title: A DNA aptamer as a new target-specific chiral selector for HPLC.
abstract: In this paper, a DNA aptamer, known to bind stereospecifically the D-enantiomer of an oligopeptide, i.e., arginine-vasopressin, was immobilized on a chromatographic support. The influence of various parameters (such as column temperature, eluent pH, and salt concentration) on the L- and D-peptide retention was investigated in order to provide information about the binding mechanism and then to define the utilization conditions of the aptamer column. The results suggest that dehydration at the binding interface, charge-charge interactions, and adaptive conformational transitions contribute to the specific D-peptide-aptamer complex formation. A very significant enantioselectivity was obtained in the optimal binding conditions, the D-peptide being strongly retained by the column while the L-peptide eluted in the void volume. A rapid baseline separation of peptide enantiomers was also achieved by modulating the elution conditions. Furthermore, it was established that the aptamer column was stable during an extended period of time. This work indicates that DNA aptamers, specifically selected against an enantiomer, could soon become very attractive as new target-specific chiral selectors for HPLC.
authors.affiliation: Equipe de Chimie Analytique, Département de Pharmacochimie Moléculaire (UMR 5063 CNRS-UJF), ICMG FR 2607, UFR de Pharmacie de Grenoble, Université Joseph Fourier, Avenue de Verdun, 38240 Meylan, France.
authors.forename: Mickael
authors.initials: M
authors.lastname: Michaud
authors.forename: Eric
authors.initials: E
authors.lastname: Jourdan
authors.forename: Annick
authors.initials: A
authors.lastname: Villet
authors.forename: Anne
authors.initials: A
authors.lastname: Ravel
authors.forename: Catherine
authors.initials: C
authors.lastname: Grosset
authors.forename: Eric
authors.initials: E
authors.lastname: Peyrin
mesh_terms: D001127:Arginine Vasopressin / Q000737:chemistry* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009690:Nucleic Acid Conformation; D009994:Osmolar Concentration; D013237:Stereoisomerism; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D001127:Arginine Vasopressin; D004247:DNA