The Holy Grail sought by all Bioprocess Cell Line Development (CLD) groups is achieving high yields from easily-cultured, robustly-growing cells in timelines measured in weeks rather than months. As the first bottleneck in process development, CLD must first birth its product for upstream and downstream groups to initiate their own reproductive cycles. To facilitate shortened CLD timelines, scientists have turned to new technologies and automation platforms. Emerging high-throughput instrumentation such as Clonepix and Automated MicroBioreactors (AMBR) have been enthusiastically integrated into stable cell line generation platforms; however, application of these methodologies among users is divergent.
In this study, we describe method development that is part of an ongoing effort to create a vigorous, adaptable, high-throughput cell line development platform, incorporating in-house VRC-DG44-CHO cells, expression vectors with high-activity regulatory elements, and efficient selection and evaluation methods that result in early generation of optimal-yielding, vigorous, stable cell lines. This study was aimed toward a) evaluating early incorporation of Clonepix technology to maximize transfectant pool heterogeneity, b) identifying Clonepix fluorescence attributes that increase the probability of high-productivity clone isolation, c) evaluating and optimizing AMBR operating conditions to achieve process attributes comparable to and predictive of medium-scale bioreactors, and implementation of these practices for maximal results with shortened timelines.