Immortalization of antigen selected B cells.
This paper reports the generation of monoclonal antibody producing hybridomas from a small number of antigen-specific B cells selected by panning on antigen-coated dishes and rosetting with antigen-coupled paramagnetic beads. Anti-HIV positive B cells from spleen could be recovered by panning with an efficiency of 5% and a purity of 24%. Immunobead selection of anti-HIV positive B cells from the same mice yielded a recovery of 17% and a purity of 7%. Various experimental conditions with respect to the selection of specific B cells were investigated, leading to an optimized protocol for the isolation of a limited subset of B cells. The selected cells retained their property to produce immunoglobulins and could be clonally expanded in the presence of human T cell supernatant and irradiated murine thymoma helper cells to generate sufficient cells for a mini-electrofusion with NS-1 myeloma cells. Up to 78 specific hybridomas could be generated from one anti-HIV positive B cell. An overall efficiency of specific B cell immortalization of up to 10% was obtained.
Steenbakkers
P G
PG
Department of Immunology, Organon International B.V., Oss, Netherlands.
van Wezenbeek
P M
PM
Olijve
W
W
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D004558:Electric Stimulation; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006678:HIV / Q000276:immunology*; D015483:HIV Antibodies / Q000096:biosynthesis / Q000276:immunology; D015488:HIV Antigens / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D015488:HIV Antigens; D007074:Immunoglobulin G