Enantiomeric separation using an l-RNA aptamer as chiral additive in partial-filling capillary electrophoresis

In this paper, we report the chiral resolution of arginine using an anti-arginine l-RNA aptamer chiral selector in partial-filling CE. The effects of the capillary temperature, sample load, and aptamer plug length on the enantiomeric separation were assessed. Very high chiral resolving capability was observed at low or moderate capillary temperatures (the target peak being not detected in the separation window), whereas the practical chiral resolution was achieved only at high enough temperatures (50-60 degrees C). Over this high-temperature range, the electrophoretic behavior of the target enantiomer appeared to result from a combination of binding site heterogeneity, slow desorption kinetics, and concentration overload of aptamer binding sites. From additional thermal UV melting experiments, three RNA conformations were identified for the 50-60 degrees C temperatures. It was suggested that the presence of these different RNA conformations was a plausible source of the binding site heterogeneity.