Effect of trypsin digestion on the integrity and antigenic epitopes of GII.6 norovirus virus-like particles

IF 2.5 4区医学 Q3 VIROLOGY

Archives of Virology Pub Date : 2025-01-06 DOI:10.1007/s00705-024-06213-1

Jie Ma, Jinjin Liu, Xinyi Liu, Chao Wang, Yuqi Huo

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Abstract

Trypsin digestion of the GII.6 norovirus (NoV) major capsid protein VP1 promotes its binding to histo-blood group antigens (HBGAs), which are believed to be co-receptors for NoVs. In our previous study, we found that trypsin digestion led to the disassembly of GII.6 NoV virus-like particles (VLPs). In this study, we investigated the effect of trypsin digestion on the integrity of GII.6 NoV VLPs using a modified purification approach and determined the N- and C-terminal residues of the fragments produced by digestion, using peptide mass fingerprinting. We also characterized the antigenic epitopes that were affected by trypsin digestion using monoclonal antibodies (mAbs). Our results indicated that GII.6 VLPs remained intact even after complete cleavage at the P1-1/P2 junction. Most of the mAbs in supernatants of hybridoma cell clones showed reduced binding to trypsin-digested GII.6 VLPs. Eight mAbs that showed reduced binding to digested GII.6 VP1 were produced, and these were found primarily to recognize residues located in the P domain. Our results provide evidence of flexibility and extensive morphological changes in the antigenic epitope of GII.6 VLPs after trypsin digestion. It remains to be investigated whether this phenomenon also occurs in virions.

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胰蛋白酶酶切对GII.6诺如病毒样颗粒完整性和抗原表位的影响

胰蛋白酶消化GII.6诺如病毒（NoV）主要衣壳蛋白VP1促进其与组织血型抗原（HBGAs）的结合，HBGAs被认为是NoVs的共受体。在我们之前的研究中，我们发现胰蛋白酶消化导致GII.6 NoV病毒样颗粒（VLPs）的分解。在本研究中，我们使用改进的纯化方法研究了胰蛋白酶消化对GII.6 NoV VLPs完整性的影响，并使用肽质量指纹图谱测定了消化产生的片段的N端和c端残基。我们还利用单克隆抗体（mab）鉴定了受胰蛋白酶消化影响的抗原表位。我们的研究结果表明，即使在P1-1/P2连接处完全切割后，GII.6 VLPs仍然保持完整。杂交瘤细胞克隆上清中大多数单克隆抗体与胰蛋白酶消化的GII.6 VLPs结合减少。8个单抗显示与消化GII.6 VP1结合减少，发现这些单抗主要识别位于P结构域的残基。我们的研究结果提供了GII.6 VLPs在胰蛋白酶消化后抗原表位的灵活性和广泛形态变化的证据。这种现象是否也发生在病毒粒子中还有待研究。

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

Archives of Virology 医学-病毒学

CiteScore

5.10

自引率

7.40%

发文量

324

审稿时长

4.5 months

期刊介绍： Archives of Virology publishes original contributions from all branches of research on viruses, virus-like agents, and virus infections of humans, animals, plants, insects, and bacteria. Coverage spans a broad spectrum of topics, from descriptions of newly discovered viruses, to studies of virus structure, composition, and genetics, to studies of virus interactions with host cells, organisms and populations. Studies employ molecular biologic, molecular genetics, and current immunologic and epidemiologic approaches. Contents include studies on the molecular pathogenesis, pathophysiology, and genetics of virus infections in individual hosts, and studies on the molecular epidemiology of virus infections in populations. Also included are studies involving applied research such as diagnostic technology development, monoclonal antibody panel development, vaccine development, and antiviral drug development.Archives of Virology wishes to publish obituaries of recently deceased well-known virologists and leading figures in virology.