The immortal B-lymphocyte does not detectably express endogenous immunoglobulin heavychain and may contain, in certain embodiments, an altered immunoglobulin heavy chain-encoding gene. The invention provides a rabbit-derived immortal B-lymphocyte capable of fusion with a rabbit splenocyte to produce a hybrid cell that produces an antibody. References of interest include Spieker-Polet et al., Proc. Hybridomas were produced from lymphocytes ofrabbits immunized using plasma proteins or whole cells, and were shown to secrete IgG antibodies.Īn improved fusion partner cell line for the production of monoclonal antibodies from rabbits is provided. They isolated a plasmacytoma cell line, termed 240E-1, from these tumors, and showed that fusions of 240E-1 cells with rabbit lymphocytes produced hybridomas. 1995 92:9348-52), who succeeded in producing a myeloma-like tumor in transgenic rabbits expressing oncogenes under the control of theimmunoglobulin heavy and light chain enhancers. Knight and colleagues (Spieker-Polet et al., Proc. Attempts have been made to use mouse myeloma cell lines as fusion partners (e.g., Raybould & Takahashi, 1988, supra), or to use in vitro-transformed lymphoid cell lines from rabbits, but in all cases these approaches were hampered by the instability of the resulting hybridomas.Ī novel approach was taken by K. Since myeloma-like tumors are unknown in the rabbit, it has been impossible to use thisapproach for producing rabbit monoclonal antibodies. This method involves immortalization of antibody-producing lymphocytes by fusion with myeloma cells. Monoclonal antibodies are predominantly produced by using the hybridoma technology originallydeveloped by Koehler and Milstein (Nature 1975 256:495-7). In spite of these advantages, until recently it has been impossible to raise monoclonal antibodies in the rabbit. Third, because most monoclonal antibodies are generated in mice and rats, relatively few monoclonal antibodies are availablethat react with mouse or rat immunogens. Second, rabbit antibodies are generally of high affinity. For example, Bystryn et al (Hybridoma 1982 1:465-72) directly compared rabbit and mouse antibodies directed against humanmelanoma cells and showed that they recognize different epitopes. First, rabbits are known to produce antibodies to many antigens that are poorly immunogenic in mice (e.g., Norrby et al., Proc. It has long been recognized that rabbit antibodies have favorable properties compared to mouse antibodies.
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