A competitive enzyme-linked immunosorbent assay (ELISA) to detect retronecine and monocrotaline in vitro.
Antibodies to the nonesterified pyrrolizidine nucleus, retronecine (155 mol.wt), were produced in rabbits and detected using an avidin-biotin antibody ELISA. A competitive ELISA for the detection of retronecine and the cyclic diester monocrotaline was also developed using the antiserum produced against the hapten conjugate, retronecine-bovine serum albumin. Retronecine was obtained by hydrolysis of monocrotaline, succinylated and directly coupled to bovine serum albumin or ovalbumin. Antibodies to the pyrrolizidine nucleus, retronecine, can be detected within 5 min after the addition of substrate using the avidin-biotin ELISA. Competitive inhibition of antibodies to retronecine is obtained by the addition of known amounts (0-11.42 micrograms/microliters) of either the homologous antigen, retronecine, or the heterologous antigen, monocrotaline, however, retronecine acts as the better competitor.
Bober
M A
MA
Department of Chemistry, University of California, Davis 95616.
Milco
L A
LA
Miller
R B
RB
Mount
M
M
Wicks
B
B
Kurth
M J
MJ
D000818:Animals; D003429:Cross Reactions; D003956:Dialysis; D004797:Enzyme-Linked Immunosorbent Assay; D016686:Monocrotaline; D010047:Ovalbumin / Q000276:immunology; D011763:Pyrrolizidine Alkaloids / Q000032:analysis*; D011817:Rabbits
D011763:Pyrrolizidine Alkaloids; C014795:retronecine; D016686:Monocrotaline; D010047:Ovalbumin