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PMID: 8785280 Published · ppublish English Comparative Study Journal Article

Kinetics of ligand binding to receptor immobilized in a polymer matrix, as detected with an evanescent wave biosensor. I. A computer simulation of the influence of mass transport.

Biophysical journal ·Vol. 70 ·No. 3 ·1996-03-00 ·Pages 1230-49

Schuck P

Abstract

The influence of mass transport on ligand binding to receptor immobilized in a polymer matrix, as detected with an evanescent wave biosensor, was investigated. A one-dimensional computer model for the mass transport of ligand between the bulk solution and the polymer gel and within the gel was employed, and the influence of the diffusion coefficient, the partition coefficient, the thickness of the matrix, and the distribution of immobilized receptor were studied for a variety of conditions. Under conditions that may apply to many published experimental studies, diffusion within the matrix was found to decrease the overall ligand transport significantly. For relatively slow reactions, small spatial gradients of free and bound ligand in the gel are found, whereas for relatively rapid reactions strong inhomogeneities of ligand within the gel occur before establishment of equilibrium. Several types of deviations from ideal pseudo-first-order binding progress curves are described that resemble those of published experimental data. Extremely transport limited reactions can in some cases be fitted with apparently ideal binding progress curves, although with apparent reaction rates that are much lower than the true reaction rates. Nevertheless, the ratio of the apparent rate constants can be semiquantitatively consistent with the true equilibrium constant. Apparently "cooperative" binding can result from high chemical on rates at high receptor saturation. Dissociation in the presence of transport limitation was found to be well described empirically by a single or a double exponential, with both apparent rate constants considerably lower than the intrinsic chemical rate constant. Transport limitations in the gel can introduce many generally unknown factors into the binding progress curve. The simulations suggest that unexpected deviations from ideal binding progress curves may be due to highly transport influenced binding kinetics. The use of a thinner polymer matrix could significantly increase the range of detectable rate constants.

MeSH Terms
Binding Sites Biological Transport, Active Biophysical Phenomena Biophysics Biosensing Techniques Computer Simulation Diffusion Gels Kinetics Ligands Models, Chemical Polymers Receptors, Cell Surface/metabolism
Chemicals
Gels Ligands Polymers Receptors, Cell Surface
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Schuck P
Laboratory of Biochemical Pharmacology, National Institute of Diabetes, Digestive and Kidney Diseases, National Institute of Health, Bethesda, Maryland 20892, USA. pschuck@helix.nih.gov
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(96)79681-9
Email
pschuck@helix.nih.gov
Published
1996-03-00
Pages
1230-49
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1225050
Subset
IM
External Links
PubMed source DOI: 10.1016/S0006-3495(96)79681-9 PMC Full Text View journal details
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