Characterization and immunogenicity of SARS-CoV-2 spike proteins with varied glycosylation. Issue 47 (8th November 2022)

• Record Type: Journal Article
• Authors: Deng, Tingting; Li, Tingting; Chen, Gege; Zhu, Yuhe; Xu, Lang; Lin, Yanlin; Sun, Hui; Zhang, Hui; Fang, Qianjiao; Hong, Junping; Wu, Dinghui; Gao, Shuangquan; Li, Shaoyong; Wang, Yingbin; Zhang, Tianying; Chen, Yixin; Yuan, Quan; Zheng, Qingbing; Yu, Hai; Zhao, Qinjian; Zhang, Jun; Li, Shaowei; Xia, Ningshao; Gu, Ying
• Is Part Of: Vaccine. Volume 40:Issue 47(2022)
• Page Start: 6839
• Page End: 6848
• Publication Date: 2022-11-08
• Subjects: SARS-CoV-2 -- Spike -- 293 cells -- Glycosylation -- Immunogenicity; Vaccines -- Periodicals; 615.372
• DOI: 10.1016/j.vaccine.2022.09.057
• Languages: English
• ISSNs: 0264-410X

Abstract

Highlights: Optimized the expression of the S-2P protein in 293F cells, and obtained a 5-times higher yield. Different glycosylated S-2P proteins varied in physiochemical properties, antigenicity and immunogenicity. S-2P proteins from 293 cells induced a more potent nAb response than did proteins produced from insect cells. Priming with 293S S-2P and then boosting with Bac S-2P and then 293F S-2P can elicit a higher nAb response than with a single S-2P alone. Abstract: The ongoing coronavirus disease-19 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has drastically changed our way of life and continues to have an unmitigated socioeconomic impact across the globe. Research into potential vaccine design and production is focused on the spike (S) protein of the virus, which is critical for virus entry into host cells. Yet, whether the degree of glycosylation in the S protein is associated with vaccine efficacy remains unclear. Here, we first optimized the expression of the S protein in mammalian cells. While we found no significant discrepancy in purity, homogeneity, or receptor binding ability among S proteins derived from 293F cells (referred to as 293F S-2P), 293S GnTI- cells (defective in N -acetylglucosaminyl transferase I enzyme; 293S S-2P), or TN-5B1-4 insect cells (Bac S-2P), there was significant variation in the glycosylation patterns and thermal stability of the proteins. Compared with the partially glycosylated 293S S-2P or Bac S-2P, the fully glycosylated 293F S-2P exhibited higher binding reactivity to convalescent sera. In addition, 293F S-2P induced higher IgG and neutralizing antibody titres than 293S or Bac S-2P in mice. Furthermore, a prime-boost-boost regimen, using a combined immunization of S-2P proteins with various degrees of glycosylation, elicited a more robust neutralizing antibody response than a single S-2P alone. Collectively, this study provides insight into ways to design a more effective SARS-CoV-2 immunogen.