An improved immunoassay for chlorpyrifos‐methyl (Reldan) in grain

Edward, Simone L.; Skerritt, John H.; Hill, A. V.; McAdam, David P.

doi:10.1080/09540109309354792

Cited by 23 publications

(13 citation statements)

References 17 publications

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“…16,17 This method was applied to the synthesis of haptens for the development of ELISAs of several OP pesticides. [4][5][6][7][8][9][10][11][12] This method requires a synthetic route involving seven steps including protection and deprotection of both amino and carboxyl groups. In an effort to simplify the synthetic process for this class of haptens, we developed a simpler general method, which requires only two steps.…”

Section: Resultsmentioning

confidence: 99%

“…One type of hapten for OP pesticides is the one with an aminocarboxylic acid bridge at the thiophosphate group, which has been used for the development of ELISA for several OP pesticides. [4][5][6][7][8][9][10][11][12] We have developed a simple method for the synthesis of such haptens, which requires far fewer steps and can provide a wider range of hapten structures. [4][5][6] We previously applied this method to the synthesis of several OP pesticides including chlorpyrifos, 4 isofenphos, 5 and cyanophos.…”

Section: Introductionmentioning

confidence: 99%

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Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Fungicide Tolclofos-methyl

2003

Bulletin of the Korean Chemical Society

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A simple synthetic method for haptens of organophosphorus (OP) pesticides with a spacer arm (aminocarboxylic acid) attached at the pesticide thiophosphate group was developed and was applied to the synthesis of haptens for the OP fungicide tolclofos-methyl. Using the haptens, a selective enzyme-linked immunosorbent assay (ELISA) for tolclofos-methyl was developed. One of the haptens was coupled to BSA to use as an immunogen. Rabbits were immunized with this conjugate to obtain polyclonal antibodies to tolclofos-methyl. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the serum with highest specificity, an antigen-coated ELISA was developed, which showed an IC 50 of 160 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivity with other OP pesticides. An antibody-coated ELISA was also developed, which showed an IC 50 of 410 ng/mL with a detection limit of 130 ng/mL.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Fungicide Tolclofos-methyl

2003

Bulletin of the Korean Chemical Society

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“…Although antibodies for detecting organophosphorus compounds in vitro are known [26-30], antibodies to OP-adducted proteins have not been widely reported [30-32]. Indirectly, immunoprecipitation of OP-protein targets using antibodies to butyrylcholinesterase followed by digestion and mass spectral characterization of the OP-modified peptide has been applied to address the problem [20, 33-37].…”

Section: Introductionmentioning

confidence: 99%

Preparation and characterization of diethoxy- and monoethoxy phosphylated (‘aged’) serine haptens and use in the production of monoclonal antibodies

Belabassi¹,

Chao²,

Holly³

et al. 2014

Chemico-Biological Interactions

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In this study, the first mechanism-based monoclonal antibodies have been produced that recognize and differentiate diethoxy- and monoethoxyphosphorylated serine residues. Haptens were synthesized as the stable phosphonate form of phosphoserine esters to improve the immunoresponse. Following condensation with a glutaric anhydride to link the phosphoserine moieties to carrier protein, the hapten densities attached to bovine serum albumin and keyhole limpet henocyananin were determined by partial trypsin digestion and MALDI mass spectrometry, and confirmed using a fluorescent assay (FITC) to quantify unmodified lysine residues. The conjugation reactions were pH optimized to improve hapten density. Screening of subclones led to the identification of two monoclonal antibodies: (a) N257/25.11 that specifically recognizes (EtO)2P(O)-Ser as the phosphylated or inhibited form, and (b) N262/16 that recognizes (EtO)(HO)P(O)-Ser as the ‘aged’ form. Analysis of blood samples treated with paraoxon (EtO)2P(O)-OPhNO2 showed a concentration dependent recognition of the phosphylated form.

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“…Since OPs are all small molecules, hapten must be synthesized and coupled to carrier protein to induce animals to produce antibody. The haptens derived through the thiophosphate moiety using an amino carboxylic acid as a spacer arm have been reported in several articles to obtain highly sensitive antibodies for paraoxon, [7] fenitrothion, [8] chlorpyrifos, [9,10] chlorpyrifos-methyl, [11,12] and pirimiphos-methyl. [8] The aim of this work is to develop an ELISA for the determination of some OPs involving…”

Section: Introductionmentioning

confidence: 99%

Development of IC-ELISA for detection of organophosphorus pesticides in water

Tang

Zhang

Cheng

et al. 2008

Journal of Environmental Science and Health, Part B

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Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC50 of 0.428 micro g/mL with a detection limit of 0.0125 micro g/mL to parathion. The assay also indicated that the IC50 values of pirimiphos and dichlorvos were 0.331 micro g/mL and 1.25 micro g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 micro g/mL and 0.048 micro g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples.

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An improved immunoassay for chlorpyrifos‐methyl (Reldan) in grain

Cited by 23 publications

References 17 publications

Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Fungicide Tolclofos-methyl

Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Fungicide Tolclofos-methyl

Preparation and characterization of diethoxy- and monoethoxy phosphylated (‘aged’) serine haptens and use in the production of monoclonal antibodies

Development of IC-ELISA for detection of organophosphorus pesticides in water

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