Engineered single chain variable fragments (scFvs) with improved pH-dependent kinetics for use in continuous biosensor applications.

ABSTRACT: One challenge in the continuous monitoring of insulin lies with the unavailability of a high-affinity biological recognition element (BRE) with kinetic parameters sufficient to track fluctuating concentrations of insulin in vivo. An approach to overcome this limitation is to engineer a high-affinity BRE to selectively modulate its binding kinetics in response to an external signal. Herein, we design and evaluate changes in the pH-dependent binding kinetics of an anti-insulin single chain variable fragment (scFv) we previously employed in a point-of-care insulin sensor. We predicted the scFv structure in complex with human insulin and selected scFv residues directly involved in insulin binding for histidine substitution. We identify one mutation, T32H, that improves the pH-sensitivit

SUBMITTER: Wilson ED 

PROVIDER: S-EPMC12268258 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature