Enzyme immunoassay of two nicergoline metabolites, 10 alpha-methoxy-9, 10-dihydrolysergol (MDL) and 1-methyl-10 alpha-methoxy-9, 10-dihydrolysergol (MMDL).
Specific and sensitive enzyme immunoassays for two nicergoline metabolites, 10 alpha-methoxy-9, 10-dihydrolysergol (MDL) and 1-methyl-10 alpha-methoxy-9, 10-dihydrolysergol (MMDL) have been developed. The hydroxyl group of hydroxymethyl at position 8 of either MDL or MMDL was carboxymethylated to introduce a carboxyl group for protein conjugation. Antibodies generated from O-carboxymethyl MDL or MMDL recognized the spacer arm between the hapten and the carrier protein and the molecular domain near the conjugation site as well. A heterologous bridge strategy was used to improve the affinity of the hapten-enzyme conjugate to the antibodies. The sensitivity of both assays was greatly increased by using such an approach. Both antibodies are specific for their own haptens. Little cross reactivity was observed with nicergoline and other metabolites. Determination of MDL and MMDL from both spiked plasma and urine showed nearly quantitative recovery. Detection of MDL and MMDL can be as sensitive as 10 pg/ml.
Chen
P
P
Division of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Kentucky, Lexington 40536-0082, USA.
Tian
Z
Z
Digenis
G A
GA
Tai
H H
HH
D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D006241:Haptens / Q000276:immunology; D007124:Immunoenzyme Techniques; D008237:Lysergic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D009530:Nicergoline / Q000276:immunology / Q000493:pharmacokinetics*; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C104370:1-methyl-10-methoxy-9,10-dihydrolysergol; D006241:Haptens; C074961:10-methoxy-9,10-dihydrolysergol; D008237:Lysergic Acid; D009530:Nicergoline