Ultrahigh‐Throughput FACS‐Based Screening for Directed Enzyme Evolution |
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| Authors: | Guangyu Yang Dr Stephen G Withers Prof |
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| Affiliation: | 1. Centre for High‐Throughput Biology (CHiBi) and Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, B.C. V6T 1Z1 (Canada), Fax: (+1)?604‐822‐8869;2. Key Laboratory of Molecular Enzymology and Engineering of Ministry of Education, Jilin University, 2519 Jiefang Road, Changchun 130023 (China) |
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| Abstract: | Directed enzyme evolution has proven to be a powerful tool for improving a range of properties of enzymes through consecutive rounds of diversification and selection. However, its success depends heavily on the efficiency of the screening strategy employed. Fluorescence‐activated cell sorting (FACS) has recently emerged as a powerful tool for screening enzyme libraries due to its high sensitivity and its ability to analyze as many as 108 mutants per day. Applications of FACS screening have allowed the isolation of enzyme variants with significantly improved activities, altered substrate specificities, or even novel functions. This review discusses FACS‐based screening for enzymatic activity and its potential application for the directed evolution of enzymes, ribozymes, and catalytic antibodies. |
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| Keywords: | directed evolution enzymes fluorescence fluorescence‐activated cell sorting high‐throughput screening |
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