(1127-D) 2^nd gen UP.SIGHT: All-in-one solution for the generation of monoclonal cell lines, from single cell cloning to clone selection

Abstract: There is a constant pressure to reduce timelines in mammalian cell line development (CLD) for biotherapeutic protein production and gene and cell therapies. To ensure the consistency, quality, and safety of biologics, producing cell lines must be derived from a single-cell progenitor. Demonstration of clonal derivation of the generated cell lines is key for health authorities’ approval. CLD is time and resource intensive as large numbers of clones must be screened for productivity, stability, and product quality before finding promising candidates for scaling up and production. Therefore, there is a continuous need for faster clone generation while providing assurance of monoclonality. To meet these regulatory and process-oriented demands, single-cell dispensers and plate imagers have become vital instruments in any CLD laboratory.

Here we present UP.SIGHT 2nd Gen, Cytena’s next generation of single cell dispenser with integrated plate imaging capabilities. The UP.SIGHT 2nd Gen is the all-in-one solution for the generation of monoclonal cell lines that saves months of integration effort & validation time. The full workflow from single cell cloning to selection of the best clones with comprehensive documentation for IND/BLA submissions can be performed with the UP.SIGHT 2nd Gen , given its key features:
• Single-cell cloning with documented day-0 clonality, either by our next generation 3D Volume Imaging for 384-well plates and/or traditional plate bottom imaging for 384 and 96-well plates.
• Plate imaging in 48-, 24-, 12- and 6-well SBS plate formats enables confluency analysis to monitor growth of both suspension and adherent cell lines.
• Integrated cell count & cell viability analysis tool
• High-throughput titer quantification of antibodies and Fc fusion proteins
• Software-guided selection of best producer clones based on growth, titer, and external assays (e.g. ELISA or NGS for target integration).
• Clone tracking with comprehensive data logs and report generation for IND submission ready reports

Here we report validation experiments to determine this instrument’s single-cell dispensing efficiency (SCDE) and probability of clonal derivation (p(clonal)). Process optimization to maximize clone recovery with several cell lines was also performed, focusing on cloning media and plate type. We provide examples of imaging of several plate formats and the corresponding confluence analysis with the analysis software. Results show that UP.SIGHT 2nd Gen enables fast and efficient CLD workflows, with full documentation of clonal derivation.