An empirical modeling platform to evaluate the relative control discrete CHO cell synthetic processes exert over recombinant monoclonal antibody production process titer.
An empirical modeling platform to evaluate the relative control discrete CHO cell synthetic processes exert over recombinant monoclonal antibody production process titer.
复制标题
一个经验建模平台，用于评估相对控制离散 CHO 细胞合成过程对重组单克隆抗体生产过程滴度的影响。
DOI：
10.1002/bit.23146
复制
发表时间：
2011
影响因子：
3.8
通讯作者：
McLeod J
中科院分区：
文献类型：
--
作者：
McLeod J
在本研究中，我们结合了经验推导的细胞内Mab合成数学模型，以定量比较在整个最先进的工业流加培养过程中，各个细胞过程限制GS-CHO细胞产生重组IgG 4单克隆抗体的程度。基于细胞内Mab合成和分泌途径每个阶段的生产过程控制系数的计算，我们确定了整个培养过程中Mab生产的主要细胞限制是重组重链基因转录和重链mRNA翻译。令人惊讶的是，尽管培养期间细胞生物质合成速率显著下降，伴随着能量密集型Mab合成过程（Mab折叠/组装和分泌）的计算速率常数下降，但这些在任何生产阶段都没有对Mab合成产生显著控制。相反，通过增加Mab基因转录来维持细胞特异性Mab生产，这抵消了细胞生物合成速率的下降。重要的是，这项研究表明，这种全过程预测建模策略的应用应该合理地先于细胞工程方法并为细胞工程方法提供信息，以增加哺乳动物宿主细胞重组蛋白的产量，其中生产力的控制本质上是蛋白质产物和细胞系特异性的。Biotechnol. Bioeng. 2011;108：2193-2204.© 2011 Wiley Periodicals，Inc.
In this study we have combined empirically derived mathematical models of intracellular Mab synthesis to quantitatively compare the degree to which individual cellular processes limit recombinant IgG4monoclonal antibody production by GS‐CHO cells throughout a state‐of‐the‐art industrial fed‐batch culture process. Based on the calculation of a production process control coefficient for each stage of the intracellular Mab synthesis and secretion pathway, we identified the major cellular restrictions on Mab production throughout the entire culture process to be recombinant heavy chain gene transcription and heavy chain mRNA translation. Surprisingly, despite a substantial decline in the rate of cellular biomass synthesis during culture, with a concomitant decline in the calculated rate constants for energy‐intensive Mab synthetic processes (Mab folding/assembly and secretion), these did not exert significant control of Mab synthesis at any stage of production. Instead, cell‐specific Mab production was maintained by increased Mab gene transcription which offset the decline in cellular biosynthetic rates. Importantly, this study shows that application of this whole‐process predictive modeling strategy should rationally precede and inform cell engineering approaches to increase production of a recombinant protein by a mammalian host cell—where control of productivity is inherently protein product and cell line specific. Biotechnol. Bioeng. 2011;108:2193–2204. © 2011 Wiley Periodicals, Inc.