Regeneration of His-tagged Streptavidin on OpenSPR® NTA Sensors Sensors


Capture coupling techniques are advantageous to investigate biomolecular interactions as they provide specificity for immobilization on the OpenSPR real-time, label-free biosensor platform. Streptavidin sensors offer a streptavidin coated sensor surface which can capture biotin-tagged ligands with high affinity. However, the biotin-streptavidin interaction is irreversible, limiting researchers to one biotinylated ligand per sensor. In this application note, Eliza Hanson, a member of Rebecca Whelan’s research team at the University of Kansas explored the use of His-streptavidin on OpenSPR NTA sensors to create a regeneratable surface and measure binding interactions for multiple biotinylated aptamer ligands on a single sensor.

Thrombin (analyte) binding to immobilized 29-mer (ligand) on OpenSPR. Analyte was injected at concentrations of 5, 10, 25, 50, 75, 100, 200, and 300 nM. The surface was regenerated with 2M NaCl between each thrombin injection. Curves were double reference subtracted to obtain the resultant corrected curves.