Binding Assays for the Quantitative Detection of P. brevis Polyether Neurotoxins in Biological Samples and Antibodies as Therapeutic Aids for Polyether Marine Intoxication
Final rept. 1 Dec 1986-30 Nov 1989
MIAMI UNIV CORAL GABLES FL
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The polyether lipid-soluble toxins isolated from the marine dinoflagellate Ptychodiscus brevis can be detected using immunoassays. Brevetoxins can be covalently linked to proteins to produce fully antigenic materials. Antigen can be used to raise antibodies in goats using the toxin-protein conjugates. Both radioimmunoassays and enzyme-linked immunoassays can be developed using the goat antibodies. Toxin derivatives in which enzymes are covalently attached are unstable and lose enzymatic activity under all conditions examined. Toxin antibody purification can be aided using covalent toxin affinity columns employing aminohexyl Sepharose as solid support. Immunoassays show no crossreactivity with okadaic acid or the organisms responsible for its biosynthesis cells or in the adsorbed cells themselves in enzyme-linked formats. Ciguatoxin or fish flesh toxins can be detected as well, although the quantitative nature of the interaction is still unknown. Antibrevetoxin antibodies can be used in vitro to either compete for toxin with synaptosomes, or to remove toxin from synaptosomes once they have adsorbed. Successful immunoprophylaxis can be anticipated using appropriate regimes. Fully developed enzyme-linked immunoassays for the detection and quantification of brevetoxin-like polyether toxins in biological samples is anticipated within a year.