Preparation and characterization of an immunoaffinity column for the selective extraction of azaspiracids.

Autor: Samdal IA; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway. Electronic address: ingunn.samdal@vetinst.no., Sandvik M; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway., Vu J; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway; Oslo Metropolitan University, P.O. Box 4, St. Olavs plass, N-0130 Oslo, Norway., Sukenthirarasa MS; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway; Oslo Metropolitan University, P.O. Box 4, St. Olavs plass, N-0130 Oslo, Norway., Kanesamurthy S; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway; Oslo Metropolitan University, P.O. Box 4, St. Olavs plass, N-0130 Oslo, Norway., Løvberg KLE; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway., Kilcoyne J; Marine Institute, Rinville, Oranmore H91 R673, County Galway, Ireland., Forsyth CJ; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43220, United States., Wright EJ; Biotoxin Metrology, National Research Council of Canada, 1411 Oxford Street, Halifax, Nova Scotia B3H 3Z1, Canada., Miles CO; Norwegian Veterinary Institute, P.O. Box 64, 1431 Ås, Norway; Biotoxin Metrology, National Research Council of Canada, 1411 Oxford Street, Halifax, Nova Scotia B3H 3Z1, Canada.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2022 Sep 01; Vol. 1207, pp. 123360. Date of Electronic Publication: 2022 Jul 07.
DOI: 10.1016/j.jchromb.2022.123360
Abstrakt: The presence of azaspiracids (AZAs) in shellfish may cause food poisoning in humans. AZAs can accumulate in shellfish filtering seawater that contains marine dinoflagellates such as Azadinium and Amphidoma spp. More than 60 AZA analogues have been identified, of which AZA1, AZA2 and AZA3 are regulated in Europe. Shellfish matrices may complicate quantitation by ELISA and LC-MS methods. Polyclonal antibodies have been developed that bind specifically to the C-26-C-40 domain of the AZA structure and could potentially be used for selectively extracting compounds containing this substructure. This includes almost all known analogues of AZAs, including AZA1, AZA2 and AZA3. Here we report preparation of immunoaffinity chromatography (IAC) columns for clean-up and concentration of AZAs. The IAC columns were prepared by coupling polyclonal anti-AZA IgG to CNBr-activated sepharose. The columns were evaluated using shellfish extracts, and the resulting fractions were analyzed by ELISA and LC-MS. The columns selectively bound over 300 ng AZAs per mL of gel without significant leakage, and did not retain the okadaic acid, cyclic imine, pectenotoxin and yessotoxin analogues that were present in the applied samples. Furthermore, 90-92% of the AZAs were recovered by elution with 90% MeOH, and the columns could be re-used without significant loss of performance.
(Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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