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

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• 作者：Zeck ; Anne ; Eikenberg ; Anja ; Weller ; Michael G. ; Niessner ; Reinhard
• 关键词：Adda ; (2S ; 3S ; 8S ; 9S)-3-amino-9-methoxy-2 ; 6 ; 8-trimethyl-10-phenyldeca-4 ; 6-dienoic acid ; BSA ; bovine serum albumin ; cOVA ; cationized ovalbumin ; EDC ; N-(3-dimethylaminopropyl)-N&prime ; -ethylcarbodiimide hydrochloride ; EDTA ; ethylenediaminetetraacetic acid ; El
• 刊名：Analytica Chimica Acta
• 出版年：2001
• 出版时间：August 16, 2001
• 年：2001
• 卷：441
• 期：1
• 页码：1-13
• 全文大小：139 K

文摘

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 0.06μgl⁻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 1μgl⁻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 0.1μgl⁻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.