Design of Hapten Synthesis and Antibody Characterization of G-group Aflatoxins


  • Yanan Wang College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, China https://orcid.org/0000-0001-5412-6731
Keywords: G-group Aflatoxins, Hapten, Artificial Antigen, Polyclonal Antibody, Specificity Analysis

Abstract

Food and feed contamination with Aflatoxins pose a serious threat to human health and animal husbandry development and has caused widespread concern, among them, G-group Aflatoxins as the main pollutant has attracted more and more attention. In order to establish a rapid, sensitive, specific and efficient immunoassay method for G-group aflatoxins, this study aimed to designed to synthesize 3 immunogens and coating antigens and identified by UV and SDS-PAGE. Then used to immunize Balb/c mice with prepared of three immunogen the titers were determined by indirect ELISA and the sensitivity was determined by competitive indirect ELISA (icELISA), the specificity was assessed by the cross-reaction test (CR). The results of UV and SDS-PAGE showed that the three immunogens and the corresponding coated antigens were successfully synthesized and the best one was SA method among three synthesis methods of G-group AF artificial antigen and its conjugation ratio of AFG1 to BSA was about 5.641. The immune efficacy of SA method was the best, its AFG1 pAb had high titers of 1(6.4×103) by indirect ELISA, a good sensitivity with the 50 % inhibition concentration(IC50) of 13.6 μg/kg1 to AFG1 by blocking ELISA and a high CR to AFG2 of 82.19 %, it showed high specificity for other aflatoxins. The experimental results not only obtained the ideal G group aflatoxin antibody, but also established a substance and technology foundation for G group aflatoxin immunization methods, and can be referenced in the similar tests.

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References

Aiko, V., & Mehta, A. (2015). Occurrence, detection and detoxification of mycotoxins. Journal of Biosciences, 40(5), 943–954. doi: 10.1007/s12038-015-9569-6.

China, N. h. a. f. p. c. a. s. f. a. d. a. o. (2017). Determination of aflatoxin B1, B2, G1, G2 in grain:ultra-high performance liquid chromatography. In (Vol. LS/T 6128-2017(2017)).

Chu, F. S., Steinert, B., & Gaur, P. (1985). Production and characterization of antibody against aflatoxin G1. Journal of Food Safety, 7(3), 161–170. doi: 10.1007/bf01949970.

EC. (2006). (EC) No 1881/2006 setting maximum levels for certain contaminants in foodstuffs.

FDA. (2018). Adulteration with Aflatoxin-CPG Sec 555.400 Foods.

Gefen, T., Vaya, J., Khatib, S., Rapoport, I., Lupo, M., Barnea, E., ... Pitcovski, J. (2015). The effect of haptens on protein-carrier immunogenicity. Immunology, 144(1), 116–126. doi: 10.1111/imm.12356.

Guan, L. U., Zhang, Y. Z., Yang, T. T., You, J. M., Hong-Mei, L. I., & Sun, X. L. (2013). Synthesis and Characterization of the Hapten for Paraquat. Journal of Food ence & Biotechnology, 32(5), 524–528. doi: 10.1016/S0926-860X(97)00036-7.

Iyer, R. S., & Harris, T. M. (1993). Preparation of aflatoxin B1 8,9-epoxide using m-chloroperbenzoic acid. Chemical Research in Toxicology, 6(3), 313–316. doi: 10.1021/tx00033a010.

Japan (2011). Management of food containing aflatoxin. No.0331-5.

Kim, S. H., Cha, S. H., Karyn, B., Park, S. W., & Kang, H. G. (2011). Production of Group Specific Monoclonal Antibody to Aflatoxins and its Application to Enzyme-linked Immunosorbent Assay. Toxicological Research, 27(2), 125–131. doi: 10.5487/TR.2011.27.2.125.

Kortkhonjia, E., Brandman, R., Zhou, J. Z., Voelz, V. A., Chorny, I., Kabakoff, B., ... Swartz, T. E. (2013). Probing antibody internal dynamics with fluorescence anisotropy and molecular dynamics simulations. Microbiol Spectr, 5(2), 306–322. doi: 10.4161/mabs.23651.

