Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins using High-Resolution Mass Spectrometry

Yong Zhang; Wanjun Zhao; Yonghong Mao; Shisheng Wang; Yi Zhong; Tao Su; Meng Gong; Xiaofeng Lu; Jingqiu Cheng; Hao Yang

The global outbreak and rapid spread of COVID-19 caused by SARS-CoV-2 urgently call for specific prevention and intervention measures(22). The development of preventative vaccines and neutralizing antibodies remains a chief goal in the efforts to control viral spread and stockpile candidates for future use. However, this work greatly relies on the understanding of the antigen structure and state of glycosylation for the rational determination of accessible epitopes. The S protein is posited to be the main or even the only antigen on viral surfaces for priming the immune system to produce an effective response (15, 18). Previous studies have revealed the structural information of the SARS-CoV-2 S protein and found the coverage of N-glycans (4, 10) . In this study, we profiled the site-specific N-glycosylation of the recombinant . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.03.28.013276 doi: bioRxiv preprint SARS-CoV-2 S protein. All potential N-glycosites on the S protein were identified experimentally. The N-glycan compositions and types on S protein subunits were revealed among different host cells. Our data provide a large-scale N-glycosylation information of the S protein and present a promising prospect for developing a vaccine and therapeutic antibodies.