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Aptamer-Based Molecular Recognition of Lysergamine, Metergoline and Small Ergot Alkaloids
Laboratory of Systemic Physiological and Ecotoxicological Research (SPHERE), Department of Biology, University of Antwerp, Groenenborgerlaan 171, Antwerp B-2020, Belgium
Institute of Agricultural and Fisheries Research (ILVO), Ankerstraat 1, Oostende B-8400, Belgium
Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, Ghent B-9000, Belgium
Polymer Chemistry and Biomaterials Group, Ghent University, campus Sterre S4, Krijgslaan 281, Ghent B-9000, Belgium
Department of Chemistry, University of Antwerp, Groenenborgerlaan 171, Antwerp B-2020, Belgium
Structural Biology and Bioinformatics Center, Free University of Brussels (ULB), campus de la Plaine, CP206/02, boulevard du Triomphe, Brussels B-1050, Belgium
Mycothèque de l'Université catholique de Louvain (BCCMTM/MUCL), Mycology, Applied Microbiology, Earth and Life Institute (ELI), Université catholique de Louvain (UCL), Croix du Sud 2 boîte L7.05.06, Louvain-la-Neuve B-1348, Belgium
* Author to whom correspondence should be addressed.
Received: 18 September 2012; in revised form: 26 November 2012 / Accepted: 5 December 2012 / Published: 14 December 2012
Abstract: Ergot alkaloids are mycotoxins produced by fungi of the genus Claviceps, which infect cereal crops and grasses. The uptake of ergot alkaloid contaminated cereal products can be lethal to humans and animals. For food safety assessment, analytical techniques are currently used to determine the presence of ergot alkaloids in food and feed samples. However, the number of samples which can be analyzed is limited, due to the cost of the equipment and the need for skilled personnel. In order to compensate for the lack of rapid tests for the detection of ergot alkaloids, the aim of this study was to develop a specific recognition element for ergot alkaloids, which could be further applied to produce a colorimetric reaction in the presence of these toxins. As recognition elements, single-stranded DNA ligands were selected by using an iterative selection procedure named SELEX, i.e., Systematic Evolution of Ligands by EXponential enrichment. After several selection cycles, the resulting aptamers were cloned and sequenced. A surface plasmon resonance analysis enabled determination of the dissociation constants of the complexes of aptamers and lysergamine. Dissociation constants in the nanomolar range were obtained with three selected aptamers. One of the selected aptamers, having a dissociation constant of 44 nM, was linked to gold nanoparticles and it was possible to produce a colorimetric reaction in the presence of lysergamine. This system could also be applied to small ergot alkaloids in an ergot contaminated flour sample.
Keywords: single-stranded nucleic acid; ergot alkaloids; SELEX procedure; surface plasmon resonance
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Rouah-Martin, E.; Mehta, J.; van Dorst, B.; de Saeger, S.; Dubruel, P.; Maes, B.U.W.; Lemiere, F.; Goormaghtigh, E.; Daems, D.; Herrebout, W.; van Hove, F.; Blust, R.; Robbens, J. Aptamer-Based Molecular Recognition of Lysergamine, Metergoline and Small Ergot Alkaloids. Int. J. Mol. Sci. 2012, 13, 17138-17159.
Rouah-Martin E, Mehta J, van Dorst B, de Saeger S, Dubruel P, Maes BUW, Lemiere F, Goormaghtigh E, Daems D, Herrebout W, van Hove F, Blust R, Robbens J. Aptamer-Based Molecular Recognition of Lysergamine, Metergoline and Small Ergot Alkaloids. International Journal of Molecular Sciences. 2012; 13(12):17138-17159.
Rouah-Martin, Elsa; Mehta, Jaytry; van Dorst, Bieke; de Saeger, Sarah; Dubruel, Peter; Maes, Bert U.W.; Lemiere, Filip; Goormaghtigh, Erik; Daems, Devin; Herrebout, Wouter; van Hove, François; Blust, Ronny; Robbens, Johan. 2012. "Aptamer-Based Molecular Recognition of Lysergamine, Metergoline and Small Ergot Alkaloids." Int. J. Mol. Sci. 13, no. 12: 17138-17159.