Synthesis of novel europium-labeled estradiol derivatives for time-resolved fluoroimmunoassays.
The O-(5-carboxypentyl)-, O-(4-aminobutyl)-, O-(6-aminohexyl)oximes of 2- and 4-formylestradiol as well as the 4-carboxyethylthioether derivative of estradiol were synthesized. These estradiol derivatives were characterized using IR-, 1H-, and 13C NMR spectroscopy. All synthesized estradiol derivatives were labeled with europium chelates. These "tracers" were purified and tested in a competitive time-resolved fluoroimmunoassay with monoclonal antibody (SSI 57-2) raised against the 6-O-(carboxymethyl)oxime-bovine serum albumin derivative of estradiol. All the tested europium-labeled estradiol-4-derivatives were bound by the antibody, whereas tracers linked via position 2 were not recognized by this antibody. It was observed that tracers conjugated via C-4 gave more sensitive standard curves than tracers conjugated via C-6. Especially, the estradiol-4-thioether derivative was found to be highly useful in time-resolved fluoroimmunoassays of estradiol while using this antibody.
Meltola
N
N
University of Turku, Medical Physics and Chemistry and Department of Biotechnology, P.O. Box 123, FIN-20521 Turku, Finland. niko.meltola@utu.fi
Jauria
P
P
Saviranta
P
P
Mikola
H
H
D000911:Antibodies, Monoclonal / Q000276:immunology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D005063:Europium / Q000737:chemistry; D015200:Fluoroimmunoassay / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy; D013997:Time Factors
D000911:Antibodies, Monoclonal; D005063:Europium; D004958:Estradiol