期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 66, 期 4, 页码 950-956出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.7b04923
关键词
Cry1Ab toxin; anti-idiotypic camel nanobody; competitive chemiluminescent immunoassay
资金
- Ministry of Science and Technology of people's Republic of China [2016YFD0200803-3]
- NIEHS Superfund Research Program of USA [P42 ES004699]
- National Natural Science Foundation of China [31371778]
- Jiangsu Province Natural Science Foundation [BK20131333]
- Department of Science and technology of Jiangsu Province [BE2014722]
- Jiangsu Planned Projects for Postdoctoral Research Funds [1701030A]
- Department of Finance of Jiangsu Province [CX (15) 1011]
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.
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