Production of mouse monoclonal antibodies against Helicobacter pylori Catalase and mapping the antigenic epitope by phage display library

被引:20
|
作者
Li, Yan [1 ]
Ning, Yun-Shan [1 ]
Wang, Yun-Dan [1 ]
Luo, Jun [2 ]
Wang, Wenjing [1 ]
Dong, Wen-Qi [1 ]
Li, Ming [1 ]
机构
[1] So Med Univ, Sch Biotechnol, Guangzhou 510515, Guangdong, Peoples R China
[2] So Med Univ, Dept Microbiol, Guangzhou 510515, Guangdong, Peoples R China
基金
国家高技术研究发展计划(863计划);
关键词
Helicobacter pylori; Catalase; monoclonal antibody; phage display library; epitope;
D O I
10.1016/j.vaccine.2007.11.055
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
The Catalase of Helicobacter pylori (H. pylori) helps bacteria to protect themselves from oxygen toxicity and damage and have been identified an immunodominant antigen. To obtain mouse monoclonal antibodies (mAbs) against Catalase and to map its antigenic epitope is potentially to develop a vaccine for prevention and treatment of H. pylori infection. In our study, MAbs were produced by the hybridoma technique using recombinant Catalase-GST as the immunogen and were immunoscreened against phage-displayed random dodecapeptide library (Ph.D.-12). After three rounds of biopanning, 34 phage clones were randomly selected and their specificity to mAb was verified by sandwich and competitive inhibition ELISA. Fifteen phage clones were sequenced and their amino acids were deduced. One mimotope (SVSLPYANLATH) showed good match with Catalase protein at 394-405aa and the serum of mice induced by the phage clone clearly recognized Catalase protein. Our work suggests that the antigenic epitope could be mapped through screening the phage-displayed peptide libraries with mAb and a mimotope of Catalase would provide an alternative approach for the development of a vaccine for H. pylori. (C) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1263 / 1269
页数:7
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