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志贺样毒素Ⅱ变异体(SLT-Ⅱe)是猪水肿病的主要致病因子之一,为了制备SLT-Ⅱe单克隆抗体(mAb),我们从TB1菌中用多粘菌素诱导提取了SLT-Ⅱe蛋白,并且用硫酸铵盐析沉淀法对该蛋白进行了提纯,以提纯蛋白免疫BALB/c小鼠,利用杂交瘤细胞技术制备mAb。细胞融合后,培养上清经间接ELISA检测,共得到3株阳性杂交瘤细胞2E11、2H9和4F3,ELISA效价分别为1∶213、1∶213和1∶28,3株杂交瘤细胞培养上清分泌的mAb在Westernblot试验中均与提纯的毒素蛋白反应,在Vero细胞中和试验中均能够中和毒素对Vero细胞的毒性作用,从而为临床分离株SLT-Ⅱe产生的免疫学检测方法奠定了一定的基础。
Shiga-like toxin II variant (SLT-IIe) is one of the major pathogenic factors of swine edema disease. In order to prepare SLT-IIe monoclonal antibody (mAb), we induced polymyxin-induced SLT from TB1 strain -Ie protein, and the protein was purified by ammonium sulfate precipitation method. Purified protein was used to immunize BALB / c mice and mAb was prepared by hybridoma cell technology. After cell fusion, the culture supernatants were detected by indirect ELISA. Three positive hybridoma cells, 2E11, 2H9 and 4F3, were obtained. The ELISA titers were 1: 213, 1: 21 and 1:28, The purified secreted mAbs reacted with the purified toxin protein in the Western blotting assay and were able to neutralize the toxic effects of the toxin on the Vero cells both in the Vero cells and in the assay so as to lay down the immunological detection method for the clinical isolates SLT-IIe A certain basis.