Preparation and characterization of a single-domain antibody specific for the porcine epidemic diarrhea virus spike protein
Preparation and characterization of a single-domain antibody specific for the porcine epidemic diarrhea virus spike protein
复制标题
猪流行性腹泻病毒刺突蛋白特异性单域抗体的制备及表征
DOI：
10.1186/s13568-019-0834-1
复制
发表时间：
2019-07-12
期刊：
影响因子：
3.7
通讯作者：
Du, Yanan
中科院分区：
文献类型：
--
作者：
Bao, Fuxiang;Wang, Lixin;Du, Yanan
猪流行性腹泻（Porcine epidemic diarrhea，PED）是由猪流行性腹泻病毒（Porcine epidemic diarrhea virus，PEDV）引起的一种传染性疾病。其特征是所有年龄猪的急性水样腹泻、脱水和呕吐，尤其是新生和断奶后仔猪的致命性。PEDV的刺突蛋白在介导病毒与靶细胞的粘附和融合中起着重要作用，最近的研究也报道了刺突蛋白的中和表位主要位于S1亚基，这使其成为疫苗开发和临床诊断的候选者。本研究成功构建了一个免疫噬菌体展示单域抗体库，库容量为3.4 × 106。从噬菌体展示单域抗体库中筛选到一株特异性抗PEDV刺突蛋白的单域抗体S7。S7在E. coli中表达的重组质粒，ELISA检测与PEDV纤突蛋白结合，Vero细胞染色，但对PEDV无中和活性。这些结果表明S7抗体作为成像探针或作为诊断测定开发的候选物的有效应用。
Porcine epidemic diarrhea (PED) is a diarrheal disease of swine caused by porcine epidemic diarrhea virus (PEDV). It is characterized by acute watery diarrhea, dehydration and vomiting in swine of all ages and is especially fatal for neonatal and postweaning piglets. The spike protein of PEDV plays an important role in mediating virus attachment and fusion to target cells, and recent studies also reported that the neutralizing epitopes of the spike protein were mainly located in the S1 subunit, which makes it a candidate for vaccine development and clinical diagnosis. In this study, we successfully constructed an immune phage display single-domain antibody library with a library size of 3.4 × 106. A single-domain antibody, named S7, specific for the spike protein of PEDV was identified from the phage display single-domain antibody library. S7 could be expressed in a soluble form inE. coli, bound to the spike protein of PEDV in ELISA and stained the PEDV virus in Vero cells, but it showed no neutralization activity on PEDV. These results indicated the potent application of the S7 antibody as an imaging probe or as a candidate for the development of a diagnostic assay.