Challenges and Solutions for Leave-One-Out Biosensor Design in the Context of a Rugged Fitness Landscape.

ABSTRACT: The leave-one-out (LOO) green fluorescent protein (GFP) approach to biosensor design combines computational protein design with split protein reconstitution. LOO-GFPs reversibly fold and gain fluorescence upon encountering the target peptide, which can be redefined by computational design of the LOO site. Such an approach can be used to create reusable biosensors for the early detection of emerging biological threats. Enlightening biophysical inferences for nine LOO-GFP biosensor libraries are presented, with target sequences from dengue, influenza, or HIV, replacing beta strands 7, 8, or 11. An initially low hit rate was traced to components of the energy function, manifesting in the over-rewarding of over-tight side chain packing. Also, screening by colony picking required a low library

SUBMITTER: Banerjee S 

PROVIDER: S-EPMC11478963 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature