The structure of HBsAg particles is not modified upon their adsorption on aluminium hydroxide gel.
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.
Laboratoire de Biophotonique et Pharmacologie, UMR 7213 CNRS, Université de Strasbourg, Faculté de Pharmacie, 74 Route du Rhin, 67401 Illkirch, France.
Vanille J
VJ
Greiner
Frédéric
F
Ronzon
Eric
E
Larquet
Bernard
B
Desbat
Catherine
C
Estèves
Julie
J
Bonvin
Frédéric
F
Gréco
Catherine
C
Manin
Andrey S
AS
Klymchenko
Yves
Y
Mély
D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry*; D002942:Circular Dichroism; D005419:Flavonoids / Q000737:chemistry; D006514:Hepatitis B Surface Antigens / Q000737:chemistry*; D017325:Hepatitis B Vaccines / Q000737:chemistry; D008055:Lipids; D008854:Microscopy, Electron; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D013055:Spectrophotometry, Infrared; D020349:Surface Plasmon Resonance; D058425:Vaccines, Virus-Like Particle / Q000737:chemistry
D005419:Flavonoids; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D008055:Lipids; D058425:Vaccines, Virus-Like Particle; D000536:Aluminum Hydroxide; C041477:3-hydroxyflavone