Portable optical immunosensor for highly sensitive detection of microcystin-LR in water samples.

Fast and sensitive detection of microcystin-LR (MC-LR) was conducted with a portable trace organic pollutant analyzer (TOPA) based on the principle of immunoassay and total internal reflection fluorescence. The reusable fiber optic probe surface was produced by covalently immobilizing a MC-LR-ovalbumin (MC-LR-OVA) conjugate onto a self-assembled thiol-silane monolayer of fiber optic probe through a heterobifunctional reagent. It has been established that the MC-LR-OVA immobilized fiber optic probe is highly resistive to non-specific binding of proteins, while bound anti-MC-LR antibody can easily be eluted from the immunosurface with high recoveries (more than 150 assay cycles) in the way of pepsin solution, without any damage to the surface-immobilized MC-LR derivatives. One assay cycle including surface regeneration was less than 20 min. For the standard curve, the limit of detection (LOD) of 0.03 microg/L and the quantitative detection range of 0.1-10.1 microg/L was obtained when the concentration of antibody labeled by Cy5.5 was 0.28 microg/mL. Cross-reactivity against a few compounds structurally similar to MC-LR was little. The developed immunosensor method was applied to the monitoring of MC-LR in various types of water. The recovery of MC-LR added to water samples at different concentrations ranged from 80 to 110% with relative standard deviation (R.S.D.) values less than 5%. The immunoassay performance of the TOPA was validated with respect to conventional high-performance liquid chromatography, and the correlation between methods was in good agreement (r(2)=0.9978). The TOPA is a portable, easy-to-use, and robust immunoassay system and commercially obtained from the company JQ-environ Co. Ltd. (China).

[1]  L M Lechuga,et al.  Determination of carbaryl in natural water samples by a surface plasmon resonance flow-through immunosensor. , 2006, Biosensors & bioelectronics.

[2]  Volker Schurig,et al.  Chiral discrimination using an immunosensor , 1999, Nature Biotechnology.

[3]  Kiyoshi Toko,et al.  Fabrication of a novel immunosensor using functionalized self-assembled monolayer for trace level detection of TNT by surface plasmon resonance. , 2007, Talanta.

[4]  M. Hennion,et al.  Determination of some physicochemical parameters of microcystins (cyanobacterial toxins) and trace level analysis in environmental samples using liquid chromatography. , 1998, Journal of chromatography. A.

[5]  David W. Conrad,et al.  Detection of TNT in Water Using an Evanescent Wave Fiber-Optic Biosensor , 1995 .

[6]  M. He,et al.  Development of evanescent wave all-fiber immunosensor for environmental water analysis. , 2008, Biosensors & bioelectronics.

[7]  Lise,et al.  LIVER FAILURE AND DEATH AFTER EXPOSURE TO MICROCYSTINS AT A HEMODIALYSIS CENTER IN BRAZIL LIVER FAILURE AND DEATH AFTER EXPOSURE TO MICROCYSTINS AT A HEMODIALYSIS CENTER IN BRAZIL , 2000 .

[8]  Vicki S. Thompson,et al.  Fiber-Optic Immunosensor for the Detection of Fumonisin B1 , 1996 .

[9]  Norio Miura,et al.  A simple and versatile self-assembled monolayer based surface plasmon resonance immunosensor for highly sensitive detection of 2,4-D from natural water resources , 2008 .

[10]  M. Hennion,et al.  Development of a bioanalytical phosphatase inhibition test for the monitoring of microcystins in environmental water samples , 1999 .

[11]  K. Beattie,et al.  Colorimetric protein phosphatase inhibition assay of laboratory strains and natural blooms of cyanobacteria: comparisons with high-performance liquid chromatographic analysis for microcystins. , 1997, FEMS microbiology letters.

[12]  C. MacKintosh,et al.  Purification of type 1 protein (serine/threonine) phosphatases by microcystin‐Sepharose affinity chromatography , 1994, FEBS letters.

