Zhou Jing, Yan Guoying Grace, Cluckey David, Meade Caryl, Ruth Margaret, Sorm Rhady, Tam Amy S, Lim Sean, Petridis Constantine, Lin Laura, D'Antona Aaron M, Zhong Xiaotian
BioMedicine Design, Medicinal Sciences, Pfizer Worldwide R&D, 610 Main Street, Cambridge, MA 02139, USA.
Antibodies (Basel). 2023 Aug 10;12(3):53. doi: 10.3390/antib12030053.
Rapidly producing drug-like antibody therapeutics for lead molecule discovery and candidate optimization is typically accomplished by large-scale transient gene expression technologies (TGE) with cultivated mammalian cells. The TGE methodologies have been extensively developed over the past three decades, yet produce significantly lower yields than the stable cell line approach, facing the technical challenge of achieving universal high expression titers for a broad range of antibodies and therapeutics modalities. In this study, we explored various parameters for antibody production in the TGE cell host Expi293F and ExpiCHO-S with the transfection reagents ExpiFectamine and polyethylenimine. We discovered that there are significant differences between Expi293F and ExpiCHO-S cells with regards to DNA complex formation time and ratio, complex formation buffers, DNA complex uptake trafficking routes, responses to dimethyl sulfoxide and cell cycle inhibitors, as well as light-chain isotype expression preferences. This investigation mechanistically dissected the TGE processes and provided a new direction for future transient antibody production optimization.
利用培养的哺乳动物细胞通过大规模瞬时基因表达技术（TGE）快速生产类药物抗体疗法，用于先导分子发现和候选药物优化，这一过程通常得以实现。在过去三十年里，TGE方法得到了广泛发展，但与稳定细胞系方法相比，其产量显著较低，面临着为广泛的抗体和治疗方式实现普遍高表达滴度的技术挑战。在本研究中，我们用转染试剂ExpiFectamine和聚乙烯亚胺，探索了TGE细胞宿主Expi293F和ExpiCHO-S中抗体生产的各种参数。我们发现，Expi293F和ExpiCHO-S细胞在DNA复合物形成时间和比例、复合物形成缓冲液、DNA复合物摄取运输途径、对二甲基亚砜和细胞周期抑制剂的反应以及轻链同种型表达偏好方面存在显著差异。这项研究从机制上剖析了TGE过程，并为未来瞬时抗体生产优化提供了新方向。
