Optimization and qualification of a quantitative reversed-phase HPLC method for hemagglutinin in influenza preparations and its comparative evaluation with biochemical assays

Particle quantification of influenza viruses by high performance liquid chromatography

“…The presence of the three aromatic amino acids namely tryptophan, tyrosine and phenylalanine but particularly tryptophan when excited at wavelengths between 280 and 295 nm contributes to the strong signal. It has been demonstrated that the HA1 detection of influenza vaccines was significantly improved by up to 2.7 logs when using nFL instead of the classical 280 nm UV absorbance [29,30]. For quantification, a calibration curve is plotted using an in-house standard with known VP/ml.…”

Preclinical evaluation of Vaxfectin®-adjuvanted Vero cell-derived seasonal split and pandemic whole virus influenza vaccines

“…17 Although the SRD assay is currently the reference release assay for influenza vaccines, alternative, assays such as HPLC or others are in development and may ultimately be preferable to the SRD assay. 37,38 Influenza viruses with pandemic potential including H5N1, H7N7, H9N2 and other novel subtypes pose continuous threats to humans due to lack of preexisting immunity and the attendant specter of high fatality rates, as exemplified by the continuous infections of humans by H5N1 subtypes since 1997 (WHO). The vast array of different influenza virus subtypes (as many as 170) circulating in and between avian and other natural reservoirs with ample opportunity for reassortment clearly indicates the continuous need for both surveillance as well as proactive development and testing of pandemic vaccine candidates.…”

Standardisation of inactivated influenza vaccines—Learning from history

“…Reverse‐phase high‐performance liquid chromatography (RP‐HPLC) is another commonly used technique for quantifying HA,30, 31 particularly during manufacturing in‐process testing. The methodology is fairly common, rapid and accurate and was used to formulate vaccine for some H1N1pdm09 clinical trials in the United States in 2009, although it was not used for vaccine approval or release.…”