31. The BIND™ System and the High-Throughput Ranking of Macromolecular-Complex Stabilities.

GA CANZIANI AND M BRIGHAM-BURKE

Phoenix, AR, USA

Simposio; DiPIA (Development in Protein Interaction Analysis); 2007

GE HEALTH SCIENCE

ABSTRACT: Early in therapeutics development, the high-throughput screening of biomolecules to targets of biopharmaceutical interest consists in driving biomolecular interaction analyses to their maximum speed without sacrificing resolution, precision and reproducibility. Many emerging detection methods that promise a high-throughput analysis of multiple interactions have eliminated the use of tags or labels. However, sensitivity of these methods is often lower than that offered by the application of fluorescence and chemioluminescence reporters. A new interactions detector that deserves attention for high-throughput screening assays is the BIND™ (Biomolecular Interaction Detection) Reader, an optical biosensor that relies on the dielectric permittivity of detector-bound biomolecules to produce a shift in reflected light wavelength at the sensor surface as a function of mass. The surface of the BIND biosensor is manufactured from continuous sheets of plastic film and incorporated into standard 96- and 384-well microtiter plate formats. This format ensures low cost and compatibility with standard robotic-handling systems used in many screening applications. We used model molecular systems to assess the resolution in ranking of off-rates of multiple complexes. Our goal was to optimize the assay for BIND interaction analysis in order to minimize artifacts and non-specific interactions to maximize the binding response. The experimental procedures and the sensitivity limits of this high-throughput platform will be documented.