Generation of stable Chinese hamster ovary pools yielding antibody titers of up to 7.6 g/L using the piggyBac transposon system
Generation of stable Chinese hamster ovary pools yielding antibody titers of up to 7.6 g/L using the piggyBac transposon system
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DOI：
10.1002/btpr.2307
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发表时间：
2016-09
影响因子：
2.9
通讯作者：
Yashas Rajendra;R. Peery;Gavin C Barnard
中科院分区：
文献类型：
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作者：
Yashas Rajendra;R. Peery;Gavin C Barnard
中国仓鼠卵巢（CHO）细胞仍然是许多生物制药药物，特别是单克隆抗体（mAb）的默认生产宿主。克和千克数量的蛋白质的生产通常需要产生稳定的CHO克隆。不幸的是，这个过程需要几个月的时间，大大减缓了药物的发现和开发过程。因此，需要改进的技术来加速生物制药药物的发现和最终药物的生产。在这项研究中，我们描述了使用piggyBac转座子系统产生稳定的CHO池。我们使用四种模型抗体分子（3种mAb和1种双特异性Ab）评估了该系统。在7-12天内分离稳定的CHO合并物。使用简单的16天补料分批工艺，我们测量了四种模型抗体的滴度范围为2.3至7.6 g/L。这表示相对于对照组增加了4至12倍。此外，我们分离了稳定的CHO克隆。我们发现，从piggyBac转座子池中分离的稳定CHO克隆产生的滴度比对照克隆高2至3倍。总之，这些结果表明，稳定的CHO库和克隆产生可以通过使用piggyBac转座子系统显著改善。© 2016美国化学工程师学会生物技术。程序：32：1301-1307，2016
Chinese hamster ovary (CHO) cells remain the default production host for many biopharmaceutical drugs, particularly monoclonal antibodies (mAb). Production of gram and kilogram quantities of protein typically requires the generation of stable CHO clones. Unfortunately, this process takes several months, significantly slowing down the drug discovery and development process. Therefore, improved technologies are needed to accelerate biopharmaceutical drug discovery and final drug substance manufacturing. In this study, we describe the generation of stable CHO pools using the piggyBac transposon system. We evaluated the system using four model antibody molecules (3 mAbs and 1 bispecific Ab). Stable CHO pools were isolated in 7–12 days. Using a simple 16‐day fed‐batch process, we measured titers ranging from 2.3 to 7.6 g/L for the four model antibodies. This represented a 4‐ to 12‐fold increase relative to the controls. Additionally, we isolated stable CHO clones. We found that the stable CHO clones isolated from the piggyBac transposon pools yielded titers two to threefold higher relative to the control clones. Taken together, these results suggest that stable CHO pool and clone generation can be significantly improved by using the piggyBac transposon system. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1301–1307, 2016