Detection of chemicals by a reporter immunoassay: application to fluoride.
This report describes a concept in which an immunoassay is used indirectly to quantify a nonantigenic very low molecular weight compound participating in a chemical reaction with a haptenic reporter. The detection limit of each reagent is, therefore, governed only by the affinity of the antibodies toward the reporter. Fluoride was used as a model, and silylated estradiol was used as a reporter. Upon silylation with N-O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) or N-O-bis(dimethylterbutylsilyl) trifluoroacetamide (MTBSTFA), estradiol is no longer recognized by antibodies specific to estradiol. After reaction with hydrofluoric acid (HF) or fluoride salts (KF, CsF, NaF), its immunoreactivity is restored, and native estradiol is formed and is detected by immunoassay. The level of synthesized estradiol is dependent on the concentration of fluoride. A fluoride detection limit of 0.3 microg/L (15 nM) is obtained. Potential interference with other acids has been eliminated by choosing the silyl group (trimethylsilyl vs tert-butyldimethylsilyl) and by selecting optimal reaction conditions for the desilylation. The method has been applied to the detection of fluoride salts in natural waters (range 0.28-9.0 mg/L) and in an atmosphere artificially contaminated with HF between 8 and 160 microg/m(3) in the parts-per-billion range. This indirect immunoassay combines simplicity and high sensitivity and, therefore, can be used in field monitoring. Finally, the extension of the concept to other chemicals is discussed.
CEA, Service de Pharmacologie et d'Immunologie, UMR CNRS6014, 76131 Mont St-Aignan, France.
Marie-Astrid
MA
Sagot
Florence
F
Heutte
Pierre-Yves
PY
Renard
Frédéric
F
Dollé
Philippe
P
Pradelles
Eric
E
Ezan
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004958:Estradiol / Q000031:analogs & derivatives / Q000032:analysis*; D005459:Fluorides; D007118:Immunoassay / Q000379:methods; D007124:Immunoenzyme Techniques; D007202:Indicators and Reagents; D014297:Trimethylsilyl Compounds
D007202:Indicators and Reagents; D014297:Trimethylsilyl Compounds; D004958:Estradiol; C047270:N,N-bis(trimethylsilyl)-2,2,2-trifluoroacetamide; D005459:Fluorides