Simultaneous quantification of the viral antigens hemagglutinin and neuraminidase in influenza vaccines by LC–MSE

Collaborative studies on the development of national reference standards for potency determination of H7N9 influenza vaccine

“…This method has the potential to increase the accuracy determination of the reference antigen standards and to verify label claims for HA content in formulated vaccines. 19 Chun et al prepared the universal antibody against conserved epitope on HA2 subunit and proved that it can react with all subtypes of influenza HA proteins, and the Slot-Blot assay was established basing on this universal antibody. 20,21 All these efforts provide the promising tools to overcome the difficulties due to the unavailability of references in SRID method, although the extensive validations are still needed before these techniques are adopted internationally.…”

Standardisation of inactivated influenza vaccines—Learning from history

“…Mass spectrometry (MS) techniques are able to accurately quantify HA in vaccines,33, 34, 35 and the isotope‐dilution MS (IDMS) method is able to quantify the HA subtypes in a multivalent vaccine, and using common tryptic peptides is not dependent upon strain‐specific standards 34, 35. Despite these notable advantages, IDMS is not stability‐indicating and the technique must be coupled with other methods to measure only conformationally correct HA.…”

Design and expression of a QconCAT protein to validate Hi3 protein quantification of influenza vaccine antigens

“…faster methods for quality assessment of influenza vaccines[7-10] and we have published methods using 55 liquid chromatography coupled to mass spectrometry (LC-MS) to obtain both strain confirmation and 56 quantification of antigens [7,10]. For protein quantification, we employed the "Hi3" protein 57 quantification technique originally described by Silva and coworkers [11], which has proven useful for 58 many applications across multiple MS platforms [10,[12][13][14][15][16].…”

“…For protein quantification, we employed the "Hi3" protein 57 quantification technique originally described by Silva and coworkers [11], which has proven useful for 58 many applications across multiple MS platforms [10,[12][13][14][15][16]. Thorough discussions of quantitative 59 proteomics techniques using isotope-labeled strategies have also been published by Brun …”

“…Protein quantities are reported relative to BSA. Similar 276 sample concentrations and volumes have been used here as in our earlier work [10] to ensure data are 277 within the linear range of the instrument. 278…”