Development of a monoclonal antibody against the left wing of ciguatoxin CTX1B: Thiol strategy and detection using a sandwich ELISA

Takeshi Tsumuraya, Katsutoshi Takeuchi, Shuji Yamashita, Ikuo Fujii, Masahiro Hirama

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)

Abstract

Ciguatera fish poisoning (CFP) is a form of food poisoning caused by the ingestion of a variety of reef fish that have accumulated trace amounts of ciguatoxins produced by dinoflagellates of the genus Gambierdiscus through the food chain. CFP affects more than 50,000 people each year. The extremely low level of the causative neurotoxins, ciguatoxins, in fish has hampered the preparation of antibodies for detecting the toxins. In this paper, we describe a thiol strategy for synthesizing a keyhole limpet hemocyanin (KLH)-conjugate (20) of the ABCDE-ring fragment of the Pacific ciguatoxins, CTX1B (1) and 54-deoxyCTX1B (4). We succeeded in producing a monoclonal antibody (3G8) against the left wings of these ciguatoxins by immunizing mice with the hapten-KLH conjugate (20) as the synthetic antigen. The most promising mAb, 3G8, does not cross-react with other related marine toxins. Sandwich enzyme-linked immunosorbent assay (ELISA) utilizing 3G8 and the previously prepared monoclonal antibody (8H4) enabled us to detect 1 specifically at less than 0.28 ng/mL.

Original languageEnglish
Pages (from-to)348-357
Number of pages10
JournalToxicon
Volume60
Issue number3
DOIs
Publication statusPublished - 2012 Sep 1

Keywords

  • Ciguatera
  • Ciguatoxin
  • Immunization
  • Immunoassay
  • Monoclonal antibody
  • Sandwich ELISA

ASJC Scopus subject areas

  • Toxicology

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