Picoliter cell lysate assays in microfluidic droplet compartments for directed enzyme evolution

Kintses, Balint, Hein, Christopher, Mohamed, Mark F., Fischlechner, Martin, Courtois, Fabienne, Lainé, Céline and Hollfelder, Florian (2012) Picoliter cell lysate assays in microfluidic droplet compartments for directed enzyme evolution. Chemistry & Biology, 19 (8), 1001-1009. (doi:10.1016/j.chembiol.2012.06.009). (PMID:22921067)

Abstract

We demonstrate the utility of a microfluidic platform in which water-in-oil droplet compartments serve to miniaturize cell lysate assays by a million-fold for directed enzyme evolution. Screening hydrolytic activities of a promiscuous sulfatase demonstrates that this extreme miniaturization to the single-cell level does not come at a high price in signal quality. Moreover, the quantitative readout delivers a level of precision previously limited to screening methodologies with restricted throughput. The sorting of 3 × 107 monodisperse droplets per round of evolution leads to the enrichment of clones with improvements in activity (6-fold) and expression (6-fold). The detection of subtle differences in a larger number of screened clones provides the combination of high sensitivity and high-throughput needed to rescue a stalled directed evolution experiment and make it viable

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