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Miniaturized fluid array for high‐throughput protein expression
Author(s) -
Khnouf Ruba,
Olivero Daniel,
Jin Shouguang,
Fan Z. Hugh
Publication year - 2010
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.474
Subject(s) - throughput , high throughput screening , protein expression , yield (engineering) , function (biology) , gene expression , cell free protein synthesis , gene , chemistry , computational biology , protein biosynthesis , chromatography , computer science , biology , biochemistry , microbiology and biotechnology , materials science , telecommunications , metallurgy , wireless
We describe a miniaturized fluid array device for high‐throughput cell‐free protein synthesis (CFPS), aiming to match the throughput and scale of gene discovery. Current practice of using E. coli cells for production of recombinant proteins is difficult and cost‐prohibitive to implement in a high‐throughput format. As more and more new genes are being identified, there is a considerable need to have high‐throughput methods to produce a large number of proteins for studying structures and functions of the corresponding genes. The device consists of 96 units and each unit is for expression of one protein; thus up to 96 proteins can be produced simultaneously. The function of the fluid array was demonstrated by expression of a variety of proteins, with more than two orders of magnitude reduction in reagent consumption compared with a commercially available CFPS instrument. The protein expression yield in the device was up to 87 times higher for β‐glucoronidase than that in a conventional microplate. The concentration of β‐galactosidase expressed in the device was determined at 5.5 μg/μL. The feasibility of using the device for drug screening was demonstrated by measuring the inhibitory effects of mock drug compounds on synthesized β‐lactamase without the need for harvesting proteins, which enabled us to reduce the analysis time from days to hours. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010