title: Affinity capillary electrophoresis with laser induced fluorescence detection for thrombin analysis using nuclease-resistant RNA aptamers.
abstract: Aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) combines the advantages of affinity aptamer, rapid CE separation, and high sensitivity detection. Here we reported an affinity CE-LIF assay for thrombin by using a fluorophore-labeled RNA aptamer containing 2'-fluoro modification in sugar rings of pyrimidine nucleotides (C and U) as affinity ligand. This RNA aptamer has high binding affinity, specificity and biostability. Thrombin at 0.2nM was successfully detected. This RNA aptamer allowed for the detection of thrombin spiked in diluted human serum sample due to the nuclease resistance. The RNA aptamer has comparable binding affinity to a 29-mer DNA aptamer for thrombin, and the binding site of the RNA aptamer on thrombin partially overlaps with the binding site of the 29-mer DNA aptamer on thrombin. It shows the nuclease-resistant RNA aptamers are promising in assays for thrombin.
authors.affiliation: State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; College of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Hao
authors.affiliation: State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; Institute of Environmental Science, Shanxi University, Taiyuan 030006, China.
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Bai
authors.affiliation: State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Hailin
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: qiangzhao@rcees.ac.cn.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhao
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D007834:Lasers; D008024:Ligands; D013917:Thrombin / Q000032:analysis*
chemical_list: D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D008024:Ligands; C081676:thrombin aptamer; D013917:Thrombin
keywords: 2′-Fluoro modification; Affinity binding; Capillary electrophoresis; Laser induced fluorescence; RNA aptamer; Thrombin