Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc., West Point, Pennsylvania 19486, USA.
Biotechnol Bioeng. 2010 May 1;106(1):57-67. doi: 10.1002/bit.22664.
With increasing timeline pressures to get therapeutic and vaccine candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for process development. One such high-throughput system is the SimCell platform, a robotically driven, cell culture bioreactor system developed by BioProcessors Corp. This study describes the use of the SimCell micro-bioreactor technology for fed-batch cultivation of a GS-CHO transfectant expressing a model IgG4 monoclonal antibody. Cultivations were conducted in gas-permeable chambers based on a micro-fluidic design, with six micro-bioreactors (MBs) per micro-bioreactor array (MBA). Online, non-invasive measurement of total cell density, pH and dissolved oxygen (DO) was performed. One hundred fourteen parallel MBs (19 MBAs) were employed to examine process reproducibility and scalability at shake flask, 3- and 100-L bioreactor scales. The results of the study demonstrate that the SimCell platform operated under fed-batch conditions could support viable cell concentrations up to least 12 x 10(6) cells/mL. In addition, both intra-MB (MB to MB) as well as intra-MBA (MBA to MBA) culture performance was found to be highly reproducible. The intra-MB and -MBA variability was calculated for each measurement as the coefficient of variation defined as CV (%) = (standard deviation/mean) x 100. The % CV values for most intra-MB and intra-MBA measurements were generally under 10% and the intra-MBA values were slightly lower than those for intra-MB. Cell growth, process parameters, metabolic and protein titer profiles were also compared to those from shake flask, bench-top, and pilot scale bioreactor cultivations and found to be within +/-20% of the historical averages.
随着将治疗和疫苗候选物推向临床的时间压力不断增加，使用摇瓶和台式生物反应器等资源密集型方法可能会限制实验设计空间，以产生高产率的工艺。需要进行大量实验导致了使用微型化高通量 (HT) 技术进行工艺开发。一种这样的高通量系统是 SimCell 平台，这是一种由 BioProcessors Corp 开发的机器人驱动的细胞培养生物反应器系统。本研究描述了使用 SimCell 微生物反应器技术进行 fed-batch 培养表达模型 IgG4 单克隆抗体的 GS-CHO 转染子。培养在基于微流体设计的透气室中进行，每个微生物反应器阵列 (MBA) 中有六个微生物反应器 (MB)。在线进行非侵入式总细胞密度、pH 和溶解氧 (DO) 的测量。使用 114 个平行 MB（19 个 MBA）来检查摇瓶、3 升和 100 升生物反应器规模下的工艺重现性和可扩展性。该研究的结果表明，在 fed-batch 条件下运行的 SimCell 平台能够支持至少 12 x 10(6) 个细胞/mL 的活细胞浓度。此外，还发现每个 MBA 内 (MB 之间) 和每个 MBA 内 (MBA 之间) 的培养性能具有高度重现性。每个测量的 MB 内和 MBA 内变异性作为定义为 CV (%) = (标准偏差/平均值) x 100 的变异系数进行计算。大多数 MB 内和 MBA 内测量的 %CV 值通常低于 10%，而 MBA 内的值略低于 MB 内的值。细胞生长、工艺参数、代谢和蛋白质滴度谱也与摇瓶、台式和中试规模生物反应器培养物进行了比较，发现与历史平均值的差异在 +/-20% 以内。
