Rational design of an influenza subunit vaccine powder with sugar glass technology: preventing conformational changes of haemagglutinin during freezing and freeze-drying.
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.
Amorij
J-P
JP
Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. J.Amorij@rug.nl
Meulenaar
J
J
Hinrichs
W L J
WL
Stegmann
T
T
Huckriede
A
A
Coenen
F
F
Frijlink
H W
HW
D000818:Animals; D000941:Antigens / Q000276:immunology; D002241:Carbohydrates / Q000737:chemistry; D002942:Circular Dichroism; D003956:Dialysis; D015195:Drug Design; D004797:Enzyme-Linked Immunosorbent Assay; D005612:Freeze Drying; D005615:Freezing; D006385:Hemagglutination Inhibition Tests; D006388:Hemagglutinins / Q000737:chemistry* / Q000276:immunology*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D005779:Immunodiffusion; D053122:Influenza A Virus, H3N2 Subtype / Q000276:immunology; D007252:Influenza Vaccines / Q000276:immunology*; D051379:Mice; D011208:Powders; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D014357:Trypsin / Q000737:chemistry
D000941:Antigens; D002241:Carbohydrates; D006388:Hemagglutinins; D007252:Influenza Vaccines; D011208:Powders; D014357:Trypsin