Mass Activated Droplet Sorting (MADS) Enables High-Throughput Screening of Enzymatic Reactions at Nanoliter Scale.
Angew Chem Int Ed Engl
; 59(11): 4470-4477, 2020 03 09.
Article
em En
| MEDLINE
| ID: mdl-31868984
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 0.7â
samples s-1 is achieved with 98 % accuracy using a self-correcting and adaptive sorting algorithm. We use the system to screen ≈15 000â
samples in 6â
h and demonstrate its utility by sorting 25â
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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MEDLINE
Assunto principal:
Piridinas
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Microfluídica
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Ensaios Enzimáticos
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Ensaios de Triagem em Larga Escala
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Aminas
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Transaminases
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article