Development of a new broad-specific monoclonal antibody with uniform affinity for aflatoxins and magnetic beads-based enzymatic immunoassay |
| |
| Affiliation: | 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;1. Graduate Institute of Engineering Technology Doctoral, National Taipei University of Technology, Taipei, Taiwan;2. Graduate Institute of Biochemical and Biomedical Engineering, Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan;3. Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan;4. Graduate Institute of Physiology, National Defense Medical Center, Taipei, Taiwan;1. Key Laboratory of Applied Organic Chemistry (College of Jiangxi Province), Department of Chemistry, Shangrao Normal University, Shangrao 334001, Jiangxi, PR China;2. Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China;1. Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China;2. Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, People’s Republic of China;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 |
| |
| Abstract: | To reduce the incidence of false-positive and false-negative results caused by high or low cross-reactivity (CR%) values of the antibodies for total aflatoxins (AFs, AFB1+AFB2+AFG1+AFG2) detection, a new broad-specific monoclonal antibody (MAb) with uniform affinity, named 5H3, was developed. Moreover, magnetic beads (MBs) replaced microplates as immobile phase to improve the sensitivity of the enzymatic immunoassay. Then, a direct competitive enzyme-linked immunosorbent assay (ELISA) based on MBs (MBs-dcELISA) that could simultaneously detect the total AFs with similar sensitivity was developed. Following optimization of conditions, the half maximal inhibitory concentrations (IC50) of the MBs-dcELISA in buffer were 0.05 ng/mL for AFB1, 0.04 ng/mL for AFB2, 0.05 ng/mL for AFG1, 0.06 ng/mL for AFG2. The corresponding CR% values were 100%, 125%, 100% and 83.3%, respectively. The limit of detection (LOD) of the MBs-dcELISA for the total AFs was 0.21 ng/g with a working range from 0.22 ng/g to 19.8 ng/g, and the recoveries for the total AFs ranged from 74.5% to 96.5% with coefficients of variation (CV) under 12.1% in spiked maize samples. In addition, the MBs-dcELISA was more sensitive than the conventional dcELISA. Finally, the MBs-dcELISA was applied to screen 9 naturally contaminated maize samples and 6 spiked samples and the results indicated a good agreement with that obtain by HPLC-MS/MS method. |
| |
| Keywords: | Monoclonal antibody Aflatoxins Magnetic beads Direct competitive enzyme-linked immunosorbent assay |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
