Functional analysis of small-molecule and protein binding domains using monomeric streptavidin-based two-component biosensors

A monomeric streptavidin-based two-component biosensor platform enables ligand-dependent activation with biotinylated methotrexate, vitamin D3, galectin-3, and S100A7.

• Why it matters: Binding affinity and specificity alone do not reliably predict whether candidate recognition domains will function compatibly within a biosensor scaffold.
• What they did: The authors built a modular system using monomeric streptavidin (mSA2) as a surrogate recognition domain and used a chromatography-free protocol to recover functional components from Escherichia coli.
• The result: In reporter biosensors, activation depended on the presence of the tested ligands, supporting scalable rapid evaluation and prototyping of binder domains for future biosensor development.

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