In today's high-throughput searches for specific genes, proteins or protein interactions, plates containing rows of tiny wells have replaced old-fashioned test tubes. However, trawling for a gene or protein with just the right qualifications may require sorting through millions, or even billions, of possibilities. Instead of wells, the new method, developed by Dr. Dan Tawfik and Amir Aharoni of the Institute's Biological Chemistry Department and Prof. Shlomo Magdassi of the Hebrew University's Institute of Chemistry with support from the Israel Ministry of Science and Technology, relies on microscopic droplets of water suspended inside oil droplets. Using their system, millions of tests can be performed at once.
The method, which relies on a type of emulsion dubbed WOW, for water-oil-water, takes a page from living cells, which employ a fatty membrane to keep the inside and outside environments separate. The oily layer surrounding each miniscule water droplet acts as a barrier, keeping genes, proteins and other materials contained. Alternately, the team inserted harmless bacteria containing genes for testing into the drops. Confining individual tests within a cell-like bubble allowed them to employ a widely-used method for analyzing living cells. This method involves adding a fluorescent marker that lights up in color when activated by the right protein and sorting through the cells for those containing the marked proteins and their coding genes. Automated devices for sorting cells can handle many thousands of droplets per second. "Searches that now take a year to complete can be done in a matter of days," says Tawfik.
To demonstrate the efficiency of the system, the team isolated a new enzyme from a gene that was mutated artificially to produce random variations. They generated the enzymes in the droplets and sorted them according to which ones were better at cleaving a specific toxin in the bloodstream. The results from a screen completed in one afternoon were equivalent to those previously obtained through several rounds of mutation and screening – a several-month process.
Dr. Dan Tawfik's research is supported by the Y. Leon Benoziyo Institute for Molecular Medicine; the Dolfi and Lola Ebner Center for Biomedical Research; the Estelle Funk Foundation; the Dr. Ernst Nathan Fund for Biomedical Research; the Henry S. and Anne Reich Family Foundation; the Charles and M.R. Shapiro Foundation Endowed Biomedical Research Fund; the Harry and Jeanette Weinberg Fund for Molecular Genetics of Cancer; the Eugene and Delores Zemsky Charitable Foundation Inc; and Mr. and Mrs. Mordechai and Dalia Segal, Israel. Dr. Tawfik is the incumbent of the Elaine Blond Career Development Chair.
Last reviewed: By John M. Grohol, Psy.D. on 21 Feb 2009
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