Production of antibody to individual polypeptides derived from purified hepatitis B surface antigen.
Purified preparations of hepatitis B surface antigen (HBsAg) were solubilized with sodium dodecyl sulfate and urea under reducing conditions and subsequently fractionated by preparative sodium dodecyl sulfate-urea polyacrylamide gel electrophoresis (PAGE). Pools of the individual fractions eluted from the preparative PAGE were concentrated and purified further by analytical PAGE. Five purified polypeptides were isolated from HBsAg, types adw and ayw, with molecular weights of 19,000, 24,000, 27,000, 35,000, and 40,000. Each preparations was emulsified in Freund complete adjuvant and injected into guinea pigs. Antibody to each HBsAg type was measured by radioimmunoassay. The 19,000 molecular weight polypeptide derived from ayw particles and the 27,000 molecular weight subunit obtained from both types failed to elicit an antibody response. The other three polypeptides derived from the ayw particles elicited group-specific antibody responses. Similar group-specific reactivities were observed in the testing of anti-adw 35,000 and anti-adw 40,000 molecular weight polypeptide sera. However, guinea pigs immunized with the 19,000 and the 24,000 molecular weight polypeptides of the adw type produced antibody that reacted preferentially with adw particles. This indicates that either these subunits carry predominately d determinants or that, because of the low levels of material used for inoculation, no immune response or an undetectable one was elicited to the a or w components.
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000917:Antibody Formation; D003429:Cross Reactions; D000939:Epitopes; D006168:Guinea Pigs; D006511:Hepatitis B Antigens / Q000032:analysis; D008970:Molecular Weight; D010455:Peptides / Q000276:immunology* / Q000302:isolation & purification
D000914:Antibodies, Viral; D000939:Epitopes; D006511:Hepatitis B Antigens; D010455:Peptides