Antibody Microarray Immunoassay for Simultaneous Quantification of Multiple Mycotoxins in Corn Samples

Zhang, Xian; Wang, Zuohuan; Fang, Yun; Sun, Renjie; Cao, Tong; Paudyal, Narayan; Fang, Weihuan; Song, Houhui

doi:10.3390/toxins10100415

Cited by 26 publications

(12 citation statements)

References 45 publications

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“…e immunoassay methods based on antigen-antibody reaction have the common characteristics of strong selectivity and high sensitivity and have become a hot topic in ZEN detection research, such as flexible pressure sensor (FPS) [16], self-powered temperature sensor (SPTS) [17], nano-hybrid-mediated photoelectrochemical immunoassay (NMPECIA) [18], and quantum dot-based photoelectrochemical immunoassay (QDPECIA) [19]. In addition, the established immunoassay methods, such as enzyme-linked immunosorbent assay (ELISA) [20], gold immunochromatographic assay (GICA) [21], and the time-resolved fluoroimmunoassay (TRFIA) [22], also have the advantages of fast, low cost, and on-site detection [15], fluorescence polarization immunoassay (FPIA) [23], immunosensor (IS) [24], and antibody microarray (AMA) [25] have the advantage of the highthroughput detection. ese immunoassay techniques play an important role in the rapid detection of ZEN.…”

Section: Introductionmentioning

confidence: 99%

Synthesis of Zearalenone Immunogen and Comparative Analysis of Antibody Characteristics

Wang

Zhang

et al. 2021

International Journal of Analytical Chemistry

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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 × 103) (OAE), 1 : (3.2 × 103) (CMA), 1 : (1.6 × 103) (FA), and 1 : (1.6 × 103) (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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Section: Introductionmentioning

confidence: 99%

Synthesis of Zearalenone Immunogen and Comparative Analysis of Antibody Characteristics

Wang

Zhang

et al. 2021

International Journal of Analytical Chemistry

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“…Some modifications included the application of urchin-like and flower-like gold nanoparticles (AuNP), which slightly increased the sensitivity when compared to a spherical particle [107,135]. [190] As an alternative to color intensity measurements, a chemiluminescent substrate could be incubated with HRP for a slight improvement of the LOD [93,127], or the application of quantum dot (QD) in which a radiometric analysis revealed a constant signal from the test line with biotin-BSA, compared to the calibration with anti-mouse IgG [134]. Nevertheless, the application of fluorescent QDs not always result in an improved sensitivity, as reported for a nitrocellulose strip for the detection of FB1 (LOD: 60 µg/L) , ZEN and OTA with a monoclonal antibody-QD probe placed on the conjugate pad, through the competitive interaction with mycotoxin-BSA at the test line [189], and a mAB-Europium fluorescent nanoparticle with FB1 (LOD: 8.26 µg/L) and FB1-BSA (Test line) [190].…”

Section: Immunosensors For Detection Of Fumonisinmentioning

confidence: 99%

Aptamer-Based Detection of Fumonisin B1: A Systematic Comparison with Conventional and Other Novel Methods

Mirón-Mérida¹,

Gong²,

Goycoolea³

2020

Preprint

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Mycotoxin contamination is a current issue affecting several crops and processed products worldwide. Among the diverse mycotoxin group, fumonisin B1 (FB1) has become a relevant compound because of its adverse effects in the food chain. Conventional analytical methods previously proposed to quantify FB1 comprise LC-MS, HPLC-FLD and ELISA, while novel approaches integrate different sensing platforms and fluorescently labelled agents in combination with antibodies. Nevertheless, such methods could be expensive, time-consuming and require experience. Aptamers (ssDNA) are promising alternatives to overcome some of the drawbacks of conventional analytical methods, their high affinity through specific aptamer-target binding has been exploited in various designs attaining favorable limits of detection (LOD). So far, two aptamers specific to FB1 have been reported, and their modified and shortened sequences have been explored for a successful target quantification. In this critical review spanning the last eight years, we have conducted a systematic comparison based on principal component analysis of the aptamer-based techniques for FB1, compared with chromatographic, immunological and other analytical methods. We have also conducted an <i>in-silico</i> prediction of the folded structure of both aptamers under their reported conditions. The potential of aptasensors for the future development of highly sensitive FB1 testing methods is emphasized.

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“…The LC-MS method has the advantages of a high sensitivity and good repeatability, but it has the disadvantages of high cost and complex sample processing. Immunoassays have been widely performed for mycotoxin detection owing to its simplicity, specificity, and low cost [ 12 , 13 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

Research on the Mechanism of Action of a Citrinin and Anti-Citrinin Antibody Based on Mimotope X27

et al. 2020

Toxins

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Immunoassays are developed based on antigen–antibody interactions. A mimotope is an effective recognition receptor used to study the mechanism of action of antigens and antibodies, and is used for improving the sensitivity of the antibody. In this study, we built a 3D structure of the citrinin (CIT) mimotope X27 and anti-CIT single-chain antibody fragment (ScFv) through a “homologous modeling” strategy. Then, CIT and X27 were respectively docked to anti-CIT ScFv by using the “molecular docking” program. Finally, T28, F29, N30, R31, and Y32 were confirmed as the key binding sites in X27. Furthermore, the result of the phage-ELISA showed that the mutational phage lost the binding activity to the anti-CIT ScFv when the five amino acids were mutated to “alanine”, thereby proving the correctness of the molecular docking model. Lastly, a site-directed saturation strategy was adopted for the sites (T28, F29, N30, R31, and Y32). Eighteen different amino acids were introduced to each site on average. The activities of all mutants were identified by indirect competitive ELISA. The sensitivities of mutants T28F, T28I, F29I, F29V, N30T, and N30V were 1.83-, 1.37-, 1.70-, 2.96-, 1.31-, and 2.01-fold higher than that of the wild-type, respectively. In conclusion, the binding model between the CIT and antibody was elaborated for the first time based on the mimotope method, thereby presenting another strategy for improving the sensitivity of citrinin detection in immunoassays.

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Antibody Microarray Immunoassay for Simultaneous Quantification of Multiple Mycotoxins in Corn Samples

Cited by 26 publications

References 45 publications

Synthesis of Zearalenone Immunogen and Comparative Analysis of Antibody Characteristics

Synthesis of Zearalenone Immunogen and Comparative Analysis of Antibody Characteristics

Aptamer-Based Detection of Fumonisin B1: A Systematic Comparison with Conventional and Other Novel Methods

Research on the Mechanism of Action of a Citrinin and Anti-Citrinin Antibody Based on Mimotope X27

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