Crossing from poly to mono - the race to create rabbit monoclonal antibodies

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by: Gen Federico
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Word Count: 342
Date: Thu, 17 Nov 2011 Time: 6:48 AM

We at Novus Biologicals have an extensive antibody catalog of monoclonal and polyclonal antibodies. While our polyclonal antibody database includes reagents raised to a variety of hosts, our monoclonal antibodies were, until recently, murine derived. However, we now have a new option - rabbit monoclonals. Vimentin, ApoE, Oct2, Chromogranin, CD3 epsilion and Bax are just some of the immunogens for which a rabbit monoclonal antibody is available.

In vitro hybridoma production has been in use for over 35 years. B-cells from immunized mice are fused with non-secretory murine myeloma cells, and the resultant hybridomas harvested and cloned. Mouse monoclonal antibodies (MMAs) are more costly to produce than polyclonals, but offer numerous advantages in terms of purity, uniformity and continuous availability. However, not all immunogens stimulate an adequate antibody response in mice, and in these cases polyclonals from another species must be used. Mice are rarely used for polyclonal production, as their small size means they produce insufficient sera unless peritoneal ascites production is stimulated.

A polyclonal antibody (PA) targets a range of epitopes or amino acid sequences for a given antigen, and thus has higher affinity than the mouse monoclonal equivalent. The rabbit, a commonly used polyclonal host, is known to produce high titre, high affinity PAs against a range of targets, including proteins which do not produce an immunogenic response in mice.

Experiments comparing affinity of rabbit PAs and mouse MAs to short synthetic polypeptides suggested the rabbit in general may produce higher affinity antibodies than mice to human epitopes. In recent years, many groups have attempted to create rabbit monoclonal antibodies (RabMAbs) combining the best properties of MMAs and RPAs. However, plasmacytomas are unknown in rabbits, and viral transformation of rabbit B-cells is difficult in vitro. Mouse-rabbit heterohybridomas were developed, but with unsuccessful results.

Eventually, a new transgenic c-myc/v-abl rabbit model was created, which yielded rabbit plasmacytoma cells – the fusion partner that was needed. This led to the development of the rabbit monoclonal antibodies sold by antibody suppliers today.

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