【摘要】 猪瘟病毒(CSFV)囊膜表面结构糖蛋白 E2 (gp55)是诱导机体产生中和抗体及激发保护性免疫应答的主要抗原蛋白。E2 和 Erns与细胞表面受体的相互作用介导病毒对细胞的感染过程。本文采用抗 CSFV(Alfort Tübingen毒株)E2 结构蛋白的单克隆抗体 c2410 和 a18,淘选噬菌体展示的 12 肽随机肽库,对 CSFV E2 蛋白表位进行分析。研究发现:单克隆抗体 c2410 和 a18 识别同一线性表位,定位于 E2 蛋白的 832-837 位氨基酸(SPTTLR),但二者在 ELISA 和免疫印迹分析中对不同表(拟)位的反应性存在差异。自杂交瘤细胞中提取总 RNA,对单克隆抗体轻链和重链可变区 cDNA 进行序列分析。结果表明:c2410 和 a18 虽然来源于同一批次融合的杂交瘤细胞系,识别同一表位,但仍属于不同的单克隆抗体。更多还原

【Abstract】 Structural and envelope glycoprotein E2 (gp55) of Classical swine fever virus (CSFV) is the most antigenic protein being responsible for eliciting neutralizing antibodies and conferring protective immunity. Infection of cells with CSFV is mediated by the interaction of E2 and Erns with the cell surface receptor. In this paper we report the analysis of E2 epitope by screening a 12-mer random peptide phage display library using the monoclonal antibodies (MAbs), c2410 and a18, raised against CSFV strain alfort Tübingen and reacted with the E2 structural protein. MAbs, c2410 and a18 recognized the same linear epitope, located at aa832- aa 837 (SPTTLR) of E2 protein, but showed some different reactivities with the mimotopes by ELISA and Western blot analysis. Sequencing both cDNA of light chain and heavy chain variable regions of the two MAbs from total RNA of the hybridoma cells was carried out. The results showed that MAb c2410 and MAb a18 are different monoclonal antibodies, though both MAbs derived from the same fusion and recognized the same epitope.更多还原