A novel linear B cell epitope of the porcine circovirus type 3 capsid protein identified by phage display technology

IF 1.6 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2025-04-01 Epub Date: 2024-12-05 DOI:10.1016/j.jviromet.2024.115080
Shu-qing Yang , Ke Yang , Xin-ran Li , Yi Zheng , San-jie Cao , Qi-gui Yan , Xiao-bo Huang , Yi-ping Wen , Qin Zhao , Sen-yan Du , Yi- fei Lang , Shan Zhao , Chun Li , Rui Wu
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Abstract

Porcine circovirus type 3 (PCV3) is endemic in swine worldwide and causes reproductive disorders, dermatitis and nephrotic syndrome, and multi-organ inflammation. PCV3 capsid protein (Cap) can self-assemble into virus-like particles (VLPs), and is an ideal candidate for vaccines and diagnostic reagents.In this study, the recombinant PCV3 Cap protein was successfully expressed in E. coli by deleting the nuclear localization sequence (NLS). The PCV3 VLPs were observed by transmission electron microscopy, and its immunogenicity was evaluated in six-week-old female BALB/c mice. A monoclonal antibody was named mAb 2D6, and demonstrated strong reactivity and specificity to PCV3 Cap. The purified mAb 2D6 was further used for bio-panning to select phage expressing specific epitopes from phage-displayed 7 mer-peptide library. A novel linear B-cell epitope, recognized by mAb 2D6, was identified at the amino acid region 47–53 of Cap. The phage peptide sequences were analyzed using multiple sequence alignment and evaluated by peptide ELISA. These results provide insights for developing diagnostic tools and potential vaccines for PCV3.
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利用噬菌体展示技术鉴定猪圆环病毒3型衣壳蛋白线性B细胞表位。
猪圆环病毒3型(PCV3)在世界各地的猪中流行,可引起生殖障碍、皮炎和肾病综合征以及多器官炎症。PCV3衣壳蛋白(Cap)可以自组装成病毒样颗粒(vlp),是疫苗和诊断试剂的理想候选物。本研究通过删除核定位序列(NLS),在大肠杆菌中成功表达了重组PCV3 Cap蛋白。通过透射电镜观察PCV3 VLPs,并在6周龄雌性BALB/c小鼠中评价其免疫原性。该单克隆抗体被命名为mAb 2D6,对PCV3 Cap具有较强的反应性和特异性。纯化后的mAb 2D6进一步用于生物筛选,从噬菌体显示的7 mer-peptide文库中选择表达特异性表位的噬菌体。在Cap的47-53氨基酸区发现一个新的线性b细胞表位,可被mAb 2D6识别。利用多序列比对分析噬菌体肽序列,并利用肽酶联免疫吸附试验(ELISA)对其进行评价。这些结果为开发PCV3的诊断工具和潜在疫苗提供了见解。
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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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