Rapid Manufacture and Release of a GMP Batch of Avian Influenza A(H7N9) Virus-Like Particle Vaccine Made Using Recombinant Baculovirus-Sf9 Insect Cell Culture Technology

Abstract

ovavax reported that, in 28 days following public release of the avian influenza A/Anhui/1/2013 (H7N9) virus gene sequences, its recombinant DNA and baculovirus-Spodoptera frugiperda cell culture-based technology was used to produce a virus-like particle vaccine to avian influenza A(H7N9) virus, and murine animal challenge studies were initiated. [1] This report describes Novavax's manufacturing process and the coordinated timing of critical activities necessary to produce and release a clinical batch of avian influenza A(H7N9) virus virus-like particle vaccine, under current good manufacturing practices, within three months from the time that the virus genomic sequences for this potential pandemic influenza virus were reported. The key enabling factors were: • A detailed, integrated project plan and daily coordination meetings • Advanced use of the baculovirus master virus seed to bypass production of the passage 3 virus stock • Successful functional testing of the master virus seed to establish process parameters • Drug substance quantitation with an alternate method prior to availability of single radial immunodiffusion assay reagents • Forward processing of intermediates prior to completion of quality control testing • A process that uses single-use manufacturing technology This project has been funded in whole or in part with federal funds from the Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority (BARDA), Department of Health and Human Services, under Contract No. HHSO100201100012C.

Membrane‐Based Approach for the Downstream Processing of Influenza Virus‐Like Particles

“…Although higher impurity removal steps are described for chromatography processes, the recovery yields are usually lower and/or the purification design is strain‐dependent . Complex purification processes were developed for clinical purposes combining membrane filtration, centrifugation, and ultracentrifugation as well as chromatography, highly impacting the product recovery, process time, and cost.…”

Single‐use wave‐mixed versus stirred bioreactors for insect‐cell/BEVS‐based protein expression at benchtop scale

“…Werner et al (2010) ascribe this fact to the more homogeneous energy dissipation and reduced foaming detected in these bioreactor types in comparison to stirred systems [19]. Nevertheless, single-use stirred bioreactors used, for example, by Novavax for its Sf-9/BEVS-based VLP vaccine productions are also suitable and even scalable up to 2 m 3 [18,20]. Nevertheless, single-use stirred bioreactors used, for example, by Novavax for its Sf-9/BEVS-based VLP vaccine productions are also suitable and even scalable up to 2 m 3 [18,20].…”

Bioprocess decision support tool for scalable manufacture of extracellular vesicles

“…Yet another emerging method is ultrafiltration. 8 L; Hahn et al, 2013), continuous chromatography (e.g. UF1 represents a published protocol employing a series of dead-end and tangential-flow filters (Heinemann et al, 2014).…”

“…In general, largerscale versions of each method may be possible with continuous-flow ultracentrifuges (e.g. 8 L; Hahn et al, 2013), continuous chromatography (e.g. 16 L; Bisschops, Frick, & Levison, 2016), and other industrial equipment, but feasibility of their use for EVs has not been reported.…”