Determination of Rate Constants and Equilibrium Constants for Solution-Phase Drug–Protein Interactions by Ultrafast Affinity Extraction
Autor: | Zhao Li, David S. Hage, Maria Podariu, Xiwei Zheng |
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Jazyk: | angličtina |
Rok vydání: | 2014 |
Předmět: |
Kinetics
Serum albumin Plasma protein binding 010402 general chemistry 01 natural sciences Article Chromatography Affinity Analytical Chemistry Reaction rate constant Computational chemistry medicine Humans Solubility Equilibrium constant Serum Albumin Chromatography biology Chemistry 010401 analytical chemistry Extraction (chemistry) Human serum albumin 0104 chemical sciences Pharmaceutical Preparations biology.protein medicine.drug Protein Binding |
Zdroj: | Analytical Chemistry |
ISSN: | 1520-6882 0003-2700 |
Popis: | A method was created on the basis of ultrafast affinity extraction to determine both the dissociation rate constants and equilibrium constants for drug-protein interactions in solution. Human serum albumin (HSA), an important binding agent for many drugs in blood, was used as both a model soluble protein and as an immobilized binding agent in affinity microcolumns for the analysis of free drug fractions. Several drugs were examined that are known to bind to HSA. Various conditions to optimize in the use of ultrafast affinity extraction for equilibrium and kinetic studies were considered, and several approaches for these measurements were examined. The dissociation rate constants obtained for soluble HSA with each drug gave good agreement with previous rate constants reported for the same drugs or other solutes with comparable affinities for HSA. The equilibrium constants that were determined also showed good agreement with the literature. The results demonstrated that ultrafast affinity extraction could be used as a rapid approach to provide information on both the kinetics and thermodynamics of a drug-protein interaction in solution. This approach could be extended to other systems and should be valuable for high-throughput drug screening or biointeraction studies. |
Databáze: | OpenAIRE |
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