A novel neutralizing monoclonal antibody targeting the N-terminal domain of the MERS-CoV spike protein

Chen, Yingzhu; Lu, Shuai; Jia, Hao; Deng, Yao; Zhou, Jianfang; Huang, Baoying; Yu, Yueyang; Lan, Jiaming; Wang, Wenling; Lou, Yongliang; Qin, Kun; Tan, Wenjie

doi:10.1038/emi.2017.18

Cited by 48 publications

(52 citation statements)

References 49 publications

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“…Previous work demonstrated that an anti-PEDV-S2 mAb recognizing the linear epitope GPRLQPY at the carboxy-terminal of the S2 subunit exhibited neutralizing activity against PEDV (Cruz et al, 2008). Similarly, a neutralizing mAb that specifically recognizes the S1-NTD of MERS-CoV S protein has been identified (Chen et al, 2017). In our study, we also found that regions S2 and S1-NTD can induce neutralizing antibody, consistent with previous reports (Chen et al, 2017;Cruz et al, 2008).…”

Section: Discussionsupporting

confidence: 92%

“…Similarly, a neutralizing mAb that specifically recognizes the S1-NTD of MERS-CoV S protein has been identified (Chen et al, 2017). In our study, we also found that regions S2 and S1-NTD can induce neutralizing antibody, consistent with previous reports (Chen et al, 2017;Cruz et al, 2008). In addition, all CoVs use the S1-CTD or S1-NTD regions to interact with cellular receptors or co-receptors for viral attachment.…”

Section: Discussionsupporting

confidence: 91%

“…In addition, all CoVs use the S1-CTD or S1-NTD regions to interact with cellular receptors or co-receptors for viral attachment. The main functional RBD regions of most α-CoVs and some β-CoVs, including SARS-CoV, MERS-CoV, and HCoV-HKU1, are located in the S1-CTD (Chen et al, 2017;Deng et al, 2016). In contrast, the RBDs of human CoV OC43, MHV, and avian γ-CoV are located in the S1-NTD region.…”

Section: Discussionmentioning

confidence: 99%

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Identification of the immunodominant neutralizing regions in the spike glycoprotein of porcine deltacoronavirus

Chen

et al. 2020

Virus Research

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A B S T R A C TPorcine deltacoronavirus (PDCoV), is an emerging enteropathogenic coronavirus in pigs, that poses a novel threat to swine husbandry worldwide. Crucial to halting PDCoV transmission and infection is the development of effective therapies and vaccines. The spike (S) protein of coronavirus is the major target of host neutralizing antibodies, however the immunodominant neutralizing region in the S protein of PDCoV has not been defined.Here, three truncations of the PDCoV S protein were generated, the N-terminal domain of the S1 subunit (NTD, amino acids (aa) 50-286), the C-terminal domain of the S1 subunit (CTD, aa 278-616), and S2 subunit (aa 601-1087). The proteins were expressed using an E. coli expression system. Polyclonal antisera against the three recombinant proteins were produced in rabbits and mice. All three antisera were able to inhibit PDCoV infection in vitro, as determined by virus neutralization assay, fluorescent focus neutralization assay, and plaque-reduction neutralization. The CTD-specific antisera had the most potent PDCoV-neutralizing effect, indicating that the CTD region may contain the major neutralizing epitope(s) in the PDCoV S protein. Based on these findings, CTD may be a promising target for development of an effective vaccine against PDCoV infection in pigs.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

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Identification of the immunodominant neutralizing regions in the spike glycoprotein of porcine deltacoronavirus

