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Anal Biochem. 2002 Jul 15;306(2):228-36.

Compensation for loss of ligand activity in surface plasmon resonance experiments.

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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.


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.

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