擬態弧菌OmpU蛋白B細胞表位的篩選鑒定及其免疫活性研究

李槿年

摘 要：擬態弧菌是一種人和水產動物共患病病原菌，不僅引起水產動物嚴重的腹水病，而且可經水體和水產品感染人類，引起腹瀉和食物中毒。擬態弧菌OmpU蛋白是一種具有良好抗原性和保守性的首選診斷抗原和疫苗候選分子，但其抗原表位仍不清楚。對此，該項目擬開展“擬態弧菌OmpU蛋白B細胞表位及其免疫活性研究”。應用生物信息軟件預測OmpU蛋白B細胞線性表位，進而利用合成肽ELISA鑒定預測表位，確定優勢B細胞線性表位。再以OmpU多克隆抗體和單克隆抗體為靶標從噬菌體隨機肽庫中篩選鑒定優勢B細胞模擬表位。最后，對表（擬）位噬菌體克隆進行免疫活性檢測，分析不同表位的免疫活性差異。本項目預期研究成果不僅為建立以表位為基礎的特異快速的診斷方法和研制表位疫苗奠定基礎。

關鍵詞：擬態弧菌 OmpU蛋白 B細胞表位 免疫活性

Abstract： Vibrio mimicus （V. mimicus） is a zoonisis pathogens. It not only cause ascitic fluid disease of aquatic animals， but also cause human gastroenteritis and food poisoning through contacting aquatic environment and aquatic products. The out membrance protein U （OmpU） of V. mimicus with higher antigenicity and conservative property is taken as a primary diagnostic antigen and good candidate protein for developing vaccine， but B-cell epitopes of OmpU protein are not clear now. In order to design new vaccine and diagnosing reagent in the future， the B-cell eptopes of OmpU protein of V. mimicus and their immune characteristics were researched. Firstly， B-cell epitopes of OmpU protein were analyzed with bio-softwares Biosun and Hopp-wood. The putative epitopes peptides were synthesized and were defined by peptide enzyme-linked immunosorbent assay. Secondly， in order to define B-cell mimotopes of OmpU protein， the phage clones binding to purified polyclonal antibody and monoclonal antibody against OmpU protein were selected from the 12-mer random peptide library. Then positive monoclonal phages were sequenced and analyzed. Finally， the immunogenicity and immunoreactivity of differnt phage clones containing B-cell epitopes were determined with immunoassay to compare their immune characteristics.

Key Words： Vibrio mimicus； OmpU protien； B cells epitopes； Immune characteristics

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