title: Approach to the profiling and characterization of influenza vaccine constituents by the combined use of size-exclusion chromatography, gel electrophoresis and mass spectrometry.
abstract: A combination of separation and identification techniques was used to rapidly and reproducibly analyze influenza vaccine constituents. Size-exclusion HPLC analysis reduced significantly the complexity by providing a constituents profile according to size. Significantly, no sample treatment was required prior to analysis thus eliminating a potential source of artifacts and degradation. Distinct profiles were associated with influenza strains as well as with vaccines from different manufacturers. Samples analyzed over several years allowed evaluation of method performance and provided stability-indicating data relating to the structural integrity of separated components. Collected chromatographic peaks were identified by gel electrophoresis and MALDI/MS of tryptic digests from excised gel bands. The challenge in obtaining high quality analytical data from complex mixtures clearly demonstrated the value of separation steps prior to MS identification. The method presented here is not intended to replace existing methodology; it is intended to provide a product specific profile to be used as a rapid screen for manufacturer, year (for annual influenza vaccines), stability or counterfeit product. It is a new screening method that provides a rapid and robust indication of products which require further investigation as a result of a deviation in their characteristic profile. Until now this tool did not exist.
authors.affiliation: Centre for Biologics Research, Health Canada, Banting Building, Tunney's Pasture, Ottawa, Ontario, Canada.
authors.forename: Virginia
authors.initials: V
authors.lastname: Garcia-Cañas
authors.forename: Barry
authors.initials: B
authors.lastname: Lorbetskie
authors.forename: Terry D
authors.initials: TD
authors.lastname: Cyr
authors.forename: Mary A
authors.initials: MA
authors.lastname: Hefford
authors.forename: Sophie
authors.initials: S
authors.lastname: Smith
authors.forename: Michel
authors.initials: M
authors.lastname: Girard
mesh_terms: D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004353:Drug Evaluation, Preclinical; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D007252:Influenza Vaccines / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D055442:Metabolome; D011499:Protein Processing, Post-Translational / Q000502:physiology; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013997:Time Factors; D014764:Viral Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism
chemical_list: D007252:Influenza Vaccines; D014764:Viral Proteins