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Xenoprotein engineering via synthetic libraries

Author(s)
Gates, Zachary P; Vinogradov, Alexander Alexandrovich; Quartararo, Anthony James; Bandyopadhyay, Anupam; Choo, Zi-Ning; Evans, Ethan Daniel; Halloran, Kathryn; Mijalis, Alexander James; Mong, Surin Khai; Simon, Mark; Standley, Eric Alan; Styduhar, Evan; Tasker, Sarah Zinnen; Touti, Faycal; Weber, Jessica Marie; Wilson, Jessica Laura; Jamison, Timothy F; Pentelute, Bradley L.; ... Show more Show less
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Abstract
Chemical methods have enabled the total synthesis of protein molecules of ever-increasing size and complexity. However, methods to engineer synthetic proteins comprising noncanonical amino acids have not kept pace, even though this capability would be a distinct advantage of the total synthesis approach to protein science. In this work, we report a platform for protein engineering based on the screening of synthetic one-bead one-compound protein libraries. Screening throughput approaching that of cell surface display was achieved by a combination of magnetic bead enrichment, flow cytometry analysis of on-bead screens, and high-throughput MS/MS-based sequencing of identified active compounds. Direct screening of a synthetic protein library by these methods resulted in the de novo discovery of mirror-image miniprotein-based binders to a ∼150-kDa protein target, a task that would be difficult or impossible by other means. Keywords: xenoprotein; mirror-image miniprotein; D-protein; protein engineering; flow cytometry
Date issued
2018-05
URI
http://hdl.handle.net/1721.1/120509
Department
Massachusetts Institute of Technology. Department of Chemistry
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences (U.S.)
Citation
Gates, Zachary P. et al. “Xenoprotein Engineering via Synthetic Libraries.” Proceedings of the National Academy of Sciences 115, 23 (May 2018): E5298–E5306 © 2018 National Academy of Sciences
Version: Final published version
ISSN
0027-8424
1091-6490

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