Wyatt Technology Corporation, a leader in absolute macromolecular characterization instrumentation and software, announces the availability of a new application note demonstrating the advanced capability of Composition Gradient Multi-angle Light Scattering (CG-MALS) technology. The experiment assessed the performance of Wyatt Technology’s Calypso automated composition gradient light scattering system for the evaluation of the inhibition efficacy of AEBSF, a small-molecule, irreversible inhibitor, on the self-association of the digestive enzyme ?-chymotrypsin. The Calypso is able to perform accurate and rapid analysis of inhibition kinetics of protein-protein binding as well as equilibrium association properties. The new application note, entitled “Inhibition Kinetics of Protein-Protein Binding with Calypso”, is available to download free-of-charge via www.wyatt.com.
CG-MALS is a powerful, non-destructive method for efficient characterization of macromolecular interactions without labeling, immobilization or additional biochemical substrates. It employs a series of samples of different composition or concentration in order to characterize reversible self- and hetero-association of proteins, reaction rates and affinities of aggregation or dissociation and non-specific, repulsive or attractive macromolecular interactions. The Calypso offers superior analytical capabilities, simplifying and automating CG-MALS measurements. The system’s automation enhances productivity by improving repeatability and reliability, while minimizing time and effort. The Calypso offers many advantages over other biophysical techniques for characterizing protein interactions, providing fast and accurate results.
The new application note describes the CG-MALS method using the Calypso automated composition gradient system. The instrument is used in conjunction with Wyatt Technology’s DAWN-HELEOS and miniDAWN-TREOS MALS detectors and an optional dRI or UV absorption on-line concentration detector. Experimental results demonstrate that the dissociation rate is dependent on the concentrations of protein and inhibitor, determining both the protein-inhibitor kinetics parameter kM and protein homodimer dissociation constant Kd.
Filed Under: Drug Discovery
