Why is RabMab Needed?
The immune system of the rabbit host is highly developed and produces a number of very sensitive antibodies which can recognize a great variety of antigens. This has made it the system of choice to generate antibodies which are hard to produce in rabbits 1,2.
Two decades ago, scientists recognized that the demand for rabbit monoclonal antibodies was great since polyclonal antibodies could not provide signals of consistent quality, but instead possessed non-specificity and high background signal. Monoclonals, on the other hand, recognize a single epitope consistently from separate lots, and are sourced in a sustainable manner.
Weimin Zhu, who is the co-inventor of RabMAb® technology, Abcam, states: “With increasing demands from scientific communities and healthcare industries for antibodies with better sensitivity and specificity, rabbit monoclonals were the answer.”
To meet the need, researchers tried repeatedly to produce rabbit monoclonal antibodies using mouse hybridomas, that is, a hybrid cell made by fusing myeloma cells with mouse spleen cells3.
The earliest was the mouse-rabbit heterohybridomas, but these presented obstacles in the cloning process, were not stable, and could be produced from only a few species4.
It was in 1995 that the double transgenic rabbit was developed by Katherine Knight and her research team at Loyola University, Chicago. They produced overexpression of the oncogenes v-abl and c-myc in the rabbit which were regulated by the heavy and light immunoglobulin chain enhancers. As a result a tumor resembling a myeloma was formed in the rabbit.
This allowed a plasmacytoma cell line named 240E-1 to be isolated. This was the earliest fusion partner cell line to produce rabbit monoclonal antibodies. When this was fused with rabbit lymphocytes, hybridomas were formed that produced rabbit monoclonals on a steady basis, but the fusion partner was found to be unstable5.
In 1996, at the University of California San Francisco, 240E-1 obtained from Dr. Knight’s laboratory was used by Weimin Zhu and Robert Pytela to refine the process still more6.
The 240E-1 cells were subjected to subcloning a number of times and then the best cells were selected to obtain the fusion cell line with the best characteristics. Some of the parameters for selection included efficiency of fusion, growth and morphologic attributes like a bright appearance when viewed using phase contrast microscopy.
The selected subclones were then examined in detail to make sure that they gave rise to hybridomas which were both stable and capable of efficient monoclonal antibody generation. Several subcloning rounds were performed and finally a new cell line emerged, named 240E-W that had impressively stable and effective fusion characteristics.
The next step was the development of the patented Abcam process, named RabMAb, aimed at generating very specific monoclonals with great sensitivity.
The 240E-W underwent further refining after endogenous IgG production became a concern. This has now become 240E-W2, the latest fusion partner, which has been at the heart of today’s RabMAb technology since 2006, to generate rabbit monoclonals suitable for commercial as well as research settings.
The Situation Today
RabMAb is today used to produce monoclonal antibodies of high quality for use in research, diagnostic kits and therapeutic models. Over the last 16 years, the scientific community has accepted RabMAb as being the best technology for many and various needs.
Along with the enormous catalog of RabMAb products, numbering more than 6000 altogether, Abcam also provides this advanced RabMAb technology for the generation of custom antibodies with high sensitivity and specificity.
- Krause RM. Experimental approaches to homogenous antibody populations: factors controlling the occurrence of antibodies with uniform properties. Fed Proc. 1970;29:59-65.
- Weller A, Meek J, Adamson ED. Preparation and properties of monoclonal and polyclonal antibodies to mouse epidermal growth factor (EGF) receptors: evidence for cryptic EGF receptors in embryonal carcinoma cells. Development. 1987;100:351-363.
- Raybould TJ, Takahashi M. Production of stable rabbit-mouse hybridomas that secrete rabbit mAb of defined specificity. Science. 1988;240:1788-1790.
- Verbanac KM, Gross UM, Rebellato LM, et al. Production of stable rabbit-mouse heterohybridomas: characterization of a rabbit monoclonal antibody recognizing a 180 kDa human lymphocyte membrane antigen. Hybridoma. 1993;12:285-295.
- Spieker-Polet H, Sethupathi P, Yam PC, et al. Rabbit monoclonal antibodies: generating a fusion partner to produce rabbit-rabbit hybridomas. Proc Natl Acad Sci U S A. 1995;92:9348-9352.
- Liguori MJ, Hoff-Velk JA, Ostrow DH. Recombinant human interleukin-6 enhances the immunoglobulin secretion of a rabbit-rabbit hybridoma. Hybridoma. 2001 Jun;20(3):189-98.
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