Binding Assays for the Quantitative Detection of P. brevis Polyether Neurotoxins in Biological Samples and Antibodies as Therapeutic Aids for Polyether Marine Intoxication

Abstract

The polyether lipid-soluble toxins isolated from the marine dinoflagellate Ptychodiscus brevis (formerly Gymnodinium breve bind to a unique site, Site V, associated with voltage-dependent sodium channels in rat brain synaptosomes. Using tritiated PbTx-3 as a specific probe for binding a Site V, a K sub d of 2.9 nM and a Bmax of 6.8 pmoles/mg synaptosomal protein has been determined. Binding equilibria and displacement by unlabeled PbTx-3 occur in a comparable concentration range to that of saxitoxin (site I). Labeled toxin can be displaced in a competitive manner by any of the other 5 naturally-occuring toxins; the quantitative displacement ability of each appears to reflect individual potency in fish bioassay. Preliminary K sub i calculations have been made for four of the toxins. A comparison of ED50 in radioimmunoassay and ED50 in synaptosome binding for detection of toxins which possess the structural backbone of PbTx-3, the immunizing hapten. Thus, the two assays have quantitative applicability; the former with respect to potency and the latter with respect to structure. Preliminary experiments involving conversion of the radioimmunoassay to urease enzyme linked form have been successful. Keywords: Antibodies; Brevetoxin.

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Document Details

Document Type
Technical Report
Publication Date
Dec 01, 1987
Accession Number
ADA191072

Entities

People

  • Daniel G. Baden

Organizations

  • University of Miami

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  • Biomedical

DTIC Thesaurus Topics

  • Acid-Base Indicators
  • Albumins
  • Alcohols
  • Alkanes
  • Animals
  • Assays
  • Bioassay
  • Chemical Synthesis
  • Chemistry
  • Detection
  • Immune Serums
  • Immunoassay
  • Liquid Chromatography
  • Organic Chemistry
  • Pharmacology
  • Radioimmunoassay
  • Stationary

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  • Biology
  • Chemistry

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  • Molecular and Cellular Biochemistry