Grating-coupled interferometry reveals binding kinetics and affinities of Ni ions to genetically engineered protein layers.

ABSTRACT: Reliable measurement of the binding kinetics of low molecular weight analytes to their targets is still a challenging task. Often, the introduction of labels is simply impossible in such measurements, and the application of label-free methods is the only reliable choice. By measuring the binding kinetics of Ni(II) ions to genetically modified flagellin layers, we demonstrate that: (1) Grating-Coupled Interferometry (GCI) is well suited to resolve the binding of ions, even at very low protein immobilization levels; (2) it supplies high quality kinetic data from which the number and strength of available binding sites can be determined, and (3) the rate constants of the binding events can also be obtained with high accuracy. Experiments were performed using a flagellin variant incorporating

SUBMITTER: Jankovics H 

PROVIDER: S-EPMC7746762 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature