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TitleSARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates.
Journal, issue, pagesEmerg Microbes Infect, Vol. 9, Issue 1, Page 2076-2090, Year 2020
Publish dateOct 5, 2020
AuthorsTingting Li / Qingbing Zheng / Hai Yu / Dinghui Wu / Wenhui Xue / Hualong Xiong / Xiaofen Huang / Meifeng Nie / Mingxi Yue / Rui Rong / Sibo Zhang / Yuyun Zhang / Yangtao Wu / Shaojuan Wang / Zhenghui Zha / Tingting Chen / Tingting Deng / Yingbin Wang / Tianying Zhang / Yixin Chen / Quan Yuan / Qinjian Zhao / Jun Zhang / Ying Gu / Shaowei Li / Ningshao Xia /
PubMed AbstractThe current coronavirus disease 2019 (COVID-19) pandemic was the result of the rapid transmission of a highly pathogenic coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for ...The current coronavirus disease 2019 (COVID-19) pandemic was the result of the rapid transmission of a highly pathogenic coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for which there is no efficacious vaccine or therapeutic. Toward the development of a vaccine, here we expressed and evaluated as potential candidates four versions of the spike (S) protein using an insect cell expression system: receptor binding domain (RBD), S1 subunit, the wild-type S ectodomain (S-WT), and the prefusion trimer-stabilized form (S-2P). We showed that RBD appears as a monomer in solution, whereas S1, S-WT, and S-2P associate as homotrimers with substantial glycosylation. Cryo-electron microscopy analyses suggested that S-2P assumes an identical trimer conformation as the similarly engineered S protein expressed in 293 mammalian cells but with reduced glycosylation. Overall, the four proteins confer excellent antigenicity with convalescent COVID-19 patient sera in enzyme-linked immunosorbent assay (ELISA), yet show distinct reactivities in immunoblotting. RBD, S-WT and S-2P, but not S1, induce high neutralization titres (>3-log) in mice after a three-round immunization regimen. The high immunogenicity of S-2P could be maintained at the lowest dose (1 μg) with the inclusion of an aluminium adjuvant. Higher doses (20 μg) of S-2P can elicit high neutralization titres in non-human primates that exceed 40-times the mean titres measured in convalescent COVID-19 subjects. Our results suggest that the prefusion trimer-stabilized SARS-CoV-2 S-protein from insect cells may offer a potential candidate strategy for the development of a recombinant COVID-19 vaccine.
External linksEmerg Microbes Infect / PubMed:32897177 / PubMed Central
MethodsEM (single particle)
Resolution4.4 Å
Structure data

EMDB-30506:
Cryo-EM structure of SARS-CoV-2 spike trimer expressed in insect cells
Method: EM (single particle) / Resolution: 4.4 Å

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