Polarization fluoroimmunoassays for the specific determination of amphetamine and methamphetamine in urine with the abbott TDx-analyzer

Еремин, С. А.; Zaitchev, S.V.; Egorov, A. M.; Ermakov, A. N.; Smirnov, A.V.; Izotov, Boris N.

doi:10.1016/s0003-2670(00)82669-0

Cited by 10 publications

(3 citation statements)

References 6 publications

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“…The fluorescence polarization assay (FPIA) is the most widely used homogeneous assay with applications mainly in therapeutic and drug abuse monitoring. − In recent years a lot of work has been performed for FPIA development and application in environmental analysis. Pesticides such as 2,4D, 2,4,5T, atrazine, simazine, isoproturone, methabenzthiazurone, dichlorprop, and propazine have been used as antigens. ,,, Despite the apparent advantages, the relatively low sensitivity remains as the major limitation of the assay .…”

mentioning

confidence: 99%

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

et al. 2002

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Fluorescence polarization immunoassay (FPIA) is a convenient homogeneous assay, the use of which is restricted in environmental analysis by low sensitivity and matrix effects. We selected the herbicides 2,4D and 2,4,5T to synthesize new L-lysine-based fluorescent tracers using solid-phase chemistry. In addition, three different immunogens of 2,4,5T were prepared for immunization and antibody production. The new tracers and antibodies were adapted to FPIA. Tracers with the hapten attached to the alpha-aminogroup of L-lysine and fluorescein to the e-amino group exhibited at least a 5-fold increased sensitivity when compared to the previously reported ethylenediamine-based tracer (2,4D-EDA-F). The isomeric structure (hapten attached to the e-amino and fluorescein to the alpha-amino group) appeared 7.6 times less sensitive, and all other alternative structures exhibited even lower sensitivities. This observation was confirmed against the monoclonal anti-2,4D antibody E2/G2 and polyclonal anti-2,4,5T antibodies. The affinity constant of 2,4D-EDA-F with E2/G2 was 8.1 times higher when compared with the new tracer, suggesting the more specific nature of the L-lysine-based tracer, the use of which leads to a more sensitive assay. This type of tracer could improve performance and lower substantially the detection limits of FPIAs.

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confidence: 99%

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

et al. 2002

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“…Most of the applications of homogeneous PFIA are for analytes present in substantial concentrations, for example, drugs (Eremin et al, 1989(Eremin et al, ,1992aNithipatikom et al, 1989;Colbertet al, 1991). PFIA has been scarcely applied to the determination of pesticide contaminants.…”

Section: Polarization Fluoroimmunoassay Of the Herbicide Dichlorpropmentioning

confidence: 99%

“…The advantages of this technique as a means of measuring analytes in serum were outlined in original publications on this subject (Dandliker etal., 1973;Spencer etal., 1973). Most of the applications of homogeneous PFIA are for analytes present in substantial concentrations, for example, drugs (Eremin et al, 1989(Eremin et al, ,1992aNithipatikom et al, 1989;Colbert etal., 1991). PFIA has been scarcely applied to the determination of pesticide contaminants.…”

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confidence: 99%

Polarization fluoroimmunoassay of the herbicide dichlorprop

Sánchez¹,

Navas²,

Alonso³

et al. 1993

J. Agric. Food Chem.

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A polarization fluorescence immunoassay has been developed for the screening of the herbicide dichlorprop [(f)-2-(2,4-dichlorophenoxy)propanoic acid]. The assay is based on the difference in fluorescence polarization between free and antibody-bound fluoresceinamine-labeled dichlorprop. Antibodies toward dichlorprop were produced by immunizing a rabbit with conjugates of dichlorprop to bovine serum albumin. Polarization measurements were performed by using two spectrofluorometers which use different optic configurations. Calibration curves fitted to an IC50 four-parameter logistic model gives chi-square 10-5. The upper end of the standard curve for polarization fluorescence immunoassay is 0.01 pg/mL and the lower end 100 pg/mL. The method offers good analytical specifications and little crossreactivity against similar compounds. The proposed method is compared with enzyme-linked immunosorbent assay using the same set of reagents. The effectiveness of the method on real samples was checked in spiked apple samples.

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Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules

2008

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Fluorescence polarization immunoassay (FPIA) is a homogeneous (without separation) competitive immunoassay method based on the increase in fluorescence polarization (FP) of fluorescent-labeled small antigens when bound by specific antibody. The minimum detectable quantity of FPIAs with fluorescein label (about 0.1 ng analyte) is comparable with chromatography and ELISA methods, although this may be limited by sample matrix interference. Because of its simplicity and speed, FPIA is readily automated and therefore suitable for high-throughput screening (HTS) in a variety of application areas. Systems that involve binding of ligands to receptor proteins are also susceptible to analysis by analogous FP methods employing fluorescent-labeled ligand and HTS applications have been developed, notably for use in candidate drug screening.

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Polarization fluoroimmunoassays for the specific determination of amphetamine and methamphetamine in urine with the abbott TDx-analyzer

Cited by 10 publications

References 6 publications

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

Use of l-Lysine Fluorescence Derivatives as Tracers To Enhance the Performance of Polarization Fluoroimmunoassays. A Study Using Two Herbicides as Model Antigens

Polarization fluoroimmunoassay of the herbicide dichlorprop

Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules

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