| Title | Identification of three conserved linear B cell epitopes on the SARS-CoV-2 spike protein |
| Authors | Wang, Aiping Tian, Yuanyuan Liu, Hongliang Ding, Peiyang Chen, Yumei Liang, Chao Du, Yongkun Jiang, Dawei Zhu, Xifang Yin, Jiajia Zhang, Gaiping |
| Affiliation | Zhengzhou Univ, Sch Life Sci, Zhengzhou 450001, Peoples R China Longhu Lab Adv Immunol, Zhengzhou, Peoples R China Henan Key Lab Immunobiol, Zhengzhou, Peoples R China Henan Agr Univ, Coll Vet Med, Zhengzhou, Peoples R China Peking Univ, Sch Adv Agr Sci, Beijing, Peoples R China |
| Keywords | ANTIBODIES |
| Issue Date | 31-Dec-2022 |
| Publisher | EMERGING MICROBES & INFECTIONS |
| Abstract | 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 6x 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. |
| URI | http://hdl.handle.net/20.500.11897/654560 |
| DOI | 10.1080/22221751.2022.2109515 |
| Indexed | SCI(E) |
| Appears in Collections: | 现代农学院 |
