K. Kawatsu et al., RAPID AND HIGHLY SENSITIVE ENZYME-IMMUNOASSAY FOR QUANTITATIVE-DETERMINATION OF TETRODOTOXIN, Japanese Journal of Medical Science & Biology, 50(3), 1997, pp. 133-150
A monoclonal antibody against tetrodotoxin (TTX) was obtained from Bal
b/c mice immunized with TTX-bovine serum albumin (BSA) conjugate. The
monoclonal antibody was highly specific for TTX and had no cross-react
ion to tetrodonic acid, which is a TTX derivative, or gonyautoxins, al
though a minor cross-reaction to anhydro-tetrodotoxin was observed. Th
e monoclonal antibody neutralized the lethal activity of TTX. By using
the monoclonal antibody, a rapid and highly sensitive competitive enz
yme immunoassay (EIA) for quantitative analysis of TTX was developed.
By the competitive EIA system, TTX can be determined quantitatively in
about 30 min (90 min are required if the time for preparation of the
solid-phase antigen was included), and the working range for quantitat
ive analysis of TTX was 2-100 ng/ml. In recovery tests and examination
s of TTX samples, results of the mouse bioassay and EIA analyses corre
lated well (r = 0.987). Moreover, it was demonstrated that low concent
rations of TTX, which could not be detected by the mouse bioassay, cou
ld be determined quantitatively by the competitive EIA.