Compensation for loss of ligand activity in surface plasmon resonance experiments
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.
BIAcore, Equilibrium analysis, Loss of activity, Surface plasmon resonance
228-236
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
15 July 2002
Ober, Raimund J.
31f4d47f-fb49-44f5-8ff6-87fc4aff3d36
Ward, E. Sally
b31c0877-8abe-485f-b800-244a9d3cd6cc
Ober, Raimund J. and Ward, E. Sally
(2002)
Compensation for loss of ligand activity in surface plasmon resonance experiments.
Analytical Biochemistry, 306 (2), .
(doi:10.1006/abio.2002.5716).
Abstract
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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e-pub ahead of print date: 11 July 2002
Published date: 15 July 2002
Keywords:
BIAcore, Equilibrium analysis, Loss of activity, Surface plasmon resonance
Identifiers
Local EPrints ID: 424924
URI: http://eprints.soton.ac.uk/id/eprint/424924
ISSN: 0003-2697
PURE UUID: 88e06bcf-dc0b-48bf-a1bc-4cf0935c445d
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Date deposited: 05 Oct 2018 16:30
Last modified: 18 Mar 2024 03:48
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