Rapid screening of small ligand affinity to human serum albumin by an optical biosensor.
Here we report the use of IAsys biosensor technology for determining the binding parameters of low molecular weight compounds, such as warfarin and bilirubin, to surface immobilized human serum albumin. The protein was covalently immobilized on the surface of the biosensor cuvette, bearing a carboxymethyl dextran layer, through a condensing reaction between the carboxyl groups of the biosensor surface and epsilon-amine groups of protein lysine residues. This system detects and quantifies the changes in refractive index in the vicinity of the surface of the sensor chip to which the protein is immobilized. The changes in the refractive index are proportional to the change in the absorbed mass, thus the analysis allows the monitoring of the interaction process and the determination of the binding parameters. Optical biosensor analysis, most suited for studying protein/protein or protein/nucleic acid interactions, was sensitive enough to monitor the binding of low molecular weight compounds to human serum albumin and then suitable for a rapid screening of libraries of potential drugs when bioavailability is the research target.
Bertucci
Carlo
C
Department of Pharmaceutical Sciences, University of Bologna, via Belmeloro 6, Bologna, Italy. bertucci@unibo.it
Cimitan
Samanta
S
D001663:Bilirubin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008024:Ligands; D055095:Optics and Photonics; D011485:Protein Binding / Q000502:physiology; D012709:Serum Albumin / Q000032:analysis* / Q000378:metabolism*; D014859:Warfarin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism
D008024:Ligands; D012709:Serum Albumin; D014859:Warfarin; D001663:Bilirubin