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Protein engineering by highly parallel screening of computationally designed variants.

Authors :
Sun MG
Seo MH
Nim S
Corbi-Verge C
Kim PM
Source :
Science advances [Sci Adv] 2016 Jul 20; Vol. 2 (7), pp. e1600692. Date of Electronic Publication: 2016 Jul 20 (Print Publication: 2016).
Publication Year :
2016

Abstract

Current combinatorial selection strategies for protein engineering have been successful at generating binders against a range of targets; however, the combinatorial nature of the libraries and their vast undersampling of sequence space inherently limit these methods due to the difficulty in finely controlling protein properties of the engineered region. Meanwhile, great advances in computational protein design that can address these issues have largely been underutilized. We describe an integrated approach that computationally designs thousands of individual protein binders for high-throughput synthesis and selection to engineer high-affinity binders. We show that a computationally designed library enriches for tight-binding variants by many orders of magnitude as compared to conventional randomization strategies. We thus demonstrate the feasibility of our approach in a proof-of-concept study and successfully obtain low-nanomolar binders using in vitro and in vivo selection systems.

Details

Language :
English
ISSN :
2375-2548
Volume :
2
Issue :
7
Database :
MEDLINE
Journal :
Science advances
Publication Type :
Academic Journal
Accession number :
27453948
Full Text :
https://doi.org/10.1126/sciadv.1600692