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A protein A-binding, polyethylene glycol precipitation-based immunoradiometric assay. Application to the detection of immune complexes and C3 in human sera and of private antigens in cross-reacting parasite extracts.

Authors :
Pontes-de-Carvalho LC
Lannes-Vieira J
Giovanni-de-Simone S
Galvão-Castro B
Source :
Journal of immunological methods [J Immunol Methods] 1986 May 01; Vol. 89 (1), pp. 27-35.
Publication Year :
1986

Abstract

An immunoradiometric assay, based on the precipitation of antigen-antibody complexes by polyethylene glycol (PEG) and on the subsequent binding of PEG-soluble radiolabelled staphylococcal protein A to the PEG-insoluble complexes, is described. The assay can be applied to the detection of naturally occurring, circulating immune complexes, and of complexes artificially created by mixing antigen and antibody solutions, which makes it of potential use for the detection of either antigen or antibody in several situations. Pre-treatment of the antibody-containing sera with 3% PEG greatly reduced the background values and increased the sensitivity of the assay. The assay was also applied to the detection and isolation of Leishmania donovani antigens that did not cross-react with antigens of the related parasite Trypanosoma cruzi (private antigens) and private antigens of insect-derived metacyclic trypomastigotes of T. cruzi in relation to culture-derived metacyclic trypomastigotes of T. cruzi. A simple and extremely effective procedure for washing precipitates with just one centrifugation is also described.

Details

Language :
English
ISSN :
0022-1759
Volume :
89
Issue :
1
Database :
MEDLINE
Journal :
Journal of immunological methods
Publication Type :
Academic Journal
Accession number :
3517172
Full Text :
https://doi.org/10.1016/0022-1759(86)90028-1