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Validation of IgA1 and IgA2 measurements by a solid-phase immunoradiometric assay in serum and secretions

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Depelchin, Suzanne [UCL] Dehennin, Jean-Pierre [UCL] Bottaro, A. Carbonara, A. Vaerman, Jean-Pierre [UCL] Sibille, Yves [UCL]

We describe specific, sensitive and reproducible immunoradiometric assays to measure total IgA and IgA subclass levels in biological fluids, which take into account the problem that polymeric forms are differently recognized in immunoassays. Sera from subjects totally deficient in one of the IgA subclasses allowed us to ensure the specificity of the subclass assays and to define the proportions of IgA1 (84%) and IgA2 (16%) in the normal pooled serum (from 30 blood donors) used as standard. With purified milk 11-S secretory IgA1 and 11-S secretory IgA2, we determined a correction factor for the corresponding polymeric forms using, respectively, monomeric IgA1 and monomeric IgA2 from pooled serum as standards. With the monoclonal antibodies used, purified 11-S secretory IgA1 was similarly recognized by both the total IgA assay and the IgA1 assay; both total IgA and IgA1 concentrations were underestimated compared with monomeric IgA or monomeric IgA1. In contrast, 11-S secretory IgA2 was better recognized by the IgA2 assay than by the total IgA assay and the values were thus overestimates. Considering this problem of recognition, we fractionated saliva and lung secretions by sucrose density gradient ultracentrifugation before measuring their IgA1 and IgA2 levels.

Document type Article de périodique (Journal article) – Article de recherche
Access type Accès restreint
Publication date 1994
Journal information "International Journal of Clinical and Laboratory Research" - Vol. 24, no. 3, p. 154-161 (1994)
Peer reviewed yes
Publisher Springer Italia Srl
issn 0940-5437
e-issn 1434-4467
Publication status Publié
Affiliations UCL - MD/MINT - Département de médecine interne
UCL - MD/MIGE - Département de microbiologie, d'immunologie et de génétique
UCL - (MGD) Service de pneumologie
MESH Subject Adult ; Body Fluids - immunology ; Bronchoalveolar Lavage Fluid - immunology ; Female ; Humans ; Immunoglobulin A - analysis - blood ; Immunoradiometric Assay ; Male ; Middle Aged ; Molecular Weight ; Reproducibility of Results ; Saliva - immunology ; Sensitivity and Specificity
Keywords IgA subclasses ; IgA molecular size ; Immunoradiometric assay ; Exocrine secretions ; Monoclonal antibodies
Links
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Bibliographic reference Depelchin, Suzanne ; Dehennin, Jean-Pierre ; Bottaro, A. ; Carbonara, A. ; Vaerman, Jean-Pierre ; et. al. Validation of IgA1 and IgA2 measurements by a solid-phase immunoradiometric assay in serum and secretions. In: International Journal of Clinical and Laboratory Research, Vol. 24, no. 3, p. 154-161 (1994)
Permanent URL http://hdl.handle.net/2078.1/24595