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

Wen Hsiang Chen, Lanying Du, Shivali M. Chag, Cuiqing Ma, Nancy Tricoche, Xinrong Tao, Christopher A. Seid, Elissa M. Hudspeth, Sara Lustigman, Chien-Te Tseng, Maria Elena Bottazzi, Peter J. Hotez, Bin Zhan, Shibo Jiang

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Pages (from-to)648-658
Number of pages11
JournalHuman Vaccines and Immunotherapeutics
Volume10
Issue number3
DOIs
StatePublished - 2014

Fingerprint

Severe Acute Respiratory Syndrome
Recombinant Proteins
Protein Binding
Vaccines
Yeasts
Proteins
Neutralizing Antibodies
Antibody Formation
Carrier Proteins
Coronavirus
Antigen Receptors
Pandemics
Glycosylation

Keywords

  • Deglycosylation
  • Receptor-binding domain
  • SARS-CoV
  • Vaccine
  • Yeast expression

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology

Cite this

Yeast-expressed recombinant protein of the receptor-binding domain in SARS-CoV spike protein with deglycosylated forms as a SARS vaccine candidate. / Chen, Wen Hsiang; Du, Lanying; Chag, Shivali M.; Ma, Cuiqing; Tricoche, Nancy; Tao, Xinrong; Seid, Christopher A.; Hudspeth, Elissa M.; Lustigman, Sara; Tseng, Chien-Te; Bottazzi, Maria Elena; Hotez, Peter J.; Zhan, Bin; Jiang, Shibo.

In: Human Vaccines and Immunotherapeutics, Vol. 10, No. 3, 2014, p. 648-658.

Research output: Contribution to journalArticle

Chen, WH, Du, L, Chag, SM, Ma, C, Tricoche, N, Tao, X, Seid, CA, Hudspeth, EM, Lustigman, S, Tseng, C-T, Bottazzi, ME, Hotez, PJ, Zhan, B & Jiang, S 2014, 'Yeast-expressed recombinant protein of the receptor-binding domain in SARS-CoV spike protein with deglycosylated forms as a SARS vaccine candidate', Human Vaccines and Immunotherapeutics, vol. 10, no. 3, pp. 648-658. https://doi.org/10.4161/hv.27464
Chen, Wen Hsiang ; Du, Lanying ; Chag, Shivali M. ; Ma, Cuiqing ; Tricoche, Nancy ; Tao, Xinrong ; Seid, Christopher A. ; Hudspeth, Elissa M. ; Lustigman, Sara ; Tseng, Chien-Te ; Bottazzi, Maria Elena ; Hotez, Peter J. ; Zhan, Bin ; Jiang, Shibo. / Yeast-expressed recombinant protein of the receptor-binding domain in SARS-CoV spike protein with deglycosylated forms as a SARS vaccine candidate. In: Human Vaccines and Immunotherapeutics. 2014 ; Vol. 10, No. 3. pp. 648-658.
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AU - Chen, Wen Hsiang

AU - Du, Lanying

AU - Chag, Shivali M.

AU - Ma, Cuiqing

AU - Tricoche, Nancy

AU - Tao, Xinrong

AU - Seid, Christopher A.

AU - Hudspeth, Elissa M.

AU - Lustigman, Sara

AU - Tseng, Chien-Te

AU - Bottazzi, Maria Elena

AU - Hotez, Peter J.

AU - Zhan, Bin

AU - Jiang, Shibo

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N2 - 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.

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