Physicochemical studies on the stability of influenza haemagglutinin in vaccine bulk material

Dion M A M Luykx (Corresponding Author), Marco G. Casteleijn, Wim Jiskoot, Janny Westdijk, Peter M J M Jongen

Research output: Contribution to journalArticleScientificpeer-review

33 Citations (Scopus)

Abstract

The relative unknown conformational stability of monovalent bulks of influenza virus haemagglutinin (HA) from three different strains (B/Guangdong, A/New Caledonia and A/Panama) was investigated with fluorescence and circular dichroism (CD) spectroscopy. Various stress conditions (concentration of denaturant, freeze-thawing, pH and temperature) affected the spectroscopic properties of the haemagglutinin proteins differently. Unfolding experiments revealed a poor stability of Guangdong haemagglutinin (GD-HA) in comparison with New Caledonia (NC-HA) and Panama haemagglutinin (P-HA). Freeze-thawing altered the secondary and tertiary structure of Guangdong haemagglutinin and only the tertiary structure of Panama haemagglutinin. From pH 4.6-9.2 the tertiary structures of Guangdong, New Caledonia and Panama haemagglutinin were all affected to a different extent. The secondary structure was only altered at low pH. Incubation of haemagglutinin at 60°C resulted in denaturation of the protein and a dramatic change of the fluorescence spectrum, indicative of oxidised tryptophan (Trp). In conclusion, fluorescence and circular dichroism spectroscopy are highly suitable techniques to monitor the stability of haemagglutinin in a straightforward and fast way.
Original languageEnglish
Pages (from-to)65-75
Number of pages11
JournalEuropean Journal of Pharmaceutical Sciences
Volume23
Issue number1
DOIs
Publication statusPublished - Sep 2004
MoE publication typeA1 Journal article-refereed

Fingerprint

Hemagglutinins
Human Influenza
Vaccines
Panama
New Caledonia
Fluorescence
Circular Dichroism
Spectrum Analysis
Protein Denaturation
Orthomyxoviridae
Tryptophan
Temperature

Keywords

  • Circular dichroism spectroscopy
  • Fluorescence spectroscopy
  • Influenza virus haemagglutinin
  • Physicochemical characterisation
  • Stability

Cite this

Luykx, Dion M A M ; Casteleijn, Marco G. ; Jiskoot, Wim ; Westdijk, Janny ; Jongen, Peter M J M. / Physicochemical studies on the stability of influenza haemagglutinin in vaccine bulk material. In: European Journal of Pharmaceutical Sciences. 2004 ; Vol. 23, No. 1. pp. 65-75.
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Physicochemical studies on the stability of influenza haemagglutinin in vaccine bulk material. / Luykx, Dion M A M (Corresponding Author); Casteleijn, Marco G.; Jiskoot, Wim; Westdijk, Janny; Jongen, Peter M J M.

In: European Journal of Pharmaceutical Sciences, Vol. 23, No. 1, 09.2004, p. 65-75.

Research output: Contribution to journalArticleScientificpeer-review

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