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National Institute of Environmental Health Sciences

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Your Environment. Your Health.

Publication Detail

Title: Phage-Mediated Competitive Chemiluminescent Immunoassay for Detecting Cry1Ab Toxin by Using an Anti-Idiotypic Camel Nanobody.

Authors: Qiu, Yulou; Li, Pan; Dong, Sa; Zhang, Xiaoshuai; Yang, Qianru; Wang, Yulong; Ge, Jing; Hammock, Bruce D; Zhang, Cunzheng; Liu, Xianjin

Published In J Agric Food Chem, (2018 Jan 31)

Abstract: Cry toxins have been widely used in genetically modified organisms for pest control, raising public concern regarding their effects on the natural environment and food safety. In this work, a phage-mediated competitive chemiluminescent immunoassay (c-CLIA) was developed for determination of Cry1Ab toxin using anti-idiotypic camel nanobodies. By extracting RNA from camels' peripheral blood lymphocytes, a naive phage-displayed nanobody library was established. Using anti-Cry1Ab toxin monoclonal antibodies (mAbs) against the library for anti-idiotypic antibody screening, four anti-idiotypic nanobodies were selected and confirmed to be specific for anti-Cry1Ab mAb binding. Thereafter, a c-CLIA was developed for detection of Cry1Ab toxin based on anti-idiotypic camel nanobodies and employed for sample testing. The results revealed a half-inhibition concentration of developed assay to be 42.68 ± 2.54 ng/mL, in the linear range of 10.49-307.1 ng/mL. The established method is highly specific for Cry1Ab recognition, with negligible cross-reactivity for other Cry toxins. For spiked cereal samples, the recoveries of Cry1Ab toxin ranged from 77.4% to 127%, with coefficient of variation of less than 9%. This study demonstrated that the competitive format based on phage-displayed anti-idiotypic nanobodies can provide an alternative strategy for Cry toxin detection.

PubMed ID: 29293334 Exiting the NIEHS site

MeSH Terms: Animals; Antibodies, Anti-Idiotypic*; Antibodies, Monoclonal; Bacterial Proteins/analysis*; Bacterial Proteins/immunology; Bacteriophages*; Camelus/blood; Camelus/immunology*; Edible Grain/chemistry; Endotoxins/analysis*; Endotoxins/immunology; Food Contamination/analysis; Hemolysin Proteins/analysis*; Hemolysin Proteins/immunology; Luminescent Measurements/methods*; Lymphocytes/chemistry; Peptide Library; RNA/isolation & purification; Single-Domain Antibodies*/genetics

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