Rajendra Yashas, Balasubramanian Sowmya, Peery Robert B, Swartling James R, McCracken Neil A, Norris Dawn L, Frye Christopher C, Barnard Gavin C
Biotechnology Discovery Research, Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, 46225.
Bioprocess Research and Development, Eli Lilly and Company, LTC-North, 1200 Kentucky Avenue, Indianapolis, IN, 46221.
Biotechnol Prog. 2017 Mar;33(2):534-540. doi: 10.1002/btpr.2447. Epub 2017 Mar 7.
Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies, and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. We recently described the generation of CHO pools yielding mAb titers as high as 7.6 g/L in a 16 day bioprocess using piggyBac transposon-mediated gene integration. In this study, we wanted to understand why the piggyBac pool titers were significantly higher (2-10 fold) than the control CHO pools. Higher titers were the result of a combination of increased average gene copy number, significantly higher messenger RNA levels and the homogeneity (i.e. less diverse population distribution) of the piggyBac pools, relative to the control pools. In order to validate the use of piggyBac pools to support preclinical drug development, we then performed an in-depth product quality analysis of purified protein. The product quality of protein obtained from the piggyBac pools was very similar to the product quality profile of protein obtained from the control pools. Finally, we demonstrated the scalability of these pools from shake flasks to 36L bioreactors. Overall, these results suggest that gram quantities of therapeutic protein can be rapidly obtained from piggyBac CHO pools without significantly changing product quality attributes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:534-540, 2017.
中国仓鼠卵巢（CHO）细胞仍然是生产生物制药药物最受欢迎的宿主，特别是单克隆抗体（mAb）、双特异性抗体和Fc融合蛋白。创建和表征生产这些治疗性蛋白质所需的稳定CHO克隆衍生细胞系（CDCL）是一个漫长而费力的过程。因此，CHO细胞池越来越多地被用于快速生产蛋白质，以支持和推动临床前药物开发。我们最近描述了使用piggyBac转座子介导的基因整合在16天的生物过程中产生的CHO细胞池，其mAb滴度高达7.6 g/L。在本研究中，我们想了解为什么piggyBac细胞池的滴度比对照CHO细胞池显著更高（2-10倍）。更高的滴度是平均基因拷贝数增加、信使RNA水平显著更高以及piggyBac细胞池相对于对照细胞池的同质性（即群体分布多样性更低）共同作用的结果。为了验证使用piggyBac细胞池来支持临床前药物开发，我们随后对纯化蛋白进行了深入的产品质量分析。从piggyBac细胞池获得的蛋白质的产品质量与从对照细胞池获得的蛋白质的产品质量概况非常相似。最后，我们证明了这些细胞池从摇瓶到36L生物反应器的可扩展性。总体而言，这些结果表明，可以从piggyBac CHO细胞池中快速获得克级量的治疗性蛋白质，而不会显著改变产品质量属性。© 2017美国化学工程师学会生物技术进展，33:534 - 540，2017。
