Detection of 13,19-didesmethyl C spirolide by fluorescence polarization using Torpedo electrocyte membranes

Fonfría, Eva S.; Vilariño, Natalia; Molgó, Jordi; Aráoz, Rómulo; Otero, Paz; Espiña, Begoña; Louzao, M. Carmen; Alvarez, Mercedes; Botana, Luís M.

doi:10.1016/j.ab.2010.04.006

Cited by 26 publications

(18 citation statements)

References 17 publications

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“…However, radioactive debris and radio decomposition ( 125 I -α -BgTx halflife = 59.6 days) are serious drawbacks that hamper the use of radioactive ligand binding assays for routine monitoring of marine toxins. On the other hand, the microplate-receptor binding assay is more sensitive than the nonradioactive fluorescence polarization receptor assay 27,33 or solid-phase receptor-based assay 34 for the detection of cyclic imine toxins (Table 1). …”

Section: Resultsmentioning

confidence: 99%

Coupling theTorpedoMicroplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

et al. 2012

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Cyclic imine neurotoxins constitute an emergent family of neurotoxins of dinoflagellate origin that are potent antagonists of nicotinic acetylcholine receptors. We developed a target-directed functional method based on the mechanism of action of competitive agonists/antagonists of nicotinic acetylcholine receptors for the detection of marine cyclic imine neurotoxins. The key step for method development was the immobilization of Torpedo electrocyte membranes rich in nicotinic acetylcholine receptors on the surface of microplate wells and the use of biotinylated-α-bungarotoxin as tracer. Cyclic imine neurotoxins competitively inhibit biotinylated-α-bungarotoxin binding to Torpedo-nicotinic acetylcholine receptors in a concentration-dependent manner. The microplate-receptor binding assay allowed rapid detection of nanomolar concentrations of cyclic imine neurotoxins directly in shellfish samples. Although highly sensitive and specific for the detection of neurotoxins targeting nicotinic acetylcholine receptors as a class, the receptor binding assay cannot identify a given analyte. To address the low selectivity of the microplate-receptor binding assay, the cyclic imine neurotoxins tightly bound to the coated Torpedo nicotinic receptor were eluted with methanol, and the chemical nature of the eluted ligands was identified by mass spectrometry. The immobilization of Torpedo electrocyte membranes on the surface of microplate wells proved to be a high-throughput format for the survey of neurotoxins targeting nicotinic acetylcholine receptors directly in shellfish matrixes with high sensitivity and reproducibility.

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Section: Resultsmentioning

confidence: 99%

Coupling theTorpedoMicroplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

et al. 2012

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“…Gymnodimine A, 13-desmethyl spirolide C, and 13,19-didesmethyl spirolide C inhibited tracer binding in the nanomolar region using certified toxin standards and shellfish extracts. 134,135,136,137 Okadaic acid, yessotoxin, and brevetoxin-2 did not interfere with this assay. 134 The matrix effect of mussels, clams, cockles and scallops on the competitive fluorescence polarization assay was also assessed.…”

Section: Current Methods For Detection Of CI Toxinsmentioning

confidence: 83%

Synthesis and biology of cyclic imine toxins, an emerging class of potent, globally distributed marine toxins

et al. 2015

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From a small group of exotic compounds isolated only two decades ago, Cyclic Imine (CI) toxins have become a major class of marine toxins with global distribution. Their distinct chemical structure, biological mechanism of action, and intricate chemistry ensures that CI toxins will continue to be the subject of fascinating fundamental studies in the broad fields of chemistry, chemical biology, and toxicology. The worldwide occurrence of potent CI toxins in marine environments, their accumulation in shellfish, and chemical stability are important considerations in assessing risk factors for human health. This review article aims to provide an account of chemistry, biology, and toxicology of CI toxins from their discovery to the present day.

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“…Later on, the matrix effect of several mollusc species was analyzed, in a quantification range for 13-desMeC of 85-700 g kg −1 of shellfish meat [36]. Subsequent, this method was checked with 13,19-didesMeC [37]. The basis of all these previous assays is that the presence of the cyclic imines in solution inhibited the interaction of fluorescent-labelled alpha-bungarotoxin with membrane-enriched nAChRs in a concentration-dependent manner [20].…”

Section: Discussionmentioning

confidence: 99%

First direct fluorescence polarization assay for the detection and quantification of spirolides in mussel samples

Otero

Alfonso

Alfonso³

et al. 2011

Analytica Chimica Acta

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Detection of 13,19-didesmethyl C spirolide by fluorescence polarization using Torpedo electrocyte membranes

Cited by 26 publications

References 17 publications

Coupling theTorpedoMicroplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

Coupling theTorpedoMicroplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

Synthesis and biology of cyclic imine toxins, an emerging class of potent, globally distributed marine toxins

First direct fluorescence polarization assay for the detection and quantification of spirolides in mussel samples

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