Micropurification and two-dimensional polyacrylamide gel electrophoresis of immunoglobulins for studying the clonal diversity of antigen-specific antibodies

Détails

ID Serval
serval:BIB_2876137F9B50
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Micropurification and two-dimensional polyacrylamide gel electrophoresis of immunoglobulins for studying the clonal diversity of antigen-specific antibodies
Périodique
Journal of Immunological Methods
Auteur⸱e⸱s
Layer  A., Tissot  J. D., Schneider  P., Duchosal  M. A.
ISSN
0022-1759 (Print)
Statut éditorial
Publié
Date de publication
07/1999
Volume
227
Numéro
1-2
Pages
137-148
Langue
anglais
Notes
Journal Article Research Support, Non-U.S. Gov't --- Old month value: Jul 30
Résumé
Presented here is a simplified method for purifying antigen-specific antibodies and analysing them by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). For this procedure, specific immunoglobulins (Ig) are bound to resin beads coated with antigen and, in the presence of sodium dodecyl sulphate and dithioerythritol, loaded directly onto isoelectric focusing strips for 2D-PAGE analysis. This technique bypasses antigen-specific Ig elution and concentration and therefore minimises antibody loss and simplifies sample preparation. Because of its high sensitivity, this technique permits clonal analysis of samples containing small quantities of Ig. We studied Ig anti-tetanus toxoid (TT) and IgG in sera from several sources. The resulting oligoclonal Ig anti-TT patterns contrasted with the polyclonal patterns of total IgG visualised after 2D-PAGE analysis of the respective serum samples.
Mots-clé
Animals *Antibody Diversity Electrophoresis, Gel, Two-Dimensional Humans Immunoglobulin G/blood Immunoglobulins/analysis/immunology/*isolation & purification Mice Mice, SCID Tetanus Toxoid/immunology
Pubmed
Web of science
Création de la notice
25/01/2008 16:34
Dernière modification de la notice
20/08/2019 14:07
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