Microdialysis sampling method for evaluation of binding kinetics of small molecules to macromolecules. Export

@article{citeulike:2857529, abstract = {Here we present an application of microdialysis sampling for evaluation of the binding kinetics of small molecules to macromolecules. It is label-free, and no immobilization of any interaction partner is required. The method was established by the coupling of a binding reaction with a membrane transport in a miniature and dynamic microdialysis sampling system. A theoretical model was established to describe the quantitative relationship between the binding kinetics of small ligands to macromolecules and the enhanced mass transport of small ligands and was applied to estimate the binding kinetics. To demonstrate the proof-of-principle, we examined the binding kinetics of an abundant plasma protein human serum albumin (HSA) and a representative drug ketoprofen as an example. The primary binding constant of ketoprofen to HSA was estimated as 1.63 (+/-0.12) x 10(6) M(-1). The estimated association and dissociation rate constants (k1 and k(-1)) were about 3.71 x 10(5) M(-1) s(-1) and 0.227 s(-1), respectively. The results suggest a fast binding of ketoprofen to HSA and a fast dissociation of the formed complex, which are consistent with the reversible binding property of drug and HSA (k(-1) in the order of s(-1)). This is the first report on binding-kinetics measurement using microdialysis sampling.}, address = {State Key Laboratory of Environmental Chemistry and Eco-toxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China. hlwang@rcees.ac.cn}, author = {Wang, H. and Wang, Z. and Lu, M. and Zou, H.}, citeulike-article-id = {2857529}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/ac702066x}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/ac702066x}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/18355058}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=18355058}, day = {15}, doi = {10.1021/ac702066x}, issn = {1520-6882}, journal = {Analytical chemistry}, keywords = {dialysis, human, kinetic\_modeling}, month = {April}, number = {8}, pages = {2993--2999}, posted-at = {2008-06-02 18:07:15}, priority = {2}, title = {Microdialysis sampling method for evaluation of binding kinetics of small molecules to macromolecules.}, url = {http://dx.doi.org/10.1021/ac702066x}, volume = {80}, year = {2008} }

TY - JOUR ID - citeulike:2857529 L3 - citeulike-article-id:2857529 N2 - Here we present an application of microdialysis sampling for evaluation of the binding kinetics of small molecules to macromolecules. It is label-free, and no immobilization of any interaction partner is required. The method was established by the coupling of a binding reaction with a membrane transport in a miniature and dynamic microdialysis sampling system. A theoretical model was established to describe the quantitative relationship between the binding kinetics of small ligands to macromolecules and the enhanced mass transport of small ligands and was applied to estimate the binding kinetics. To demonstrate the proof-of-principle, we examined the binding kinetics of an abundant plasma protein human serum albumin (HSA) and a representative drug ketoprofen as an example. The primary binding constant of ketoprofen to HSA was estimated as 1.63 (+/-0.12) x 10(6) M(-1). The estimated association and dissociation rate constants (k1 and k(-1)) were about 3.71 x 10(5) M(-1) s(-1) and 0.227 s(-1), respectively. The results suggest a fast binding of ketoprofen to HSA and a fast dissociation of the formed complex, which are consistent with the reversible binding property of drug and HSA (k(-1) in the order of s(-1)). This is the first report on binding-kinetics measurement using microdialysis sampling. IS - 8 JF - Analytical chemistry SN - 1520-6882 AD - State Key Laboratory of Environmental Chemistry and Eco-toxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China. hlwang@rcees.ac.cn EP - 2999 TI - Microdialysis sampling method for evaluation of binding kinetics of small molecules to macromolecules. VL - 80 SP - 2993 KW - dialysis KW - human KW - kinetic_modeling AU - Wang, H AU - Wang, Z AU - Lu, M AU - Zou, H PY - 2008/04/15/ UR - http://dx.doi.org/10.1021/ac702066x ER -