骆驼血清中不同抗体亚型对除草剂麦草畏的性能研究

Identification of the performance of different antibody subtypes in camel serum to herbicide dicamba

为构建麦草畏快速检测技术,本研究通过rProtein A/G Beads 4FF重力柱和rProtein G Beads 4FF重力柱对经免疫后的双峰驼体内3种亚型的IgG抗体进行分离纯化,得到常规抗体IgG1和天然缺失轻链的重链抗体IgG2和IgG3,并检测了3种不同抗体亚型对麦草畏的亲和性和特异性的差异。结果显示,IgG1、IgG2和IgG3的IC_(50)值分别为0.11、0.10和0.19μg/mL,线性范围分别为0.002~5.47,0.012~2.97和0.013~2.98μg/mL。特异性检测试验结果显示,IgG1对麦草畏的结构类似物2,3,6-三氯苯甲酸具有一定的交叉反应,交叉反应率为52%,而IgG2和IgG3无交叉反应。IgG2和IgG3是获得高灵敏性和特异性纳米抗体的重要基础,本研究将为双峰驼体内麦草畏纳米抗体的筛选工作奠定了重要基础。

In order to develop the rapid detection technology of dicamba,three subtypes of IgG antibodies in immunized bactrian camels were separated and purified using the rProtein A/G Beads 4FF column and rProtein G Beads 4FF column.Conventional antibodies IgG1 and natural heavy-chain antibodies IgG2 and IgG3 were obtained.The affinity and specificity of the three subtypes against dicamba were detected.The IC_(50) of IgG1,IgG2 and IgG3 were 0.11,0.10 and 0.19μg/mL,respectively.And the linear ranges were 0.002-5.47,0.012-2.97 and 0.013-2.98μg/mL,respectively.The cross-reactivity of IgG1 was 52%to 2,3,6-trichlorobenzoic acid,a structural analogue of dicamba,while IgG2 and IgG3 showed no cross-reactivity.IgG2 and IgG3 are important for obtaining high sensitivity and specificity of nanobodies.This study lays an important foundation for the screening of nanobodies against dicamba in bactrian camels.