Rabbit monoclonal antibodies: generating a fusion partner to produce rabbit-rabbit hybridomas.
During the last 15 years several laboratories have attempted to generate rabbit monoclonal antibodies, mainly because rabbits recognize antigens and epitopes that are not immunogenic in mice or rats, two species from which monoclonal antibodies are usually generated. Monoclonal antibodies from rabbits could not be generated, however, because a plasmacytoma fusion partner was not available. To obtain a rabbit plasmacytoma cell line that could be used as a fusion partner we generated transgenic rabbits carrying two transgenes, c-myc and v-abl. These rabbits developed plasmacytomas, and we obtained several plasmacytoma cell lines from which we isolated hypoxanthine/aminopterin/thymidine-sensitive clones. One of these clones, when fused with spleen cells of immunized rabbits, produced stable hybridomas that secreted antibodies specific for the immunogen. The hybridomas can be cloned and propagated in nude mice, and they can be frozen without change in their ability to secrete specific monoclonal antibodies. These rabbit-rabbit hybridomas will be useful not only for production of monoclonal antibodies but also for studies of immunoglobulin gene rearrangements and isotype switching.
Spieker-Polet
H
H
Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University Chicago, Maywood, IL 60153, USA.
Sethupathi
P
P
Yam
P C
PC
Knight
K L
KL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000941:Antigens / Q000032:analysis; D000951:Antigens, Neoplasm / Q000032:analysis; D001798:Blood Proteins / Q000276:immunology; D002459:Cell Fusion; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007158:Immunologic Techniques; D008198:Lymph Nodes / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D008223:Lymphoma / Q000276:immunology; D051379:Mice / Q000276:immunology; D010047:Ovalbumin / Q000276:immunology; D010581:Peyer's Patches / Q000276:immunology; D010954:Plasmacytoma; D011817:Rabbits / Q000276:immunology*; D051381:Rats / Q000276:immunology; D013154:Spleen / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000941:Antigens; D000951:Antigens, Neoplasm; D001798:Blood Proteins; D000939:Epitopes; D010047:Ovalbumin