Analysis of human chorionic gonadotropin-monoclonal antibody interaction in BIAcore

Ashish, Banerjee and Satyanarayana Murthy, Gundlupet (2004) Analysis of human chorionic gonadotropin-monoclonal antibody interaction in BIAcore. In: Journal of Biosciences, 29 (1). pp. 57-66.

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Abstract

Kinetic studies of macromolecular ligand-ligate interaction have generated ample interest since the advent of plasmon resonance based instruments like BIAcore. Most of the studies reported in literature assume a simple 1 : 1 Langmuir binding and complete reversibility of the system. However we observed that in a high affinity antigen-antibody system [human chorionic gonadotropin-monoclonal antibody (hCG-mAb)] dissociation is insignificant and the sensogram data cannot be used to measure the equilibrium and kinetic parameters. At low concentrations of mAb the complete sensogram could be fitted to a single exponential. Interestingly we found that at higher mAb concentrations, the binding data did not conform to a simple bimolecular model. Instead, the data fitted a two-step model, which may be because of surface heterogeneity of affinity sites. In this paper, we report on the global fit of the sensograms. We have developed a method by which a single two-minute sensogram can be used in high affinity systems to measure the association rate constant of the reaction and the functional capacity of the ligand (hCG) immobilized on the chip. We provide a rational explanation for the discrepancies generally observed in most of the BIAcore sensograms

Item Type:	Journal Article
Publication:	Journal of Biosciences
Publisher:	springer
Additional Information:	Copyright for this article belongs to springer.
Department/Centre:	Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Date Deposited:	23 Oct 2009 10:01
Last Modified:	19 Sep 2010 05:51
URI:	http://eprints.iisc.ac.in/id/eprint/24744

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