Measurement of Microcystin and Nodularin Activity in Human Urine by Immunocapture-Protein Phosphatase 2A Assay

Autor:	Sophia M. Guldberg, Elizabeth I. Hamelin, Rudolph C. Johnson, Rebekah E. Wharton, Brady R. Cunningham, Adam M. Schaefer
Rok vydání:	2019
Předmět:	Cyanobacteria Health Toxicology and Mutagenesis Microcystin Urine Nod Toxicology Peptides Cyclic Algal bloom Article Microbiology 03 medical and health sciences chemistry.chemical_compound Humans Protein Phosphatase 2 Quantitation Range harmful algal bloom 030304 developmental biology Immunoassay chemistry.chemical_classification 0303 health sciences biology protein phosphatase 2A 030302 biochemistry & molecular biology microcystins biology.organism_classification Nodularin chemistry nodularin biology.protein immunocapture Antibody
Zdroj:	Toxins Volume 11 Issue 12
ISSN:	2072-6651
DOI:	10.3390/toxins11120729
Popis:	Microcystins (MC) and nodularin (NOD) are toxins released by cyanobacteria during harmful algal blooms. They are potent inhibitors of protein phosphatases 1 and 2A (PP1 and PP2A) and cause a variety of adverse symptoms in humans and animals if ingested. More than 250 chemically diverse congeners of MCs have been identified, but certified reference materials are only available for a few. A diagnostic test that does not require each reference material for detection is necessary to identify human exposures. To address this need, our lab has developed a method that uses an antibody to specifically isolate MCs and NOD from urine prior to detection via a commercially available PP2A kit. This assay quantitates the summed inhibitory activity of nearly all MCs and NOD on PP2A relative to a common MC congener, microcystin-LR (MC-LR). The quantitation range for MC-LR using this method is from 0.050&ndash 0.500 ng/mL. No background responses were detected in a convenience set of 50 individual urines. Interday and intraday % accuracies ranged from 94%&ndash 118% and relative standard deviations were 15% or less, meeting FDA guidelines for receptor binding assays. The assay detected low levels of MCs in urines from three individuals living in close proximity to harmful algal blooms (HABs) in Florida.
Databáze:	OpenAIRE
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