Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, Connecticut, USA.
Janssen Biotherapeutics, Spring House, Pennsylvania, USA.
Biotechnol J. 2021 Apr;16(4):e2000251. doi: 10.1002/biot.202000251. Epub 2021 Jan 25.
Transient gene expression (TGE) using mammalian cells is an extensively used technology for the production of antibodies and recombinant proteins and has been widely adopted by both academic and industrial labs. Chinese Hamster Ovary (CHO) cells have become one of the major workhorses for TGE of recombinant antibodies due to their attractive features: post-translational modifications, adaptation to high cell densities, and use of serum-free media. In this study, we describe the optimization of parameters for TGE for antibodies from CHO cells. Through a matrix evaluation of multiple factors including inoculum, transfection conditions, amount and type of DNA used, and post-transfection culture conditions, we arrived at an uniquely optimized process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply. We further investigated the amount of coding DNA used in TGE and the influence of kinetics and size of the transfection complex on the in vitro efficiency of the transfection. We present here the first report of an optimized TGE platform using Filler DNA in an early drug discovery setting for the screening and production of therapeutic mAbs.
瞬时基因表达 (TGE) 利用哺乳动物细胞是一种广泛用于生产抗体和重组蛋白的技术,已被学术和工业实验室广泛采用。由于其吸引人的特点,中国仓鼠卵巢 (CHO) 细胞已成为 TGE 重组抗体的主要工作载体之一:翻译后修饰、适应高细胞密度和使用无血清培养基。在这项研究中,我们描述了优化 CHO 细胞中抗体 TGE 的参数。通过对包括接种物、转染条件、使用的 DNA 量和类型以及转染后培养条件在内的多个因素的矩阵评估,我们得到了一个独特的优化工艺,具有更高的滴度和更低的成本和时间,从而提高了早期抗体材料供应的整体效率。我们进一步研究了 TGE 中使用的编码 DNA 量以及转染复合物的动力学和大小对转染体外效率的影响。我们在这里首次报道了在早期药物发现环境中使用 Filler DNA 的优化 TGE 平台,用于筛选和生产治疗性 mAb。