Identification of a dominant linear epitope on the VP2 capsid protein of porcine parvovirus and characterization of two monoclonal antibodies with neutralizing abilities

Liu, Yunchao; Wang, Jucai; Chen, Yumei; Wang, Aiping; Wei, Qiang; Yang, Suzhen; Feng, Hao; Chai, Shujun; Liu, Dongmin; Zhang, Gaiping

doi:10.1016/j.ijbiomac.2020.09.055

Cited by 7 publications

(8 citation statements)

References 23 publications

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“…Not only, our results but also threonine have been reported in other areas such as Brazil, China, and Germany. The resultant 320 residues occurred as a consequence of its epitope location; thus, upon closer examination, the sites identified in this research study were found to be congruent with the previously identified B cell epitopes (Sun et al, 2015;Liu et al, 2020). Thus, amino acid changes of VP2 capsid protein may be the result of immune evasion.…”

Section: Discussionsupporting

confidence: 71%

“…It has been hypothesized that the VP2 capsid protein of the two surrogate strains could modify certain biological properties e.g. viral tissue tropism while stimulating the host immune system to produce a tentative neutralizing antibody (Vasudevacharya and Compans, 1992;Sun et al, 2015;Liu et al, 2020). This would likely be achieved by stimulating the host immune system to produce a tentative neutralizing antibody and through viral tissue tropism.…”

Section: Introductionmentioning

confidence: 99%

“…This would likely be achieved by stimulating the host immune system to produce a tentative neutralizing antibody and through viral tissue tropism. Epitopes that generate neutralizing antibodies are found in the VP2 capsid protein have been identified according to the antigenic structure of PPV using synthetic peptides, monoclonal antibodies, and viral-like particles (Zhou et al, 2010;Guo et al, 2014;Sun et al, 2015;Hua et al, 2020;Liu et al, 2020;Streck et al, 2022). The VP2 capsid protein plays a role in cell-virus interaction before viral replication because a change in residue on the VP2 capsid protein has been known to contribute to viral replication.…”

Section: Introductionmentioning

confidence: 99%

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Molecular characterization of porcine parvovirus 1 based on partial VP2 gene in the ovaries of Thai pigs

Saekhow¹,

Sriphannam

Yamsakul

2022

VIS

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Porcine parvovirus 1 (PPV1) is the causative agent of swine reproductive disease, particularly in naive gilts and sows. This study aimed to investigate the prevalence and genetic diversity of the partial nucleotide sequence of the VP2 gene and to compare the substitution of amino acid residues that affect relevant biological properties. The prevalence of PPV1 was found to be 12% (12/100) when the viral genome was detected using polymerase chain reaction (PCR). Determination of the genetic diversity of a partial nucleotide sequence of the VP2 gene through phylogenetic analysis indicated that a single cluster of Thai PPV1s was allocated on the phylogenetic tree. According to a comparison of the substitution of amino acid residues that affected the biological properties at 378, 383, 365, and 436 of the VP2 capsid protein between the 12 Thai PPV1s, the Kresse strain (a surrogate pathogenic strain), and the NADL-2 strain (a surrogate nonpathogenic strain). It was determined that the substitution of amino acid residues at 378, 383, and 436 of 12 Thai PPV2s was identical to those of the Kresse strains. The substitution of amino acid residues at 436 of the 12 Thai PPV1s was similar to that of a proven virulent strain in vivo. Additionally, substituting amino acid residue at 320 of the VP2 capsid protein revealed that seven Thai PPV1s were associated with isoleucine PPV1s and identical to that of both surrogate strains, whereas five Thai PPV1s were associated with threonine. This outcome was similar to what had been deposited in GenBank. Our data suggest that Thai PPV1s isolated from the ovaries of pigs raised in Chiang Mai may have originated from the Kresse strains. Based on a change of VP2 capsid protein that occurred amongst the substitution amino acid residue at 320 of the VP2 capsid protein, viruses found in this region were determined to be similar to those found in other areas. This was likely because the viruses had adapted to evade the immune systems of animals.

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Section: Discussionsupporting

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular characterization of porcine parvovirus 1 based on partial VP2 gene in the ovaries of Thai pigs

Saekhow¹,

Sriphannam

Yamsakul

2022

VIS

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“…SE131, China) was used to detect the peptide-bound mAbs at room temperature for 1 h. Finally, 100 µl/well TMB substrate was added to develop and 50 µl/well 2 M H 2 SO 4 was used as the stop buffer. The OD value at 450 nm was performed using the microplate reader [ 21 ] (Omega, Germany).…”

Section: Methodsmentioning

confidence: 99%

Identification of three conserved linear B cell epitopes on the SARS-CoV-2 spike protein

Wang

Tian

Liu

et al. 2022

Emerging Microbes & Infections

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Spike (S) glycoprotein is the most significant structural protein of SARS-CoV-2 and a key target for neutralizing antibodies. In light of the on-going SARS-CoV-2 pandemic, identification and screening of epitopes of spike glycoproteins will provide vital progress in the development of sensitive and specific diagnostic tools. In the present study, NTD, RBD, and S2 genes were inserted into the pcDNA3.1(+) vector and designed with N-terminal 6× His-tag for fusion expression in HEK293F cells by transient transfection. Six monoclonal antibodies (4G, 9E, 4B, 7D, 8F, and 3D) were prepared using the expressed proteins by cell fusion technique. The characterization of mAbs was performed by indirect -ELISA, western blot, and IFA. We designed 49 overlapping synthesized peptides that cover the extracellular region of S protein in which 6 amino acid residues were offset between adjacent (S1–S49). Peptides S12, S19, and S49 were identified as the immunodominant epitope regions by the mAbs. These regions were further truncated and the peptides S12.2 286 TDAVDCALDPLS 297 , S19.2 464 FERDISTEIYQA 475 , and S49.4 1202 ELGKYEQYIKWP 1213 were identified as B- cell linear epitopes for the first time. Alanine scans showed that the D 467, I 468, E 471, Q 474, and A 475 of the epitope S19.2 and K 1205, Q 1208, and Y 1209 of the epitope S49.4 were the core sites involved in the mAbs binding. The multiple sequence alignment analysis showed that these three epitopes were highly conserved among the variants of concern (VOCs) and variants of interest (VOIs). Taken together, the findings provide a potential material for rapid diagnosis methods of COVID-19.

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“…Mice with high antibody titers were lastly boosted by intraperitoneal injection with 50 µg rVP3 pro for hybridoma production. Mice were euthanized three days later, and spleen cells were obtained and then fused with SP2/0 cells by PEG 1500 (Solarbio, Beijing, China) as previously described [18]. The fused cells were plated into 96-well plates and grew in a hypoxanthine-aminopterin-thymidine (HAT) selection medium.…”

Section: Preparation Of Monoclonal Antibodies Against Vp3mentioning

confidence: 99%

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

Chen

Wang

et al. 2021

Viruses

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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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Identification of a dominant linear epitope on the VP2 capsid protein of porcine parvovirus and characterization of two monoclonal antibodies with neutralizing abilities

Cited by 7 publications

References 23 publications

Molecular characterization of porcine parvovirus 1 based on partial VP2 gene in the ovaries of Thai pigs

Molecular characterization of porcine parvovirus 1 based on partial VP2 gene in the ovaries of Thai pigs

Identification of three conserved linear B cell epitopes on the SARS-CoV-2 spike protein

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

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