimized for cell growth and a different amino acid mixture for mAb production at high density.
机译：培养基设计对于优化单克隆抗体（mAb）的生产至关重要。尽管目前尚不存在用于优化培养基的一般策略，但基于简单物质平衡的统计设计与实验和策略分析的结合使用可以促进培养基设计。在这项研究中，我们评估选定的氨基酸对中国仓鼠卵巢DG-44（CHO-DG44）细胞系的生长速率和单克隆抗体产生的影响。这些氨基酸是根据其在本研究中产生的特定mAb中的相对质量分数，在生物反应器实验中的消耗率以及通过文献综述选择的。进行了Plackett-Burman统计设计，以最大程度地减少获得统计上相关的信息所需的实验次数。在指数细胞培养（将活细胞浓度和比生长速率视为主要输出变量）和高细胞密度阶段（将mAb终浓度和比产率作为相关输出变量）期间评估了这组氨基酸的作用。对于这种特定的细胞系，亮氨酸（Leu）和精氨酸（Arg）分别对细胞活力具有最高的负面和正面影响。 Leu和苏氨酸（Thr）对生长速率的负面影响最大，而缬氨酸（Vol）和Arg对mAb最终浓度的阳性影响最大。结果表明，针对氨基酸补充的两阶段策略具有相关性，其中一种混合物针对细胞生长进行了优化，而另一种氨基酸混合物则用于以高密度生产mAb。

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