耶尔森氏菌YadA蛋白单克隆抗体的研制与鉴定

The Preparation of The Monoclonal Antibody to Yersinia Adherence Protein A (YadA) and Its Identification

１．以纯化的ＹａｄＡ蛋白〔１、３〕免疫普通小鼠４只，获得小鼠ＹａｄＡ多克隆抗血清；２．利用上述免疫血清建立了ＹａｄＡ多克隆抗体的ＥＬＩＳＡ检测技术，并对其最适条件进行了观察；３．用纯化的ＹａｄＡ蛋白免疫ＢＡＬＢ／Ｃ小鼠２只，以免疫ＢＡＬＢ／Ｃ小鼠的脾细胞与骨髓瘤细胞系ＳＰ２／０融合，获得１７株分泌ＹａｄＡ－ＭｃＡｂ杂交瘤细胞株，将其中４株进行了再克隆，经过半年左右的反复传代及冷冻、复苏测试，抗体分泌稳定；４．Ｗｅｓｔｅｒｎ－ｂｌｏｔｉｎｇ分析表明瘤株所分泌的单克隆抗体特异；５．

The purification method of YadA was improved on the basis of primary procedure. (2) 4 white mice were immunized with the purified YadA and the polyclonal antibody to YadA was obtained. (3)ELISA detecting polyclonal antibody to YadA had established and its optimal reacting conditions been observed. (4) 2 BALB/C mice were immunized by purified YadA and the spleen cells of the mice were fused with plasmacytoma SP2/0, Obtained 17 fused cell strains secred YadA- McAb, cloning of 4 strains of them were carried out. Testing results shown the above strains could stably secret the YadA-monoclonal antibody.(5) The result of the Western Blotting analysis proved that the monoclonal antibody secreted by the fused cells was special. (6) The YadA-McAb will be possiblly possessed of the good applied prospects for coagnosis differentiating Yersinia enterocolitica from Brucella.