Ochratoxins A, B, and C (OTA, OTB, and OTC) can be found in cereals and feeds; the simultaneous detection of these ochratoxins holds a great need in food safety. In this study, four antibodies raised from two ochrotoxin haptens and two coating antigens were compared, and then a sensitive and broad-specificity enzyme-linked immunosorbent assay (ELISA) was established for the simultaneous determination of three ochratoxins, where the detection limits were 0.005, 0.001, and 0.001 ng/mL for OTA, OTB, and OTC, respectively, and recoveries of three ochratoxins were between 84.3% and 111.7%. This developed method had been successfully applied to detect ochratoxins in both millet and maize. Molecular modeling revealed that the broad-specificity was related with the chlorine electronegativity on OTA and OTC and the potential of the acetyl ester group on OTC. The proposed ELISA can be used for simultaneous detection of three ochratoxins.
Antibody-based immunoassay methods have been important tools for monitoring drug residues in animal foods. However, because of limited knowledge about the quantitative structure-activity relationships between a hapten and its resultant antibody specificity, antibody production with the desired specificity is still a huge challenge. In this study, the three-dimensional quantitative structure-activity relationship (3D QSAR) was analyzed in accordance with the cross-reactivity of quinolone drugs reacting with the antibody raised by pipemidic acid as the immunizing hapten and compared with the reported cross-reactivity data and their hapten structures. It was found that the specificity of a quinolone antibody was strongly related to the conformation of the hapten used and that hapten conformations shaped like the letters "I", "P", and "Φ" were essential for the desired high specificity with low cross-reactivity, but that the hapten conformation shaped like the letter "Y" led to an antibody with broad specificity and high cross-reactivity. Almost all of the antibodies against quinolones could result from these four hapten conformations. It was first found that the concrete conformations dominated the specificity of the antibody to quinolone, which will be of significance for the accurate hapten design, predictable antibody specificity, and better understanding the recognition mechanism between haptens and the antibodies for immunoassays.
Immunoassay methods are important for monitoring β-agonists illegally used for reducing animal fat deposition in livestock. However, there is no simultaneous screening surveillance immunoassay for detecting various β-agonist chemicals that are possibly present in food. In this study, through the use of an R-(-)-salbutamol derivative as the immunizing hapten, an antibody recognizing 31 β-agonists and analogues was generated for the first time. Three-dimensional quantitative structure-activity relationship (3D QSAR) revealed that strong steric and hydrophobic fields around the hapten spacer near C-2, as well as a chirality at C-1', dominantly modulated the class specificity of the raised antibody. However, a hapten spacer linked at C-2' or C-1 would lead to a narrow specificity, and the spacer charge at C-6 could affect the raised antibody specificity spectrum. A class specificity competitive indirect enzyme-linked immunosorbent assay (ciELISA) was established with an ideal recovery ranging from 81.8 to 118.3% based on the obtained antibody. With a good agreement to the HPLC/MS method, the proposed ciELISA was confirmed to be reliable for the rapid surveillance screening assay of β-agonists in urine. This investigation will contribute to the rational design and control of the immunoassay specificity.
Environmental hormones, also called environmental endocrine-disrupting chemicals (EDCs), are produced by various human activities and environment pollution, and they are composed of industrial chemicals and environmental pollutants. They play a harmful role in breaking the balance of normal physiological metabolism, causing serious diseases and disrupting reproductive development in wildlife and human beings by inhibiting or imitating the action of normal gonadal hormones. Strict controls of maximum residue levels to some environmental hormones in foodstuff have been set up by many countries. Traditional detection methods, due to their high accuracy and maturity, are used extensively, such as HPLC, HPLC-MS, and GC-MS. However, these methods are time consuming and require extraordinary skills of operators, and sometimes they cannot meet the requirements of field testing. Immunoassay has the advantages of high specificity, sensitivity, simplicity, convenience, and the ability to achieve high throughput, playing an irreplaceable role in the field of rapid detection. This review focuses on the antibody production and the development of immunoassays to detect environmental hormones in food and environment. Therefore, estrogen, progesterone, and testosterone are selected as examples to introduce a series of procedures for the detection of environmental hormones including antigen synthesis, antibody production, and antibody-based detection methods.
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