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非洲猪瘟病毒p17在CHO细胞中的表达及利用单克隆抗体鉴定一个新表位

Expression of ASFV p17 in CHO cells and identification of one novel epitope using a monoclonal antibody.

作者信息

Li Liwei, Qiao Sina, Wang Shumao, Liu Jiachen, Zhao Kuan, Zhou Yanjun, Li Guoxin, Jiang Yifeng, Liu Changlong, Tong Guangzhi, Tong Wu, Gao Fei

机构信息

Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.

Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China; College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China.

出版信息

Virus Res. 2023 Oct 15;336:199194. doi: 10.1016/j.virusres.2023.199194. Epub 2023 Aug 23.

Abstract

As a highly pathogenic large DNA virus, African swine fever virus (ASFV) has huge particles and numerous encoded proteins. At present, few of the existing studies on ASFV proteins have investigated the function of p17. Specific antibodies against p17 to promote the development of prevention techniques against African swine fever (ASF) are urgently needed. Herein, we successfully expressed ASFV p17 in CHO cells using a suspension culture system and generated a monoclonal antibody (mAb) against p17. The mAb recognized a novel linear epitope (LLSHNLSTREGIK) and exhibited specific reactivity, which was conducive to the identification of ectopically expressed p17, the recombinant porcine reproductive and respiratory syndrome virus expressing p17, and the ASFV-SY18. The epitope was conservative among genotype I and genotype II ASFV strains. Overall, the mAb against p17 revealed efficient detection and promising application prospects, making it a useful tool for future vaccine research on ASF. Determination of the conserved linear epitope of p17 would contribute to the in-depth exploration of the biological function of ASFV antigen protein.

摘要

作为一种高致病性大型DNA病毒,非洲猪瘟病毒(ASFV)具有巨大的颗粒和众多编码蛋白。目前,现有的关于ASFV蛋白的研究中很少有对p17功能进行探究的。迫切需要针对p17的特异性抗体以促进非洲猪瘟(ASF)预防技术的发展。在此,我们利用悬浮培养系统在CHO细胞中成功表达了ASFV p17,并制备了针对p17的单克隆抗体(mAb)。该mAb识别一个新的线性表位(LLSHNLSTREGIK)并表现出特异性反应性,这有利于鉴定异位表达的p17、表达p17的重组猪繁殖与呼吸综合征病毒以及ASFV-SY18。该表位在I型和II型ASFV毒株中是保守的。总体而言,针对p17的mAb显示出高效检测和良好的应用前景,使其成为未来ASF疫苗研究的有用工具。确定p17的保守线性表位将有助于深入探究ASFV抗原蛋白的生物学功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae21/10470389/36852255b6b3/gr1.jpg

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