Mass Activated Droplet Sorting (MADS) Enables High-Throughput Screening of Enzymatic Reactions at Nanoliter Scale |
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| Authors: | Daniel A Holland-Moritz Michael K Wismer Dr Benjamin F Mann Dr Iman Farasat Dr Paul Devine Dr Erik D Guetschow Dr Ian Mangion Dr Christopher J Welch Dr Jeffrey C Moore Dr Shuwen Sun Prof Robert T Kennedy |
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| Institution: | 1. Dept. of Chemistry, University of Michigan, 930 N University, Ann Abor, MI, 48109 USA;2. Scientific Engineering and Design, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ, 07033 USA;3. Process Research and Development, Merck & Co., Inc., 126 E. Lincoln Ave, Rahway, NJ, 07065 USA;4. Janssen R&D, 1400 McKean Rd., Spring House, PA, 19477 USA;5. Indiana Consortium Analyt Sci & Engn, Indianapolis, IN, 46202 USA |
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| Abstract: | Microfluidic droplet sorting enables the high-throughput screening and selection of water-in-oil microreactors at speeds and volumes unparalleled by traditional well-plate approaches. Most such systems sort using fluorescent reporters on modified substrates or reactions that are rarely industrially relevant. We describe a microfluidic system for high-throughput sorting of nanoliter droplets based on direct detection using electrospray ionization mass spectrometry (ESI-MS). Droplets are split, one portion is analyzed by ESI-MS, and the second portion is sorted based on the MS result. Throughput of 0.7 samples s−1 is achieved with 98 % accuracy using a self-correcting and adaptive sorting algorithm. We use the system to screen ≈15 000 samples in 6 h and demonstrate its utility by sorting 25 nL droplets containing transaminase expressed in vitro. Label-free ESI-MS droplet screening expands the toolbox for droplet detection and recovery, improving the applicability of droplet sorting to protein engineering, drug discovery, and diagnostic workflows. |
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| Keywords: | Biokatalyse Tröpfchenmikrofluidik Hochdurchsatz-Screening Massenspektrometrie Mikroreaktoren |
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