Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal |
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| Authors: | Mei Shu Yang Xu Xing Liu Yanping Li Qinghua He Zhui Tu Jinheng Fu Shirley J Gee Bruce D Hammock |
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| Institution: | 1. State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People''s Republic of China;2. Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People''s Republic of China;3. Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616, USA;4. College of Food Science and Technology, Hainan University, No. 58 Renmin Avenue, Haikou 570228, China |
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| Abstract: | A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB1 was developed. The anti-idiotypic nanobody–alkaline phosphatase (Ab2β−Nb−AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB1 were 2.69 and 0.35 ng mL−1, respectively, with a linear range of 0.93–7.73 ng mL−1. The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL−1, and the IC50 was 0.89 ± 0.09 ng mL−1 with a linear range of 0.29–2.68 ng mL−1. Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β−Nb−AP fusion protein based one-step competitive immunoassay for monitoring FB1 contamination in cereals. The Ab2β−Nb−AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems. |
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| Keywords: | Anti-idiotypic antibody Nanobody One-step immunoassay |
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