An engineered redox-switchable streptavidin mutein enables the high-affinity capture and efficient elution of biotinylated ligands.

ABSTRACT: Although the high affinity and specificity of the interaction between streptavidin and biotin are widely utilized to isolate biotinylated ligands, their elution over practical timescales requires the use of strongly denaturing conditions that are incompatible with many target biomolecules. To overcome this limitation, we previously engineered a disulfide bond into a critical loop of streptavidin to create the redox-sensitive M88 mutein, which releases biotinylated ligands ~19,000-fold faster in the reduced state when compared with the oxidized state. To optimize the speed and efficiency of ligand recovery, we describe how multiple mechanisms like disulfide bond reduction, increasing temperature, elevating pH, and adding organic cosolvents can dramatically increase the rates of dissociation

SUBMITTER: Marangoni JM 

PROVIDER: S-EPMC12267106 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature