Labeling of steroids on solid phase.
Up to four tetra-tert-butyl-1-[4-aminoacetamido)benzyl]diethylenetriaminetetrakis(acetato) derivatives of Fmoc glutamic acid (1) were attached to two steroids (17alpha-hydroxyprogesterone-3-O-carboxymethyloxime 2 and 1,3,5(10)-estratriene-3,16alpha,17beta-triol-6-one-6-O-carboxymethyloxime, 3)) on solid phase using an oligopeptide synthesizer. Upon deprotection and conversion to the corresponding europium(III) chelates, these steroid conjugates were used in DELFIA-based competitive fluoroimmunoassays. The more chelates conjugated to 17-alpha-hydroxyprogesterone, the more diluted antiserum could be used in an immunoassay for 17-alpha-hydroxyprogesterone, without any alteration of the measurement range. Hence, 17-alpha-hydroxyprogesterone tracers with several chelates are useful when a high serum dilution factor is desired i.e., when only a limited quantity of antiserum is available. The result demonstrates the suitability and usefulness of lanthanide(III) chelates as multilabels in bioaffinity assays.
Peuralahti
Jari
J
PerkinElmer Life and Analytical Sciences, Turku Site, P.O. Box 10, FIN-20101, Finland.
Suonpää
Katriina
K
Blomberg
Kaj
K
Mukkala
Veli-Matti
VM
Hovinen
Jari
J
D002851:Chromatography, High Pressure Liquid; D007118:Immunoassay / Q000295:instrumentation*; D015394:Molecular Structure; D013194:Staining and Labeling / Q000379:methods*; D013256:Steroids / Q000138:chemical synthesis / Q000737:chemistry*
D013256:Steroids