title: Open-tubular capillary electrochromatography of bovine beta-lactoglobulin variants A and B using an aptamer stationary phase.
abstract: DNA aptamers that form a G-quartet conformation were covalently attached to a capillary surface for open-tubular capillary electrochromatographic separation of bovine beta-lactoglobulin variants A and B, which vary by 2 of their 162 amino acid residues. Separation was achieved using a 4-plane, G-quartet aptamer stationary phase with tris(hydroxymethyl)aminomethane (Tris) or phosphate buffer as the mobile phase. In control experiments, separation did not occur using either an oligonucleotide of similar base composition but which does not form a G-quartet structure, or using capillary zone electrophoresis on a bare capillary under similar experimental conditions. Separation was achieved using a capillary coated only with the covalent linker molecule. In phosphate buffer, the separations were similar for aptamer-coated and linker-only stationary phases, while in Tris buffer, retention times were almost doubled for the linker-only capillary. When Tris buffer is the mobile phase, there appears to be weaker interactions between the proteins and the stationary phase that may result in a gentler, less denaturing separation than is commonly achieved using hydrocarbon-based stationary phases.
authors.affiliation: Department of Chemistry, Duke University, Durham, NC 27705, USA.
authors.forename: M A
authors.initials: MA
authors.lastname: Rehder
authors.forename: L B
authors.initials: LB
authors.lastname: McGown
mesh_terms: D000818:Animals; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods; D014644:Genetic Variation; D007202:Indicators and Reagents; D007782:Lactoglobulins / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D007202:Indicators and Reagents; D007782:Lactoglobulins