Synthesis of conjugates for a barbiturate screening assay.
Novel derivatives of barbiturates functionalized with free carboxylic acids were designed and synthesized. Coupling of 5-cyclopentyl-5-carboxycrotylbarbituric acid via its active ester to an aminofluorescein derivative produced a fluorescent tracer. Conjugation of the 5-cyclopentenyl-5-carboxyethylbarbituric acid via its mixed anhydride to thyroglobulin allowed for subsequent development of a polyclonal antibody which was evaluated for binding in a fluorescence polarization immunoassay format with various barbiturates. The binding studies showed good cross-reactivity of a variety of barbiturates containing both aromatic and aliphatic 5-substituents with the tested antisera. The relationship between the immunogen architecture, the chemical structure of the binding analytes, and the characteristics of the antisera is also presented.
Adamczyk
M
M
Department of Chemistry (D9NM), Abbott Diagnostics Division, Abbott Laboratories, abbott Park, Illinois 60064-3500, USA. adamczykm@apmac.abbott.com
Grote
J
J
Douglas
J
J
Dubler
R
R
Harrington
C
C
D000818:Animals; D001463:Barbiturates / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D007106:Immune Sera / Q000276:immunology; D007118:Immunoassay; D012756:Sheep; D013329:Structure-Activity Relationship
D001463:Barbiturates; D007106:Immune Sera