Ever since Kohler and Milstein’s ground-breaking somatic cell hybridization experiments of 1975, the mouse has remained the host animal for monoclonal antibody production. Polyclonal antibodies, on the other hand, can be derived from many host species, the rabbit being the one most commonly used owing to its size and availability.
Recently, antibody researchers began to look at methods of producing rabbit monoclonal antibodies, using the same techniques as for mice. The result is that a handful of antibody suppliers – among them us at Novus Biologicals – are now able to provide validated rabbit monoclonal antibodies for research.
The start point for any monoclonal antibody product is the creation, culture and cloning of hybridoma cells specific to that antigen. These are created by fusing spleen B-cells from immunized mice, with fusion partner cells – screened, cultured myeloma cells which secrete no antibodies of their own. This technique has been successfully in use for over 20 years. However, as biotechnology has advanced, scientists have seen disadvantages in using mouse monoclonals, and turned their attention to other species.
An obvious choice would be the rabbit, which has already proven its worth as a host on the polyclonal antibody database. It has several advantages over the mouse as a host, producing higher affinity antibodies to a more diverse range of immunogens – including phospho-peptides and other antigens not immunogenic in mice. However, production of rabbit monoclonal antibodies was hampered by the lack of fusion partners, i.e. those with the qualities demonstrated by murine myeloma cells.
In 1995, Katherine Knight et al developed a double transgenic rabbit myeloma model, which yielded a plasmacytoma cell line fusible with rabbit splenocytes. The resultant hybridomas secreted rabbit monoclonals but, like the early mouse myeloma lines, they were unstable. However, from this a more robust myeloma line, 240E-W, was developed, resulting in stable hybridomas capable of sustained monoclonal antibody secretion. This has now progressed to the next stage, with 240E-W further developed and refined to create a rapidly growing antibody catalog of high affinity rabbit monoclonal antibodies.
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