title: Detection and isolation of anti-hapten antibody-secreting cells by cellular affinity matrix technology.
abstract: We developed a method to detect and isolate plasma cells that produce antigen-specific antibodies. An affinity matrix of hapten was constructed on a cell surface, and subsequent incubation allowed cells to secrete antibodies. Anti-hapten antibodies preferentially bound to the affinity matrix on the cells from which they were secreted. We showed that the combination of surface biotinylation and streptavidin which was conjugated with a high valence of hapten was suitable for sensitive detection of antibody binding. Using this protocol, anti-hapten plasma cells from immunized mouse spleen were detected and enriched by flow cytometry. This method allows for isolation of intact plasma cells according to the antibody specificity and may be useful for highly efficient and precise analysis of an antibody repertoire.
authors.affiliation: Department of Immunology, Kochi Medical School, Kochi University, Oko-cho Kohasu, Nankoku, Kochi 783-8505, Japan. Electronic address: tshimizu@kochi-u.ac.jp.
authors.forename: Takeyuki
authors.initials: T
authors.lastname: Shimizu
authors.affiliation: Laboratory of Structural Immunology, Division of Bioinformatics, Research Institute for Biomedical Sciences (RIBS), Tokyo University of Science, Yamazaki, Noda, Chiba 278-0022, Japan. Electronic address: tazuma@rs.noda.tus.ac.jp.
authors.forename: Takachika
authors.initials: T
authors.lastname: Azuma
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D002478:Cells, Cultured; D005434:Flow Cytometry; D006241:Haptens / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009596:Nitrophenols / Q000276:immunology*; D010648:Phenylacetates / Q000276:immunology*; D010950:Plasma Cells / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology
chemical_list: D000888:Antibodies, Anti-Idiotypic; D006241:Haptens; D009596:Nitrophenols; D010648:Phenylacetates; C026242:4-hydroxy-5-nitrophenyl acetic acid
keywords: Affinity matrix; Antibody; Flow cytometry; Hapten; Plasma cells