Abstract
A novel polyclonal antibody against tetrodotoxin (TTX) was raised using its haptenic antigen, where 4,9-anhydroTTX was reacted with 1,2-ethanedithiol and this derivative was further reacted with keyhole limpet hemocyanin (KLH). This newly designed antigen (KLH-TTX) was inoculated into rabbits, resulting in the production of the specific polyclonal antibody, which reacted well with TTX and its analogs, 4-epiTTX, 11-oxoTTX and 5,6,11-trideoxyTTX, except for 4,9-anhydroTTX. The enzyme-linked immunosorbent assay (ELISA) system using this specific antibody was also developed in the present study. This newly developed polyclonal antibody with analytical procedures using direct one-step ELISA is useful to detect TTX and its analogs in toxic organisms and also disclose the mechanisms involved in their metabolic pathways and accumulation of TTX.
Highlights
Tetrodotoxin (TTX), one of the most potent natural neurotoxins [1,2], has been first detected in pufferfish [3,4] and later determined for its unique structure by Tsuda et al [5], Woodward [6]and Goto et al [7]
TTX and various TTX analogs have been found in toxic organisms such as pufferfish [18,19]
Yasumoto et al [11] reported that some marine bacteria produce TTX and 4,9-anhydroTTX, but the amount produced by these bacteria cannot well explain a high level of TTX accumulated in toxic organisms
Summary
Tetrodotoxin (TTX), one of the most potent natural neurotoxins [1,2], has been first detected in pufferfish [3,4] and later determined for its unique structure by Tsuda et al [5], Woodward [6]and Goto et al [7]. Recent studies revealed that various TTX analogs, such as 4-epiTTX, 11-oxoTTX and 5,6,11-trideoxyTTX, exist together with TTX in diverse animals [18], their concentrations in toxic organisms are much lower than those of TTX except 5,6,11-trideoxyTTX [19,20,21,22,23,24]. Their metabolic pathways to be converted to TTX have remained largely unknown
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