A capillary‐based microfluidic device enables primary high‐throughput room‐temperature crystallographic screening. Issue 4 (14th June 2021)

• Record Type: Journal Article
• Authors: Sui, Shuo; Mulichak, Anne; Kulathila, Raviraj; McGee, Joshua; Filiatreault, Danny; Saha, Sarthak; Cohen, Aina; Song, Jinhu; Hung, Holly; Selway, Jonathan; Kirby, Christina; Shrestha, Om K.; Weihofen, Wilhelm; Fodor, Michelle; Xu, Mei; Chopra, Rajiv; Perry, Sarah L.
• Is Part Of: Journal of applied crystallography. Volume 54:Issue 4(2021)
• Page Start: 1034
• Page End: 1046
• Publication Date: 2021-06-14
• Subjects: compound screening -- microfluidics -- structural biology -- protein crystallography -- X‐ray diffraction; Crystallography -- Periodicals; 548.05
• DOI: 10.1107/S1600576721004155
• Languages: English
• ISSNs: 0021-8898

Abstract

Abstract : 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. Abstract : 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.