Rational design of an influenza subunit vaccine powder with sugar glass technology: preventing conformational changes of haemagglutinin during freezing and freeze-drying

Vaccine. 2007 Aug 29;25(35):6447-57. doi: 10.1016/j.vaccine.2007.06.054. Epub 2007 Jul 16.

Abstract

The development of a stable influenza subunit vaccine in the dry state was investigated. The influence of various carbohydrates, buffer types and freezing rates on the integrity of haemagglutinin after freeze-thawing or freeze-drying was investigated with a range of analytical and immunological methods. The use of fast freezing, Hepes buffer and carbohydrates (trehalose, inulin or dextran) as cryo- and lyoprotectants resulted in a significant reduction or even absence of conformational changes of HA as revealed by the used methods. The subunit vaccine in the powder was shown to remain immunogenic in an in vivo study in mice, using reconstituted powder. Moreover, the HA potency of the influenza subunit vaccine powder was stable for at least 26 weeks at room temperature.

MeSH terms

  • Animals
  • Antigens / immunology
  • Carbohydrates / chemistry
  • Circular Dichroism
  • Dialysis
  • Drug Design
  • Enzyme-Linked Immunosorbent Assay
  • Freeze Drying
  • Freezing
  • Hemagglutination Inhibition Tests
  • Hemagglutinins / chemistry*
  • Hemagglutinins / immunology*
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Immunodiffusion
  • Influenza A Virus, H3N2 Subtype / immunology
  • Influenza Vaccines / immunology*
  • Mice
  • Powders
  • Protein Conformation
  • Spectrometry, Fluorescence
  • Trypsin / chemistry

Substances

  • Antigens
  • Carbohydrates
  • Hemagglutinins
  • Influenza Vaccines
  • Powders
  • Trypsin