Yeast-expressed recombinant protein of the receptor-binding domain in SARS-CoV spike protein with deglycosylated forms as a SARS vaccine candidate.

Abstract

Development of vaccines for preventing a future pandemic of severe acute respiratory syndrome (SARS) caused by SARS coronavirus (SARS-CoV) and for biodefense preparedness is urgently needed. Our previous studies have shown that a candidate SARS vaccine antigen consisting of the receptor-binding domain (RBD) of SARS-CoV spike protein can induce potent neutralizing antibody responses and protection against SARS-CoV challenge in vaccinated animals. To optimize expression conditions for scale-up production of the RBD vaccine candidate, we hypothesized that this could be potentially achieved by removing glycosylation sites in the RBD protein. In this study, we constructed two RBD protein variants: 1) RBD193-WT (193-aa, residues 318-510) and its deglycosylated forms (RBD193-N1, RBD193-N2, RBD193-N3); 2) RBD219-WT (219-aa, residues 318-536) and its deglycosylated forms (RBD219-N1, RBD219-N2, and RBD219-N3). All constructs were expressed as recombinant proteins in yeast. The purified recombinant proteins of these constructs were compared for their antigenicity, functionality and immunogenicity in mice using alum as the adjuvant. We found that RBD219-N1 exhibited high expression yield, and maintained its antigenicity and functionality. More importantly, RBD219-N1 induced significantly stronger RBD-specific antibody responses and a higher level of neutralizing antibodies in immunized mice than RBD193-WT, RBD193-N1, RBD193-N3, or RBD219-WT. These results suggest that RBD219-N1 could be selected as an optimal SARS vaccine candidate for further development.

• Authors: Wen-Hsiang Chen, Lanying Du, Shivali M Chag, Cuiqing Ma, Nancy Tricoche, Xinrong Tao, Christopher A Seid, Elissa M Hudspeth, Sara Lustigman, Chien-Te K Tseng, Maria Elena Bottazzi, Peter J Hotez, Bin Zhan, Shibo Jiang
• Journal: Human vaccines & immunotherapeutics (Hum Vaccin Immunother) Vol. 10 Issue 3 Pg. 648-58 ( 2014) ISSN: 2164-554X [Electronic] United States
• PMID: 24355931 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Adjuvants, Immunologic
• Alum Compounds
• Recombinant Proteins
• Spike Glycoprotein, Coronavirus
• Vaccines, Synthetic
• Viral Vaccines
• aluminum sulfate

Topics

• Adjuvants, Immunologic (administration & dosage)
• Alum Compounds (administration & dosage)
• Animals
• Female
• Gene Expression
• Glycosylation
• Mice, Inbred BALB C
• Pichia (genetics)
• Recombinant Proteins (genetics, immunology)
• Severe acute respiratory syndrome-related coronavirus (immunology)
• Spike Glycoprotein, Coronavirus (genetics, immunology)
• Vaccines, Synthetic (administration & dosage, genetics, immunology)
• Viral Vaccines (administration & dosage, genetics, immunology)