INTEGRATION OF SHORT-END INJECTION PROCEDURE INTO EMMA METHODOLOGY
| Authors | |
|---|---|
| Year of publication | 2006 |
| Type | Article in Proceedings |
| Conference | Book of abstract 15th International Symposium on Capillary Electroseparation Techniques (ITP-2006) |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | short-end; EMMA |
| Description | Fourteen years ago a new application for the evaluation of enzymatic reactions in capillary electrophoresis was proposed and developed by Bao and Regnier. In this method called electrophoretically mediated microanalysis (EMMA), the capillary is used not only as a separation medium but also as a reaction chamber. Since its discovery, the EMMA methodology has been utilized in a number of enzyme and non-enzyme systems. The posibility of using the short-end injection procedure in the EMMA methodology is proposed in this work. This combination preserved the benefits of both these instrumental approaches. Whereas the short-end injection procedure brought reducing analysis time, increasing sensitivity, and decreasing buffer depletion, the EMMA methodology allowed full automatization of the assay with the dramatic reduction in consumption of enzyme preparations. Moreover the repeatability of retention times and peak areas was comparably to that obtained using the classic EMMA set-up. The integration of both approaches was demonstrated on the kinetic studies of mono- and two substrates enzymatic reactions. |
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