Rabbit Hybridoma Development

Rabbit Hybridoma Development

Rabbits belong to the order Lagomorpha, which is evolutionary distinct from the order Rodentia, to which mice and rats belong. Rabbit monoclonal antibodies possess several key advantages over rodent monoclonal antibodies. First, rabbit monoclonal antibodies are able to recognize epitopes on human antigens that are not immunogenic in rodents, therefore increasing the total number of targetable epitopes. Second, owing to their larger body size, rabbits can provide 50 times more spleen B cells than mice. Higher total B cell number warrants higher antibody diversity. Third, rabbit monoclonal antibodies can be more readily humanized by grafting into human antibody framework or Fc domain.

Rabbit hybridomas, however, are difficult to make. Both rabbit-mouse hetero-hybridomas, derived from fusion of rabbit splenocytes with mouse myeloma cells, and rabbit homo-hybridomas, derived from fusion of rabbit splenocytes with rabbit plasmacytoma cells, showed low growth rates, low antibody yields, and gradual losses of antibody secretion.

BiCell Scientific Inc has developed a serum-free culturing method that supports the rapid growth of rabbit hybridomas and the stable production of rabbit monoclonal antibodies. Our rabbit hybridoma culturing approach is included into rabbit monoclonal antibody service in Custom Antibodies.

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Rabbit-mouse hetero-hybridoma development. Rabbit splenocytes were fused with mouse myeloma cells and cultured in HAT containing medium. Stable hybridoma clones started to show on Day 7.

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Hybridoma cell plating and screening. Antibody secreting rabbit hybridoma cells were cultured in serum-free medium and their supernatants screened by ELISA in 96-well plates.

"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

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