Use of a partial-filling technique in affinity capillary electrophoresis for determining binding constants of ligands to receptors |
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Authors: | Joseph Heintz Monica Hernandez Frank A Gomez |
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Institution: | Department of Chemistry and Biochemistry, California State University, Los Angeles, 5151 State University Drive, Los Angeles, CA 90032-8202, USA |
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Abstract: | This work evaluates the concept of a partial-filling technique in affinity capillary electrophoresis (ACE) using two model systems: vancomycin from Streptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). In this technique the capillary is first partially-filled with ligand followed by a sample of receptor and non-interacting standard and electrophoresed. Analysis of the change in the mobility ratio, M, of the receptor, relative to the non-interacting standard, as a function of the concentration of the ligand, yields a value for the binding constant. These values agree well with those estimated using other binding and ACE techniques. Data demonstrating the quantitative potential of this method is presented. |
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Keywords: | Vancomycin Carbonic anhydrase Enzymes Antibiotics |
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