Multiple-plug binding assays using affinity capillary electrophoresis electrophoresis

This work evaluates the concept of a multiple-plug binding assay to estimate binding constants of proteins to ligands using affinity capillary electrophoresis (ACE). This concept is demonstrated using two model systems: carbonic anhydrase B (CAB, EC 4.2.1.1) and vancomycin from Streptomyces oriental...

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Bibliographic Details
Published in:Journal of Chromatography A 1996-03, Vol.727 (2), p.291-299
Main Authors: Gomez, Frank A., Mirkovich, Joseph N., Dominguez, Victor M., Liu, Kok W., Macias, Doreen M.
Format: Article
Language:English
Subjects:
Affinity electrophoresis
Binding constants
Carbonic anhydrase B
Mesityl oxide
Multiple-plug binding assays
Myoglobin
Protein-ligand interactions
Proteins
Vancomycin
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Summary:This work evaluates the concept of a multiple-plug binding assay to estimate binding constants of proteins to ligands using affinity capillary electrophoresis (ACE). This concept is demonstrated using two model systems: carbonic anhydrase B (CAB, EC 4.2.1.1) and vancomycin from Streptomyces orientalis. Multiple plugs of protein, and non-interacting neutral and protein standards, are injected and analysis of the electrophoretic mobilities of the individual protein plugs, relative to the non-interacting neutral standard, as a function of the concentration of ligand yields values for their binding constants to the protein. These values agree well with those estimated using other assay and ACE techniques. This technique offers a new and expeditions approach to estimating binding constants of ligands to proteins.
ISSN:0021-9673
DOI:10.1016/0021-9673(95)01155-2