tetano
Editor, Senior Moderator
Int J Biol Macromol
. 2021 Dec 3;S0141-8130(21)02597-6.
doi: 10.1016/j.ijbiomac.2021.11.192. Online ahead of print.
Precise location of two novel linear epitopes on the receptor-binding domain surface of MERS-CoV spike protein recognized by two different monoclonal antibodies
Pan Wang[SUP] 1 [/SUP], Peiyang Ding[SUP] 2 [/SUP], Qiang Wei[SUP] 3 [/SUP], Aiping Wang[SUP] 2 [/SUP], Hongliang Liu[SUP] 2 [/SUP], Yunchao Liu[SUP] 3 [/SUP], Qingmei Li[SUP] 3 [/SUP], Yunrui Xing[SUP] 3 [/SUP], Ge Li[SUP] 3 [/SUP], Enmin Zhou[SUP] 4 [/SUP], Gaiping Zhang[SUP] 5 [/SUP]
Affiliations
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) is a coronavirus which can cause severe human respiratory diseases with a fatality rate of almost 36%. In this study, we report the generation, characterization and epitope mapping of several monoclonal antibodies against the spike receptor-binding domain (RBD) of MERS-CoV. Two monoclonal antibodies (4C7 and 6E8) that can react with linearized RBD have been selected for subsequent identification of RBD mAb-binding epitopes. Two distinct novel linear epitopes, [SUP]423[/SUP]FTCSQIS[SUP]429[/SUP] and [SUP]546[/SUP]SPLEGGGWL[SUP]554[/SUP],were precisely located at the outermost surface of RBD by dot-blot hybridization and ELISAs. Multiple sequence alignment analysis showed that these two peptides were highly conserved. Alanine (A)-scanning mutagenesis demonstrated that residues 423F, 428I, and 429S are the crucial residues for the linear epitope [SUP]423[/SUP]FTCSQIS[SUP]429[/SUP] while residues 548L, 550G, 553W, 554L for epitope [SUP]546[/SUP]SPLEGGGWL[SUP]554[/SUP]. These findings may be helpful for further understanding of the function of RBD protein and the development of subsequent diagnosis and detection methods.
Keywords: MERS-CoV RBD; Monoclonal antibody; Novel linear B cell epitopes.
. 2021 Dec 3;S0141-8130(21)02597-6.
doi: 10.1016/j.ijbiomac.2021.11.192. Online ahead of print.
Precise location of two novel linear epitopes on the receptor-binding domain surface of MERS-CoV spike protein recognized by two different monoclonal antibodies
Pan Wang[SUP] 1 [/SUP], Peiyang Ding[SUP] 2 [/SUP], Qiang Wei[SUP] 3 [/SUP], Aiping Wang[SUP] 2 [/SUP], Hongliang Liu[SUP] 2 [/SUP], Yunchao Liu[SUP] 3 [/SUP], Qingmei Li[SUP] 3 [/SUP], Yunrui Xing[SUP] 3 [/SUP], Ge Li[SUP] 3 [/SUP], Enmin Zhou[SUP] 4 [/SUP], Gaiping Zhang[SUP] 5 [/SUP]
Affiliations
- PMID: 34871658
- PMCID: PMC8641979
- DOI: 10.1016/j.ijbiomac.2021.11.192
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) is a coronavirus which can cause severe human respiratory diseases with a fatality rate of almost 36%. In this study, we report the generation, characterization and epitope mapping of several monoclonal antibodies against the spike receptor-binding domain (RBD) of MERS-CoV. Two monoclonal antibodies (4C7 and 6E8) that can react with linearized RBD have been selected for subsequent identification of RBD mAb-binding epitopes. Two distinct novel linear epitopes, [SUP]423[/SUP]FTCSQIS[SUP]429[/SUP] and [SUP]546[/SUP]SPLEGGGWL[SUP]554[/SUP],were precisely located at the outermost surface of RBD by dot-blot hybridization and ELISAs. Multiple sequence alignment analysis showed that these two peptides were highly conserved. Alanine (A)-scanning mutagenesis demonstrated that residues 423F, 428I, and 429S are the crucial residues for the linear epitope [SUP]423[/SUP]FTCSQIS[SUP]429[/SUP] while residues 548L, 550G, 553W, 554L for epitope [SUP]546[/SUP]SPLEGGGWL[SUP]554[/SUP]. These findings may be helpful for further understanding of the function of RBD protein and the development of subsequent diagnosis and detection methods.
Keywords: MERS-CoV RBD; Monoclonal antibody; Novel linear B cell epitopes.