Evidence for the denaturation of recombinant hepatitis B surface antigen on aluminium hydroxide gel.
Despite the complexity of the subject of protein-alum interactions, a valuable information can be obtained by analyzing the adsorbed and desorbed protein by common physico-chemical methods. In the present work, to approach the adsorption of hepatitis B surface antigen (HBsAg) on alum, the experimental data were supported by complementary analyses of the adsorbed protein by immunoelectron microscopy and the desorbed protein by denaturing size-exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. First, the depletion of HBsAg was investigated. The aspects assessed were the conditions, recovery and chromatographic performance of the desorbed protein. The results obtained strongly suggested the loss of particulate structure of HBsAg after adsorption on alum. This conclusion was further reinforced by direct immunoelectron microscopic visualization of HBsAg in the adsorbed state.
Tleugabulova
D
D
Quality Control Department, National Center for Bioproducts, Havana, Cuba.
Falcón
V
V
Pentón
E
E
D000536:Aluminum Hydroxide / Q000737:chemistry*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000737:chemistry*; D016253:Microscopy, Immunoelectron; D008956:Models, Chemical; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry
D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins; D000536:Aluminum Hydroxide