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Mar. Drugs 2015, 13(12), 7433-7445; doi:10.3390/md13127066

Generation of Internal-Image Functional Aptamers of Okadaic Acid via Magnetic-Bead SELEX

1
Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, Jilin University, Changchun 130062, China
2
Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Science, Jilin University, Changchun 130062, China
3
Institute of Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, Jilin, Changchun 130062, China
*
Authors to whom correspondence should be addressed.
Academic Editors: Angela Capper and Cherie Motti
Received: 31 March 2015 / Revised: 9 October 2015 / Accepted: 5 November 2015 / Published: 17 December 2015
(This article belongs to the Special Issue Okadaic Acid and Dinophysis Toxins)
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Abstract

Okadaic acid (OA) is produced by Dinophysis and Prorocentrum dinoflagellates and primarily accumulates in bivalves, and this toxin has harmful effects on consumers and operators. In this work, we first report the use of aptamers as novel non-toxic probes capable of binding to a monoclonal antibody against OA (OA-mAb). Aptamers that mimic the OA toxin with high affinity and selectivity were generated by the magnetic bead-assisted systematic evolution of ligands by exponential enrichment (SELEX) strategy. After 12 selection rounds, cloning, sequencing and enzyme-linked immunosorbent assay (ELISA) analysis, four candidate aptamers (O24, O31, O39, O40) were selected that showed high affinity and specificity for OA-mAb. The affinity constants of O24, O31, O39 and O40 were 8.3 × 108 M−1, 1.47 × 109 M−1, 1.23 × 109 M−1 and 1.05 × 109 M−1, respectively. Indirect competitive ELISA was employed to determine the internal-image function of the aptamers. The results reveal that O31 has a similar competitive function as free OA toxin, whereas the other three aptamers did not bear the necessary internal-image function. Based on the derivation of the curvilinear equation for OA/O31, the equation that defined the relationship between the OA toxin content and O31 was Y = 2.185X − 1.78. The IC50 of O31 was 3.39 ng·mL−1, which was close to the value predicted by the OA ELISA (IC50 = 4.4 ng·mL−1); the IC10 was 0.33 ng·mL−1. The above data provides strong evidence that internal-image functional aptamers could be applicable as novel probes in a non-toxic assay. View Full-Text
Keywords: okadaic acid; SELEX; internal-image function okadaic acid; SELEX; internal-image function
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Lin, C.; Liu, Z.-S.; Wang, D.-X.; Li, L.; Hu, P.; Gong, S.; Li, Y.-S.; Cui, C.; Wu, Z.-C.; Gao, Y.; Zhou, Y.; Ren, H.-L.; Lu, S.-Y. Generation of Internal-Image Functional Aptamers of Okadaic Acid via Magnetic-Bead SELEX. Mar. Drugs 2015, 13, 7433-7445.

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