Wang Yuzhen, Zhang Xiaohua, Xiao Nan, Zhang Min, Dai Heping
Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Aug;24(8):1387-94.
White spot syndrome virus (WSSV) is one of the most important pathogens in shrimp farm throughout the world. Many researches on WSSV have been done, but no efficient approach has been gained to protect and cure the disease. In this study, we constructed a single-chain fragment variable (scFv) antibody library displayed on phage using spleen cells from mice immunized with denatured WSSV. After several rounds of panning respectively against purified intact WSSV virions and purified VP28 expressed in Escherichia coli, five novel scFv antibodies specifically against WSSV were selected, one of which, clone P75E8, recognized a linear epitope. The location in virions of the epitopes recognized by the five scFv clones was determined by immunoelectron microscopy. This study provides a new way to obtain more different antibodies specifically binding to WSSV, and especially provides a new strategy to obtain scFvs against linear epitopes.
白斑综合征病毒（WSSV）是全球对虾养殖中最重要的病原体之一。针对WSSV已开展了许多研究，但尚未找到有效的防治该疾病的方法。在本研究中，我们利用经变性WSSV免疫的小鼠脾细胞构建了展示于噬菌体上的单链抗体可变区（scFv）抗体文库。分别经过几轮针对纯化的完整WSSV病毒粒子和在大肠杆菌中表达的纯化VP28的淘选，筛选出了五种特异性针对WSSV的新型scFv抗体，其中克隆P75E8识别一个线性表位。通过免疫电镜确定了五个scFv克隆所识别表位在病毒粒子中的位置。本研究为获得更多特异性结合WSSV的不同抗体提供了一条新途径，尤其为获得针对线性表位的scFv提供了一种新策略。
