ANYL 307 |
| Determination of the dissociation constant of the thrombin-aptamer complex was done using microchip capillary electrophoresis (CE). A microchip CE method was optimized by choosing an appropriate injection scheme, the buffer, the detection length and by minimizing protein adsorption. The choice of the detection point along the separation channel in microchip CE is advantageous over traditional CE when rapid analysis is required. Protein adsorption on bare silica walls may be reduced by coating or by adding the appropriate separation buffer additives. The consequences of adsorption can also be reduced by using a short detection length when a sticky protein, such as thrombin, is involved in the analysis. The rapidly dissociating thrombin-aptamer complex was detected in less than 10 s using a modified gated injection scheme. The dissociation constant was determined to be 23 nM which is consistent with previously reported results. The calibration curve showed good linearity. |
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Analytical Approaches: Separation Science
8:30 AM-12:00 PM, Thursday, 14 September 2006 Moscone Center -- Room 130, Oral
Division of Analytical Chemistry |