Characterization and epitope mapping of monoclonal antibodies against PEDV N protein

Zhao, Yongxiang; Fan, Baochao; Xue, Jun‐biao; Guo, Ren; Li, Jizhong; Zhou, Jinzhu; Xu, Song; Zhang, Xuehan; Tao, Sheng-Ce; Li, Bin

doi:10.1016/j.virol.2022.12.011

Cited by 5 publications

(2 citation statements)

References 37 publications

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“…The slides were subjected to H&E-stained and immunohistochemistry (IHC) assay. The monoclonal antibody against the PEDV N protein (made in our laboratory) was used as the primary antibody ( 28 ), and an Alexa Fluor 555-labeled Donkey anti-mouse IgG (Beyotime) was used as the secondary antibody to perform IHC.…”

Section: Methodsmentioning

confidence: 99%

PEDV-spike-protein-expressing mRNA vaccine protects piglets against PEDV challenge

Zhao,

Fan,

Song

et al. 2024

mBio

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Porcine epidemic diarrhea virus (PEDV), a swine enteropathogenic coronavirus, causes severe diarrhea in neonatal piglets, which is associated with a high mortality rate. Thus, developing effective and safe vaccines remains a top priority for controlling PEDV infection. Here, we designed two lipid nanoparticle (LNP)-encapsulated mRNA (mRNA-LNP) vaccines encoding either the full-length PEDV spike (S) protein or a multiepitope chimeric spike (Sm) protein. We found that the S mRNA-LNP vaccine was superior to the Sm mRNA-LNP vaccine at inducing antibody and cellular immune responses in mice. Evaluation of the immunogenicity and efficacy of the S mRNA vaccine in piglets confirmed that it induced robust PEDV-specific humoral and cellular immune responses in vivo . Importantly, the S mRNA-LNP vaccine not only protected actively immunized piglets against PEDV but also equipped neonatal piglets with effective passive anti-PEDV immunity in the form of colostrum-derived antibodies after the immunization of sows. Our findings suggest that the PEDV-S mRNA-LNP vaccine is a promising candidate for combating PEDV infection. IMPORTANCE Porcine epidemic diarrhea virus (PEDV) continues to harm the global swine industry. It is important to develop a highly effective vaccine to control PEDV infection. Here, we report a PEDV spike (S) mRNA vaccine that primes a potent antibody response and antigen-specific T-cell responses in immunized piglets. Active and passive immunization can protect piglets against PED following the virus challenge. This study highlights the efficiency of the PEDV-S mRNA vaccine and represents a viable approach for developing an efficient PEDV vaccine.

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Section: Methodsmentioning

confidence: 99%

PEDV-spike-protein-expressing mRNA vaccine protects piglets against PEDV challenge

Zhao,

Fan,

Song

et al. 2024

mBio

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“…Heretofore, mAbs targeting PEDV have predominantly focused on the N and S proteins [ 12 , 13 , 14 , 15 ]. The N protein is comparatively conservative across diverse PEDV strains, rendering it a promising candidate as an early diagnostic indicator for PEDV infection [ 16 ]. Meanwhile, the S protein has the capacity to stimulate the production of neutralizing antibodies in the host [ 17 , 18 ].…”

Section: Introductionmentioning

confidence: 99%

Identification of Three Novel Linear B-Cell Epitopes in Non-Structural Protein 3 of Porcine Epidemic Diarrhea Virus Using Monoclonal Antibodies

Ye,

Zhu,

Yang

et al. 2024

Viruses

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Porcine epidemic diarrhea virus (PEDV) is a highly pathogenic swine coronavirus that causes diarrhea and high mortality in piglets, resulting in significant economic losses within the global swine industry. Nonstructural protein 3 (Nsp3) is the largest in coronavirus, playing critical roles in viral replication, such as the processing of polyproteins and the formation of replication-transcription complexes (RTCs). In this study, three monoclonal antibodies (mAbs), 7G4, 5A3, and 2D7, targeting PEDV Nsp3 were successfully generated, and three distinct linear B-cell epitopes were identified within these mAbs by using Western blotting analysis with 24 truncations of Nsp3. The epitope against 7G4 was located on amino acids 31-TISQDLLDVE-40, the epitope against 5A3 was found on amino acids 141-LGIVDDPAMG-150, and the epitope against 2D7 was situated on amino acids 282-FYDAAMAIDG-291. Intriguingly, the epitope 31-TISQDLLDVE-40 recognized by the mAb 7G4 appears to be a critical B-cell linear epitope due to its high antigenic index and exposed location on the surface of Nsp3 protein. In addition, bioinformatics analysis unveiled that these three epitopes were highly conserved in most genotypes of PEDV. These findings present the first characterization of three novel linear B-cell epitopes in the Nsp3 protein of PEDV and provide potential tools of mAbs for identifying host proteins that may facilitate viral infection.

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A novel double antibody sandwich quantitative ELISA for detecting porcine epidemic diarrhea virus infection

Han,

Ma,

et al. 2024

Appl Microbiol Biotechnol

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Porcine epidemic diarrhea (PED), a contagious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV), has caused significant economic losses to the global pig farming industry due to its rapid course and spread and its high mortality among piglets. In this study, we prepared rabbit polyclonal antibody and monoclonal antibody 6C12 against the PEDV nucleocapsid (N) protein using the conserved and antigenic PEDV N protein as an immunogen. A double-antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-qELISA) was established to detect PEDV using rabbit polyclonal antibodies as capture antibodies and horseradish peroxidase (HRP)-labeled 6C12 as the detection antibody. Using DAS-qELISA, recombinant PEDV N protein, and virus titer detection limits were approximately 0.05 ng/mL and 10 3.02 50% tissue culture infective dose per mL (TCID 50 /mL), respectively. There was no cross-reactivity with porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (PoRV), porcine pseudorabies virus (PRV), porcine deltacoronavirus (PDCoV), or porcine circovirus (PCV). The reproducibility of DAS-qELISA was verified, and the coefficient of variation (CV) for intra- and inter-batch replicates was less than 10%, indicating good reproducibility. When testing anal swab samples from PEDV-infected piglets using DAS-qELISA, the coincidence rate was 92.55% with a kappa value of 0.85 when using reverse transcription-polymerase chain reaction (RT-PCR) and 94.29% with a kappa value of 0.88 when using PEDV antigen detection test strips, demonstrating the reliability of the method. These findings provide fundamental material support for both fundamental and practical studies on PEDV and offer a crucial diagnostic tool for clinical applications. Key points • A new anti-PEDV N protein monoclonal antibody strain was prepared • Establishment of a more sensitive double antibody sandwich quantitative ELISA • DAS-qELISA was found to be useful for controlling the PEDV spread

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Characterization and epitope mapping of monoclonal antibodies against PEDV N protein

Cited by 5 publications

References 37 publications

PEDV-spike-protein-expressing mRNA vaccine protects piglets against PEDV challenge

PEDV-spike-protein-expressing mRNA vaccine protects piglets against PEDV challenge

Identification of Three Novel Linear B-Cell Epitopes in Non-Structural Protein 3 of Porcine Epidemic Diarrhea Virus Using Monoclonal Antibodies

A novel double antibody sandwich quantitative ELISA for detecting porcine epidemic diarrhea virus infection

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