Design of nanobody-based bispecific constructs by in silico affinity maturation and umbrella sampling simulations.

ABSTRACT: Random mutagenesis is the natural opportunity for proteins to evolve and biotechnologically it has been exploited to create diversity and identify variants with improved characteristics in the mutant pools. Rational mutagenesis based on biophysical assumptions and supported by computational power has been proposed as a faster and more predictable strategy to reach the same aim. In this work we confirm that substantial improvements in terms of both affinity and stability of nanobodies can be obtained by using combinations of algorithms, even for binders with already high affinity and elevated thermal stability. Furthermore, in silico approaches allowed the development of an optimized bispecific construct able to bind simultaneously the two clinically relevant antigens TNF-α and IL-23

SUBMITTER: Bai Z 

PROVIDER: S-EPMC9822835 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature