Development of a Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Analysis of the Organophosphorous Pesticide Fenthion in Real Samples Based on Monoclonal Antibody

A new specific monoclonal antibody against the organophosphorous pesticide fenthion was produced based on our previous study. A sensitive and specific Enzyme-Linked Immunosorbent Assay (ELISA) for fenthion based on the new immunizing/coating hapten combination was developed. In this study, the H1 which attempts to expose the aromatic ring group was conjugated with bovine serum albumin (BSA) for the immunogen. All of the haptens that have been synthesized were conjugated with ovalbumin (OVA) for the coating antigen. The efficient antibody/coating conjugated combinations were selected, and the optimized ELISA was developed. In the optimized system, the IC50 value was 5.8 ng · mL−1 with the detection limit (IC20) of 0.028 ng · mL−1. The cross reactivity with all of the tested pesticides was lower than the 0.5%. Also, the system can detect the fenthion addition to the real samples such as soil, water, rice, and Chinese cabbages.

[1]  J. Skerritt,et al.  Mono- and polyclonal antibodies to the organophosphate fenitrothion. 1. Approaches to hapten-protein conjugation , 1992 .

[2]  J. H. Méndez,et al.  Determination of the pesticides fenthion and fenitrothion by flow injection with amperometric detection , 1988 .

[3]  B. Hammock,et al.  Development of a microtiter plate ELISA and a dipstick ELISA for the determination of the organophosphorus insecticide fenthion. , 2003, Journal of agricultural and food chemistry.

[4]  B. Hammock,et al.  Synthesis of haptens for immunoassay of organophosphorus pesticides and effect of heterology in hapten spacer arm length on immunoassay sensitivity , 2003 .

[5]  M. Tutudaki,et al.  Three-year study of fenthion and dimethoate pesticides in olive oil from organic and conventional cultivation , 2003, Food additives and contaminants.

[6]  J. M. Ortiz,et al.  Determination of fenthion and fenthion-sulfoxide, in olive oil and in river water, by square-wave adsorptive-stripping voltammetry. , 2008, Talanta.

[7]  C. Milstein,et al.  Continuous cultures of fused cells secreting antibody of predefined specificity , 1975, Nature.

[8]  Á. Maquieira,et al.  Enzyme-linked immunosorbent assay for the organophosphorus insecticide fenthion. Influence of hapten structure. , 2004, Journal of immunological methods.

[9]  Hye-Sung Lee,et al.  Oxidation of organophosphorus pesticides for the sensitive detection by a cholinesterase-based biosensor. , 2002, Chemosphere.

[10]  A. Garrido-Frenich,et al.  Reduction of analysis time in gas chromatography. Application of low-pressure gas chromatography-tandem mass spectrometry to the determination of pesticide residues in vegetables. , 2003, Journal of chromatography. A.

[11]  Guonian Zhu,et al.  Development of a McAb-based immunoassay for parathion and influence of the competitor structure , 2009 .

[12]  Y. Kim,et al.  Investigation of the effect of hapten heterology on immunoassay sensitivity and development of an enzyme-linked immunosorbent assay for the organophosphorus insecticide fenthion , 2003 .

[13]  J. Marty,et al.  Identification of fenthion and temephos and their transformation products in water by high-performance liquid chromatography with diode array detection and atmospheric pressure chemical ionization mass spectrometric detection , 1997 .

[14]  Xiao Liang,et al.  Development of a sensitive ELISA for the analysis of the organophosphorous insecticide fenthion in fruit samples , 2008 .

[15]  Yang Liu,et al.  A monoclonal antibody-based sensitive enzyme-linked immunosorbent assay (ELISA) for the analysis of the organophosphorous pesticides chlorpyrifos-methyl in real samples , 2009 .

[16]  K. Ahn,et al.  Hapten heterology for a specific and sensitive indirect enzyme-linked immunosorbent assay for organophosphorus insecticide fenthion. , 2007, Analytica chimica acta.

[17]  G. Raab Comparison of a logistic and a mass-action curve for radioimmunoassay data. , 1983, Clinical chemistry.

[18]  Peiwu Li,et al.  Effect of hapten structures on specific and sensitive enzyme-linked immunosorbent assays for N-methylcarbamate insecticide metolcarb. , 2008, Analytica chimica acta.