Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen.
J Immunol Methods
; 202(2): 193-204, 1997 Mar 28.
Article
em En
| MEDLINE
| ID: mdl-9107308
ABSTRACT
A novel cell display system was developed for cloning the variable region (V) genes of antigen-specific human antibodies. The system is based on an antibody library displayed on the surface of COS cells, using a plasmid vector designed to direct expression of membrane-bound antibodies. COS cells expressing antigen-specific antibodies were separated using a flow cytometer for their binding to a fluorescent dye-labeled antigen. To test the performance of this system. We cloned V genes of 4 antibodies directed against hepatitis B surface antigen (HBsAg) from a library prepared from peripheral blood lymphocytes of a vaccinated donor. These membrane-bound anti-HBsAg antibodies were easily converted to soluble forms, all of which showed a size similar to human serum IgG in SDS-PAGE and the same specific binding to HBsAg as membrane-bound forms in ELISA. All VH and VK gene segments of the 4 clones isolated in this study belonged to VHIII and VKI subgroups, respectively. These findings demonstrate the potential and selection capabilities of our cell display system for cloning the V genes of antigen-specific human antibodies.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Região Variável de Imunoglobulina
/
Genes de Imunoglobulinas
/
Anticorpos Anti-Hepatite B
/
Antígenos de Superfície da Hepatite B
Limite:
Humans
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Japão