Development of an immunoassay for the detection of carbaryl in cereals based on a camelid variable heavy‐chain antibody domain
- 9 March 2019
- journal article
- research article
- Published by Wiley in Journal of the Science of Food and Agriculture
- Vol. 99 (9), 4383-4390
- https://doi.org/10.1002/jsfa.9672
Abstract
BACKGROUND The variable domain of camelid heavy‐chain antibodies (VHH) is increasingly being adapted to detect small molecules in various matrices. The insecticide carbaryl is widely used in agriculture while its residues have posed a threat to food safety and human health. RESULTS VHHs specific for carbaryl were generated from an alpaca immunized with the hapten CBR1 coupled to keyhole limpet hemocyanin. An enzyme‐linked immunosorbent assay (ELISA) based on the VHH C1 and the coating antigen CBR2‐BSA was developed for the detection of carbaryl in cereals. This assay, using an optimized assay buffer (pH 6.5) containing 10% methanol and 0.8% NaCl, has a half‐maximum signal inhibition concentration of 5.4 ng mL−1 and a limit of detection (LOD) of 0.3 ng mL−1 for carbaryl, and shows low cross reactivity (≤0.8%) with other tested carbamates. The LOD of carbaryl using the VHH‐based ELISA was 36 ng g−1 in rice and maize and 72 ng g−1 in wheat. Recoveries of carbaryl in spiked rice, maize and wheat samples were in the range of 81–106%, 96–106% and 83–113%, respectively. Relative standard deviations of repeatability and intra‐laboratory reproducibility were in the range of 0.8–9.2% and 2.9–9.7%, respectively. CONCLUSION The VHH‐based ELISA was highly effective in detecting carbaryl in cereal samples after simple sample extraction and dilution. © 2019 Society of Chemical IndustryKeywords
This publication has 39 references indexed in Scilit:
- Nanobodies as novel agents for disease diagnosis and therapyInternational Journal of Nanomedicine, 2013
- Nanobodies: Natural Single-Domain AntibodiesAnnual Review of Biochemistry, 2012
- A solid-phase microextraction-gas chromatographic approach combined with triple quadrupole mass spectrometry for the assay of carbamate pesticides in water samplesJournal of Chromatography A, 2012
- Isolation of Alpaca Anti-Hapten Heavy Chain Single Domain Antibodies for Development of Sensitive ImmunoassayAnalytical Chemistry, 2011
- An anti‐hapten camelid antibody reveals a cryptic binding site with significant energetic contributions from a nonhypervariable loopProtein Science, 2011
- Biotechnological applications of recombinant single-domain antibody fragmentsMicrobial Cell Factories, 2011
- Development of a sensitivity-improved immunoassay for the determination of carbaryl in food samplesJournal of the Science of Food and Agriculture, 2010
- Single domain antibodies: promising experimental and therapeutic tools in infection and immunityMedical Microbiology and Immunology, 2009
- Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs)Journal of Immunological Methods, 2007
- Validation of two immunoassay methods for environmental monitoring of carbaryl and 1-naphthol in ground water samplesAnalytica Chimica Acta, 1995