Wu Bing, Zhu Bibo, Luo Yuling, Wang Min, Lu Yuanan, Wang Weimin, Cao Shengbo, Huang Fengtao, Liu Xueqin
College of Fisheries, Huazhong Agricultural University, Wuhan, China.
Hybridoma (Larchmt). 2011 Dec;30(6):555-8. doi: 10.1089/hyb.2011.0063.
Three monoclonal antibodies (MAbs) against channel catfish virus (CCV) were generated from mice immunized with purified CCV. Western blot analysis revealed that the MAb 3G12 reacted with three CCV proteins of 94 kDa, 130 kDa, and 170 kDa; the MAb 4C4 reacted with two CCV proteins of 130 kDa and 170 kDa; and the MAb 4D4 reacted with two CCV proteins of 94 kDa and 98 kDa. Indirect immunofluorescence assay showed intense fluorescence in the CCV-infected channel catfish ovary (CCO) cells in areas corresponding to the location of granular structures. In addition, the three MAbs could completely neutralize CCV at a dilution of 1:500. This study demonstrated that these MAbs could recognize CCV specifically and will be useful in the development of diagnostic methods for the detection of fish CCV infection.
用纯化的斑点叉尾鮰病毒(CCV)免疫小鼠,制备了三种抗CCV的单克隆抗体(MAb)。蛋白质免疫印迹分析显示,单克隆抗体3G12与94 kDa、130 kDa和170 kDa的三种CCV蛋白发生反应;单克隆抗体4C4与130 kDa和170 kDa的两种CCV蛋白发生反应;单克隆抗体4D4与94 kDa和98 kDa的两种CCV蛋白发生反应。间接免疫荧光试验显示,在CCV感染的斑点叉尾鮰卵巢(CCO)细胞中,对应颗粒结构位置的区域有强烈荧光。此外,这三种单克隆抗体在1:500稀释度下可完全中和CCV。本研究表明,这些单克隆抗体能够特异性识别CCV,将有助于开发检测鱼类CCV感染的诊断方法。