Monteil Dominique T, Kuan Jeffrey
Cell Culture, Process Science, Boehringer Ingelheim, Fremont, CA, USA.
Contingent worker providing support to Cell Culture, Process Science, Boehringer Ingelheim, Fremont, CA, USA.
Methods Mol Biol. 2018;1850:133-145. doi: 10.1007/978-1-4939-8730-6_10.
Most pharmaceutical biotechnology companies use stirred-tank bioreactors (STR) for recombinant protein manufacturing. These bioreactors are used at a variety of different scales ranging from bench to production scales, with working volumes from 10 mL to 25,000 L. Bench-scale STRs are commonly used to culture mammalian cells for process development, to troubleshoot production scale bioreactors using scale-down models (SDM), or to conduct fundamental research. In this chapter, we describe the operations of a bench-scale STR for the production of recombinant proteins with suspension-adapted Chinese hamster ovary (CHO-DG44) cells. These operations include bioreactor setup and configuration, batching media, inoculation of the seed cell culture, production phase, and harvest of cell-free fluids.
大多数制药生物技术公司使用搅拌罐生物反应器(STR)来生产重组蛋白。这些生物反应器在从实验室规模到生产规模的各种不同规模下使用,工作体积从10毫升到25000升不等。实验室规模的STR通常用于培养哺乳动物细胞以进行工艺开发,使用缩小模型(SDM)对生产规模的生物反应器进行故障排除,或进行基础研究。在本章中,我们描述了使用适应悬浮培养的中国仓鼠卵巢细胞(CHO-DG44)生产重组蛋白的实验室规模STR的操作。这些操作包括生物反应器的设置和配置、培养基配制、种子细胞培养物的接种、生产阶段以及无细胞流体的收获。