Streamlined stable CHO cell line development using droplet microfluidics with monoclonality assurance

Abstract Rapid generation of monoclonal CHO cell lines remains a key bottleneck in mammalian cell line development, often requiring months of iterative screening and cloning. Here, we present an integrated droplet microfluidic workflow for high-throughput screening and isolation of antibody-producing ExpiCHO-S cells using a FRET-based secretion assay. The platform enables picodroplet-based detection of secreted antibodies and monoclonality-verified single-cell sorting, followed by sequential enrichment and clonal isolation. Application of this approach yielded stable monoclonal cell lines with consistent antibody production and maintained expression over extended culture in the absence of selection pressure. By reducing cell line development timelines from 6–12 months to approximately five weeks, this workflow addresses key limitations of conventional approaches and provides a scalable strategy for streamlining early cell line development workflows in biomanufacturing.