Detection and isolation of anti-hapten antibody-secreting cells by cellular affinity matrix technology.
J Immunol Methods
; 422: 80-6, 2015 Jul.
Article
em En
| MEDLINE
| ID: mdl-25896213
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fenilacetatos
/
Plasmócitos
/
Anticorpos Anti-Idiotípicos
/
Haptenos
/
Especificidade de Anticorpos
/
Nitrofenóis
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
2015
Tipo de documento:
Article