Chen

et al. 2020

Virus Research

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“…The mAbs developed by other groups, CDC-C2, m336, and REGN3051, also bind to RBD and potently neutralize MERS-CoV (de Wit et al, 2018;Pascal et al, 2015;Wang et al, 2018;Ying et al, 2014). Meanwhile, it has also been reported that non-RBD S1-specific mAbs, G2 and 5F9, and S2-specific mAb G4 neutralized MERS-CoV (Chen et al, 2017;Pallesen et al, 2017;Wang et al, 2015;Zhang et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein

et al. 2020

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Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes severe pulmonary infection, with ∼35 % mortality. Spike glycoprotein (S) of MERS-CoV is a key target for vaccines and therapeutics because S mediates viral entry and membrane-fusion to host cells. Here, four different S subunit proteins, receptor-binding domain (RBD; 358-606 aa), S1 (1-751 aa), S2 (752-1296 aa), and SΔTM (1-1296 aa), were generated using the baculoviral system and immunized in mice to develop neutralizing antibodies. We developed 77 hybridomas and selected five neutralizing mAbs by immunization with SΔTM against MERS-CoV EMC/2012 strain S-pseudotyped lentivirus. However, all five monoclonal antibodies (mAb) did not neutralize the pseudotyped V534A mutation. Additionally, one mAb RBD-14F8 did not show neutralizing activity against pseudoviruses with amino acid substitution of L506 F or D509 G (England1 strain, EMC/2012 L506 F, and EMC/2012 D509 G), and RBD-43E4 mAb could not neutralize the pseudotyped I529 T mutation, while three other neutralizing mAbs showed broad neutralizing activity. This implies that the mutation in residue 506-509, 529, and 534 of S is critical to generate neutralization escape variants of MERS-CoV. Interestingly, all five neutralizing mAbs have binding affinity to RBD, although most mAbs generated by RBD did not have neutralizing activity. Additionally, chimeric antibodies of RBD-14F8 and RBD-43E4 with human Fc and light chain showed neutralizing effect against wild type MERS-CoV KOR/KNIH/002, similar to the original mouse mAbs. Thus, our mAbs can be utilized for the identification of specific mutations of MERS-CoV.

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“…The isolation of potent neutralizing monoclonal antibodies has been reported shortly after the identification of MERS-CoV. This was achieved by using various technological platforms such as phage or yeast display of antibody library (Jiang et al, 2014;Tang et al, 2014;Ying et al, 2014), immunization of experimental animals (Chen et al, 2017a;Du et al, 2014;Li et al, 2015;Pascal et al, 2015;Wang et al, 2015a), and direct isolation from human survivors of MERS-CoV infection (Chen et al, 2017b;Corti et al, 2015;Wang et al, 2018). Currently, close to 20 different neutralizing monoclonal antibodies have been characterized in cell culture and experimental animal models.…”

Section: Introductionmentioning

confidence: 99%

Structural Definition of a Unique Neutralization Epitope on the Receptor-Binding Domain of MERS-CoV Spike Glycoprotein

et al. 2018

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The major mechanism of antibody-mediated neutralization of the Middle East respiratory syndrome coronavirus (MERS-CoV) involves competition with the cellular receptor dipeptidyl peptidase 4 (DPP4) for binding to the receptor-binding domain (RBD) of the spike (S) glycoprotein. Here, we report a unique epitope and unusual neutralizing mechanism of the isolated human antibody MERS-4. Structurally, MERS-4 approached the RBD from the outside of the RBD-DPP4 binding interface. Such binding resulted in the folding of the β5-β6 loop toward a shallow groove on the RBD interface critical for accommodating DPP4. The key residues for binding are identified through site-directed mutagenesis. Structural modeling revealed that MERS-4 binds to RBD only in the "up" position in the S trimer. Furthermore, MERS-4 demonstrated synergy with several reported antibodies. These results indicate that MERS-4 neutralizes MERS-CoV by indirect rather than direct competition with DPP4. This mechanism provides a valuable addition for the combined use of antibodies against MERS-CoV infection.

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A novel neutralizing monoclonal antibody targeting the N-terminal domain of the MERS-CoV spike protein

Cited by 48 publications

References 49 publications

Identification of the immunodominant neutralizing regions in the spike glycoprotein of porcine deltacoronavirus

Identification of the immunodominant neutralizing regions in the spike glycoprotein of porcine deltacoronavirus

Characterization of novel monoclonal antibodies against MERS-coronavirus spike protein

Structural Definition of a Unique Neutralization Epitope on the Receptor-Binding Domain of MERS-CoV Spike Glycoprotein

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