Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for the Organophosphorus Insecticide <i>O</i>-Ethyl <i>O</i>-4-Nitrophenyl Phenylphosphonothioate (EPN)

et al. 2011

Analytica Chimica Acta

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An immunochromatographic assay (ICA) based on competitive antigen-coated format using colloidal gold as the label was developed for the detection of the organophosphorus insecticide chlorpyrifos. The ICA test strip consisted of a membrane with a detection zone, a sample pad and an absorbent pad. The membrane was separately coated with chlorpyrifos hapten-OVA conjugate (test line) and anti-mouse IgG (control line). Based on the fact that the competition is between the migrating analyte in the sample and the analyte hapten immobilized on the test strip for the binding sites of the antibody-colloidal gold (Ab-CG) conjugate migrating on the test strip, this study suggests that the relative migration speed between the two migrating substances is a critically important factor for the sensitive detection by competitive ICA. This criterion was utilized for the confirmation of appropriateness of a nitrocellulose (NC) membrane for chlorpyrifos ICA. The detection limit of the ICA for chlorpyrifos standard and chlorpyrifos spiked into agricultural samples were 10 and 50 ng mL−1, respectively. The assay time for the ICA test was less than 10 min, suitable for rapid on-site testing of chlorpyrifos.

“…Procedures for the production of monoclonal antibodies against the hapten-KLH conjugates were similar to those previously described in an earlier paper [43]. …”

Section: Methodsmentioning

confidence: 99%

“…The concentrations of the antibodies and the chosen coating antigen were further optimized. Additionally, optimization of the concentration of methanol and phosphate ion in assay mixture was carried out as described in previous papers [42, 43]. …”

Section: Methodsmentioning

confidence: 99%

Competitive immunochromatographic assay for the detection of the organophosphorus pesticide chlorpyrifos

et al. 2011

Analytica Chimica Acta

Self Cite

“…The capture antigen (EPN hapten-OVA) and the monoclonal anti-EPN antibody used were those prepared previously in this laboratory (Shim, Kim, Lee, Lee, & Lee, 2008;Shim, Kim, Lee, Hammock, & Lee, 2010). The haptens used for immunisation and capture antigen were Hapten A and Hapten B, respectively, presented in Figure 2.…”

Section: Capture Antigen and Antibodymentioning

confidence: 99%

Competitive immunochromatographic assay for the detection of the organophosphorus pesticide EPN

Food and Agricultural Immunology

et al. 2013

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An immunochromatographic assay (ICA) based on competitive antigen-coated format and colloidal gold label was developed for the detection of the organophosphorus insecticide O-ethyl O-4-nitrophenyl phenylphosphonothioate (EPN). The ICA test strip consisted of a membrane with a detection zone, a sample pad and an absorbent pad. The membrane was separately coated with EPN hapten-OVA conjugate (test line) and anti-mouse IgG (control line). As the competition is between the migrating analyte and the immobilised analyte hapten for the binding sites of the migrating antibodyÁcolloidal gold (AbÁCG) conjugate, this study suggests that the relative migration speed between the two migrating substances is a critically important factor for the sensitive detection by competitive ICA. Based on this criterion, a nitrocellulose (NC) membrane was chosen as the most suitable for EPN ICA. The detection limit of the ICA for EPN standard and EPN spiked into agricultural samples were 10 Á2 and 5 )10 Á2 mg mL (1 , respectively.

“…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).…”

Section: Introductionmentioning

confidence: 99%

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

Guo

Sanders

Galvita

et al. 2014

WMJ

(2014). Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues. World Mycotoxin Journal AbstractHapten heterology was introduced into the steps of hybridoma selection for the development of monoclonal antibodies (MAbs) against deoxynivalenol (DON). Firstly, a novel heterologous DON hapten was synthesized and covalently coupled to proteins (i.e., bovine serum albumin (BSA), ovalbumin and horseradish peroxidase) using the linkage of cyanuric chloride (CC). respectively. This is the first broad-specific MAb against DON and its two acetylated forms and thus it can be used for simultaneous detection of the three mycotoxins.