In vitro immunization of mouse spleen cells for the production of monoclonal IgG1 antibodies using an antigen-specific T helper cell clone (D.10.G4.1).
The in vitro immunization of mouse spleen cells for the production of antigen-specific monoclonal antibodies has been investigated in detail. Using various, previous published in vitro immunization protocols with and without antigen, we were able to show that IgM antibody production was not induced by the antigen. Nevertheless we developed a new in vitro immunization protocol for eliciting antigen-specific monoclonal IgG1 antibodies. Specific antibodies were generated by an antigen-driven immune response which included an immunoglobulin class switch. Co-culture of mouse spleen cells with an activated conalbumin specific T helper cell clone (TH2 D.10.G4.1) in the presence of conalbumin elicited three monoclonal IgG1 antibodies that bound specifically to conalbumin. We conclude that for the in vitro generation of monoclonal antibodies against T cell dependent proteins the addition of antigen activated specific TH2 cell clones is essential.
Uthoff
S
S
Gesellschaft für Biotechnologische Forschung, Department of Genetics, Braunschweig, Germany.
Böldicke
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology* / Q000494:pharmacology; D002478:Cells, Cultured; D002999:Clone Cells; D003207:Conalbumin / Q000494:pharmacology; D003470:Culture Media; D000939:Epitopes; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000276:immunology*; D007075:Immunoglobulin M / Q000276:immunology; D008222:Lymphokines / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008934:Mitogens / Q000494:pharmacology; D016685:Mitomycin / Q000494:pharmacology; D010047:Ovalbumin / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000941:Antigens; D003470:Culture Media; D000939:Epitopes; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008222:Lymphokines; D008934:Mitogens; D003207:Conalbumin; D016685:Mitomycin; D010047:Ovalbumin