title: Nonequilibrium capillary electrophoresis of equilibrium mixtures: a universal tool for development of aptamers.
abstract: Aptamers are DNA (or RNA) ligands selected from large libraries of random DNA sequences and capable of binding different classes of targets with high affinity and selectivity. Both the chances for the aptamer to be selected and the quality of the selected aptamer are largely dependent on the method of selection. Here we introduce selection of aptamers by nonequilibrium capillary electrophoresis of equilibrium mixtures (NECEEM). The new method has a number of advantages over conventional approaches. First, NECEEM-based selection has exceptionally high efficiency, which allows aptamer development with fewer rounds of selection. Second, NECEEM can be equally used for selecting aptamers and finding their binding parameters. Finally, due to its comprehensive kinetic capabilities, the new method can potentially facilitate selection of aptamers with predefined K(d), k(off), and k(on) of the aptamer-target interaction. In this proof-of-principle work, we describe the theoretical bases of the method and demonstrate its application to a one-step selection of DNA aptamers with nanomolar affinity for protein farnesyltransferase.
authors.affiliation: Department of Chemistry, York University, Toronto, Ontario M3J 1P3, Canada.
authors.forename: Maxim
authors.initials: M
authors.lastname: Berezovski
authors.forename: Andrei
authors.initials: A
authors.lastname: Drabovich
authors.forename: Svetlana M
authors.initials: SM
authors.lastname: Krylova
authors.forename: Michael
authors.initials: M
authors.lastname: Musheev
authors.forename: Victor
authors.initials: V
authors.lastname: Okhonin
authors.forename: Alexander
authors.initials: A
authors.lastname: Petrov
authors.forename: Sergey N
authors.initials: SN
authors.lastname: Krylov
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D051231:Farnesyltranstransferase; D015723:Gene Library; D007700:Kinetics; D009841:Oligonucleotides / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*
chemical_list: D009841:Oligonucleotides; D012313:RNA; D004247:DNA; D019883:Alkyl and Aryl Transferases; D051231:Farnesyltranstransferase