Production of monoclonal antibodies for sandwich immunoassay detection of Pacific ciguatoxins.
Ciguatoxins are the major causative toxins of ciguatera seafood poisoning. Limited availability of ciguatoxins has hampered the development of a reliable and specific immunoassay for detecting these toxins in contaminated fish. Monoclonal antibodies (mAbs) specific against both ends of Pacific ciguatoxins CTX3C and 51-hydroxyCTX3C were prepared by immunization of mice with the protein conjugates of rationally designed synthetic haptens in place of the natural toxin. Haptenic groups that possess a surface area larger than 400 A(2) were required to produce mAbs that can bind strongly to CTX3C or 51-hydroxyCTX3C. A direct sandwich enzyme-linked immunosorbent assay (ELISA) using these mAbs was established to detect CTX3C and 51-hydroxyCTX3C at the ppb level with no cross-reactivity against the other marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, or maitotoxin.
Tsumuraya
Takeshi
T
Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Sakai, Osaka 599-8570, Japan. tsumu@b.s.osakafu-u.ac.jp
Fujii
Ikuo
I
Hirama
Masahiro
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002922:Ciguatoxins / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006241:Haptens; D051379:Mice
D000911:Antibodies, Monoclonal; D006241:Haptens; D002922:Ciguatoxins