title: Development of a monoclonal antibody against the left wing of ciguatoxin CTX1B: thiol strategy and detection using a sandwich ELISA.
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.
authors.affiliation: Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Osaka 599-8531, Japan. tsumu@b.s.osakafu-u.ac.jp
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tsumuraya
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Takeuchi
authors.forename: Shuji
authors.initials: S
authors.lastname: Yamashita
authors.forename: Ikuo
authors.initials: I
authors.lastname: Fujii
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hirama
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000915:Antibody Affinity; D036841:Ciguatera Poisoning / Q000517:prevention & control; D002922:Ciguatoxins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D003429:Cross Reactions; D004141:Dinoflagellida / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005399:Fishes; D005506:Food Contamination; D005513:Food Inspection / Q000379:methods*; D006241:Haptens / Q000737:chemistry; D006433:Hemocyanins / Q000737:chemistry; D057230:Limit of Detection; D017747:Seafood / Q000032:analysis*
chemical_list: D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D006241:Haptens; C454647:ciguatoxin 1B (CTX 1B); D002922:Ciguatoxins; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin