Sodium dodecylsulfate polyacrylamide gel electrophoresis of recombinant hepatitis B surface antigen particles.
To investigate the factors leading to broadening of the recombinant hepatitis B surface antigen (HBsAg) peak in size-exclusion chromatography, the HBsAg particles eluting in different regions of the peak were subjected here to electrophoretic analysis. In nonreduced samples, the 24-kD band corresponding to the S monomer was detected when excessively large amounts of HBsAg were loaded onto the gel. Hence, some monomers are not disulfide-crosslinked in assembled particles. On the other hand, the results of alkylation experiments indicated the presence of free sulfhydryl group(s) in a little portion of freshly-purified HBsAg which was retarded on the size-exclusion chromatographic column and had significant antigenicity. This fraction of HBsAg was shown to be oligomeric and capable of spontaneous assembly into higher-order structures during aging.
Tleugabulova
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Quality Control Department, National Center for Bioproducts, Havana, Cuba.
D000478:Alkylation; D015153:Blotting, Western; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013501:Surface-Active Agents
D003432:Cross-Linking Reagents; D004220:Disulfides; D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins; D013438:Sulfhydryl Compounds; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate