Abstract In this solid-phase two-site enzyme immunoassay for hepatitis B surface antigen (HBsAg), three monoclonal anti-HBs are used: 5D3 (IgM) is immobilized on plastic beads; 5C3 (IgG2a) and 5C 11 (IgG1), labeled with biotin, are used as the first conjugate. Horseradish peroxidase covalently linked to avidin is the second conjugate. First, serum or plasma is incubated with the antibody-coated bead and biotin-labeled antibodies, simultaneously, at 45 degrees C for 1 h ("stat" procedure), 3 h ("standard" procedure), or 18 h ("overnight" procedure), during which HBsAg forms a complex with the solid-phase antibody and the biotinylated antibodies. The enzyme-conjugated avidin is then bound to the biotin on the antigen-antibody complex at 45 degrees C for 15 min ("stat") or 30 min (standard and overnight procedures). The beads are incubated with enzyme-substrate solution (H2O2 and o-phenylenediamine). Color developed is measured at 492 nm. All procedures satisfied third-generation HBsAg tests required by the FDA Office of Biologics, being sensitive both to ad and ay subtypes in subnanogram amounts. The assay is reactive with adw2, adw4, adr, ayw2, ayw3, and ayr subtypes and can detect viral determinants in HBsAg-anti-HBs immune complex form. Thus it provides a sensitive, simple, and reproducible alternative to radioimmunoassay. © 1985 The American Association for Clinical Chemistry « Previous | Next Article » Table of Contents This Article Clinical Chemistry February 1985 vol. 31 no. 2 202-205 » Abstract PDF Services Email this article to a friend Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Download to citation manager Responses No responses published Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Liu, Y. S. Articles by Green, A. Search for related content PubMed PubMed citation Articles by Liu, Y. S. Articles by Green, A. Related Content Load related web page information Follow Us Clinical Chemistry Trainee Council Register Today! www.traineecouncil.org Information for Authors Submit a Manuscript Editorial Board Clinical Case Studies Clinical Chemistry Guide to Scientific Writing Journal Club Podcasts Translated Content Annual Meeting Abstracts Permissions and Reprints Advertising Copyright © 2012 by the American Association for Clinical Chemistry
Clinical Chemistry – American Association for Clinical Chemistry
Published: Feb 1, 1985
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