Inventors:
Devendra Kalonia - Ashford CT, US
Bajaj Harminder - Millbrae CA, US
Sharma Vikas - Millbrae CA, US
International Classification:
G01N 21/00
Abstract:
Apparatus, systems and methods for measurement of Bvalues of proteins in aqueous solutions in flow-mode utilize a dual-detector cell are provided. Simultaneous measurement of protein concentration and scattered light intensity is facilitated as the protein elutes from a size-exclusion column. Each data point on the resulting chromatograms is converted to Rayleigh's ratio, R, and to concentration c, respectively. The Bvalue is calculated from the slope of the Debye plot (Kc/R versus c) generated from a range of the concentrations obtained from these chromatograms for a single protein injection. Measurements may be analyzed using modeling data derived from a predetermined modeling equation to quantify self-association of molecules. The apparatus and method provide a reliable means for determining Bvalues for such proteins as lysozyme, chymotrypsinogen, and chymotrypsin in various solution conditions.