Experimental and in silico modelling analyses of the gene expression pathway for recombinant antibody and by-product production in NS0 cell lines.
Experimental and in silico modelling analyses of the gene expression pathway for recombinant antibody and by-product production in NS0 cell lines.
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DOI：
10.1371/journal.pone.0047422
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
von der Haar T
中科院分区：
文献类型：
--
作者：
Mead EJ;Chiverton LM;Spurgeon SK;Martin EB;Montague GA;Smales CM;von der Haar T
单克隆抗体是商业上重要的、高价值的用于治疗多种疾病的生物活性药物。这些复杂分子由通过二硫键共价连接的两条重链和两条轻链多肽组成。它们通常表达为来自培养的哺乳动物细胞的重组蛋白，其能够正确地修饰、折叠和组装多肽链成天然四级结构。此类重组细胞系通常在产生的产物量和分泌产物的异质性方面有所不同。这种变异的生物学机制尚未完全确定。在这里，我们利用实验和建模策略来验证和定义表现出不同抗体表达水平的细胞系中的生物学基础产品异质性，然后通过实验验证这些模型。在进行这些研究时，我们应用并验证了抗体表达的生化（基于速率常数）和工程（非线性）模型，以获得来自具有不同IgG4分泌速率的四种NS 0细胞系的实验数据。该模型预测，完整抗体及其片段的输出是内在联系的，因此不能在分泌机制的水平上单独操纵。相反，这些模型强调了在前体物种水平上操作的策略，以增加高产和低产细胞系中的重组蛋白产量。该模型还突出了抗体表达途径中的细胞系特异性限制。
Monoclonal antibodies are commercially important, high value biotherapeutic drugs used in the treatment of a variety of diseases. These complex molecules consist of two heavy chain and two light chain polypeptides covalently linked by disulphide bonds. They are usually expressed as recombinant proteins from cultured mammalian cells, which are capable of correctly modifying, folding and assembling the polypeptide chains into the native quaternary structure. Such recombinant cell lines often vary in the amounts of product produced and in the heterogeneity of the secreted products. The biological mechanisms of this variation are not fully defined. Here we have utilised experimental and modelling strategies to characterise and define the biology underpinning product heterogeneity in cell lines exhibiting varying antibody expression levels, and then experimentally validated these models. In undertaking these studies we applied and validated biochemical (rate-constant based) and engineering (nonlinear) models of antibody expression to experimental data from four NS0 cell lines with different IgG4 secretion rates. The models predict that export of the full antibody and its fragments are intrinsically linked, and cannot therefore be manipulated individually at the level of the secretory machinery. Instead, the models highlight strategies for the manipulation at the precursor species level to increase recombinant protein yields in both high and low producing cell lines. The models also highlight cell line specific limitations in the antibody expression pathway.
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DOI：
10.1084/jem.134.5.1316
发表时间：
1971-11-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Baumal R;Potter M;Scharff MD
通讯作者：
Scharff MD
影响因子：
3.8
作者：
Doolan, Padraig;Meleady, Paula;Clynes, Martin
通讯作者：
Clynes, Martin
影响因子：
100.3
作者：
Beck, Alain;Wurch, Thierry;Corvaia, Nathalie
通讯作者：
Corvaia, Nathalie
影响因子：
3.8
作者：
Bi, JX;Shuttleworth, J;Ai-Rubeai, M
通讯作者：
Ai-Rubeai, M
影响因子：
4.1
作者：
Abe, Yuki;Gor, Jayesh;Dalby, Paul A.
通讯作者：
Dalby, Paul A.