A capillary-based microfluidic device enables primary high-throughput room-temperature crystallographic screening.

Autor: Sui S; Department of Chemical Engineering, University of Massachusetts Amherst, Amherst, MA, USA., Mulichak A; IMCA-CAT, Argonne National Laboratory, Lemont, IL, USA., Kulathila R; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., McGee J; Department of Chemical Engineering, University of Massachusetts Amherst, Amherst, MA, USA., Filiatreault D; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Saha S; Department of Chemical Engineering, University of Massachusetts Amherst, Amherst, MA, USA., Cohen A; Macromolecular Crystallography Group, Stanford Synchrotron Radiation Lightsource, Menlo Park, CA, USA., Song J; Macromolecular Crystallography Group, Stanford Synchrotron Radiation Lightsource, Menlo Park, CA, USA., Hung H; SPT Labtech Ltd, Boston, MA, USA., Selway J; Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, MA, USA., Kirby C; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Shrestha OK; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Weihofen W; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Fodor M; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Xu M; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Chopra R; Novartis Institutes for BioMedical Research, Cambridge, MA, USA., Perry SL; Department of Chemical Engineering, University of Massachusetts Amherst, Amherst, MA, USA.
Jazyk: angličtina
Zdroj: Journal of applied crystallography [J Appl Crystallogr] 2021 Jun 14; Vol. 54 (Pt 4), pp. 1034-1046. Date of Electronic Publication: 2021 Jun 14 (Print Publication: 2021).
DOI: 10.1107/S1600576721004155
Abstrakt: A novel capillary-based microfluidic strategy to accelerate the process of small-molecule-compound screening by room-temperature X-ray crystallography using protein crystals is reported. The ultra-thin microfluidic devices are composed of a UV-curable polymer, patterned by cleanroom photolithography, and have nine capillary channels per chip. The chip was designed for ease of sample manipulation, sample stability and minimal X-ray background. 3D-printed frames and cassettes conforming to SBS standards are used to house the capillary chips, providing additional mechanical stability and compatibility with automated liquid- and sample-handling robotics. These devices enable an innovative in situ crystal-soaking screening workflow, akin to high-throughput compound screening, such that quantitative electron density maps sufficient to determine weak binding events are efficiently obtained. This work paves the way for adopting a room-temperature microfluidics-based sample delivery method at synchrotron sources to facilitate high-throughput protein-crystallography-based screening of compounds at high concentration with the aim of discovering novel binding events in an automated manner.
(© Shuo Sui et al. 2021.)
Databáze: MEDLINE
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