Measuring Protein-Protein and Protein-Nucleic Acid Interactions by Bio-Layer Interferometry

Sultana A, Lee J, 79:19.25.1-19.25.26, Curr Protoc Protein Sci, 2015

This is a detailed write-up explaining the stepwise procedure of BLI-based advanced kinetic analyses of DNA-protein and protein-protein interactions using Pall ForteBio BLItz and the Octet systems. The preparation of the biotinylated ligands, choices for controls, assay optimization approaches, strategies applicable to regeneration of Streptavidin-based biosensors, and assay troubleshooting approaches are described. Authors recommend BLI as a versatile platform providing accurate measurements of binding affinities (KD), rates of association, and rates of dissociation (kon and koff) of protein-protein and protein-nucleic acid interactions in minutes by utilizing relatively small quantities of sample.

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