Influenza Vaccine Manufacturing: Effect of Inactivation, Splitting and Site of Manufacturing. Comparison of Influenza Vaccine Production Processes

Abstract

The aim of this study was to evaluate the impact of different inactivation and splitting procedures on influenza vaccine product composition, stability and recovery to support transfer of process technology. Four split and two whole inactivated virus (WIV) influenza vaccine bulks were produced and compared with respect to release criteria, stability of the bulk and haemagglutinin recovery. One clarified harvest of influenza H3N2 A/Uruguay virus prepared on 25.000 fertilized eggs was divided equally over six downstream processes. The main unit operation for purification was sucrose gradient zonal ultracentrifugation. The inactivation of the virus was performed with either formaldehyde in phosphate buffer or with beta-propiolactone in citrate buffer. For splitting of the viral products in presence of Tween®, either Triton™ X-100 or di-ethyl-ether was used. Removal of ether was established by centrifugation and evaporation, whereas removal of Triton-X100 was performed by hydrophobic interaction chromatography. All products were sterile filtered and subjected to a 5 months real time stability study. In all processes, major product losses were measured after sterile filtration; with larger losses for split virus than for WIV. The beta-propiolactone inactivation on average resulted in higher recoveries compared to processes using formaldehyde inactivation. Especially ether split formaldehyde product showed low recovery and least stability over a period of five months.

Inactivated or damaged? Comparing the effect of inactivation methods on influenza virions to optimize vaccine production

“…As all our observations are based on in vitro tests, the correlation with the different vaccine potencies in vivo should be explored in future experiments. The use of single radial immunodiffusion assay as an estimate for protection has been shown to correlate both with in vivo responses [35] and, more recently, with in vitro results of differently manufactured influenza vaccine [36]. However, while the SRID assay would have been an alternative method to measure HA integrity, it would have given information on antibody binding only.…”

Vaccines to Prevent Infectious Diseases in the Older Population: Immunological Challenges and Future Perspectives

“…Split virus vaccines contain disrupted viral envelopes that lost infectivity but retained immunogenicity. Subunit virus vaccines are produced by further purification steps that remove the nucleocapsid from the split virus (34). One of the reasons for developing split and subunit vaccines was to decrease reactogenicity (35), however, in light of different adjuvants that may be added this effect is not always evident (36).…”

Optimization of cell culture‐derived influenza A virus particles purification using sulfated cellulose membrane adsorbers

“…Furthermore, the downstream processing (DSP) should be scalable to cope with increasing market demands for prevention of seasonal epidemics (requiring trivalent or tetravalent formulations) as well as enable a fast response in cases of pandemics. Finally, advances in upstream virus production techniques, like the use of different cell culture systems and high cell density culture systems present new challenges in DSP of influenza viruses that need to be addressed .…”