An immunogen synthesis strategy for the development of specific anti-deoxynivalenol monoclonal antibodies
An immunogen synthesis strategy for the development of specific anti-deoxynivalenol monoclonal antibodies
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DOI：
10.1080/19440049.2014.955887
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发表时间：
2014-10-03
影响因子：
2.9
通讯作者：
De Saeger, Sarah
中科院分区：
文献类型：
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作者：
Sanders, Melanie;Guo, Yirong;De Saeger, Sarah
设计了一种免疫原合成策略，以开发对结构相似的多烯类具有低交叉反应性的抗脱氧雪腐镰刀菌烯醇（DON）单克隆抗体。总共合成了八种不同的DON免疫原，其在DON分子上的接头的类型和位置不同。免疫后，检查来自用不同DON免疫原免疫的小鼠的抗血清中相关抗体的存在。然后，进行同源和异源酶联免疫吸附试验（ELISA）用于杂交瘤筛选。最后，三个单克隆抗体的DON及其类似物的产生。单克隆抗体13 H1对DON及其类似物的识别顺序为HT-2毒素>15-乙酰脱氧雪腐镰刀菌烯醇（15-ADON）>DON，IC_（50）为1.14 ~ 2.13 μ g/ml。单克隆抗体10 H10对DON、3-乙酰脱氧雪腐镰刀菌烯醇（3-ADON）和15-ADON敏感性相近，IC_（50）分别为22、15和34 ngml（-1）。使用间接ELISA形式将10 H10对15-ADON的灵敏度降低92%。第三株单克隆抗体2A 9对3-ADON具有高度的特异性和敏感性，IC_（50）为0.38ngml ~（-1）。使用2A 9和10 H10单克隆抗体允许确定唯一的DON污染。
An immunogen synthesis strategy was designed to develop anti-deoxynivalenol (DON) monoclonal antibodies with low cross-reactivity against structurally similar trichothecenes. A total of eight different DON immunogens were synthesised, differing in the type and position of the linker on the DON molecule. After immunisation, antisera from mice immunised with different DON immunogens were checked for the presence of relevant antibodies. Then, both homologous and heterologous enzyme-linked immunosorbent assays (ELISAs) were performed for hybridoma screening. Finally, three monoclonal antibodies against DON and its analogues were generated. In addition, monoclonal antibody 13H1 could recognise DON and its analogues in the order of HT-2 toxin>15-acetyldeoxynivalenol (15-ADON)>DON, with IC50 ranging from 1.14 to 2.13 mu gml(-1). Another monoclonal antibody 10H10 manifested relatively close sensitivities to DON, 3-acetyldeoxynivalenol (3-ADON) and 15-ADON, with IC50 values of 22, 15 and 34ngml(-1), respectively. Using an indirect ELISA format decreases the 10H10 sensitivity to 15-ADON with 92%. A third monoclonal antibody 2A9 showed to be very specific and sensitive to 3-ADON, with IC50 of 0.38ngml(-1). Using both 2A9 and 10H10 monoclonal antibodies allows determining sole DON contamination.