title: Hybridoma-free generation of monoclonal antibodies.
abstract: Production of monoclonal antibodies requires immortalization of splenocytes by somatic fusion to a myeloma cell line partner (hybridomas). Although hybridomas can be immortal, they may depend on a feeder cell layer and may be genetically unstable. Since the inception of hybridoma technology, efforts to improve efficiency and stability of monoclonal antibody-producing cell lines have not brought about substantial progress. Moreover, suitable human multiple myeloma-derived cell lines for the production of human antibodies have been very difficult to develop. Here we report a strategy that greatly simplifies the generation of antibodies and eliminates the need for hybridomas. We show that splenocytes derived from transgenic mice harboring a mutant temperature-sensitive simian virus 40 large tumor antigen under the control of a mouse major histocompatibility promoter are conditionally immortal at permissive temperatures and produce monoclonal antibodies. This simple approach may become a method of choice for generation and production of both polyclonal and monoclonal antibodies with advantages in high-throughput discovery and antibody-based immunotherapy.
authors.affiliation: University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. rpasqual@mdanderson.org
authors.forename: Renata
authors.initials: R
authors.lastname: Pasqualini
authors.forename: Wadih
authors.initials: W
authors.lastname: Arap
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002999:Clone Cells / Q000276:immunology; D006183:H-2 Antigens / Q000235:genetics; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D000952:Antigens, Polyomavirus Transforming; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse