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猪流行性腹泻病毒刺突蛋白特异性单域抗体的制备与表征

Preparation and characterization of a single-domain antibody specific for the porcine epidemic diarrhea virus spike protein.

作者信息

Bao Fuxiang, Wang Lixin, Zhao Xinxin, Lu Ting, Na A Mi, Wang Xuefei, Cao Jinshan, Du Yanan

机构信息

College of Veterinary Medicine, Inner Mongolia Agricultural University, No. 306, Zhaowuda Road, Saihan District, Huhhot, 010018, China.

Key Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture (LDTA), Huhhot, China.

出版信息

AMB Express. 2019 Jul 12;9(1):104. doi: 10.1186/s13568-019-0834-1.

Abstract

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 × 10. 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 in E. 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.

摘要

猪流行性腹泻(PED)是一种由猪流行性腹泻病毒(PEDV)引起的猪腹泻疾病。其特征是所有年龄段的猪都会出现急性水样腹泻、脱水和呕吐,对新生仔猪和断奶后仔猪尤其致命。PEDV的刺突蛋白在介导病毒与靶细胞的附着和融合中起重要作用,最近的研究还报道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对象。在本研究中,我们成功构建了一个库容量为3.4×10的免疫噬菌体展示单域抗体库。从噬菌体展示单域抗体库中鉴定出一种对PEDV刺突蛋白具有特异性的单域抗体,命名为S7。S7能在大肠杆菌中以可溶形式表达,在酶联免疫吸附测定(ELISA)中与PEDV的刺突蛋白结合,并在非洲绿猴肾细胞(Vero细胞)中对PEDV病毒进行染色,但对PEDV没有中和活性。这些结果表明S7抗体作为一种成像探针或作为诊断检测开发候选对象具有潜在应用价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b73/6626092/a5ddf551bf2b/13568_2019_834_Fig1_HTML.jpg

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