A general method to perform a noncompetitive immunoassay for small molecules

被引:31
作者
Giraudi, G [1 ]
Anfossi, L [1 ]
Rosso, I [1 ]
Baggiani, C [1 ]
Giovannoli, C [1 ]
Tozzi, C [1 ]
机构
[1] Univ Turin, Dipartimento Chim Analit, I-10125 Turin, Italy
关键词
D O I
10.1021/ac981282c
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A new general method to perform a noncompetitive immunoassay for low-molecular-mass analytes (less than 6000 Da) is described and checked using cortisol as a model system. The method is based on the use of a "polydentate ligand" (cortisol-poly(L-lysine) conjugate) able to block the antibody sites unoccupied by the analyte, followed by the replacement of an antibody-bound analyte by an enzyme-labeled analyte (cortisol-horseradish peroxidase), and permits the direct measurement of the analyte bound sites. The observed signal shows a near-linear correlation with the analyte concentration. The characteristics of interactions between the analyte and polydentate ligand with the specific antibody were studied to perform a preliminary evaluation of the noncompetitive immunoassay for cortisol, The noncompetitive assay was compared with a competitive immunoassay obtained under the same conditions and using the same reagents. The results of the experiments showed a lower detection limit for the noncompetitive model (0.15 ng mL(-1) rather than 0.72 ng mL(-1)), emphasizing that the model is successful. Moreover, as the polydentate ligand is prepared from the same hapten used for the immunogen synthesis, this type of noncompetitive immunoassay appears generally applicable to all small molecules for which antibodies have been obtained.
引用
收藏
页码:4697 / 4700
页数:4
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