[13]  K. O’Shea,et al.  Mechanistic study and the influence of oxygen on the photosensitized transformations of microcystins (cyanotoxins). , 2007, Environmental science & technology.

[14]  C. J. Hastie,et al.  Inhibition of several protein phosphatases by a non-covalently interacting microcystin and a novel cyanobacterial peptide, nostocyclin. , 2005, Biochimica et biophysica acta.

[15]  W. Carmichael,et al.  Use of a colorimetric protein phosphatase inhibition assay and enzyme linked immunosorbent assay for the study of microcystins and nodularins. , 1994, Toxicon : official journal of the International Society on Toxinology.

[16]  Jamie Bartram,et al.  Toxic Cyanobacteria in Water: a Guide to Their Public Health Consequences, Monitoring and Management Chapter 2. Cyanobacteria in the Environment 2.1 Nature and Diversity 2.1.1 Systematics , 2022 .

[17]  M. He,et al.  Sensitive and rapid detection of 2,4-dicholorophenoxyacetic acid in water samples by using evanescent wave all-fiber immunosensor. , 2008, Biosensors & bioelectronics.

[18]  C. Fanali,et al.  Monitoring algal toxins in lake water by liquid chromatography tandem mass spectrometry. , 2006, Environmental science & technology.

[19]  María Marazuela,et al.  Fiber-optic biosensors – an overview , 2002, Analytical and bioanalytical chemistry.

[20]  Miao He,et al.  A comprehensive immunoassay for the detection of microcystins in waters based on polyclonal antibodies. , 2006, Analytica chimica acta.

[21]  B. Nicholson,et al.  Determination of cyanobacterial hepatotoxins directly in water using a protein phosphatase inhibition assay. , 2001, Water research.

[22]  L. Lawton,et al.  Comparative assessment of the specificity of the brine shrimp and microtox assays to hepatotoxic (microcystin‐LR‐containing) cyanobacteria , 1994 .

[23]  S. Suzuki,et al.  A clean-up method for analysis of trace amounts of microcystins in lake water. , 1994, Toxicon : official journal of the International Society on Toxinology.

[24]  F. Yu,et al.  Monitoring of microcystin-protein phosphatase adduct formation with immunochemical methods. , 2000, Toxicon : official journal of the International Society on Toxinology.

[25]  Guoqing Hu,et al.  Modeling micropatterned antigen-antibody binding kinetics in a microfluidic chip. , 2007, Biosensors & bioelectronics.

[26]  J M Calvert,et al.  Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces. , 1989, Analytical biochemistry.

[27]  Jean-Louis Marty,et al.  Highly sensitive amperometric immunosensors for microcystin detection in algae. , 2007, Biosensors & bioelectronics.

[28]  Hans Wolf,et al.  Development of a highly sensitive inhibition immunoassay for microcystin-LR , 2004 .

[29]  C. J. Lewis,et al.  Cyanine dye labeling reagents: sulfoindocyanine succinimidyl esters. , 1993, Bioconjugate chemistry.

[30]  Wolfgang Schade,et al.  Fiber-optic evanescent-field laser sensor for in-situ gas diagnostics. , 2002, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[31]  Liqiang Xie,et al.  Determination of microcystins in fish tissues using HPLC with a rapid and efficient solid phase extraction , 2007 .

[32]  L. Lawton,et al.  Detection of the cyanobacterial hepatotoxins microcystins. , 2005, Toxicology and applied pharmacology.

[33]  J. Homola.  Present and future of surface plasmon resonance biosensors , 2003, Analytical and bioanalytical chemistry.

[34]  D Barceló,et al.  Analysis of bisphenol A in natural waters by means of an optical immunosensor. , 2005, Water research.

[35]  W. Carmichael,et al.  Human Fatalities from Cyanobacteria: Chemical and Biological Evidence for Cyanotoxins , 2001 .