Details

Autor(en) / Beteiligte

• Gates, Zachary P.
• Vinogradov, Alexander A.
• Quartararo, Anthony J.
• Bandyopadhyay, Anupam
• Choo, Zi-Ning
• Evans, Ethan D.
• Halloran, Kathryn H.
• Mijalis, Alexander J.
• Mong, Surin K.
• Simon, Mark D.
• Standley, Eric A.
• Styduhar, Evan D.
• Tasker, Sarah Z.
• Touti, Faycal
• Weber, Jessica M.
• Wilson, Jessica L.
• Jamison, Timothy F.
• Pentelute, Bradley L.

Titel

Xenoprotein engineering via synthetic libraries

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 2018-06, Vol.115 (23), p.E5298-E5306

Ort / Verlag

United States: National Academy of Sciences

Erscheinungsjahr

2018

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.