Abstract
The newly-emerging Middle East respiratory syndrome coronavirus (MERS-CoV) can cause severe and fatal acute respiratory disease in humans. Despite global efforts, the potential for an associated pandemic in the future cannot be excluded. The development of effective counter-measures is urgent. MERS-CoV-specific anti-viral drugs or vaccines are not yet available. Using the spike receptor-binding domain of MERS-CoV (MERS-RBD) to immunize mice, we identified two neutralizing monoclonal antibodies (mAbs) 4C2 and 2E6. Both mAbs potently bind to MERS-RBD and block virus entry in vitro with high efficacy. We further investigated their mechanisms of neutralization by crystallizing the complex between the Fab fragments and the RBD, and solved the structure of the 4C2 Fab/MERS-RBD complex. The structure showed that 4C2 recognizes an epitope that partially overlaps the receptor-binding footprint in MERS-RBD, thereby interfering with the virus/receptor interactions by both steric hindrance and interface-residue competition. 2E6 also blocks receptor binding, and competes with 4C2 for binding to MERS-RBD. Based on the structure, we further humanized 4C2 by preserving only the paratope residues and substituting the remaining amino acids with the counterparts from human immunoglobulins. The humanized 4C2 (4C2h) antibody sustained similar neutralizing activity and biochemical characteristics to the parental mouse antibody. Finally, we showed that 4C2h can significantly abate the virus titers in lungs of Ad5-hCD26-transduced mice infected with MERS-CoV, therefore representing a promising agent for prophylaxis and therapy in clinical settings.
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Acknowledgements
This work was supported by the National Basic Research Program of China (973 Program; 2015CB910503), the National Science and Technology Major Project of China (2013ZX10004608-002 and 2014ZX10004001-006), the National Natural Science Foundation of China (NSFC; 31400154 and 81461168030), the National Institutes of Health, USA (AI60699) and Hong Kong Research Grants Council (NHKU728/14). We acknowledge Dr Kwok-Hung Chan and Dr Jasper FW Chan for their technical support and the staff at the Shanghai Synchrotron Radiation Facility (SSRF-beam line 17U). We would be grateful to Yuanyuan Chen (Institute of Biophysics, CAS) for technical help with Biacore experiments. GFG is a leading principal investigator of the NSFC Innovative Research Group (8132106).
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Li, Y., Wan, Y., Liu, P. et al. A humanized neutralizing antibody against MERS-CoV targeting the receptor-binding domain of the spike protein. Cell Res 25, 1237–1249 (2015). https://doi.org/10.1038/cr.2015.113
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DOI: https://doi.org/10.1038/cr.2015.113
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