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 valua ble information can be obtained by analyzing the adsorbed and desorbed prot ein by common physico-chemical methods. In the present work to approach the adsorption of hepatitis B surface antigen (HBsAg) on alum, the experimenta l data were supported by complementary analyses of the adsorbed protein by immunoelectron microscopy and the desorbed protein by denaturing size-exclu sion chromatography and sodium dodecyl sulfate-polyacrylamide gel electroph oresis under reducing conditions. First, the depletion of HBsAg was investi gated. The aspects assessed were the conditions, recovery and chromatograph ic performance of the desorbed protein. The results obtained strongly sugge sted the loss of particulate structure of HBsAg after adsorption on alum. T his conclusion was further reinforced by direct immunoelectron microscopic visualization of HBsAg in the adsorbed state. (C) 1998 Published by Elsevie r Science B.V. All rights reserved.