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      Synthesis of Zearalenone Immunogen and Comparative Analysis of Antibody Characteristics

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          Abstract

          Background

          This study aimed to explore the zearalenone (ZEN) immunogen synthesis method, immunogenicity, and antibody characteristics and to lay a foundation for the establishment of immunoassay methods for ZEN single residue and ZEN and its analogs total residue.

          Methods

          Based on the molecular structure and active sites of ZEN, oxime active ester (OAE), condensation mixed anhydride (CMA), formaldehyde (FA), and 1,4-butanediol diglycidyl ether method (BDE) were designed and used for immunogen (ZEN-BSA) synthesis. The immunogens were identified by infrared (IR) and ultraviolet (UV) spectra and gel electrophoresis (SDS-PAGE) and were then used to immunize Balb/c mice to prepare ZEN polyclonal antibody (ZEN pAb). The titers and sensitivity of the ZEN pAb were determined by indirect noncompetitive ELISA (inELISA) and indirect competitive ELISA (icELISA), respectively, and its specificity was assessed by the cross-reaction test (CR).

          Results

          ZEN-BSA was successfully synthesized, and the molecular binding ratios of ZEN to BSA were 17.2 : 1 (OAE), 14.6 : 1 (CMA), 9.7 : 1 (FA), and 8.3 : 1 (BDE), respectively. The highest inELISA titers of ZEN pAb of each group were 1 : (6.4 × 10 3) (OAE), 1 : (3.2 × 10 3) (CMA), 1 : (1.6 × 10 3) (FA), and 1 : (1.6 × 10 3) (BDE), respectively. The 50% inhibition concentrations (IC50) for ZEN by icELISA of each group were 11.67  μg/L (OAE), 16.29  μg/L (CMA), 20.92  μg/L (FA) and 24.36  μg/L (BDE), respectively. ZEN pAb from the mice immunized with ZEN-BSA (OAE) and ZEN-BSA (CMA) had class broad specificity to ZEN and its analogs. The CRs of ZEN pAb with α-ZAL, β-ZAL, α-ZOL, β-ZOL, and ZON were 36.53%, 16.98%, 64.33%, 20.16%, and 10.66%, respectively. ZEN pAb from the mice immunized with ZEN-BSA (FA) and ZEN-BSA (BDE) had high specificity for ZEN. The CRs of ZEN pAb with its analogs were all less than 1.0%.

          Conclusion

          This study demonstrated that the preparation of the class broad-specificity antibodies of ZEN and its analogs can be achieved by immunizing animals with the immunogen ZEN-BSA prepared by the OAE method, while the preparation of highly specific antibodies can be achieved by immunizing animals with the immunogen ZEN-BSA prepared by the FA method. These findings lay the material and technical foundation for immunoassay of ZEN single residue and ZEN and its analogs total residue.

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          Most cited references54

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          Review on the toxicity, occurrence, metabolism, detoxification, regulations and intake of zearalenone: an oestrogenic mycotoxin.

          Zearalenone (ZEA) is a mycotoxin produced mainly by fungi belonging to the genus Fusarium in foods and feeds. It is frequently implicated in reproductive disorders of farm animals and occasionally in hyperoestrogenic syndromes in humans. There is evidence that ZEA and its metabolites possess oestrogenic activity in pigs, cattle and sheep. However, ZEA is of a relatively low acute toxicity after oral or interperitoneal administration in mice, rat and pig. The biotransformation for ZEA in animals involves the formation of two metabolites alpha-zearalenol (alpha-ZEA) and beta-zearalenol (beta-ZEA) which are subsequently conjugated with glucuronic acid. Moreover, ZEA has also been shown to be hepatotoxic, haematotoxic, immunotoxic and genotoxic. The exact mechanism of ZEA toxicity is not completely established. This paper gives an overview about the acute, subacute and chronic toxicity, reproductive and developmental toxicity, carcinogenicity, genotoxicity and immunotoxicity of ZEA and its metabolites. ZEA is commonly found on several foods and feeds in the temperate regions of Europe, Africa, Asia, America and Oceania. Recent data about the worldwide contamination of foods and feeds by ZEA are considered in this review. Due to economic losses engendered by ZEA and its impact on human and animal health, several strategies for detoxifying contaminated foods and feeds have been described in the literature including physical, chemical and biological process. Dietary intakes of ZEA were reported from few countries from the world. The mean dietary intakes for ZEA have been estimated at 20 ng/kgb.w./day for Canada, Denmark and Norway and at 30 ng/kgb.w./day for the USA. The Joint FAO/WHO Expert Committee on Food Additives (JECFA) established a provisional maximum tolerable daily intake (PMTDI) for ZEA of 0.5 microg/kg of body weight.
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            Risk assessment of the mycotoxin zearalenone

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              Current Advances in Quantum-Dots-Based Photoelectrochemical Immunoassays.

              As a newly developed technique, photoelectrochemical (PEC) immunoassays have attracted great attention in recent years because of their low cost and desirable sensitivity. Because the detection signal originates from the photoelectric conversion of photoelectric materials, the appearance and application of quantum dots (QDs), which possess unique photophysical properties and regulated optoelectronic characteristics, has taken the development of PEC immunoassays to new heights. This review concisely introduces the general mechanism of QDs-based photoelectric conversion for immunoassays and summarizes the current advances in QD applications in immunoassays. Given that signal strategies and photoactive materials are the key elements in PEC biosensor systems, we comprehensively highlight the state-of-the-art signaling strategies and various applications of QDs in PEC immunoassays to introduce advances in QDs-based PEC immunoassays. Finally, challenges and future developmental trends are briefly discussed.
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                Author and article information

                Contributors
                Journal
                Int J Anal Chem
                Int J Anal Chem
                ijac
                International Journal of Analytical Chemistry
                Hindawi
                1687-8760
                1687-8779
                2021
                26 July 2021
                : 2021
                : 7109383
                Affiliations
                1College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 453003, China
                2Faculty of Veterinary Medicine, Sumy National Agrarian University, Sumy 40021, Ukraine
                3Xinke College, Henan Institute of Science and Technology, Xinxiang 453003, China
                Author notes

                Academic Editor: Adil Denizli

                Author information
                https://orcid.org/0000-0001-9537-2947
                https://orcid.org/0000-0002-4092-3665
                https://orcid.org/0000-0001-6260-414X
                https://orcid.org/0000-0002-9596-9804
                https://orcid.org/0000-0002-0761-3681
                Article
                10.1155/2021/7109383
                8328739
                34349801
                b8464bcb-2b6c-49c9-8077-45497eae4665
                Copyright © 2021 Yanan Wang et al.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 6 May 2021
                : 20 June 2021
                : 12 July 2021
                Funding
                Funded by: National Natural Science Foundation of China
                Award ID: 31702263
                Funded by: 13th Five-Year National Key Research and Development Plan Food Safety Technology Research and Development Major Project
                Award ID: 2019YFC1605705
                Funded by: Henan University
                Award ID: 20IRTSTHN025
                Categories
                Research Article

                Analytical chemistry
                Analytical chemistry

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