溶藻弧菌脂多糖单克隆抗体的制备、鉴定及初步应用  

Development, Characterization and Application of Monoclonal Antibody against Vibrio alginolyticus Lipopolysaccharide

作　　者：陈晓燕[1] 胡超群[1] 任春华[1] 陈偿[1] 

机构地区：[1]中国科学院南海海洋研究所,广州510301

出　　处：《高技术通讯》2002年第11期90-95,共6页Chinese High Technology Letters

基　　金：973 (G19990 12 0 0 3 )资助项目

摘　　要：用细菌颗粒性抗原多次免疫Balb/c小鼠 ,首次制备了一株鱼类病原溶藻弧菌VAF0 1的LPS的单克隆抗体 4D5。 4D5识别VAF0 1和VAF0 2 (另一株病原溶藻弧菌 )的LPS的核心多糖抗原。除了与测试的溶藻弧菌发生免疫识别外 ,4D5还与解蛋白弧菌、哈维氏弧菌、灿烂弧菌和海藻施万氏菌发生免疫交叉反应 ,但不与副溶血弧菌、鳗弧菌、嗜水气单胞菌等发生交叉反应。用温和高碘酸氧化法对LPS处理后 ,VAF0 1的LPS不再与 4D5识别 ,VAF0 2的LPS与 4D5的反应部分减弱 ,推测VAF0 1及VAF0 2的LPS有不同的组分和结构。用抑制ELISA法半定量检测溶藻弧菌培养上清的LPS ,测得培养4 8小时LPS释放量不超过 5 7μg/ml,大大低于使鱼类致死所需的LPS的剂量。A monoclonal antibody (Mab) 4D5 against lipopolysaccharide (LPS) from a pathogenic V. alginolyticus VAF01 was produced and characterized. 4D5 reacted with V. proteolyticus, V. harveyi, V. splendidus,Shewanella algae and 5 V. alginoliticus isolates among 13 bacteria species tested. Immunoblotting after SDS PAGE shows that 4D5 is specific for core oligosaccharide of VAF01 and VAF02 (another pathogenic V. alginolyticus). 4D5 did not recognize VAF01 LPS, but reacted with VAF02 LPS after LPS oxidation by mild periodate acid, implying that some components of VAF01 and VAF02 LPS may be different. The LPS released into cultured medium in 48 hours was not more than 57μg/ml semiquantitated by inhibition ELISA, which is by far less than the amount necessary for fish lethality.

关 键 词：制备 鉴定 溶藻弧菌 脂多糖 单克隆抗体 核心多糖 温和高碘酸氧化反应 抑制ELISA法 鱼类病原菌 

分 类 号：Q813.2[生物学—生物工程]