Development of Methods to Produce SARS CoV‐2 Virus‐Like Particles at Scale

• Published in: Biotechnology and bioengineering Vol. 122; no. 5; pp. 1118 - 1129
• Main Authors: Edeling, Melissa A., Earnest, Linda, Carrera Montoya, Julio, Yap, Ashley Huey Yiing, Mumford, Jamie, Roberts, Jason, Wong, Chinn Yi, Hans, Dhiraj, Grima, Joseph, Bisset, Nicole, Bodle, Jesse, Rockman, Steven, Torresi, Joseph
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.05.2025; John Wiley and Sons Inc
• Subjects: Animals; Anion exchange; Anion exchanging; Buffers; COVID-19 - prevention & control; COVID-19 - virology; COVID-19 Vaccines; COVID‐19; Electron microscopy; Enzyme-linked immunosorbent assay; Humans; Membranes; molecular platform; Outbreaks; Pandemics; protein purification; Quality control; SARS-CoV-2 - isolation & purification; SARS‐CoV‐2; scalability; Severe acute respiratory syndrome; Spike Glycoprotein, Coronavirus - immunology; Spike Glycoprotein, Coronavirus - isolation & purification; Toll roads; vaccine; Vaccines, Virus-Like Particle - isolation & purification; Viral diseases; Viral envelope proteins; virus‐like particle; VLP; virus‐like particle; vaccine; COVID‐19; SARS‐CoV‐2; protein purification; VLP; molecular platform; scalability
• ISSN: 0006-3592; 1097-0290; 1097-0290
• DOI: 10.1002/bit.28937

ABSTRACT The devastating global toll precipitated by the SARS CoV‐2 outbreak and the profound impact of vaccines in stemming that outbreak has established the need for molecular platforms capable of rapidly delivering effective, safe and accessible medical interventions in pandemic preparedness. We describe a simple, efficient and adaptable process to produce SARS CoV‐2 virus‐like particles (VLPs) that can be readily scaled for manufacturing. A rapid but gentle method of tangential flow filtration using a 100 kDa semi‐permeable membrane concentrates and buffer exchanges 0.5 L of SARS CoV‐2 VLP containing supernatant into low salt and optimal pH for anion exchange chromatography. VLPs are washed, eluted under high salt, dialyzed into physiological buffer, sterile filtered and aliquoted for storage at –80°C. Purification is completed in less than 2 days. A simple quality control process includes Western blot for coincident detection of Spike, Membrane and Envelope protein as a proxy for intact VLP, ELISA to detect conformationally sensitive Spike using readily available anti‐Spike and/or anti‐RBD antibodies, and negative stain and immunogold electron microscopy to validate particulate, Spike crowned VLPs. This process to produce SARS CoV‐2 VLPs for preclinical studies serves as a roadmap for preparation of more distantly related VLPs for pandemic preparedness.