A highly sensitive competitive enzyme immunoassay of broad specificity quantifying microcystins and nodularins in water samples

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• 作者：Nathalie Khreich ; Patricia Lamourette ; Pierre-Yves Renard ; Guillaume Clavé ; Franç ; ois Fenaille ; Christophe Cré ; minon ; Hervé ; Voll
• 关键词：Microcystins ; Nodularins ; Monoclonal antibody ; Enzyme Immunoassay ; HPLC ; LC-MS/MS
• 刊名：Toxicon
• 出版年：2009
• 出版时间：April 2009
• 年：2009
• 卷：53
• 期：5
• 页码：551-559
• 全文大小：285 K

文摘

Microcystins (MCs) form a group of cyclic heptapeptides produced by common cyanobacteria (blue green algae) and cause both acute and chronic toxicity. For immunization purposes, an amino derivative of MC-LR was prepared before coupling to BSA. Among the different monoclonal antibodies produced, mAb MC159 was selected due to its broad specificity to develop a sensitive enzyme immunoassay (EIA). This method measures MC-LR, MC-YR, MC-LA, nodularins in a similar way and exhibits an important recognition (cross reactivity up to 69 % ) for Adda analogues. Using MC-LR as standard, the present EIA proved to be very sensitive with a limit of detection close to 10 fmol/ml, largely below the provisional guideline level for drinking water proposed by the WHO (1 pmol/ml for MC-LR). This assay showed a high accuracy (CV %  < 12) and a high recovery rate for MC-LR in spiked surface water (up to 96.5 % ). Moreover due to its broad spectrum of recognition, this method allows a real quantification of the sum of MCs in water bloom and cyanobacteria culture samples. Indeed, in parallel analysis of these samples using HPLC, EIA shows a good relationship between both measurements while LC-MS/MS demonstrates the presence of different variants of MCs whose heterogeneity did not impair EIA measurement.