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Cell line development: 99.99% Probability of Monoclonality

Combined Single-cell Dispensing and 3D Full Well Imaging for Cell Lines with 99.99 % Probability of Monoclonality. Elevate your cell line development workflow with the UP.SIGHT.

Clonal cell line development is a crucial step for biopharmaceutical generation (e.g., the production of monoclonal antibodies). To ensure a consistent product, the cell lines used in production must originate from a single cell. Common cloning workflows consist of one to two rounds of single cell cloning followed by imaging of the well plate to confirm single cells were dispensed. However, since there is no confirmation of a single cell dispensed in methods such as limiting dilution or FACS, the detection of errors in the dispensing process is solely dependent on the accuracy of well-plate imaging. Plate imaging has its own error in cell detection and is dependent on centrifugation to ensure all cells are in the imaging focal plane. In combination, these errors impact the probability of monoclonality that can be achieved with a given workflow.

The UP.SIGHT™ addresses these problems by providing double assurance that a single cell was dispensed into a well. Single-cell isolation is documented through a series of nozzle images. Directly after dispensing, the novel technology 3D Full Well Imaging captures images of the liquid volume as a second verification that a single cell was dispensed. As 3D Full Well Imaging captures the full liquid volume, centrifuging after cloning is not required. The UP.SIGHT also performs colony growth monitoring post cloning. The UP.SIGHT is wellsuited for high-throughput workflows since it is fast, compatible with 96-, 384- and 1536-well plates and has a high single-cell dispensing efficiency of >98 %. In this study, 4000 single cells and almost 1,000 growing colonies were analyzed, and the data demonstrates that the combination of nozzle images and 3D Full Well Imaging results in cell lines with a probability of monoclonality of >99.99 %, without the need for fluorescent cell staining or imaging

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