Solid Phase Radioimmunoassay for Quantitation of Antigen-Specific IgG in Human Sera with 125I-Protein A from Staphylococcus Aureus
- 28 February 1979
- journal article
- research article
- Published by The American Association of Immunologists in The Journal of Immunology
- Vol. 122 (3), 1073-1079
- https://doi.org/10.4049/jimmunol.122.3.1073
Abstract
Radiolabeled protein A from Staphylococcus aureus (Staph A) has been used to develop a solid phase, noncompetitive radioimmunoassay for quantitation of specific IgG antibody. The assay involves two incubations: First, agarose-insolubilized antigen is mixed with serum samples for 1 to 4 hr during which specific antibody is bound; second, after a washing procedure, the solid phase immune complexes are incubated for 4 to 18 hr with 125I-Staph A, during which the radiolabeled detection protein binds to the insolubilized specific IgG antibody. In a comparative study of the IgG antiphospholipase A antibody content of 23 human sera drawn from honeybee venom-sensitive patients, results of the Staph A assay correlated highly (r = 0.981, p < 0.001, N = 23) with those obtained from a liquid phase, competitive radioimmunoprecipitation (double antibody) assay. The two assays demonstrated comparable precision, sensitivity, and reproducibility. In contrast, the use of 125I-Staph A in the solid phase radioimmunoassay was supperior to 125I rabbit anti-human IgG because of lower negative serum (blank) values, shorter time required to reach equilibrium binding, and greater precision and reproducibility. In principle, the 125I Staph A assay may be applied to IgG quantitation for crude allergen extracts as well as purified antigens. Furthermore, the sera of a number of mammalian species may be studied without further modification.This publication has 11 references indexed in Scilit:
- Use of staphylococcal protein A as an immunological reagentJournal of Immunological Methods, 1978
- A solid-phase radioimmunoassay for detection of human antibodies. I. Measurement of IgG antibody to bee venom antigensJournal of Immunological Methods, 1978
- 125I protein A: Applications to the quantitative determination of fluid phase and cell-bound IgGJournal of Immunological Methods, 1977
- Specific transplantation tolerance induced by autoimmunization against the individual's own, naturally occurring idiotypic, antigen-binding receptors.The Journal of Experimental Medicine, 1976
- MEASUREMENT OF IGG-BLOCKING ANTIBODIES - DEVELOPMENT AND APPLICATION OF A RADIOIMMUNOASSAY1976
- PROTEIN A FROM STAPHYLOCOCCUS AUREUS .V. REACTION WITH GUINEA PIG GAMMA-GLOBULINS1968
- ALLERGEN-BINDING ACTIVITY OF GAMMAE GAMMAG AND GAMMAA ANTIBODIES IN SERA FROM ATOPIC PATIENTS - IN VITRO MEASUREMENTS OF REAGINIC ANTIBODY1967
- Weight Estimates of Rabbit Anti-Human Serum Albumin Based on Antigen-Binding CapacityThe Journal of Immunology, 1966
- THE PREPARATION OF 131I-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITYBiochemical Journal, 1963
- The Role of Nonprecipitating Insulin Antibodies in Diabetes1JCI Insight, 1958