>>Published online before print December 19, 2003
Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0305834101
Immunology
Hybridoma-free generation of monoclonal antibodies
Renata Pasqualini * and Wadih Arap *
University of Texas M. D. Anderson Cancer Center, Houston, TX 77030
Communicated by Richard L. Sidman, Harvard Medical School, Boston, MA, September 11, 2003 (received for review August 11, 2003)
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.<<
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