Compensation for loss of ligand activity in surface plasmon resonance experiments.
The determination of equilibrium binding constants is an important aspect of the analysis of protein-protein interactions. In recent years surface plasmon resonance experiments (e.g., with a BIAcore instrument) have provided a valuable experimental approach to determining such constants. The standard method is based on measuring amounts of analyte bound at equilibrium for different analyte concentrations. During the course of a typical surface plasmon resonance experiment the measured equilibrium levels for a given analyte concentration often decrease. This appears to be due to a loss of activity of the protein coupled to the sensor chip or other phenomena. The loss in signal can lead to an erroneous determination of the equilibrium constant. A data analysis approach is introduced that aims to compensate for the loss of activity so that its influence on the results of the experiments is reduced.
Ober
Raimund J
RJ
Center for Systems, Communications and Signal Processing, Eric Jonsson School of Electrical Engineering and Computer Science, University of Texas at Dallas, Richardson 75083-0688, USA.
Ward
E Sally
ES
D007140:Immunoglobulin Fab Fragments; D007700:Kinetics; D008024:Ligands; D009113:Muramidase / Q000276:immunology / Q000378:metabolism*; D020349:Surface Plasmon Resonance
D007140:Immunoglobulin Fab Fragments; D008024:Ligands; D009113:Muramidase