Global Analysis of a Macromolecular Interaction Measured on BIAcore

doi:10.1006/bbrc.1996.1297

Biochemical and Biophysical Research Communications

Volume 225, Issue 3, 23 August 1996, Pages 1073-1077

https://doi.org/10.1006/bbrc.1996.1297 Get rights and content

Abstract

We demonstrate that the interaction between myoglobin and an immobilized anti-myoglobin antibody measured on BIAcore 2000 can be described by a simple bimolecular reaction mechanism. We improved the quality of the sensor data by correcting for refractive index changes and nonspecific binding using a blank sensor surface. Applying nonlinear least squares analysis, we simultaneously fitted the association and dissociation phase data generated for a range of myoglobin concentrations injected across the antibody surface. The ability to globally fit these data to a simple binding model indicates that effects related to the sensor surface, like mass transport and the dextran matrix, did not complicate the observed binding responses. These results illustrate the potential of BIAcore to monitor macromolecular interactions in real-time and the utility of global analysis to resolve the reaction kinetics.

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^☆: Abbreviations used: mAb, monoclonal antibody; B_max, maximum surface capacity;k_a, association rate constant;k_d, dissociation rate constant; RU, resonance units; STD, standard deviation; NHS,N-hydroxy-succinimide; EDC,N-ethyl-N′-(3-dimethylaminopropyl)-carbodiimide hydrochloride.

¹: Corresponding author. Fax: (801)-585-3833. E-mail: [email protected].

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Regular ArticleGlobal Analysis of a Macromolecular Interaction Measured on BIAcore☆

Abstract

Regular Article
Global Analysis of a Macromolecular Interaction Measured on BIAcore☆