Journal
ANALYTICAL BIOCHEMISTRY
Volume 447, Issue -, Pages 58-63Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2013.10.035
Keywords
Azaspiracid poisoning; Anti-azaspiracid antibody; Flow fluorimetry system
Funding
- Ministerio de Ciencia y Tecnologia, Spain (NEF) [SAF2009-12581, AGL2009 13581-CO2-01, TRA2009-0189, AGL2010-17875]
- Xunta de Galicia, Spain [GRC 2010/10, PGDIT 07MMA006261PR, PGIDIT (INCITE) 09MMA003261PR, PGDIT (INCITE) 09261080PR, 2009/XA044, 10PXIB261254 PR]
- EU VIIth Frame Program [211326-CP (COMMENCE), 265896 BAMMBO, 265409 AQUA, 262649 BEADS, 312184 PharmaSea]
- Atlantic Area Programme (Interreg IVB Trans-national), -1/117 Pharmatlantic
- National Institutes of Health (USA) [ESO13314]
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Azaspiracids (AZAs) are a group of lipophilic toxins discovered in mussels from Ireland in 1995 following a human poisoning incident. Nowadays the regulatory limit for AZAs in many countries is set at 160 mu g of azaspiracid equivalents per kilogram of shellfish meat. In this work a microsphere-based immunoassay has been developed for the detection of AZAs using a Luminex system. This method is based on the competition between AZA-2 immobilized onto the surface of microspheres and free AZAs for the interaction with a monoclonal anti-azaspiracid antibody (mAb 8F4). In this inhibition immunoassay the amount of mAb 8F4 bound to AZA-2 microspheres was quantified using a phycoerythrin-labeled anti-mouse antibody, and the fluorescence was measured with a Luminex analyzer. Simple acetate/methanol or methanol extractions yielded final extracts with no matrix interferences and adequate recovery rates of 86.5 and 75.8%, respectively. In summary, this work presents a sensitive and easily performed screening method capable of detecting AZAs at concentrations below the range of the European regulatory limit using a microsphere/flow cytometry system. (C) 2013 Elsevier Inc. All rights reserved.
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