期刊
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
卷 204, 期 -, 页码 180-187出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.saa.2018.06.036
关键词
Biosensor; Aptamer; Binding way; Saxitoxin; Paralytic shellfish toxin
类别
资金
- National Natural Science Foundation of China [31501576, 21504021, 51673056]
- Preferential Foundation for Overseas Scholars, Ministry of Personnel of China [2015LXKJZZ001]
- Natural Science Foundation for the Higher Education Institutions of Anhui Province of China [KJ2015A428]
- Science and Technology Major Project of Anhui Province, China [16030901053]
Aptamers could be used to construct simple and effective biosensor because the conformational switch of aptamer upon target binding is easy to be transferred to optical or electrochemical signals. Nevertheless, we found that the binding between saxitoxin (STX) and aptamer (M-30f) is not accompanied with conformational switch. Here, the circular dichroism spectra, fluorophore and quencher labeled aptamer, and crystal violet based assays were used to identify the binding way between STX and aptamer. The results show that the conformation of aptamer is stabilized in PBS buffer (10 mM phosphate buffer, 2.7 mM kCl, 137 mM NaCl, pH 7.4) and this conformation may provide an exactly suitable cave for SIX binding. Through the analysis of UV-melting curves and circular dichroism-melting curves, it is found that different concentrations of STX produce different unfolding extents of the aptamer under high temperature. Then, a simple temperature-assisted turn-on fluorescent aptasensor was developed to detect SIX and the application in real sample detection demonstrates its feasibility. The proposed method provides not only an alternative for SIX detection but also a strategy for simple aptasensor design using aptamers that do not switch conformation upon targets binding. (C) 2018 Elsevier B.V. All rights reserved.
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