Generation of monoclonal antibodies and epitope mapping of ApxIVA of Actinobacillus pleuropneumoniae
Generation of monoclonal antibodies and epitope mapping of ApxIVA of Actinobacillus pleuropneumoniae
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胸膜肺炎放线杆菌ApxIVA单克隆抗体的产生和表位定位
DOI：
10.1016/j.molimm.2005.12.010
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发表时间：
2006-05-01
影响因子：
3.6
通讯作者：
Chen, Huanchun
中科院分区：
文献类型：
--
作者：
Huang, Hongliang;Zhou, Rui;Chen, Huanchun
为了研究ApxIV的功能，并建立区分猪自然感染和灭活疫苗和/或亚单位疫苗免疫的诊断试验，我们尝试制备抗ApxIV的单克隆抗体。用ApxIVAN和ApxIVAC免疫Balb/c小鼠，它们是大肠杆菌产生的ApxIVA的N末端和C末端半部（分别为814个和997个氨基酸）。coliBL21（DE3）。选择了8种单克隆抗体，4种（命名为1A 8、1G 5、3E 7和4 H9）针对ApxIVAN，另外4种（命名为IB 12、2 E5、4138和4G 2）针对ApxIVAC。Western blot和ELISA相加试验表明，除1A 8外，所有单克隆抗体均对相应的免疫原具有特异性，1A 8与两种免疫原反应，两者有156个残基的重叠区。ELISA加和性试验显示，ApxIV中至少有5个表位由8个单克隆抗体定义，2个在1和866个氨基酸之间，1个在867和1022个氨基酸之间，2个在1023和1863个氨基酸之间。总之，我们已经成功地生产了八种单克隆抗体，它们与ApxIV的五种不同表位反应。(c)2006爱思唯尔有限公司版权所有。
To study functions of ApxIV, a species-specific and in vivo inducible RTX toxin identified in Actinobacillus pleuropneumoniae recently, and to develop a diagnostic trial distinguishing the pigs infected naturally and vaccinated with inactivated and/or subunit vaccines, we attempted to prepare monoclonal antibodies against ApxIV. BALB/c mice were immunized with ApxIVAN and ApxIVAC which are N- and C-terminal halvies (814 and 997 amino acids, respectively) of ApxIVA produced in E. coli BL21 (DE3), respectively. Eight monoclonal antibodies were selected, four (designated as 1A8, 1G5, 3E7 and 4H9) against ApxIVAN and another four (named as IB12, 2E5, 4138 and 4G2) against ApxIVAC. Western blot and ELISA additivity assays suggested that all monoclonal antibodies except 1A8 are specific to the corresponding immunogen, 1A8 reacts with both immunogens which have a overlapping region of 156 residues. ELISA additivity tests revealed that at least five epitopes in ApxIV are defined by eight monoclonal antibodies, two between I and 866 amino acids, one between 867 and 1022 amino acids and two between 1023 and 1863 amino acids. In conclusion, we have succeeded in producing eight monoclonal antibodies, which react with five different epitopes of ApxIV. (c) 2006 Elsevier Ltd. All rights reserved.