Identification and characterization of new B cell epitopes on the nucleocapsid protein of porcine epidemic diarrhea virus using monoclonal antibodies

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology Pub Date : 2024-08-08 DOI:10.1016/j.vetmic.2024.110200

Meng Sun , Yangyang Sun , Lujie Zhang , Yanni Gao , Zhunxuan Wang , Xianwei Wang , Ping Jiang , Juan Bai

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引用次数: 0

Abstract

Porcine epidemic diarrhea virus (PEDV) is the pathogen of Porcine epidemic diarrhea (PED) and can mainly cause acute diarrhea, vomiting, dehydration and high mortality in neonatal piglets. The nucleocapsid (N) protein of PEDV is a highly conserved structural protein. In this study, 6–8-week-old BALB/c mice were immunized with purified PEDV, and three monoclonal antibodies (mAbs) against the PEDV N protein were generated, named 3C6,4F8,4C9. Among them, three new B cell epitopes, ²³⁵IGENPDKL²⁴², ¹²KRVPLSLY¹⁹, ³⁷²DAFKTGNA³⁸⁰ were firstly identified in the viral N-protein. Among them, 4F8 and 4C9 had IgG1 isotype with Kappa light chain, while 3C6 had IgG2a isotype with Kappa light chain. Three monoclonal antibodies (mAbs) demonstrated specific reactivity with PEDV as evidenced by Western blot and indirect immunofluorescence assay. By studying the interaction between the mAbs and the N protein, we can gain insights into the protein's conformation and functional regions. This information will help develop fast and accurate PEDV diagnostic methods.

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使用单克隆抗体鉴定猪流行性腹泻病毒核壳蛋白上的新 B 细胞表位并确定其特征。

猪流行性腹泻病毒（PEDV）是猪流行性腹泻（PED）的病原体，主要引起新生仔猪急性腹泻、呕吐、脱水和高死亡率。PEDV 的核壳（N）蛋白是一种高度保守的结构蛋白。本研究用纯化的PEDV对6-8周龄的BALB/c小鼠进行免疫，产生了三种针对PEDV N蛋白的单克隆抗体（mAbs），分别命名为3C6、4F8和4C9。其中，235IGENPDKL242、12KRVPLSLY19、372DAFKTGNA380三个新的B细胞表位首次在病毒N蛋白中被发现。其中，4F8 和 4C9 为带有 Kappa 轻链的 IgG1 同工型，3C6 为带有 Kappa 轻链的 IgG2a 同工型。通过 Western 印迹和间接免疫荧光检测，三种单克隆抗体（mAbs）显示出与 PEDV 的特异性反应。通过研究 mAbs 与 N 蛋白之间的相互作用，我们可以深入了解该蛋白的构象和功能区。这些信息将有助于开发快速准确的 PEDV 诊断方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Veterinary microbiology 农林科学-兽医学

CiteScore

5.90

自引率

6.10%

发文量

221

审稿时长

52 days

期刊介绍： Veterinary Microbiology is concerned with microbial (bacterial, fungal, viral) diseases of domesticated vertebrate animals (livestock, companion animals, fur-bearing animals, game, poultry, fish) that supply food, other useful products or companionship. In addition, Microbial diseases of wild animals living in captivity, or as members of the feral fauna will also be considered if the infections are of interest because of their interrelation with humans (zoonoses) and/or domestic animals. Studies of antimicrobial resistance are also included, provided that the results represent a substantial advance in knowledge. Authors are strongly encouraged to read - prior to submission - the Editorials (''Scope or cope'' and ''Scope or cope II'') published previously in the journal. The Editors reserve the right to suggest submission to another journal for those papers which they feel would be more appropriate for consideration by that journal. Original research papers of high quality and novelty on aspects of control, host response, molecular biology, pathogenesis, prevention, and treatment of microbial diseases of animals are published. Papers dealing primarily with immunology, epidemiology, molecular biology and antiviral or microbial agents will only be considered if they demonstrate a clear impact on a disease. Papers focusing solely on diagnostic techniques (such as another PCR protocol or ELISA) will not be published - focus should be on a microorganism and not on a particular technique. Papers only reporting microbial sequences, transcriptomics data, or proteomics data will not be considered unless the results represent a substantial advance in knowledge. Drug trial papers will be considered if they have general application or significance. Papers on the identification of microorganisms will also be considered, but detailed taxonomic studies do not fall within the scope of the journal. Case reports will not be published, unless they have general application or contain novel aspects. Papers of geographically limited interest, which repeat what had been established elsewhere will not be considered. The readership of the journal is global.