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Congener-independent immunoassay for microcystins and nodularins

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2001

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Fischer, Werner Jürgen
Garthwaite, Ian
Miles, Christopher O.
Ross, Kathryn Marie
Aggen, James B.
Chamberlin, A. Richard
Towers, Neale R.

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Environmental Science & Technology. 2001, 35(24), pp. 4849-4856. ISSN 0013-936X. eISSN 1520-5851. Available under: doi: 10.1021/es011182f

Zusammenfassung

Cyanobacteria (blue-green algae) (e.g., Microcystis and Nodularia spp.) capable of producing toxic peptides are found in fresh and brackish water worldwide. These toxins include the microcystin (MC) heptapeptides (>60 congeners) and the nodularin pentapeptides (ca. 5 congeners). Cyanobacterial cyclic peptide toxins are harmful to man, other mammals, birds, and fish. Acute exposure to high concentrations of these toxins causes liver damage, while subchronic or chronic exposure may promote liver tumor formation. The detection of cyclic peptide cyanobacterial toxins in surface and drinking waters has been hampered by the low limits of detection required and that the present routine detection is restricted to a few of the congeners. The unusual β-amino acid ADDA (4E,6E-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid) is present in most (>80%) of the known toxic penta- and heptapeptide toxin congeners. Here, we report the synthesis of two ADDA-haptens, the raising of antibodies to ADDA, and the development of a competitive indirect ELISA for the detection of microcystins and nodularins utilizing these antibodies. The assay has a limit of quantitation of 0.02-0.07 ng/mL (depending on which congeners are present), lower than the WHO-proposed guideline (1 ng/mL) for drinking water, irrespective of the sample matrix (raw water, drinking water, or pure toxin in PBS). This new ELISA is robust, can be performed without sample preconcentration, detects toxins in freshwater samples at lower concentrations than does the protein phosphatase inhibition assay, and shows very good cross-reactivity with all cyanobacterial cyclic peptide toxin congeners tested to date (MC-LR, -RR, -YR, -LW, -LF, 3-desmethyl-MC-LR, 3-desmethyl-MC-RR, and nodularin).

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570 Biowissenschaften, Biologie

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ISO 690FISCHER, Werner Jürgen, Ian GARTHWAITE, Christopher O. MILES, Kathryn Marie ROSS, James B. AGGEN, A. Richard CHAMBERLIN, Neale R. TOWERS, Daniel R. DIETRICH, 2001. Congener-independent immunoassay for microcystins and nodularins. In: Environmental Science & Technology. 2001, 35(24), pp. 4849-4856. ISSN 0013-936X. eISSN 1520-5851. Available under: doi: 10.1021/es011182f
BibTex
@article{Fischer2001Conge-6543,
  year={2001},
  doi={10.1021/es011182f},
  title={Congener-independent immunoassay for microcystins and nodularins},
  number={24},
  volume={35},
  issn={0013-936X},
  journal={Environmental Science & Technology},
  pages={4849--4856},
  author={Fischer, Werner Jürgen and Garthwaite, Ian and Miles, Christopher O. and Ross, Kathryn Marie and Aggen, James B. and Chamberlin, A. Richard and Towers, Neale R. and Dietrich, Daniel R.}
}
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    <dcterms:abstract xml:lang="eng">Cyanobacteria (blue-green algae) (e.g., Microcystis and Nodularia spp.) capable of producing toxic peptides are found in fresh and brackish water worldwide. These toxins include the microcystin (MC) heptapeptides (&gt;60 congeners) and the nodularin pentapeptides (ca. 5 congeners). Cyanobacterial cyclic peptide toxins are harmful to man, other mammals, birds, and fish. Acute exposure to high concentrations of these toxins causes liver damage, while subchronic or chronic exposure may promote liver tumor formation. The detection of cyclic peptide cyanobacterial toxins in surface and drinking waters has been hampered by the low limits of detection required and that the present routine detection is restricted to a few of the congeners. The unusual β-amino acid ADDA (4E,6E-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid) is present in most (&gt;80%) of the known toxic penta- and heptapeptide toxin congeners. Here, we report the synthesis of two ADDA-haptens, the raising of antibodies to ADDA, and the development of a competitive indirect ELISA for the detection of microcystins and nodularins utilizing these antibodies. The assay has a limit of quantitation of 0.02-0.07 ng/mL (depending on which congeners are present), lower than the WHO-proposed guideline (1 ng/mL) for drinking water, irrespective of the sample matrix (raw water, drinking water, or pure toxin in PBS). This new ELISA is robust, can be performed without sample preconcentration, detects toxins in freshwater samples at lower concentrations than does the protein phosphatase inhibition assay, and shows very good cross-reactivity with all cyanobacterial cyclic peptide toxin congeners tested to date (MC-LR, -RR, -YR, -LW, -LF, 3-desmethyl-MC-LR, 3-desmethyl-MC-RR, and nodularin).</dcterms:abstract>
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