Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen

Mun Peak Nyon, Lanying Du, Chien Te Kent Tseng, Christopher A. Seid, Jeroen Pollet, Kevin S. Naceanceno, Anurodh Agrawal, Abdullah Algaissi, Bi Hung Peng, Wanbo Tai, Shibo Jiang, Maria Elena Bottazzi, Ulrich Strych, Peter J. Hotez

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Middle East respiratory syndrome coronavirus (MERS-CoV) has infected at least 2040 patients and caused 712 deaths since its first appearance in 2012, yet neither pathogen-specific therapeutics nor approved vaccines are available. To address this need, we are developing a subunit recombinant protein vaccine comprising residues 377–588 of the MERS-CoV spike protein receptor-binding domain (RBD), which, when formulated with the AddaVax adjuvant, it induces a significant neutralizing antibody response and protection against MERS-CoV challenge in vaccinated animals. To prepare for the manufacture and first-in-human testing of the vaccine, we have developed a process to stably produce the recombinant MERS S377-588 protein in Chinese hamster ovary (CHO) cells. To accomplish this, we transfected an adherent dihydrofolate reductase-deficient CHO cell line (adCHO) with a plasmid encoding S377-588 fused with the human IgG Fc fragment (S377-588-Fc). We then demonstrated the interleukin-2 signal peptide-directed secretion of the recombinant protein into extracellular milieu. Using a gradually increasing methotrexate (MTX) concentration to 5 μM, we increased protein yield by a factor of 40. The adCHO-expressed S377-588-Fc recombinant protein demonstrated functionality and binding specificity identical to those of the protein from transiently transfected HEK293T cells. In addition, hCD26/dipeptidyl peptidase-4 (DPP4) transgenic mice vaccinated with AddaVax-adjuvanted S377-588-Fc could produce neutralizing antibodies against MERS-CoV and survived for at least 21 days after challenge with live MERS-CoV with no evidence of immunological toxicity or eosinophilic immune enhancement. To prepare for large scale-manufacture of the vaccine antigen, we have further developed a high-yield monoclonal suspension CHO cell line.

Original languageEnglish (US)
Pages (from-to)1853-1862
Number of pages10
JournalVaccine
Volume36
Issue number14
DOIs
StatePublished - Mar 27 2018

Keywords

  • Chinese hamster ovary cells
  • Middle East respiratory syndrome coronavirus
  • Receptor binding domain

ASJC Scopus subject areas

  • Molecular Medicine
  • Immunology and Microbiology(all)
  • veterinary(all)
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

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  • Cite this

    Nyon, M. P., Du, L., Tseng, C. T. K., Seid, C. A., Pollet, J., Naceanceno, K. S., Agrawal, A., Algaissi, A., Peng, B. H., Tai, W., Jiang, S., Bottazzi, M. E., Strych, U., & Hotez, P. J. (2018). Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen. Vaccine, 36(14), 1853-1862. https://doi.org/10.1016/j.vaccine.2018.02.065