Neutralizing monoclonal antibodies against porcinophilic foot-and-mouth disease virus mapped to antigenic site 2 by utilizing novel mutagenic virus-like particles to detect the antigenic change

Lee, Heng-Wei; Deng, Ming-Chung; Pan, Chu-Hsiang; Chang, Hui-Wen; Cheng, Ivan-Chen

doi:10.1016/j.vetmic.2018.06.002

Cited by 5 publications

(9 citation statements)

References 25 publications

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“…Previously, we determined that the Q10E MAb recognized site 1 based on a synthetic GH peptide on VP1 [ 22 ] and successfully proved the P11A MAb as the site 2 antibody using mutated VLP/VP2-S72N [ 15 ]. Meanwhile, the S11B MAb, a neutralizing antibody, was found not to recognize the unprocessed protomer P1.…”

Section: Resultsmentioning

confidence: 99%

“…Meanwhile, the S11B MAb, a neutralizing antibody, was found not to recognize the unprocessed protomer P1. After processing and assembly, the capsid proteins formed a higher-order oligomer, possibly a pentamer, which could be bound to S11B [ 15 ]. To examine the binding of S11B at the neutralizing site, we expressed VLPs from putative escape mutants of O/TW/97, which included the VP1-L148R mutant for site 1, VP2-S72N and VP2-S131P mutants for site 2, VP1-P44L and VP1-K43A/P44A for site 3, VP3-D58K and VP3-H56A/D58A for site 4, and VP1-Q149H for site 5 ( Figure 1 A).…”

Section: Resultsmentioning

confidence: 99%

“…In the case of serotype O, for example, apart from linear site 1, the other four neutralizing sites are conformational epitopes, which cannot be represented by single subunit proteins [ 9 , 25 ]. For example, S11B, a site 3 MAb, could not recognize VP1 (data not provided) or the unprocessed P1 protomer [ 15 ], indicating that site 3 was formed only after the processing and assembly of P1. Similarly, Dong et al highlighted the subtle differences in the structures of a pentamer and a virion using cryo-EM, indicating that the virion package caused a structural shift in the BC and EF loops of VP2 [ 26 ].…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, with the exception of site 1, the neutralizing sites have conformational structures and cannot be presented by synthetic peptides [ 9 ]. Therefore, a platform based on virus-like particles (VLPs) established in a previous study was used to successfully identify more than six site 2 MAbs whose binding to VLPs could be abrogated or inhibited by VP2-S72N mutation on the VLPs [ 15 ]. The platform preserved the authentic antigenicity without using live or inactivated viruses.…”

Section: Introductionmentioning

confidence: 99%

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The Use of Distinctive Monoclonal Antibodies in FMD VLP- and P1-Based Blocking ELISA for the Seromonitoring of Vaccinated Swine

Lee

Yang

Lee

et al. 2022

IJMS

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The serum neutralization (SN) test has been regarded as the “gold standard” for seroconversion following foot-and-mouth disease virus (FMDV) vaccination, although a high-level biosafety laboratory is necessary. ELISA is one alternative, and its format is constantly being improved. For instance, standard polyclonal antisera have been replaced by monoclonal antibodies (MAbs) for catching and detecting antibodies, and inactive viruses have been replaced by virus-like particles (VLPs). To the best of current knowledge, however, no researchers have evaluated the performances of different MAbs as tracers. In previous studies, we successfully identified site 1 and site 2 MAbs Q10E and P11A. In this study, following the established screening platform, the VLPs of putative escape mutants from sites 1 to 5 were expressed and used to demonstrate that S11B is a site 3 MAb. Additionally, the vulnerability of VLPs prompted us to assess another diagnostic antigen: unprocessed polyprotein P1. Therefore, we established and evaluated the performance of blocking ELISA (bELISA) systems based on VLPs and P1, pairing them with Q10E, P11A, S11B, and the non-neutralizing TSG MAb as tracers. The results indicated that the VLP paired with S11B demonstrated the highest correlation with the SN titers (R2 = 0.8071, n = 63). Excluding weakly positive serum samples (SN = 16–32, n = 14), the sensitivity and specificity were 95.65% and 96.15% (kappa = 0.92), respectively. Additionally, the P1 pairing with Q10E also demonstrated a high correlation (R2 = 0.768). We also discovered that these four antibodies had steric effects on one another to varying degrees, despite recognizing distinct antigenic sites. This finding indicated that MAbs as tracers could not accurately detect specific antibodies, possibly because MAbs are bulky compared to a protomeric unit. However, our results still provide convincing support for the application of two pairs of bELISA systems: VLP:S11B-HRP and P1:Q10E-HRP.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The Use of Distinctive Monoclonal Antibodies in FMD VLP- and P1-Based Blocking ELISA for the Seromonitoring of Vaccinated Swine

