Facile Display of Homomultivalent Proteins for Selections

Overview

Journal ACS Synth Biol

Publisher American Chemical Society

Specialties Biotechnology
Molecular Biology

Date 2023 Jan 19

PMID 36655840

Authors

Ayako Ohoka

Casim A Sarkar

Affiliations

Soon will be listed here.

Abstract

Low-affinity protein binders are emerging as valuable domains for therapeutic applications because of their higher specificity when presented in multivalent ligands that increase the overall strength and selectivity of receptor binding. discovery of low-affinity binders would be enhanced by the large library sizes attainable with selection systems, but these platforms generally maximize recovery of high-affinity monovalent binders. Here, we present a facile technology that uses rolling circle amplification to create homomultivalent libraries. We show proof of principle of this approach in ribosome display with off-rate selections of a bivalent ligand against monovalent and bivalent targets, thereby demonstrating high enrichment (up to 166-fold) against a low-affinity target that is bivalent but not monovalent. This approach to homomultivalent library construction can be applied to any binder tolerant of N- and C-terminal fusions and provides a platform for performing display selections with controlled protein valency and orientation.

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