Title | Xenoprotein engineering via synthetic libraries. |
Publication Type | Journal Article |
Year of Publication | 2018 |
Authors | Gates ZP, Vinogradov AA, Quartararo AJ, Bandyopadhyay A, Choo Z-N, Evans ED, Halloran KH, Mijalis AJ, Mong SK, Simon MD, Standley EA, Styduhar ED, Tasker SZ, Touti F, Weber JM, Wilson JL, Jamison TF, Pentelute BL |
Journal | Proc Natl Acad Sci U S A |
Volume | 115 |
Issue | 23 |
Pagination | E5298-E5306 |
Date Published | 2018 06 05 |
ISSN | 1091-6490 |
Keywords | Amino Acids, Combinatorial Chemistry Techniques, Flow Cytometry, Humans, Microspheres, Peptide Library, Protein Binding, Protein Engineering, Proteins, Tandem Mass Spectrometry |
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. |
DOI | 10.1073/pnas.1722633115 |
Alternate Journal | Proc. Natl. Acad. Sci. U.S.A. |
PubMed ID | 29784819 |
Grant List | S10 OD016326 / OD / NIH HHS / United States |
Submitted by api_import on December 20, 2019 - 3:23pm