Strategy for the development of antibodies against ciguatoxins : bevetoxins as a model for polyether hydroxylated compounds.

Publication Type  Book Chapter
Year of Publication  2001
Authors  Naar, J.; Branaa, P.; Dechraoui Bottein, M. Y.; Chinain, M.; Legrand, A. M.; Pauillac, S.
Book Title  Harmful Algal Blooms
Publisher  Hallegraeff G.M., Blackburn S.I., Bolch C.J. and Lewis R.J., Eds - Intergovernmental Oceanographic Commission of UNESCO
Pages  284-7
Language  eng
Abstract  

As a model for hydroxylated polyether toxins, brevetoxin PbTx-3 was converted into a hemisuccinate derivative (PbTx-3 HS) and covalently linked to bovine serum albumin (BSA) and ovalbumin (OVA). By reacting a minute amount of PbTx-3 (0.446 µmol) with 30-fold molar excess of succinic anhydride in dry pyridine, PbTx-3 HS was synthesized with a 100% yield (as verified by HPLC), then covalently coupled to the carriers via the mixed anhydride method performed in a reversed micellar medium. Conjugate analysis, by a combination of chromatographic and radioisotopic techniques showed highly concordant results. Using the optimal conditions for BSA-hapten formation (an input hapten/carrier molar ratio of 75), epitope density of the conjugates (n = 7) was between 15-24 (24-30% yield). In competitive inhibition enzyme-linked immunosorbent assay (ELISA) experiments, immunized mice and rabbits produced PbTx-2-type specific antibodies with apparent affinity constant (KD) in the 10-6-10-8 M range. No significant crossreactivity was observed with PbTx-1-type toxin. Due to the minute hapten quantities involevd, this entire procedure appears most valuable for the preparation of immunogen with poorly available polyether toxins like ciguatoxins (CTXs).

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