events

LAMMP/BME Seminar Series

Wednesday 04th of November 2009
2:30 AM
Beckman Laser Institute Library

Interrogation of Label‐Free Biomolecular Interactions utilizing Backscattering Interferometry

Darryl J. Bornhop, Ph D, Professor of Chemistry
Vanderbilt University, Department of Chemistry and Institute for Chemical Biology Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute Molecular Sensing Inc. Nashville, Tennessee

Backscattering interferometry (BSI) is a relatively new technique used to study molecular interactions in a label‐free format that is proving to be a versatile biosensing tool. BSI consists of a simple optical train that quantifies minute changes in refractive index created by a binding event. This technique enables the interrogation of molecular interactions of numerous species, with a high dynamic range of dissociation constants and unmatched sensitivity. For example BSI has been used to pM to mM measure equilibrium KD’s at physiologically relevant concentrations for protein targets binding to ions, small molecule inhibitors, proteins, antibodies or nucleic acids, while requiring only picomole to nanomole quantities of precious protein target. In this presentation it will be shown that BSI can be used to study polyvalenency, quantify small molecule‐protein interaction where the mass difference is >1000:1, to screen libraries of disease‐related mutants, and to perform investigations in complex milieu such as 100% serum. Additionally, the utility for using BSI to investigate and quantify the equilibrium binding constants of membrane‐associated proteins and antibodies in a label‐free, solution‐based environment for a variety of ligands and over a wide range of affinities will be presented.