Antibodies conjugated with new highly luminescent Eu3+ and Tb3+ chelates as markers for time resolved immunoassays. Application to simultaneous determination of clenbuterol and free cortisol in horse urine.
Highly luminescent Eu(3+) and Tb(3+) complexes of 10-[4-(3-isothiocyanatopropoxy)benzoylmethyl]-1,4,7,10-tetraazacyclododecane-1,4,7 triacetic acid Eu(3+) is a subset of 1 and Tb(3+) is a subset of 1 were conjugated with a goat anti-rabbit IgG and a rabbit anti-mouse IgG, respectively, and applied as markers in a time resolved immunoassay for simultaneous quantitative determination of anabolic compounds clenbuterol (CL) and hydrocortisone (HC). The assay was performed in horse urine, using a monoclonal antibody specific to CL and a rabbit polyclonal antibody specific to the free HC. These lanthanide chelates are very stable and highly luminescent in aqueous solution and allowed to reach 10 microg L(-1) and 40 microg L(-1) sensitivities for CL and for HC, respectively. Application to the horse urine, that is a very complex matrix, has a considerable interest in the control of illegal use of these compounds.
Bacigalupo
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MA
Istituto di Chimica del Riconoscimento Molecolare, CNR, Milan, Italy. mariangela.bacigalupo@icrm.cnr.it
Meroni
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Secundo
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Scalera
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Quici
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D015415:Biomarkers / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry*; D002976:Clenbuterol / Q000737:chemistry / Q000652:urine; D004300:Doping in Sports / Q000517:prevention & control; D005063:Europium / Q000737:chemistry*; D006736:Horses; D006854:Hydrocortisone / Q000737:chemistry / Q000652:urine; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology; D049449:Luminescence; D008163:Luminescent Measurements; D015394:Molecular Structure; D015203:Reproducibility of Results; D013725:Terbium / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D002614:Chelating Agents; D007074:Immunoglobulin G; D013725:Terbium; D005063:Europium; D006854:Hydrocortisone; D002976:Clenbuterol