Production and characterization of a monolonal antibody to type-2 brevetoxins.

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  280-3
Language  eng

Two murine monoclonal antibodies (MAbs 1A11 and 3A12) directed against brevetoxin PbTx-3 were selected from fusion of P3-X63-Ag 8.653 myeloma cells with BALB/c mouse immune spleen cells sensitized to a PbTx-3-bovine albumin conjugate. In competitive inhibition enzyme-linked immunosorbent assay (ELISA) experiments, both MAbs exhibited strong cross-reactivity (³ 100%) to other PbTxs-2-type toxins (PbTx-2 and -9) but low cross-reactivity (9%) to a PbTx-1-type toxin (PbTx-1). The apparent dissociation constant (KD) for the interaction of these MAbs with free PbTx-2-type toxins was in the 10-6-10-7 M range. The limit of detection (» 5 ng/well) and the working range (8 - 150 ng/well) achieved with this ELISA coupled with adequate extraction methods would provide an alternative screening method to the intraperitoneal mouse bioassay for routine shellfish monitoring. Owing to the minute hapten quantities involved in conjugate preparation (0.446 µmol of PbTx-3) and the well-known advantage of monoclonal over polyclonal antibodies, this entire procedure appears most valuable for the development of immunoassays to highly potent marine polyether toxins.

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