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A high‐throughput screen for antibiotic drug discovery
Author(s) -
Scanlon Thomas C.,
Dostal Sarah M.,
Griswold Karl E.
Publication year - 2014
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25019
Subject(s) - high throughput screening , drug discovery , microfluidics , antibiotics , metagenomics , throughput , nanotechnology , computational biology , biology , microbiology and biotechnology , computer science , materials science , bioinformatics , biochemistry , telecommunications , gene , wireless
We describe an ultra‐high‐throughput screening platform enabling discovery and/or engineering of natural product antibiotics. The methodology involves creation of hydrogel‐in‐oil emulsions in which recombinant microorganisms are co‐emulsified with bacterial pathogens; antibiotic activity is assayed by use of a fluorescent viability dye. We have successfully utilized both bulk emulsification and microfluidic technology for the generation of hydrogel microdroplets that are size‐compatible with conventional flow cytometry. Hydrogel droplets are ∼25 pL in volume, and can be synthesized and sorted at rates exceeding 3,000 drops/s. Using this technique, we have achieved screening throughputs exceeding 5 million clones/day. Proof‐of‐concept experiments demonstrate efficient selection of antibiotic‐secreting yeast from a vast excess of negative controls. In addition, we have successfully used this technique to screen a metagenomic library for secreted antibiotics that kill the human pathogen Staphylococcus aureus . Our results establish the practical utility of the screening platform, and we anticipate that the accessible nature of our methods will enable others seeking to identify and engineer the next generation of antibacterial biomolecules. Biotechnol. Bioeng. 2014;111: 232–243. © 2013 Wiley Periodicals, Inc.