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

Publication Type:

Journal Article

Source:

European Journal of Pharmaceutical Sciences, Volume 23, Number 1, pp. 65-75 (2004)

ISBN:

0928-0987

DOI Name (links to online publication)

10.1016/j.ejps.2004.05.007

Keywords:

influenza virus haemagglutinin; stability; fluorescence spectroscopy; circular dichroism spectroscopy; physicochemical characterisation; radial-immunodiffusion technique; virus hemagglutinin; circular-dichroism; protein stability; b virus; subtypes; fusio

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 60degreesC 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. (C) 2004 Elsevier B.V. All rights reserved.

28/10/2009