Microcystin-LR detection based on indirect competitive enzyme-linked immunosorbent assay

2007, Volume 1, Issue 3

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Frontiers of Environmental Science & Engineering >> 2007, Volume 1, Issue 3 doi: 10.1007/s11783-007-0056-7

Microcystin-LR detection based on indirect competitive enzyme-linked immunosorbent assay

Environmental Simulation and Pollution Control (ESPC) State Key Joint Laboratory, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084, China;

Available online: 2007-09-05

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Abstract

Microcystins (MCs) are a group of closely related toxic cyclic heptapeptides produced by common cyanobacteria, which cause lots of accidents and threatens human health. In this paper, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established and used to detect microcystin-LR (MC-LR) in drinking and surface waters. The concentration of coating antigen was 5 ?g/mL, the dilution of monoclonal antibody MC10E7 was 1:3 000, the dilution of enzyme tracer (goat anti-mouse IgG-peroxidase) was 1:3 000, the standard concentration of MC-LR ranged from 0.001 μg/L to 30 μg/L, and o-phenylenediamine was used as substrate. The assay showed high relativity with high performance liquid chromatography (HPLC) with a correlation coefficient of more than 99%. The relative standard deviation was less than 10%, the detection limit was achieved down to 0.01 μg/L and up to 5.1 μg/L. The quantitative detection range was from 0.03 μg/L to 3 μg/L, and the antibody had high specificity for [4-arginine] microcystins. It performed well in spite of the influence of the real samples.

Keywords

o-phenylenediamine ; 4-arginine ; ic-ELISA ; substrate ; chromatography

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