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A monoclonal antibody that specifically binds chitosan in vitro and in situ on fungal cell walls.

Journal of microbiology and biotechnology (2010-08-28)
Max Schubert, Siham Agdour, Rainer Fischer, Yvonne Olbrich, Helga Schinkel, Stefan Schillberg
RÉSUMÉ

We report the generation of the first monoclonal antibody that specifically binds to the polysaccharide chitosan. Mice were immunized with a mixture of chitosans, and hybridoma clones were screened for specific binders resulting in the isolation of a single clone secreting a chitosan-specific IgM, mAbG7. In ELISAs, the antibody can bind to chitosans of varying composition, but demonstrates the highest affinity for chitosans with lower degrees of acetylation (DA) and very poor binding to chitin. We tested the ability of the antibody to bind to chitosan in situ, using preparations of fungal cell walls. Immunofluorescence microscopy confirmed that the antibody bound strongly to the cell walls of fungi with high levels of chitosan, whereas poor staining was observed in those species with cell walls of predominantly chitin or cellulose. The potential use of this antibody for the detection of fungal contamination and the protection of plants against fungal pathogens is discussed.

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IgM, Kappa from murine myeloma, clone TEPC 183, purified immunoglobulin, buffered aqueous solution