Monoclonal Antibodies Against Actinobacillus pleuropneumoniae TonB2 Protein Expressed in Escherichia coli
Monoclonal Antibodies Against Actinobacillus pleuropneumoniae TonB2 Protein Expressed in Escherichia coli
复制标题
大肠杆菌表达的胸膜肺炎放线杆菌 TonB2 蛋白单克隆抗体
DOI：
10.1089/hyb.2012.0031
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发表时间：
2012-10-01
期刊：
影响因子：
--
通讯作者：
Qi, Chao
中科院分区：
文献类型：
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作者：
Liu, Jinlin;Yang, Jihong;Qi, Chao
TonB是一种细菌周围质蛋白，它与ExbB和ExbD蛋白复合物将质子从内膜传导到外膜受体。胸膜肺炎放线菌TonB2蛋白是第二个对铁获取和毒力起重要作用的TonB蛋白。TonB2蛋白被证实具有免疫原性，可以部分保护动物免受致命感染。在本研究中，重组TonB2 （rTonB2）在大肠杆菌wBL21（DE3）中过表达并纯化。然后将rTonB2作为抗原免疫BALB/c小鼠产生单克隆抗体（MAb）。选择了4个tonb2特异性单抗分泌杂交瘤——2f2、2G8、3D2和6f10。单克隆抗体2F2、3D2和6F10为IgG1同型，2G8为IgG2a同型。Western blot和ELISA结果表明，单克隆抗体对rTonB2具有特异性结合活性。这里生成的单克隆抗体将用于进一步的TonB2蛋白功能分析。
TonB is known to be a bacterial periplasmic protein that transduces proton from the inner membrane to the outer membrane receptor in complex with the ExbB and ExbD proteins. Actinobacillus pleuropneumoniae TonB2 protein is the second TonB protein that is important for iron acquisition and virulence. The TonB2 protein was verified to be immunogenic and could afford partial protection for animals from lethal infection. In the present study, the recombinant TonB2 (rTonB2) was overexpressed in Escherichia coli wBL21(DE3) and purified. The rTonB2 was then used as antigen to immunize BALB/c mice for the production of monoclonal antibodies (MAb). Four clones of TonB2-specific MAb secretion hybridomas-2F2, 2G8, 3D2, and 6F10-were selected. The MAbs 2F2, 3D2, and 6F10 were classified as IgG1 isotype and 2G8 was of IgG2a isotype. Western blot and ELISA results indicated that MAbs had specific binding activity to rTonB2. The MAbs generated here will be used for further functional analyses of the TonB2 protein.