| Abstract
| - Aptamers are DNA (or RNA) ligands selected from large libraries of random DNA sequencesand capable of binding different classes of targets with high affinity and selectivity. Both the chances forthe aptamer to be selected and the quality of the selected aptamer are largely dependent on the methodof selection. Here we introduce selection of aptamers by nonequilibrium capillary electrophoresis ofequilibrium mixtures (NECEEM). The new method has a number of advantages over conventionalapproaches. First, NECEEM-based selection has exceptionally high efficiency, which allows aptamerdevelopment with fewer rounds of selection. Second, NECEEM can be equally used for selecting aptamersand finding their binding parameters. Finally, due to its comprehensive kinetic capabilities, the new methodcan potentially facilitate selection of aptamers with predefined Kd, koff, and kon of the aptamer−targetinteraction. In this proof-of-principle work, we describe the theoretical bases of the method and demonstrateits application to a one-step selection of DNA aptamers with nanomolar affinity for protein farnesyltransferase.
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