Competitive affinity capillary electrophoresis assay based on a "hybrid" pre-incubation/on-capillary mixing format using an enantioselective aptamer as affinity ligand.
In this paper, we describe an aptamer-based competitive affinity CE (ACE) assay involving (i) the pre-incubation of the target (D-arginine) and the specific ligand (anti-D-arginine-L-RNA aptamer) before (ii) the competition with the labeled target (dansylated D-arginine) through an on-capillary mixing strategy. The effects of some critical operating parameters such as the applied voltage and the sample-aptamer mixture plug length on the assay sensitivity were investigated. The ACE assay appeared particularly dependent on the plug length of the pre-incubated sample-aptamer solution. It was shown that this "hybrid" strategy significantly improved the assay sensitivity relative to that obtained with a "full" on-capillary mixing approach.
Département de Pharmacochimie Moléculaire, Institut de Chimie Moléculaire de Grenoble, Université Grenoble I (Joseph Fourier), UFR de Pharmacie, Grenoble, France.
Josephine
J
Ruta
Corinne
C
Ravelet
Isabelle
I
Baussane
Jennifer
J
Fize
Jean-Luc
JL
Décout
Eric
E
Peyrin
D000345:Affinity Labels; D052157:Aptamers, Nucleotide; D001120:Arginine / Q000032:analysis* / Q000737:chemistry*; D001483:Base Sequence; D001667:Binding, Competitive; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000706:statistics & numerical data; D008024:Ligands; D052156:SELEX Aptamer Technique; D012680:Sensitivity and Specificity; D013237:Stereoisomerism
D000345:Affinity Labels; D052157:Aptamers, Nucleotide; D008024:Ligands; D001120:Arginine