Identification and epitope mapping of anti-p72 single-chain antibody against African swine fever virus based on phage display antibody library

doi:10.1016/j.jia.2023.07.039

Journal of Integrative Agriculture

2023, Vol. 22

Issue (9): 2834-2847 DOI: 10.1016/j.jia.2023.07.039

Animal Science · Veterinary Medicine

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Identification and epitope mapping of anti-p72 single-chain antibody against African swine fever virus based on phage display antibody library

SONG Jin-xing^{1, 2}, WANG Meng-xiang^{1, 2}, ZHANG Yi-xuan^{1, 2}, WAN Bo^{1, 2}, DU Yong-kun^{1, 2}, ZHUANG Guo-qing^{1, 2}, LI Zi-bin³, QIAO Song-lin⁶, GENG Rui¹, WU Ya-nan^{1, 2#}, ZHANG Gai-ping^{1, 2, 4, 5#}

¹ College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, P.R.China

² International Joint Research Center of National Animal Immunology, Zhengzhou 450046, P.R.China

³ College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, P.R.China

⁴ Longhu Laboratory, Zhengzhou 450046, P.R.China

⁵School of Advanced Agricultural Sciences, Peking University, Beijing 100871, P.R.China

⁶ Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, P.R.China

Abstract
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摘要

非洲猪瘟病毒(ASFV)是一种致命的病原体，对全球养猪业造成灾难性的社会经济影响。到目前为止，还没有获得许可的预防性疫苗。目前对非洲猪瘟病毒主要免疫原和关键抗原表位图谱的了解有限。因此，功能性单克隆抗体(mAb)和表位图谱的研究对我们理解免疫反应和设计改进的疫苗、治疗和诊断至关重要。本研究基于自然感染ASFV的康复期PBMCs构建的ASFV抗体噬菌体展示文库，筛选出一株抗ASFV主要衣壳蛋白(p72)单链抗体，并与IgG Fc片段融合(scFv-83-Fc)，可以特异性识别ASFV Pig/HLJ/2018毒株。进一步，使用scFv-83-Fc单克隆抗体为靶点，我们从M13噬菌体展示随机肽库中鉴定到p72的一段保守表位肽（²²¹MTGYKH²²⁶）。此外，流式细胞术和细胞摄取实验表明，该表位肽在体外能显著促进BMDC成熟，并能被DC细胞有效摄取，这可能意味着其在疫苗和诊断试剂开发中的潜在应用前景。总之，本研究为确定ASFV疫苗开发的靶点提供了一个有价值的平台，并有助于亚单位疫苗和诊断试剂的优化设计。

Abstract

African swine fever virus (ASFV) is a lethal pathogen that causes severe threats to the global swine industry and it has already had catastrophic socio-economic effects. To date, no licensed prophylactic vaccine exists. Limited knowledge exists about the major immunogens of ASFV and the epitope mapping of the key antigens. As such, there is a considerable requirement to understand the functional monoclonal antibodies (mAbs) and the epitope mapping may be of utmost importance in our understanding of immune responses and designing improved vaccines, therapeutics, and diagnostics. In this study, we generated an ASFV antibody phage-display library from ASFV convalescent swine PBMCs, further screened a specific ASFV major capsid protein (p72) single-chain antibody and fused with an IgG Fc fragment (scFv-83-Fc), which is a specific recognition antibody against ASFV Pig/HLJ/2018 strain. Using the scFv-83-Fc mAb, we selected a conserved epitope peptide (²²¹MTGYKH²²⁶) of p72 retrieved from a phage-displayed random peptide library. Moreover, flow cytometry and cell uptake experiments demonstrated that the epitope peptide can significantly promote BMDCs maturation in vitro and could be effectively uptaken by DCs, which indicated its potential application in vaccine and diagnostic reagent development. Overall, this study provided a valuable platform for identifying targets for ASFV vaccine development, as well as to facilitate the optimization design of subunit vaccine and diagnostic reagents

Keywords: ASFV phage display antibody library single chain antibody p72 epitope

Received: 29 April 2023 Accepted: 19 June 2023

Fund: This work was supported by the National Natural Science Foundation of China (31941001 and 32002292), the Major Science and Technology Project of Henan Province, China (221100110600) and the Natural Science Foundation of Henan Province (202300410199).

About author: SONG Jin-xing, Tel: +86-371-56990163, E-mail: 1010746220@qq.com; #Correspondence WU Ya-nan, E-mail: wlyananjiayou@yeah.net; ZHANG Gai-ping, E-mail: zhanggaip@126.com

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	SONG Jin-xing
	WANG Meng-xiang
	ZHANG Yi-xuan
	WAN Bo
	DU Yong-kun
	ZHUANG Guo-qing
	LI Zi-bin
	QIAO Song-lin
	GENG Rui
	WU Ya-nan
	ZHANG Gai-ping

Cite this article:

SONG Jin-xing, WANG Meng-xiang, ZHANG Yi-xuan, WAN Bo, DU Yong-kun, ZHUANG Guo-qing, LI Zi-bin, QIAO Song-lin, GENG Rui, WU Ya-nan, ZHANG Gai-ping. 2023. Identification and epitope mapping of anti-p72 single-chain antibody against African swine fever virus based on phage display antibody library. Journal of Integrative Agriculture, 22(9): 2834-2847.

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