Highly sensitive immunoassay based on a monoclonal antibody specific for [4-arginine]microcystins

Anne Zeck, Anja Eikenberg, Michael G. Weller, Reinhard Niessner

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93 Scopus citations

Abstract

The production and characterization of a monoclonal antibody (clone (MC10E7)) with extraordinary sensitivity and high selectivity for [4-arginine]microcystins is described. The immunogen used for the production of the antibody was synthesized using a novel coupling chemistry to bind microcystin-LR(MC-LR) via dehydroalanine to the carrier protein. With a direct competitive enzyme-linked immunosorbent assay (ELISA) using MC10E7, IC50 values for MC-LR of 0.06 μgl-1 have been achieved. The detection limit for MC-LR was 6ngl-1. The provisional guideline value proposed by the World Health Organization (WHO) is 1 μgl-1 for drinking water. All [4-arginine]microcystins show similar IC50 values and detection limits, whereas other MCs such as MC-LA, are not recognized. The affinity constant for MC10E7 was determined to be at least 7×1010lmol-1. The antibody was tested for its robustness against interference of humic acids, pH, salt content, surfactants or organic solvents and was found to be very stable. MC-LR spiked water samples in the concentration range between 0.01 and 0.1 μg1-1 were measured and a mean recovery of 99.9 ± 16.4% was found. The antibody is well suited for sensitive analysis for MCs in drinking as well as surface water.

Original languageEnglish
Pages (from-to)1-13
Number of pages13
JournalAnalytica Chimica Acta
Volume441
Issue number1
DOIs
StatePublished - 16 Aug 2001

Keywords

  • Algal toxin
  • Cyanobacteria
  • Direct competitive enzyme immunoassay
  • ELISA
  • Microcystin
  • Monoclonal antibody

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