Development of novel monoclonal antibodies-based ultrasensitive enzyme-linked immunosorbent assay and rapid immunochromatographic strip for aflatoxin B1 detection |
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| Affiliation: | 1. School of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan;2. Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan;3. Department of Medical Research, Chung Shan Medical University Hospital, Taichung, Taiwan;1. School of Biotechnology and Food Engineering, Anhui Provincial Key Lab of Functional Materials and Devices, Hefei University of Technology, Hefei 23009, China;2. Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing 200002, China;3. Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58102, USA;1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;2. Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, China;3. Jiangxi Zodolabs Bioengineering Co, Ltd., Nanchang 330047, China;1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, 100193, Beijing, People''s Republic of China;2. Department of Genetic Toxicology, Shenzhen Center for Disease Control and Prevention, 518020, Shenzhen, People''s Republic of China;2. Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Assiut Branch, 71524 Assiut, Egypt;3. College of Applied Medical Sciences, Medical laboratories Department, Majmaah University, Kingdom of Saudi Arabia;4. Chemistry Department, Faculty of Science, Al-Azhar University, Assiut Branch, 71524 Assiut, Egypt |
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| Abstract: | Monoclonal antibodies (mAbs) that are specific to aflatoxin B1 (AFB1) were produced from hybridoma cell lines 3F6G11 and 9C7C11 by the fusion of P3/NS1/1-AG4-1 myeloma cells with spleen cells that were isolated from a BALB/c mouse that was immunized with AFB1-bovine serum albumin (BSA). Both 3F6G11 and 9C7C11 mAbs are the immunoglobulin G1 isotypes. Competitive direct enzyme-linked immunosorbent assays (cdELISA) were established to characterize these antibodies. In the 3F6G11 mAb based cdELISA, the concentrations causing 50% inhibition of binding of AFB1-horseradish peroxidase to the antibody by AFB1, AFB2, AFG1, and AFG2 were found to be 0.051, 0.050, 1.820, and 1.270 ng/ml, respectively. Using 9C7C11 mAbs, similar IC50 values for AFB1, AFB2, AFG1 and AFG2 were obtained as 0.045, 0.057, 2.530 and 2.120 ng/ml, respectively. A rapid and sensitive gold nanoparticle immunochromatographic strip (immunostrip) was also established for these antibodies. This strip has a detection limit of 1.0 ng/ml for AFB1 and the whole assay can be completed within 10 min. Extensive analysis of 20 samples by 3F6G11 mAb and 9C7C11 mAbs cdELISAs revealed that six samples were slightly contaminated by AFB1 at concentrations from 0.160 to 16.10 ng/g. Results of analyses of 20 samples with an immunostrip assay correlate well with those obtained using cdELISA. The proposed cdELISA and immunostrip methods are highly sensitive for the rapid screening of AFB1 in food samples. |
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| Keywords: | Aflatoxin B1 ELISA Gold nanoparticle immunochromatographic strip Monoclonal antibody |
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