M Pedersen2; J Walter1; S Gad3; B Sorensen1; J Ostergaard3; H Jensen1;
1 Fida Biosystems ApS, Denmark; 2 FidaBio Systems, UK; 3 Copenhagen University, Denmark
AbstractTools for protein characterization are in constant development. One of the newest and most exiting tools is the Fida 1, a revolutionary single assay platform that provides information on structure and binding affinities in solution. In this work, we introduce an approach based on Flow Induced Dispersion Analysis (FIDA) which is able to efficiently characterize chemically induced protein unfolding of Human Serum Albumin (HSA) in great detail. The platform enables local structural changes to be probed by monitoring changes in intrinsic fluorescence and loss of binding of a low-molecular weight ligand. Simultaneously, the size of the unfolding HSA is obtained by FIDA on the same samples. The integration of the methodologies enables a fully automated characterization of HSA using only a few hundred nanoliters of sample.