Cell line development is a highly-involved, multi-week workflow that involves the plating, screening, and expansion of single cell-derived clones to establish a protein expression cell line. This complex process required significant throughput of sterile cell manipulations, long-term incubations, centrifugation, cell counting, monoclonality determination and growth tracking, and protein production assessment. Automating the cell line development process on the Biomek i7 Automated Workstation enabled us to meet the throughput needed for the workflow while minimizing the manual errors that lead to costly rework. Assay data from multiple instruments and time points were used as toll gates to identify the wells that met the criteria to continue through the lengthy process and these data ultimately drove automated hit-picking and cell expansion. Further improvements on workflow efficiency and sample and data integrity can be gained by integrating the various analyzers to the liquid handler and scheduling overlapping campaigns to maximize system utilization.