Novel Polyclonal Antibody Raised against Tetrodotoxin Using Its Haptenic Antigen Prepared from 4,9-anhydrotetrodotoxin Reacted with 1,2-Ethaneditiol and Further Reacted with Keyhole Limpet Hemocyanin

Toxins (Basel). 2019 Sep 20;11(10):551. doi: 10.3390/toxins11100551.

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.

Keywords: ELISA; polyclonal antibody; pufferfish; tetrodotoxin; tetrodotoxin analogs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / immunology
  • Antigens / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Haptens / immunology*
  • Hemocyanins / immunology*
  • Rabbits
  • Tetrodotoxin / analogs & derivatives
  • Tetrodotoxin / analysis*
  • Tetrodotoxin / immunology
  • Tetrodotoxin / metabolism

Substances

  • Antibodies
  • Antigens
  • Haptens
  • Tetrodotoxin
  • Hemocyanins
  • keyhole-limpet hemocyanin