Li, P., Zhou, Q., Wang, T., Zhou, H., & Zhang, Q. (2015). Development of an Enzyme-Linked Immunosorbent Assay Method Specific for the Detection of G-Group Aflatoxins. Toxins, 8(1), 5. doi: 10.3390/toxins8010005.

Martin, C. N., & Garner, R. C. (1977). Aflatoxin B-oxide generated by chemical or enzymic oxidation of aflatoxin B1 causes guanine substitution in nucleic acids. Nature, 267(5614), 863–865. doi: 10.1038/267863a0.

Rushing, B. R., & Selim, M. I. (2019). Aflatoxin B1: A review on metabolism, toxicity, occurrence in food, occupational exposure, and detoxification methods. Food and Chemical Toxicology, 124, 81–100. doi: 10.1016/j.fct.2018.11.047.

Sevy, A. M., & Meiler, J. (2014). Antibodies: Computer-Aided Prediction of Structure and Design of Function. Microbiol Spectr, 2(6), 1–14. doi: 10.1128/ microbiolspec.AID-0024-2014.

Siahi Shadbad, M. R., Ansarin, M., Tahavori, A., Ghaderi, F., & Nemati, M. (2012). Determination of aflatoxins in nuts of Tabriz confectionaries by ELISA and HPLC methods. Adv Pharm Bull, 2(1), 123–126. doi: 10.5681/apb.2012.018.

Wang, Y., Han, D., Wang, X., Lei, Z., Niu, L., & Wang, Z. (2016). Preparation of Anti-Chromium Ion Monoclonal Antibodies and Analysis of Immunological Properties. Acta Agriculturae Boreali-Occidentalis Sinica, 25(11), 1597–1602.

Wang, Y., Wang, S., Zhang, H., & Wang, Z. (2014). Establishment of Hybridoma Cell Lines Secreting Anti-Cadmium Ion Monoclonal Antibody and Identification of Their Immunological Properties. Acta Agriculturae Boreali-Occidentalis Sinica, 23(11), 24–29.

Wu, F., Stacy, S. L., & Kensler, T. W. (2013). Global Risk Assessment of Aflatoxins in Maize and Peanuts: Are Regulatory Standards Adequately Protective? Toxicological Sciences An Official Journal of the Society of Toxicology, 135(1), 251–259. doi: 10.1093/toxsci/kft132.

Xie, L., Chen, M., & Ying, Y. (2016). Development of Methods for Determination of Aflatoxins. Crit Rev Food Sci Nutr, 56(16), 2642–2664. doi: 10.1080/10408398.2014.907234.

Xu, Z. L., Shen, Y. D., Beier, R. C., Yang, J. Y., Lei, H. T., Wang, H., & Sun, Y. M. (2009). Application of computer-assisted molecular modeling for immunoassay of low molecular weight food contaminants: A review. Anal Chim Acta, 647(2), 125–136. doi: 10.1016/j.aca.2009.06.003.

Zhang, J., Li, P. W., Zhang, W., Zhang, Q., Ding, X., Chen, X., ... Zhang, X. (2009). Production and characterization of monoclonal antibodies against aflatoxin g(1). Hybridoma (Larchmt), 28(1), 67–70. doi: 10.1089/hyb.2008.0064.

Zhang, J. Y., Pei-Wu, L. I., Zhang, W., Ding, X. X., Liang, M. A., Wen-Hua, W. U., ... Tao, L. I. (2008). Synthesis of Aflatoxin G_1 Artificial Antigen. Food Science, 29(6), 194–197.

Zhou, Q., Zhang, W., Zhang, Q., Ding, X. X., & Pei-Wu, L. I. (2010). Synthesis of aflatoxin G class artificial antigen. Chemical Reagents, 32(10), 859–872. 876. doi: 10.3788/AOS20103012.3552.

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Published
2021-06-19
How to Cite
Wang, Y. (2021). Design of Hapten Synthesis and Antibody Characterization of G-group Aflatoxins. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Veterinary Sciences, 23(102), 130-135. https://doi.org/10.32718/nvlvet10220