He Kuo, Mao Qingwen, Zang Xiuyuan, Zhang Yanyu, Li Hui, Zhang Donghao
Hebei North University, Laboratory Animal Center of Hebei North University, Zhangjiakou, 075000, China.
Hebei North University, Laboratory Animal Center of Hebei North University, Zhangjiakou, 075000, China.
Biologicals. 2017 Sep;49:57-61. doi: 10.1016/j.biologicals.2017.06.008. Epub 2017 Jul 5.
In this study, we report the production of a monoclonal broad-specificity monoclonal antibody (mAb) specific for amatoxins and development of an indirect competitive immunoassay for detection of amatoxins in mushroom samples. In the assay, the complete antigen (α-amanitin-OVA) was used as coating antigen, and amatoxins as competitor competes with coating antigen to bind with mAb. Using this approach, The half-maximum inhibition concentrations (IC) of α-amanitin, β-amanitin and γ-amanitin, and limits of detection (LODs, IC) were 66.3, 97.4, 163.1 ng/mL and 0.91, 0.98, 0.89 ng/mL, respectively. The LODs for α-amanitin, β-amanitin and γ-amanitin in mushroom samples were 4.55, 4.9, and 4.45 ng/mL. The spiked results were also confirmed by HPLC, which showed a good correlation (R = 0.996) between the two methods. The results indicated that the developed assay was reliable and suitable for the detection of amatoxins in mushroom samples.
在本研究中，我们报道了一种针对鹅膏毒素的单克隆广谱特异性单克隆抗体（mAb）的制备以及一种用于检测蘑菇样品中鹅膏毒素的间接竞争免疫测定法的开发。在该测定法中，完全抗原（α-鹅膏毒肽-卵清蛋白）用作包被抗原，鹅膏毒素作为竞争剂与包被抗原竞争结合单克隆抗体。采用这种方法，α-鹅膏毒肽、β-鹅膏毒肽和γ-鹅膏毒肽的半数抑制浓度（IC）以及检测限（LODs，IC）分别为66.3、97.4、163.1 ng/mL和0.91、0.98、0.89 ng/mL。蘑菇样品中α-鹅膏毒肽、β-鹅膏毒肽和γ-鹅膏毒肽的检测限分别为4.55、4.9和4.45 ng/mL。加标结果也通过高效液相色谱法得到证实，两种方法之间显示出良好的相关性（R = 0.996）。结果表明，所开发的测定法可靠且适用于检测蘑菇样品中的鹅膏毒素。
