Analysis of hepatitis B surface antigen components solubilized with Triton X-100.
Three glycoproteins of intact hepatitis B surface antigen (HBsAg) with mol. wt. of 32 000, 30 000 and 28 000 respectively were identified by reaction with 125I-concanavalin A (Con A) after separation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The antigen was effectively disrupted with Triton X-100 to produce a structure with a sedimentation coefficient of 3.9S. Affinity chromatography of disrupted HBsAg using concanavalin A-Sepharose 4B (Con A-Sepharose) resulted in two fractions. The first contained material which did not bind to the lectin and consisted of a single polypeptide of mol. wt. 64 000. Further studies revealed this component to be serologically identical to serum albumin and to lack any affinity for antibody to HBsAg. A comparison of the tryptic peptide map of this polypeptide with that of purified serum albumin demonstrated identical amino-acid sequences. The second fraction contained material which bound to Con A and contained two polypeptides with mol. wt. of 28000 and 23000 respectively. HBsAg reactivity was associated with this fraction. This procedure allows the prepartion of HBsAg sub-units in milligram quantities for further immunological studies.
D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006023:Glycoproteins / Q000032:analysis*; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000276:immunology; D006515:Hepatitis B virus / Q000302:isolation & purification; D006801:Humans; D008970:Molecular Weight; D002679:Pan troglodytes / Q000382:microbiology; D010455:Peptides / Q000032:analysis; D011092:Polyethylene Glycols / Q000494:pharmacology; D012709:Serum Albumin / Q000276:immunology; D012995:Solubility
D000939:Epitopes; D006023:Glycoproteins; D006514:Hepatitis B Surface Antigens; D010455:Peptides; D012709:Serum Albumin; D011092:Polyethylene Glycols