Determination of the affinity of antibodies to hepatitis B surface antigen in human sera.
The measurement of the affinity of anti-HBs antibody in human sera using 3 HBsAg-related antigens is described. The antigens used were (i) a synthetic linear peptide corresponding to amino acids 139-147 of the major polypeptide of HBsAg, (ii) a cyclical form of this same peptide and (iii) a polypeptide complex of a 28,000 MW glycoprotein and a 23,000 MW protein from purified HBsAg. The method was established with a pooled human anti-HBs immunoglobulin preparation and a monoclonal anti-HBs antibody reactive to the 'a' determinant of HBsAg. The results indicate that both these antibody preparations effectively bind the 3 antigens with affinity values of between 2 X 10(6) to 9 X 10(7) litres/mole. However, the affinity of both antibody preparations for the cyclical form of the peptide was higher than for the linear form. The level of antibody (expressed as Abt, molar antigen binding sites) in the pooled human immunoglobulin for each of the 3 antigens was similar. Measurements of anti-HBs antibodies in the sera of recovered acute hepatitis B patients and from HBsAg negative chronic liver disease patients showed that the cyclical form of the antigen was bound with a higher affinity than the linear form. Affinity values of antibody in the sera of the latter group of patients was significantly lower (3 X 10(5) to 2.7 X 10(6) litres/mole) than those observed in sera from other individuals. The implication of these results in determining the importance of the measurement of affinity in the assessment of the efficacy of vaccines is discussed.
D000595:Amino Acid Sequence; D000915:Antibody Affinity; D001666:Binding Sites, Antibody; D006509:Hepatitis B / Q000276:immunology; D006510:Hepatitis B Antibodies / Q000032:analysis; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000276:immunology; D006801:Humans; D008970:Molecular Weight; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D014764:Viral Proteins / Q000138:chemical synthesis / Q000276:immunology
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D010455:Peptides; D014764:Viral Proteins