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Matveeva, Evgenia G.; Popova, Valentina A.; Eremin, Sergei A.

doi:10.1023/a:1022517607311

Cited by 6 publications

(3 citation statements)

References 12 publications

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“…Similarly, the signal decreases when the free analyte competes for binding to the Ab. Usually these assays are less sensitive than EIAs but are very useful for sample screening [38,39,40].…”

Section: Polarization Fluorescent Immunoassays (Pfias)mentioning

confidence: 99%

Immunochemical determination of xenobiotics with endocrine disrupting effects

Estévez-Alberola

Marco

2004

Analytical and Bioanalytical Chemistry

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This paper is a review with more than 100 references discussing the immunochemical methods reported in the literature for the most important man-made chemicals with suspected endocrine disrupting activity. Details regarding immunizing hapten design, antibody production, and the features (limit of detection, dynamic range, specificity) of the most important immunochemical methods developed (ELISA, FIIA, immunosorbents, immunosensors, etc.) are presented for important environmental pollutants such as bisphenol A, phthalates, alkylphenol polyethoxylates, alkylphenols, polychlorinated biphenyl compounds, and dioxins. Availability of commercial reagents and methods is reported.

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Section: Polarization Fluorescent Immunoassays (Pfias)mentioning

confidence: 99%

Immunochemical determination of xenobiotics with endocrine disrupting effects

Estévez-Alberola

Marco

2004

Analytical and Bioanalytical Chemistry

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“…In this assay format the detection is based on a fluorophore-conjugated hapten (tracer, fluorescence probe), which is used as a competitor for the antibody binding site. Since it was observed that the fluorescence of fluorescein is changed by antibody binding, fluorescein conjugates are used in many competitive FIA. − This assay format is simple and cost-effective, because only one antibody has to be generated and no antibody labeling is required. Further homogeneous FIA for hapten detection have been reported which are based on FRET between different donor−acceptor pairs conjugated to antibody and hapten. − …”

Section: Introductionmentioning

confidence: 99%

Novel Intramolecular Energy Transfer Probe for the Detection of Benzo[a]pyrene Metabolites in a Homogeneous Competitive Fluorescence Immunoassay

2010

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The spectroscopic properties of a novel intramolecular energy transfer probe (ET probe)--consisting of 3-hydroxybenzo[a]pyrene (3OH-BaP) as the donor covalently linked to sulforhodamine B (SRB) as the acceptor--for the detection of polycyclic aromatic hydrocarbons (PAH) antibody binding were characterized. Absorption and fluorescence spectra as well as fluorescence decay curves were recorded in methanol and aqueous solution, respectively. For comparison, the parent chromophores 3OH-BaP and SRB were investigated as well. In the case of the ET probe, a very strong fluorescence quenching of the BaP-moiety-related emission due to an efficient energy transfer (energy transfer efficiency of about 0.95 for methanol) to the SRB moiety was observed. Upon addition of the PAH antibody, the fluorescence intensity and anisotropy of the BaP moiety was drastically increased. On the other hand, the fluorescence anisotropy of the SRB moiety did not change. The anisotropy results clearly indicate the binding of the antibody. On the basis of these findings, we concluded the following model: the BaP moiety is incorporated in the antibody binding site, whereas the SRB moiety sticks out from the binding site, restricting the motion of the BaP moiety, but leaving the SRB moiety uninfluenced. More important, this structure results in a disruption of the intramolecular energy transfer. The antibody-induced disruption of the intramolecular energy transfer is envisaged as a detection scheme in a future homogeneous competitive fluorescence immunoassay (FIA). This may provide a novel general detection principle for the immunodetection of low-molecular analytes (haptens) in a homogeneous competitive FIA format.

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“…The direct quenching approach has proven to be suitable only for haptens of small size, because the antibody binding site lies sufficiently close to the label to influence its signal. Competitive direct QFIAs based on the fluorescence quenching of a labeled hapten upon binding its antibody (Ab) have been developed for the detection of gentamycin 18 and cortisol 19 in serum, atrazine, 20 2,4-dichlorophenoxyacetic acid, 21 and propazine 12 in nonpolar organic media, and pyrethroid metabolites in urine. 17 More recently, fluoroimmunoassays have been combined with laser-induced fluorescence (LIF) technology to make use of sensitive optical microscope and charge-coupled device (CCD) systems.…”

mentioning

confidence: 99%

Competitive Quenching Fluorescence Immunoassay for Chlorophenols Based on Laser-Induced Fluorescence Detection in Microdroplets

et al. 2002

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An improved biomonitoring system for the analysis of 2,4,6-trichlorophenol (TCP) in urine samples has been developed. The principle of the biosensor device is the detection of laser-induced fluorescence (LIF) in single microdroplets by a homogeneous quenching fluorescence immunoassay (QFIA). The competitive immunoassay occurs in microdroplets (d = 58,4 microm) produced by a piezoelectric generator system with 10-microm-diameter orifice. A continuous Ar ion laser (488 nm) excites the fluorescent tracer; its fluorescence is detected by a spectrometer attached to a 512 x 512 cooled, charge-coupled device camera. Fluorescence is quenched by specific binding of TCP polyclonal antibodies to the fluorescent tracer (hapten A-fluorescein); the quenching effect is diminished by the presence of the analyte. Thus, an increase in the signal is produced in a positive dose-dependent manner when TCP is present in the sample. In 10 mM PBS buffer, the IC50 of the LIF-microdroplet QFIA is 0.45 microg L(-1) reaching a LOD of 0.04 microg L(-1). The QFIA with the same reagents performed in microtiter plate format achieved a LOD of 0.36 microg L(-1) in buffer solution. Performance in human urine was similar to that observed in the buffer. A LOD of 1.6 ,g L(-1), with a dynamic range between 4 and 149.5 microg L(-1) in urine, was obtained without any sample treatment other than dilution with the assay buffer. The detectability achieved is sufficient for occupational exposure risk assessment.

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Untitled

Cited by 6 publications

References 12 publications

Immunochemical determination of xenobiotics with endocrine disrupting effects

Immunochemical determination of xenobiotics with endocrine disrupting effects

Novel Intramolecular Energy Transfer Probe for the Detection of Benzo[a]pyrene Metabolites in a Homogeneous Competitive Fluorescence Immunoassay

Competitive Quenching Fluorescence Immunoassay for Chlorophenols Based on Laser-Induced Fluorescence Detection in Microdroplets

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