A monoclonal antibody against a hepatitis B e antigen epitope borne by six amino acids encoded by the precore region.
Hepatitis B e antigen (HBeAg) polypeptide in the circulation (p17e) is composed of ten amino acids (aa) coded for by the precore region and 149 aa by the core gene of hepatitis B virus. A monoclonal antibody (Y0583A) was raised against the N-terminal ten amino acids (SKLCLGWLWG) encoded by the precore region. The binding of Y0583A with a panel of 203 decapeptides on multipins, which covered the precursor of HBeAg polypeptide made of 212 aa shifting by one aa, recognized an epitope sequenced LGWLWG representing the C-terminal six aa coded for by the precore region. This HBeAg epitope was not readily accessible on HBeAg in serum, but it became exposed and bound with Y0583A by treatment with 0.2 N NaOH. Using Y0583A, an enzyme-linked immunosorbent assay was developed for specific determination of HBeAg. The test sample was incubated with the monoclonal antibody to an HBeAg determinant encoded by the core gene (904) that had been immobilized on a solid support. Captured HBeAg was treated with 0.2 N NaOH, neutralized and released into the fluid phase. The reactant was then tested for a sandwich between monoclonal antibody (C33) to the C-terminus of the HBeAg polypeptide immobilized on a solid support and Y0583A labeled with horseradish peroxidase.
Usuda
S
S
Immunology Division, Jichi Medical School, Tochigi-Ken, Japan.
Okamoto
H
H
Ohnuma
H
H
Tanaka
T
T
Machida
A
A
Miyakawa
Y
Y
Mayumi
M
M
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D001782:Blood Donors; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006512:Hepatitis B Core Antigens / Q000235:genetics / Q000276:immunology*; D006513:Hepatitis B e Antigens / Q000097:blood / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology*; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D016366:Open Reading Frames / Q000276:immunology; D011498:Protein Precursors / Q000276:immunology
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens; D016056:Immunodominant Epitopes; D009842:Oligopeptides; D011498:Protein Precursors