Production of Clinical-Grade SARS-CoV-2 Spike Ferritin Nanoparticle Protein Immunogen by Transient Transfection
Year: 2025
Type: article
Abstract: Background/Objectives: 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. Methods: Here, we describe the CGMP processes for manufacturing SpFN using transient transfection of Expi293F cells. Results: 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. Conclusions: 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.
Source: Vaccines
Authors: Agnes Hajduczki, William C. Chang, Rafael De La Barrera, James F. Wood, Wei-Hung Chen, Elizabeth J. Martinez, Jaime L. Jensen, Rajeshwer S. Sankhala, Clayton Smith, Alexander Anderson, Elaine B. Morrison, Caroline E. Peterson, Phyllis A. Rees, Sandrine Soman, Caitlin Kuklis, Aslaa Ahmed, Jocelyn King, Farooq Nasar, Courtney Corbitt, Misook Choe, Paul V. Thomas, Michelle Zemil, Lindsay Wieczorek, Victoria R. Polonis, Helen M. Dooley, John R. Mascola, Natalie de Val, Gary R. Matyas, Mangala Rao, Gregory D. Gromowski, Kayvon Modjarrad, Sandhya Vasan, Jeffrey W. Froude, Nelson L. Michael, M. Gordon Joyce, Stasya Zarling
Institutions: Henry M. Jackson Foundation, Walter Reed Army Institute of Research, National Institutes of Health, Frederick National Laboratory for Cancer Research, National Cancer Institute, Center for Cancer Research, University of Maryland, Baltimore, Institute of Marine and Environmental Technology, National Institute of Allergy and Infectious Diseases, Vaccine Research Center
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $3,258
OpenAlex ID: W4415008353