Antigen binding properties of purified immunoglobulin A and reconstituted secretory immunoglobulin A antibodies.

Détails

ID Serval
serval:BIB_935204A9CCBC
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Antigen binding properties of purified immunoglobulin A and reconstituted secretory immunoglobulin A antibodies.
Périodique
The Journal of biological chemistry
Auteur(s)
Lüllau E., Heyse S., Vogel H., Marison I., von Stockar U., Kraehenbuhl J.P., Corthésy B.
ISSN
0021-9258
Statut éditorial
Publié
Date de publication
1996
Peer-reviewed
Oui
Volume
271
Numéro
27
Pages
16300-9
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't - Publication Status: ppublish
Résumé
The hybridoma cell line ZAC3 expresses Vibrio cholerae lipopolysaccharide (LPS)-specific mouse IgA molecules as a heterogeneous population of monomeric (IgAm), dimeric (IgAd), and polymeric (IgAp) forms. We describe a gentle method combining ultrafiltration, ion-exchange chromatography, and size exclusion chromatography for the simultaneous and qualitative separation of the three molecular forms. Milligram quantities of purified IgA molecules were recovered allowing for direct comparison of the biological properties of the three forms. LPS binding specificity was tested after purification; IgAd and IgAp were found to bind strongly to LPS whereas IgAm did not. Secretory IgA (sIgA) could be reconstituted in vitro by combining recombinant secretory component (rSC) and purified IgAd or IgAp, but not IgAm. Surface plasmon resonance-based binding experiments using LPS monolayers indicated that purified reconstituted sIgA and IgA molecules recognize LPS with identical affinity (KA 1.0 x 10(8)M-1). Thus, this very sensitive assay provides the first evidence that the function of SC in sIgA complex is not to modify the affinity for the antigen. KA falls to 6.6 x 10(5) M-1 when measured by calorimetry using detergent-solubilized LPS and IgA, suggesting that the LPS environment is critical for recognition by the antibody.
Mots-clé
Animals, Antibodies, Monoclonal, Antibody Specificity, Antigen-Antibody Reactions, Calorimetry, Chromatography, Ion Exchange, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Hybridomas, Immunoblotting, Immunoglobulin A, Immunoglobulin A, Secretory, Kinetics, Lipopolysaccharides, Macromolecular Substances, Mice, Mice, Inbred BALB C, Ultrafiltration, Vibrio cholerae
Pubmed
Web of science
Création de la notice
25/01/2008 15:53
Dernière modification de la notice
03/03/2018 19:34
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