Development of a Mab-Based Heterologous Immunoassay for the Broad-Selective Determination of Organophosphorus Pesticides

Abstract

A broad-selective monoclonal antibody (Mab) for organophosphorus (OP) pesticides was raised using heterologous indirect enzyme-linked immunosorbent assay (ELISA) to screen hybridomas. On the basis of this Mab, five coating antigens were used to develop homologous and heterologous indirect competitive ELISAs. With the most suitable competitor, a sensitive and broad-selective ELISA was developed. The IC 50 values were estimated to be 20.32 ng/mL for parathion, 21.44 ng/mL for methyl-parathion, 42.15 ng/mL for fenitrothion, and 58.85 ng/mL for isocarbophos. Spike recoveries were between 70.52 and 103.27% for the detection of single pesticide residues of the four OP pesticides in purple-clayed paddy soil. Moreover, the chosen ELISA was then applied to the detection of mixtures of parathion and methyl-parathion in soil samples. The average recovery and coefficient of variation were 80.91 and 4.82%, respectively. Results proved that this broad-selective ELISA would be useful for the multiresidue determination of OP pesticides.

Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues

“…The broad-selectivity of this MAb was probably due to the involved strategy of heterologous screening for hybridomas instead of the homologous one. Previous researches Muldoon et al, 2000;Wang et al, 2010) also indicated that application of this screening strategy increased the likelihood of isolating hybridomas secreting very broad-specific MAbs.…”

“…Since hapten heterology is often employed to modify the performance of competitive immunoassay for small molecules (Kim et al, 2003;Wang et al, 2011), it can also be introduced into the steps of hybridoma selection. To date, using heterologous ELISAs for hybridoma screening is yet limited to very few kinds of MAb generation Muldoon et al, 2000;Shim et al, 2010;Wang et al, 2010).…”

Immunodiagnostics and immunosensor design (IUPAC Technical Report)

“…Some examples of applications involving small and environmentally important toxic substances are: -cadmium [206] -PAHs, PCBs [207], dioxins [208] -organophosphorus pesticides [209,210] -pyrethroid pesticides in water [211] -mycotoxins [212] and bacterial toxins [213] -the wood preservative pentachlorphenol (PCP) in soil [214] -explosives [215] -chemical warfare agents [216] …”

A highly sensitive and class-specific fluorescence polarisation assay for sulphonamides based on dihydropteroate synthase

“…To achieve this goal, the generation of an antibody with class-specificity is necessary, in addition to the development of multi-label and multiplex techniques (Franek et al, 2006). Until now, extensive effort has been devoted to developing antibodies that are class-specific to chemical contaminants, e.g., sulphonamides (Adrian et al, 2009;Pastor-Navarro et al, 2009), fluoroquinolones (Pinacho et al, 2012;Wang et al, 2007) and organophosphates (Piao et al, 2009;Wang et al, 2010).…”