Solid phase radioimmunoassay for quantitation of antigen-specific IgG in human sera with /sup 125/I-protein A from Staphylococcus aureus

Journal Article · Thu Mar 01 04:00:00 EST 1979 · J. Immunol.; (United States)

OSTI ID:5290944

Hamilton, R G; Sobotka, A K; Adkinson, Jr, N F

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 h during which specific antibody is bound; second, after a washing procedure, the solid phase immune complexes are incubated for 4 to 18 h with /sup 125/I-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 /sup 125/I-Staph A in the solid phase radioimmunoassay was superior to /sup 125/I 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 /sup 125/ 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.

• Research Organization: Johns Hopkins Univ., Baltimore, MD
• OSTI ID: 5290944
• Journal Information: J. Immunol.; (United States), Journal Name: J. Immunol.; (United States) Vol. 122:3; ISSN JOIMA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
ANIMALS
ANTIBODIES
BACTERIA
BETA DECAY RADIOISOTOPES
BLOOD SERUM
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
GLOBULINS
IMMUNOGLOBULINS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
MAN
MICROORGANISMS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOISOTOPES
STAPHYLOCOCCUS
TRACER TECHNIQUES
VERTEBRATES