Lr. Briggs et al., DEVELOPMENT OF AN ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR ANALYSIS OF SPORIDESMIN-A AND ITS METABOLITES IN OVINE URINE AND BILE, Journal of agricultural and food chemistry, 42(12), 1994, pp. 2769-2777
Monoclonal and polyclonal antibodies have been produced for the develo
pment of competitive enzyme-linked immunosorbent assays for sporidesmi
n A, the mycotoxin that causes facial eczema in grazing livestock. Hem
isuccinyl derivatives of sporidesmin conjugated to a number of protein
s were used as immunizing and coating antigens. Two immunoassays have
been developed. Each detects the presence of a different region of the
sporidesmin molecule, therefore allowing the two major sporidesmins f
ound in grass or in culture to be distinguished as well as allowing th
e detection of metabolites in ovine urine and bile. An analytical proc
edure that did not include organic solvent extraction was developed so
that water-soluble metabolites would also be detected. Matrix effects
occurring in the immunoassay of bile have been overcome by dilution o
f samples in 1% Tween 20 followed by heat treatment at 70 degrees C fo
r 30 min. Sporidesmin A in bile measured by immunoassay was compared w
ith that measured by HPLC. The coefficient of determination R(2) was 0
.97.