Identification of a B-Cell Epitope in the VP3 Protein of Senecavirus A

Senecavirus A (SVA) is a member of the genus Senecavirus of the family Picornaviridae. SVA-associated vesicular disease (SAVD) outbreaks have been extensively reported since 2014–2015. Characteristic symptoms include vesicular lesions on the snout and feet as well as lameness in adult pigs and even death in piglets. The capsid protein VP3, a structural protein of SVA, is involved in viral replication and genome packaging. Here, we developed and characterized a mouse monoclonal antibody (mAb) 3E9 against VP3. A motif 192GWFSLHKLTK201 was identified as the linear B-cell epitope recognized by mAb 3E9 by using a panel of GFP-tagged epitope polypeptides. Sequence alignments show that 192GWFSLHKLTK201 was highly conserved in all SVA strains. Subsequently, alanine (A)-scanning mutagenesis indicated that W193, F194, L196, and H197 were the critical residues recognized by mAb 3E9. Further investigation with indirect immunofluorescence assay indicated that the VP3 protein was present in the cytoplasm during SVA replication. In addition, the mAb 3E9 specifically immunoprecipitated the VP3 protein from SVA-infected cells. Taken together, our results indicate that mAb 3E9 could be a powerful tool to work on the function of the VP3 protein during virus infection.

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Benchmarking B-Cell Epitope Prediction for the Design of Peptide-Based Vaccines: Problems and Prospects

Journal of Biomedicine and Biotechnology ◽

10.1155/2010/910524 ◽

2010 ◽

Vol 2010 ◽

pp. 1-14 ◽

Cited By ~ 23

Author(s):

Salvador Eugenio C. Caoili

Keyword(s):

B Cell ◽

Protein Function ◽

Cell Epitope ◽

Epitope Prediction ◽

Cross Reactivity ◽

Sequence Alignments ◽

B Cell Epitopes ◽

B Cell Epitope ◽

Physicochemical Factors ◽

Structural Differences

To better support the design of peptide-based vaccines, refinement of methods to predict B-cell epitopes necessitates meaningful benchmarking against empirical data on the cross-reactivity of polyclonal antipeptide antibodies with proteins, such that the positive data reflect functionally relevant cross-reactivity (which is consistent with antibody-mediated change in protein function) and the negative data reflect genuine absence of cross-reactivity (rather than apparent absence of cross-reactivity due to artifactual masking of B-cell epitopes in immunoassays). These data are heterogeneous in view of multiple factors that complicate B-cell epitope prediction, notably physicochemical factors that define key structural differences between immunizing peptides and their cognate proteins (e.g., unmatched electrical charges along the peptide-protein sequence alignments). If the data are partitioned with respect to these factors, iterative parallel benchmarking against the resulting subsets of data provides a basis for systematically identifying and addressing the limitations of methods for B-cell epitope prediction as applied to vaccine design.

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Identification of a linear B-cell epitope on non-structural protein 12 of porcine reproductive and respiratory syndrome virus, using a monoclonal antibody

Archives of Virology ◽

10.1007/s00705-017-3355-8 ◽

2017 ◽

Vol 162 (8) ◽

pp. 2239-2246 ◽

Cited By ~ 3

Author(s):

Caihong Bi ◽

Zengyu Shao ◽

Yuanfeng Zhang ◽

Liang Hu ◽

Jiangnan Li ◽

...

Keyword(s):

Monoclonal Antibody ◽

B Cell ◽

Cell Epitope ◽

Respiratory Syndrome Virus ◽

Structural Protein ◽

B Cell Epitope ◽

Linear B

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Identification of Critical Residues of Linear B Cell Epitope on Goodpasture Autoantigen

PLoS ONE ◽

10.1371/journal.pone.0123277 ◽

2015 ◽

Vol 10 (4) ◽

pp. e0123277 ◽

Cited By ~ 3

Author(s):

Xiao-yu Jia ◽

Zhao Cui ◽

Jian-nan Li ◽

Shui-yi Hu ◽

Ming-hui Zhao

Keyword(s):

B Cell ◽

Cell Epitope ◽

B Cell Epitope ◽

Critical Residues ◽

Linear B

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Identification of a novel B-cell epitope of Hantaan virus glycoprotein recognized by neutralizing 3D8 monoclonal antibody

Journal of General Virology ◽

10.1099/vir.0.045302-0 ◽

2012 ◽

Vol 93 (12) ◽

pp. 2595-2600 ◽

Cited By ~ 7

Author(s):

Guolin Yan ◽

Yusi Zhang ◽

Ying Ma ◽

Jing Yi ◽

Bei Liu ◽

...

Keyword(s):

Monoclonal Antibody ◽

B Cell ◽

Binding Sites ◽

Cell Epitope ◽

Hantaan Virus ◽

Haemorrhagic Fever ◽

B Cell Epitopes ◽

B Cell Epitope ◽

The Family ◽

The World

Hantaan virus (HTNV), a member of the family Bunyaviridae, is a major agent causing haemorrhagic fever with renal syndrome, a high-mortality-rate disease threatening approximately 150 000 people around the world yearly. The 3D8 mAb displays a neutralizing activity to HTNV infection. In this study, the B-cell epitopes of HTNV glycoproteins (GPs) were finely mapped by peptide scanning. A new B-cell epitope 882GFLCPEFPGSFRKKC896 of HTNV, which locates on Gc, has been screened out from a set of 15-mer synthesized peptides covering the full-length of HTNV-GPs. It has been shown by the alanine-scanning technique that 885C, 893R, 894K, 895K and 896C are the key amino acids of the binding sites of the GPs. The implications of identifying a novel B-cell epitope for hantavirus immunology and vaccinology are discussed.

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