Production of Clinical-Grade SARS-CoV-2 Spike Ferritin Nanoparticle Protein Immunogen by Transient Transfection
<b>Background/Objectives</b>: In response to the COVID-19 pandemic, we developed a vaccine candidate against SARS-CoV-2. Spike Ferritin Nanoparticle (SpFN) comprises 24 identical prefusion-stabilized spike proteins anchored to a self-assembled nanoparticle. Organized along the three-fold axis of the ferritin particle, eight SARS-CoV-2 spike trimers are presented per nanoparticle. <b>Methods</b>: Here, we describe the CGMP processes for manufacturing SpFN using transient transfection of Expi293F cells. <b>Results</b>: The final yield of SpFN was ~10 mg per liter of media supernatant. The resulting protein is stable in cold storage for two years at -20 °C, as well as for a month at room temperature, and can withstand multiple freeze/thaw cycles. SpFN material produced using the CGMP protocols adjuvanted with Army Liposomal Formulation-QS-21 (ALFQ) elicited potent neutralizing antibodies against WA-1, Alpha, Beta, and Delta variants in mice as measured by a pseudovirus neutralization assay. <b>Conclusions</b>: This work demonstrates rapid development and scaled-up production of clinical-grade SARS-CoV-2 vaccine protein material, allowing permissive storage and transport conditions, and serves as a framework for recombinant protein production for future emergent pathogens.
Authors
- Courtney Corbitt
- Jaime L. Jensen (ORCID: https://orcid.org/0000-0002-8818-7513)
- Aslaa Ahmed (ORCID: https://orcid.org/0000-0003-3093-4982)
- Kayvon Modjarrad (ORCID: https://orcid.org/0000-0002-6514-5572)
- Jeffrey W. Froude
- Agnes Hajduczki
- Gary R. Matyas (ORCID: https://orcid.org/0000-0002-2074-2373)
- Rafael De La Barrera
- Stasya Zarling
- William C. Chang (ORCID: https://orcid.org/0000-0003-4967-6426)
- James F. Wood
- Nelson L. Michael (ORCID: https://orcid.org/0000-0001-5882-5548)
- Sandhya Vasan (ORCID: https://orcid.org/0000-0002-6378-6288)
- Gregory D. Gromowski (ORCID: https://orcid.org/0000-0003-4576-4277)
- Paul V. Thomas (ORCID: https://orcid.org/0000-0002-6772-3997)
- Victoria R. Polonis (ORCID: https://orcid.org/0000-0002-4827-9923)
- Caroline E. Peterson (ORCID: https://orcid.org/0000-0002-6087-7301)
- Elizabeth J. Martinez (ORCID: https://orcid.org/0000-0002-7755-7056)
- Michelle Zemil (ORCID: https://orcid.org/0000-0002-1700-6658)
- Michael Joyce (ORCID: https://orcid.org/0000-0002-6808-7232)
- Jocelyn King (ORCID: https://orcid.org/0000-0003-3348-1632)
- Helen Dooley (ORCID: https://orcid.org/0000-0002-2570-574X)
- Caitlin Kuklis (ORCID: https://orcid.org/0000-0002-9974-6640)
- Natalia de Val (ORCID: https://orcid.org/0000-0002-1506-2023)
- Misook Choe
- Wei-hung Chen
- Sandrine Soman
- Phyllis A. Rees (ORCID: https://orcid.org/0000-0001-9755-8403)
- Rajeshwer S. Sankhala
- Farooq Nasar (ORCID: https://orcid.org/0000-0003-3658-9649)
- Alexander Anderson (ORCID: https://orcid.org/0000-0002-0000-7734)
- Clayton Smith
- Mangala Rao (ORCID: https://orcid.org/0000-0001-9553-4969)
- John R. Mascola
- Elaine B. Morrison
- Lindsay Wieczorek
Institutions
- University of Maryland, Baltimore (US)
- Henry M. Jackson Foundation (US)
- National Institutes of Health (US)
- University of Maryland Center for Environmental Science (US)
- Frederick National Laboratory for Cancer Research (US)
- National Institute of Allergy and Infectious Diseases (US)
- National Cancer Institute (US)
- Center for Cancer Research (US)
- Walter Reed Army Institute of Research (US)
Publication Details
- Journal
- Vaccines
- Published
- 2025-10-09
- DOI
- https://doi.org/10.3390/vaccines13101041
- Primary Topic
- SARS-CoV-2 and COVID-19 Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- U.S. Department of Defense
- Henry M. Jackson Foundation
- Defense Health Agency