Generating probes for cell wall polymers

One of our research aims is to generate defined probes for the analysis of cell wall glycans in situ. Probes such as monoclonal antibodies to oligosaccharide epitopes are crucial tools to dissect the temporal and spatial occurrence of cell wall polysaccharides and their modifications during the life of a plant. Moreover, they are essential tools for determining the polymer arrangements and architecture of individual cell walls. Our probes are generally rat hybridoma monoclonal antibodies. We have also explored the capacity of recombinant his-tagged carbohydrate-binding modules (CBMs) from cell wall hydrolases to act in a similar manner as antibodies as probes for cell wall components.

The generation of monoclonal antibodies to glycans is not straightforward. The limitations include the low immunogenicity of glycans which often necessitates the preparation of neoglycoprotein immunogens. This involves the coupling of a defined oligosaccharide to an immunogenic protein such as BSA or KLH. This strategy has led to the generation of monoclonal antibodies to 1,4-galactan, 1,4-xylan, 1,5-arabinan, 1,4-mannan and xyloglucan. A compounding factor is the limited availability of appropriate defined oligosaccharides that are required both for the coupling to protein and immunogen preparation and also for the characterization of derived antibodies.

Lists of monoclonal antibodies in which we have been involved in generating and details of how samples can currently be obtained can be found here: antibodies.