Lee

Yang

Lee

et al. 2022

IJMS

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“…Their isotypes all belonged to IgG2a/κ. The puri ed monoclonal antibodies were obtained as in a previous study 55 . The hybridoma was inoculated intraperitoneally into BALB/c for ascetic uid and puri ed by Protein G (GE Healthcare).…”

Section: (Iii) Othersmentioning

confidence: 99%

Foot-and-mouth disease virus 3A hijacks Sar1 and Sec12 for ER remodeling in a COPII-independent manner

Lee

Jiang

Chang

et al. 2022

Preprint

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Positive-stranded RNA viruses modify host organelles to form replication organelles (ROs) for their replication. Enteroviral 3A protein has been demonstrated to be highly associated with the COPI pathway, in which factors work on the ER-to-Golgi intermediate and the Golgi. However, Sar1, a COPII factor exerting coordinated action at endoplasmic reticulum (ER) exit sites, rather than COPI factors, is required for foot-and-mouth disease virus (FMDV) replication. Therefore, we thought that deep understanding of FMDV 3A was the key to explaining the differences and to unlocking the secret of FMDV RO formation. In this study, FMDV 3A was confirmed as a peripheral membrane protein capable of modifying the ER into vesicle-like structures, which were neither COPII vesicles nor autophagosomes. When the C-terminus of 3A was truncated, it would be located at the ER without vesicular modification. This change was revealed by mGFP and APEX2 fusion constructs observed by fluorescence microscopy and electron tomography, respectively. Referring to other 3A truncation, the minimal region for modification was aa 42–92. Furthermore, we found that the remodeling was related to two COPII factors, Sar1 and Sec12. Both interacted with 3A, but their binding domains on 3A were different. Finally, we hypothesized that the N-terminus of 3A would interact with Sar1 as its C-terminus simultaneously interacted with Sec12, which possibly would enhance Sar1 activation. On the ER membrane, two active Sar1 were connected by 3A with regions of aa 42–59 and aa 76–92, causing curvature of the membrane. This mechanism is distinct from the traditional COPII pathway and should be crucial for FMDV RO formation.

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Identification of antigenic epitopes in the haemagglutinin protein of H7 avian influenza virus

Chen

Wang

et al. 2019

Avian Pathology

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Neutralizing monoclonal antibodies against porcinophilic foot-and-mouth disease virus mapped to antigenic site 2 by utilizing novel mutagenic virus-like particles to detect the antigenic change

Cited by 5 publications

References 25 publications

The Use of Distinctive Monoclonal Antibodies in FMD VLP- and P1-Based Blocking ELISA for the Seromonitoring of Vaccinated Swine

The Use of Distinctive Monoclonal Antibodies in FMD VLP- and P1-Based Blocking ELISA for the Seromonitoring of Vaccinated Swine

Foot-and-mouth disease virus 3A hijacks Sar1 and Sec12 for ER remodeling in a COPII-independent manner

Identification of antigenic epitopes in the haemagglutinin protein of H7 avian influenza virus

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