Magnetic Particle-Based Enzyme Assays and Immunoassays for Microcystins: From Colorimetric to Electrochemical Detection.

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• 作者：Laia Revert茅 ; Diana Garibo ; Cintia Flores ; Jorge Diog猫ne ; Josep Caixach ; M貌nica Camp脿s
• 刊名：Environmental Science & Technology (ES&T)
• 出版年：2013
• 出版时间：January 2, 2013
• 年：2013
• 卷：47
• 期：1
• 页码：471-478
• 全文大小：261K
• 年卷期：v.47,no.1(January 2, 2013)
• ISSN：1520-5851

文摘

In this work, magnetic particles (MPs) are used as supports for the immobilization of biorecognition molecules for the detection of microcystins (MCs). In one approach, a recombinant protein phosphatase 1 (PP1) has been conjugated to MPs via coordination chemistry, and MC-LR detection has been based on the inhibition of the enzyme activity. In the other approach, a monoclonal antibody (mAb) against MC-LR has been conjugated to protein G-coated MPs, and a direct competitive enzyme-linked immunoparticle assay (ELIPA) has been then performed. Conjugation of biomolecules to MPs has been first checked, and after optimization, MC detection has been performed. The colorimetric PPIA with PP1-MP and the best ELIPA strategy have provided limits of detection (LOD) of 7.4 and 3.9 渭g/L of MC-LR, respectively. The electrochemical ELIPA has decreased the LOD to 0.4 渭g/L, value below the guideline recommended by the World Health Organisation (WHO). The approaches have been applied to the analysis of a cyanobacterial culture and a natural bloom, and MC equivalent contents have been compared to those obtained by conventional assays and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results have demonstrated the viability of the use of MPs as biomolecule immobilization supports in biotechnological tools for MCs monitoring.