Preparation and development of anti-chitosan antibodies

Polyclonal antibodies directed against chitosan were produced using several immunogens, prepared by binding the polymer according to two ways (covalent and electrostatic) with a protein (bovine serum albumin or hemocyanin). It appeared that the presence of a carrier protein linked to chitosan was necessary to enhance the immune response and to obtain antibodies in a stable and reproducible way. Direct and inhibition enzyme-linked immunosorbent assay experiments were performed to assess the affinity and the specificity of the antibodies. The interactions of these antibodies with modified chitosans showed no influence of the degree of polymerization of the polymer in the range studied (from 24 to 2261), by contrast with the degree of acetylation. The affinity decreased when the degree of acetylation increased. Absence of cross-reactivity with glycosaminoglycans was observed whatever the antibody. The cationicity of the amine function along the polymer chains may have a role in the immunological recognition of the chitosan structure.