Cocaine detection by structure-switch aptamer-based capillary zone electrophoresis

• Published in: Electrophoresis Vol. 33; no. 9-10; pp. 1465 - 1470
• Main Authors: Deng, Qin-Pei, Tie, Cai, Zhou, Ying-Lin, Zhang, Xin-Xiang
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01.05.2012
• Subjects: Aptamer; Aptamers, Nucleotide - chemistry; Capillary electrophoresis; Cocaine; Cocaine - analysis; Complementary strand; DNA, Complementary - chemistry; Electrophoresis, Capillary - methods; Fluoresceins - chemistry; Fluorescent Dyes - chemistry; Magnesium - chemistry; Nucleic Acid Conformation; Reproducibility of Results; Sensitivity and Specificity
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.201100680

Aptamers, which are nucleic acid oligonucleotides that can bind targets with high affinity and specificity, have been widely applied as affinity probes in capillary electrophoresis (CE). Due to relative weak interaction between aptamers and small molecules, the application of aptamer‐based CE is still limited in certain compounds. A new strategy that is based on the aptamer structure‐switch concept was designed for small molecule detection by a novel CE method. A carboxyfluorescein (fluorescein amidite, FAM) label DNA aptamer was first incubated with partial complementary strand (CS), and then the free aptamer and the aptamer‐CS duplex were well separated and determined by metal cation mediated CE/laser‐induced fluorescence. When the target was introduced into the incubated sample, the hybridized form was destabilized, resulting in the changes of the fluorescence intensities of the free aptamer and the aptamer‐CS duplex. The length of CS was investigated and 12 mer CS showed the best sensitivity for the detection of cocaine. The presented CE‐LIF method, which combines the separation power of CE with the specificity of interactions occurring between target, aptamer, and CS, could be a universal detection strategy for other aptamer‐specified small molecules.