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SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates.
Emerg Microbes Infect. 2020 Dec; 9(1):2076-2090.EM

Abstract

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.

Authors+Show Affiliations

State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.Department of Pulmonary Medicine, The First Affiliated Hospital of Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China.State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, People's Republic of China. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University, Xiamen, People's Republic of China. The Research Unit of Frontier Technology of Structural Vaccinology of Chinese Academy of Medical Sciences, People's Republic of China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32897177

Citation

Li, Tingting, et al. "SARS-CoV-2 Spike Produced in Insect Cells Elicits High Neutralization Titres in Non-human Primates." Emerging Microbes & Infections, vol. 9, no. 1, 2020, pp. 2076-2090.
Li T, Zheng Q, Yu H, et al. SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates. Emerg Microbes Infect. 2020;9(1):2076-2090.
Li, T., Zheng, Q., Yu, H., Wu, D., Xue, W., Xiong, H., Huang, X., Nie, M., Yue, M., Rong, R., Zhang, S., Zhang, Y., Wu, Y., Wang, S., Zha, Z., Chen, T., Deng, T., Wang, Y., Zhang, T., ... Xia, N. (2020). SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates. Emerging Microbes & Infections, 9(1), 2076-2090. https://doi.org/10.1080/22221751.2020.1821583
Li T, et al. SARS-CoV-2 Spike Produced in Insect Cells Elicits High Neutralization Titres in Non-human Primates. Emerg Microbes Infect. 2020;9(1):2076-2090. PubMed PMID: 32897177.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - SARS-CoV-2 spike produced in insect cells elicits high neutralization titres in non-human primates. AU - Li,Tingting, AU - Zheng,Qingbing, AU - Yu,Hai, AU - Wu,Dinghui, AU - Xue,Wenhui, AU - Xiong,Hualong, AU - Huang,Xiaofen, AU - Nie,Meifeng, AU - Yue,Mingxi, AU - Rong,Rui, AU - Zhang,Sibo, AU - Zhang,Yuyun, AU - Wu,Yangtao, AU - Wang,Shaojuan, AU - Zha,Zhenghui, AU - Chen,Tingting, AU - Deng,Tingting, AU - Wang,Yingbin, AU - Zhang,Tianying, AU - Chen,Yixin, AU - Yuan,Quan, AU - Zhao,Qinjian, AU - Zhang,Jun, AU - Gu,Ying, AU - Li,Shaowei, AU - Xia,Ningshao, PY - 2020/9/9/pubmed PY - 2020/10/6/medline PY - 2020/9/8/entrez KW - COVID-19 KW - SARS-CoV-2 KW - immunogenicity KW - insect cell expression system KW - spike SP - 2076 EP - 2090 JF - Emerging microbes & infections JO - Emerg Microbes Infect VL - 9 IS - 1 N2 - 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. SN - 2222-1751 UR - https://www.unboundmedicine.com/medline/citation/32897177/SARS_CoV_2_spike_produced_in_insect_cells_elicits_high_neutralization_titres_in_non_human_primates_ L2 - https://www.tandfonline.com/doi/full/10.1080/22221751.2020.1821583 DB - PRIME DP - Unbound Medicine ER -