Center for Advanced Sensor Technology, University of Maryland, Baltimore County, Baltimore, MD 21250, USA.
Biotechnol Prog. 2010 Jul-Aug;26(4):1095-103. doi: 10.1002/btpr.392.
Effective clone selection is a crucial step toward developing a robust mammalian cell culture production platform. Currently, clone selection is done by culturing cells in well plates and picking the highest producers. Ideally, clone selection should be done in a stirred tank bioreactor as this would best replicate the eventual production environment. The actual number of clones selected for future evaluation in bioreactors at bench-scale is limited by the scale-up and operational costs involved. This study describes the application of miniaturized stirred high-throughput bioreactors (35 mL working volume; HTBRs) with noninvasive optical sensors for clone screening and selection. We investigated a method for testing several subclones simultaneously in a stirred environment using our high throughput bioreactors (up to 12 clones per HTBR run) and compared it with a traditional well plate selection approach. Importantly, it was found that selecting clones solely based on results from stationary well plate cultures could result in the chance of missing higher producing clones. Our approach suggests that choosing a clone after analyzing its performance in a stirred bioreactor environment is an improved method for clone selection.
有效的克隆选择是开发稳健的哺乳动物细胞培养生产平台的关键步骤。目前，通过在培养皿中培养细胞并选择产量最高的细胞来进行克隆选择。理想情况下，应该在搅拌罐生物反应器中进行克隆选择，因为这最能复制最终的生产环境。在中试规模的搅拌罐中为未来评估选择的实际克隆数量受到规模扩大和运营成本的限制。本研究描述了应用微型化搅拌高通量生物反应器（35 mL 工作体积；HTBR）和非侵入式光学传感器进行克隆筛选和选择。我们研究了一种在搅拌环境中同时测试多个亚克隆的方法，使用我们的高通量生物反应器（每个 HTBR 运行最多 12 个克隆），并将其与传统的平板选择方法进行了比较。重要的是，发现仅根据静止平板培养物的结果选择克隆可能会错过更高产的克隆。我们的方法表明，在搅拌生物反应器环境中分析其性能后选择克隆是一种改进的克隆选择方法。
