VaxArray assessment of influenza split vaccine potency and stability.

ABSTRACT

Vaccine manufacturers require more rapid and accurate tools to characterize the potency and stability of their products. Currently, the gold standard for influenza vaccine potency is the single radial immunodiffusion (SRD) assay, which has inherent disadvantages. The primary objective of this study was to investigate the ability of the VaxArray Influenza (VXI) seasonal hemagglutinin (sHA) potency assay to accurately quantify potency and stability in finished vaccines as well as to quantify hemagglutinin protein (HA) within crude in-process samples. Monobulk intermediates and mono- and multivalent vaccines were tested using VXI. Quantification of HA in crude samples was evaluated by spiking known concentrations of HA into allantoic fluid. VXI generated SRD equivalent potency measurements with high accuracy (within ±10%) and precision (CV 10±4%) for antigen components of monobulk intermediates and multivalent split vaccines. For these vaccines and vaccine intermediates, the VXI linear dynamic range was ∼0.01-0.6µg/mL, which is 12× greater than the linear range of SRD. The measured sample limit of detection (LOD) for VXI varied from 0.005 to 0.01µg/mL for the different subtypes, which in general is ≥600× lower than the LOD for SRD. VXI was able to quantify HA in crude samples where HA only accounts for 0.02% of the total protein content. Stability indication was investigated by tracking measured potency as a function of time at elevated temperature by both SRD and VXI. After 20 h at 56°C, the ratio of VXI to SRD measured potency in a quadrivalent vaccine was 76%, 125%, 60%, and 98% for H1/California, H3/Switzerland, B/Phuket and B/Brisbane, respectively. Based on the study results, it is concluded that VXI is a rapid, multiplexed immunoassay that can be used to accurately determine flu vaccine potency and stability in finished product and in crude samples from upstream processes.