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Open AccessArticle

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

1
Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA
2
Harbor Branch Oceanographic Institute, Florida Atlantic University, Ft. Pierce, FL 34946, USA
3
Oak Ridge Institute for Science, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA
*
Author to whom correspondence should be addressed.
Toxins 2019, 11(12), 729; https://doi.org/10.3390/toxins11120729
Received: 18 October 2019 / Revised: 6 December 2019 / Accepted: 11 December 2019 / Published: 13 December 2019
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 0.050–0.500 ng/mL. No background responses were detected in a convenience set of 50 individual urines. Interday and intraday % accuracies ranged from 94%–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. View Full-Text
Keywords: microcystins; nodularin; protein phosphatase 2A; immunocapture; harmful algal bloom microcystins; nodularin; protein phosphatase 2A; immunocapture; harmful algal bloom
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MDPI and ACS Style

Wharton, R.E.; Cunningham, B.R.; Schaefer, A.M.; Guldberg, S.M.; Hamelin, E.I.; Johnson, R.C. Measurement of Microcystin and Nodularin Activity in Human Urine by Immunocapture-Protein Phosphatase 2A Assay. Toxins 2019, 11, 729.

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