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Preparation and characterisation of monoclonal antibodies against deoxynivalenol
Italian Journal of Animal Science ( IF 2.5 ) Pub Date : 2020-05-25 , DOI: 10.1080/1828051x.2020.1763861
Li Han 1, 2 , Yuetao Li 2 , Jinqing Jiang 2 , Renfeng Li 2 , Guoying Fan 2 , Zhuang Lei 2 , Haojie Wang 2 , Ziliang Wang 2 , Wenju Zhang 1
Affiliation  

Abstract To prepare broad-spectrum and specific monoclonal antibodies (mAb) against deoxynivalenol (DON), DON-BSA was synthesised by the carbonyl diimidazole method as an artificial antigen to immunise Balb/C mice, and the coated DON-OVA was synthesised by the carbodiimide method to detect anti-DON antibodies. Monoclonal antibodies were screened by indirect ELISA and indirect competitive ELISA. A hybridoma cell line (4F3) capable of stably secreting DON antibody was obtained. The titre of antibody in the culture supernatant was 1:1.28 × 103, and the titre of the ascites antibody was 1:3.2 × 105. The monoclonal antibody homology was IgG1/λ. The half inhibitory concentration (IC50) of DON was 9.84 ng/mL. The cross-reaction rates with 3-Ac-DON and 15-Ac-DON were 60.44% and 52.04%, respectively, but no cross-reaction with other mycotoxins was observed. The results showed that the anti-DON monoclonal antibodies prepared in this experiment could recognise not only DON but also 3-Ac-DON and 15-Ac-DON, which could provide materials for the next step to establish a method to detect DON and its similar compounds. Highlights This study demonstrated that an artificial antigen was synthesised by carbonyl diimidazole method, and a novel broad-spectrum, high affinity monoclonal antibody was developed. The anti-DON mAb developed in this study has better sensitivity, moreover the antibody was sensitive to DON and its derivatives. Therefore, it could be used for simultaneous monitoring of three mycotoxins. Results of this study will further lay the foundation for the establishment of immunological assays for the total amount of similar compounds.

中文翻译:

脱氧雪腐镰刀菌烯醇单克隆抗体的制备和表征

摘要 为制备抗脱氧雪腐镰刀菌烯醇 (DON) 的广谱特异性单克隆抗体 (mAb),采用羰基二咪唑法合成 DON-BSA 作为人工抗原免疫 Balb/C 小鼠,并通过合成包被的 DON-OVA。碳二亚胺法检测抗 DON 抗体。通过间接ELISA和间接竞争ELISA筛选单克隆抗体。获得了能够稳定分泌DON抗体的杂交瘤细胞系(4F3)。培养上清中抗体滴度为1:1.28×103,腹水抗体滴度为1:3.2×105,单克隆抗体同源性为IgG1/λ。DON 的半抑制浓度 (IC50) 为 9.84 ng/mL。与 3-Ac-DON 和 15-Ac-DON 的交叉反应率分别为 60.44% 和 52.04%,但未观察到与其他霉菌毒素的交叉反应。结果表明,本实验制备的抗DON单克隆抗体不仅可以识别DON,还可以识别3-Ac-DON和15-Ac-DON,为下一步建立检测DON及其DON的方法提供材料。类似的化合物。亮点 本研究表明,通过羰基二咪唑法合成了一种人工抗原,并开发了一种新型广谱、高亲和力的单克隆抗体。本研究开发的抗DON mAb具有较好的敏感性,而且该抗体对DON及其衍生物敏感。因此,它可用于同时监测三种霉菌毒素。本研究的结果将进一步为建立类似化合物总量的免疫学检测方法奠定基础。
更新日期:2020-05-25
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