On-line Coupling of Immobilized Cytochrome P450 Microreactor and Capillary Electrophoresis: A Promising Tool for Drug Development
| Authors | |
|---|---|
| Year of publication | 2016 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Chromatography A |
| MU Faculty or unit | |
| Citation | |
| Doi | http://dx.doi.org/10.1016/j.chroma.2016.01.081 |
| Field | Biochemistry |
| Keywords | capillary electrophoresis; IMER; IMER; Cytochromes P450; drug developemnt |
| Description | In this work, the combination of an immobilized enzyme microreactor (IMER) based on the clinically important isoform cytochrome P450 2C9 (CYP2C9) with capillary electrophoresis (CE) is presented. The CYP2C9 was attached to magnetic SiMAG-carboxyl microparticles using the carbodiimide method. The formation of an IMER in the inlet part of the separation capillary was ensured by two permanent magnets fixed in a cassette from the CE apparatus in the repulsive arrangement. The resulting on-line system provides an integration of enzyme reaction mixing and incubation, reaction products separation, detection and quantification into a single fully automated procedure with the possibility of repetitive use of the enzyme and minuscule amounts of reactant consumption. The on-line kinetic and inhibition studies of CYP2C9’s reaction with diclofenac as a model substrate and sulfaphenazole as a model inhibitor were conducted in order to demonstrate its practical applicability. |
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