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Aptamer-based label-free method for hemin recognition and DNA assay by capillary electrophoresis with chemiluminescence detection

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Abstract

An aptamer-based label-free approach to hemin recognition and DNA assay using capillary electrophoresis with chemiluminescence detection is introduced here. Two guanine-rich DNA aptamers were used as the recognition element and target DNA, respectively. In the presence of potassium ions, the two aptamers folded into the G-quartet structures, binding hemin with high specificity and affinity. Based on the G-quartet–hemin interactions, the ligand molecule was specifically recognized with a K d ≈ 73 nM, and the target DNA could be detected at 0.1 μM. In phosphate buffer of pH 11.0, hemin catalyzed the H2O2-mediated oxidation of luminol to generate strong chemiluminescence signal; thus the target molecule itself served as an indicator for the molecule–aptamer interaction, which made the labeling and/or modification of aptamers or target molecules unnecessary. This label-free method for molecular recognition and DNA detection is therefore simple, easy, and effective.

A label-free approach to aptamer-based hemin recognition and DNA detection is introduced, which gives great potential for using a small molecule itself as the indicator for molecular recognition and DNA detection thereby avoiding any labeling or modification step

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Abbreviations

CE:

capillary electrophoresis

CL:

chemiluminescence

HRP:

horseradish peroxidase

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Acknowledgement

This work is supported by the National Natural Science Foundation of China with the Grants 20335040, 20427003 and 20675078 and Chinese Academy of Sciences KJCX2.YW.HO9.

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Authors and Affiliations

  1. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China

    Tao Li, Bingling Li & Shaojun Dong

  2. Graduate School of the Chinese Academy of Sciences, Beijing, 100039, China

    Tao Li & Bingling Li

Authors
  1. Tao Li
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  2. Bingling Li
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  3. Shaojun Dong
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Correspondence to Shaojun Dong.

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Li, T., Li, B. & Dong, S. Aptamer-based label-free method for hemin recognition and DNA assay by capillary electrophoresis with chemiluminescence detection. Anal Bioanal Chem 389, 887–893 (2007). https://doi.org/10.1007/s00216-007-1487-5

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  • DOI: https://doi.org/10.1007/s00216-007-1487-5

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