The structure of HBsAg particles is not modified upon their adsorption on aluminium hydroxide gel

July 28, 2017

Title

The structure of HBsAg particles is not modified upon their adsorption on aluminium hydroxide gel

Author

Vanille J. Greiner, Frédéric Ronzon, Eric Larquet, Bernard Desbat, Catherine Estèves, Julie Bonvin, Frédéric Gréco, Catherine Manin, Andrey S. Klymchenko, Yves Mély

Year

2012

Journal

Vaccine

Abstract

Current Hepatitis B vaccines are based on recombinant Hepatitis B surface antigen (HBsAg) virus-like particles adsorbed on aluminium (Al) gel. These particles exhibit a lipoprotein-like structure with about 70 protein S molecules in association with various types of lipids. To determine whether the adsorption on Al gel affects HBsAg structure, we investigated the effect of adsorption and mild desorption processes on the protein and lipid parts of the particles, using various techniques. Electron microscopy showed that the size and morphology of native and desorbed HBsAg particles were comparable. Moreover, infrared and Raman spectroscopy revealed that the secondary structure of the S proteins was not affected by the adsorption/desorption process. Affinity measurements with Surface Plasmon Resonance showed no difference between native and desorbed HBsAg for HBsAg-specific RF-1 monoclonal antibody. Steady-state and time-resolved fluorescence data of the intrinsic fluorescence of the S proteins further indicated that the adsorption/desorption of HBsAg particles on Al gel did not modify the environment of the most emitting Trp residues, confirming that the conformation of the S proteins remains intact. Moreover, using environment-sensitive 3-hydroxyflavone probes, no significant changes of the lipid core and lipid membrane surface of the HBsAg particles were observed during the adsorption/desorption process. Finally, the ratio between lipids and proteins in the particles was found to be similar before and after the adsorption/desorption process. Taken together, our data show that adsorption on Al gel does not affect the structure of the HBsAg particles.

Instrument

J-810

Keywords

Circular dichroism, Secondary structure, Vesicle interactions, Medicinal, Biochemistry