Conflict of Interest: The authors declare no conflict of interest.
A high-throughput screen for antibiotic drug discovery
Article first published online: 29 AUG 2013
© 2013 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 111, Issue 2, pages 232–243, February 2014
How to Cite
Scanlon, T. C., Dostal, S. M. and Griswold, K. E. (2014), A high-throughput screen for antibiotic drug discovery. Biotechnol. Bioeng., 111: 232–243. doi: 10.1002/bit.25019
- Issue published online: 19 DEC 2013
- Article first published online: 29 AUG 2013
- Accepted manuscript online: 17 AUG 2013 02:07AM EST
- Manuscript Accepted: 29 JUL 2013
- Manuscript Revised: 21 JUL 2013
- Manuscript Received: 14 MAY 2013
- National Institute of Allergy and Infectious Disease. Grant Numbers: 1R21AI094391, 1R21AI098122
- Cystic Fibrosis Foundation. Grant Number: SCANLO08F0
- drug discovery;
- in vitro compartmentalization;
- high-throughput screening;
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.