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Bioassay methods for detection of N-palmitoylbrevetoxin-B2 (BTX-B4).
- Source :
-
Toxicon : official journal of the International Society on Toxinology [Toxicon] 2010 Feb-Mar; Vol. 55 (2-3), pp. 497-506. Date of Electronic Publication: 2009 Oct 09. - Publication Year :
- 2010
-
Abstract
- Brevetoxins (BTXs) are a class of cyclic polyether toxins produced by the dinoflagellate Karenia brevis. These substances are subject to extensive conjugative metabolism in shellfish. BTX-B forms a conjugate with cysteine and is oxidized and reduced to yield BTX-B2, which is further modified by fatty acid addition via cysteine amide linkage to give biologically active brevetoxin metabolites. In this study, we evaluated the commonly used in vitro (ELISA, radioimmunoassay, receptor binding assay and N2A cytotoxicity assay) and in vivo mouse brevetoxin bioassays for the detection of the brevetoxin fatty acid conjugate N-palmitoylBTX-B2, and compared the results to those for dihydroBTX-B and BTX-B2. The receptor binding assay for N-palmitoylBTX-B2 showed comparable sensitivity to that for dihydroBTX-B, and an 11-fold higher sensitivity than for BTX-B2. Although the ELISA showed similarly high sensitivity to dihydroBTX-B and BTX-B2, with EC(50) values of ca. 0.26 ng/ml, it was 23 times less sensitive to N-palmitoylBTX-B2. On the other hand, the N2A cytotoxicity assay was highly sensitive to N-palmitoylBTX-B2, with an EC(50) of 0.15 ng/ml, but was 12- and 40-fold less sensitive to dihydroBTX-B and BTX-B2, respectively. The relative sensitivity of the N2A cytotoxicity assay for each of these metabolites paralleled that of the mouse bioassay (relative LD(50) values 1:20:30 for N-palmitoylBTX-B2:dihydroBTX-B:BTX-B2). We conclude that the most sensitive bioassay for dihydroBTX-B and BTX-B2 is the ELISA, whereas the N2A cytotoxicity assay is most sensitive for N-palmitoylBTX-B2.<br /> (Published by Elsevier Ltd.)
- Subjects :
- Acylation
Animals
Batrachotoxins chemical synthesis
Biological Assay
Cell Line, Tumor
Cell Survival drug effects
Chromatography, High Pressure Liquid
Enzyme-Linked Immunosorbent Assay
Fatty Acids analysis
Female
Magnetic Resonance Spectroscopy
Mass Spectrometry
Mice
Radioimmunoassay
Batrachotoxins analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1879-3150
- Volume :
- 55
- Issue :
- 2-3
- Database :
- MEDLINE
- Journal :
- Toxicon : official journal of the International Society on Toxinology
- Publication Type :
- Academic Journal
- Accession number :
- 19819250
- Full Text :
- https://doi.org/10.1016/j.toxicon.2009.09.022