Summary
The spike (S) glycoprotein is thought to play a complex and central role in the biology and pathogenesis of SARS coronavirus infection. In this study, a recombinant protein (rS268, corresponding to residues 268–1255 of SARS-CoV S protein) was expressed in Escherichia coli and was purified to near homogeneity. After immunization with rS268, S protein-specific BALB/c antisera and mAbs were induced and confirmed using ELISA, Western blot and IFA. Several BALB/c mAbs were found to be effectively to neutralize the infection of Vero E6 cells by SARS-CoV in a dose-dependent manner. Systematic epitope mapping showed that all these neutralizing mAbs recognized a 15-residues peptide (CB-119) corresponding to residues 1143–1157 (SPDVDLGDISGINAS) that was located to the second heptad repeat (HR2) region of the SARS-CoV spike protein. The peptide CB-119 could specifically inhibit the interaction of neutralizing mAbs and spike protein in a dose-dependent manner. Further, neutralizing mAbs, but not control mAbs, could specifically interact with CB-119 in a dose-dependent manner. Results implicated that the second heptad repeat region of spike protein could be a good target for vaccine development against SARS-CoV.
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Acknowledgments
This work was supported by a grant from the Institute of Preventive Medicine, National Defense Medical Center, Taiwan, R.O.C. awarded to Y.-C. Wang. We are very grateful to Mr. Lih-Jeng Tarn and Ms. Yu-Ying Huang for helping to take care of small animals and maintain the cell cultures, respectively. We also thank Dr. Ching-Len Liao for some material assistance. Part of this research was also supported by the National Science Council, Taiwan, Grants #SVAC12-06 (Chong, P).
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Lai, SC., Chong, PS., Yeh, CT. et al. Characterization of neutralizing monoclonal antibodies recognizing a 15-residues epitope on the spike protein HR2 region of severe acute respiratory syndrome coronavirus (SARS-CoV). J Biomed Sci 12, 711–727 (2005). https://doi.org/10.1007/s11373-005-9004-3
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DOI: https://doi.org/10.1007/s11373-005-9004-3