Time-Resolved Fluorescence of Lanthanide Probes and Applications in Biotechnology

Soini, Erkki; Lövgren, Timo; Reimer, Charles B.

doi:10.1080/10408348708542802

Cited by 91 publications

(93 citation statements)

References 121 publications

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“…The TRF offers a sensitive, nonradioactive, and rapid method for the quantification of the amplification products. 40 The QRT-PCR assay developed gave an analytical detection limit of 50 (i.e., 2 times the mean of background signal) and a functional detection limit of 10 2 (i.e., between-assay CVs under 23%) target mRNA copies with a linear range up to 10 6 mRNA copies.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Quantification of Human Glandular Kallikrein 2 and Prostate-Specific Antigen mRNAs in Peripheral Blood from Prostate Cancer Patients

Ylikoski

Karp

Pettersson

et al. 2001

The Journal of Molecular Diagnostics

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We present a multiplexed and internally calibrated quantitative reverse transcription-PCR (QRT-PCR) assay to detect human glandular kallikrein 2 (hK2) and prostate-specific antigen (PSA) transcripts in blood samples from healthy subjects and prostate cancer (PC) patients. The assay detected 50 copies of hK2 and PSA mRNA, and 1 PSA-and 10 hK2-expressing LNCaP cells in the presence of 2.5 ؋ 10 6 PSA-and hK2-negative cells. In PC patients, 20 of 25 and 19 of 25 gave detectable PSA and hK2 mRNAs, respectively. Number of hK2 mRNA copies was significantly higher than that of PSA mRNA copies in patients with biochemically progressive (P ‫؍‬ 0.02) PC, and with locally advanced and metastasized (P ‫؍‬ 0.004) PC. Patients with rapidly progressive and hormone refractory PC gave detectable hK2 mRNA only in 2 of 8 and PSA mRNA in 3 of 8 patients. Neither PSA nor hK2 mRNAs were detected in 16 healthy subjects. PSA and hK2 discriminated PC patients with biochemically progressive and advanced disease from the controls and from the aggressive distant metastatic disease. The assay provides a reliable quantification of the number of hK2 and PSA mRNA copies, allows to discriminate PC cases from healthy subjects, and offers a tool for further studies on molecular staging of PC.

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Section: Discussionmentioning

confidence: 99%

Simultaneous Quantification of Human Glandular Kallikrein 2 and Prostate-Specific Antigen mRNAs in Peripheral Blood from Prostate Cancer Patients

Ylikoski

Karp

Pettersson

et al. 2001

The Journal of Molecular Diagnostics

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“…Time gating the fluorescence detection allows reduction of such effects because no rapidly decaying fluorescence is detected and the system is inherently more sensitive. 5 TR-FRET assays are also extensively used as screening methods.…”

Section: Comparison With Other Techniquesmentioning

confidence: 99%

“…The detection relies on time-resolved luminescence, which is by far more sensitive than conventional fluorescence. 5 The unique fluorescence properties of lanthanide chemistry have led to the development of successful diagnostic and screening methods. Methods using time-resolved fluorescence resonance energy transfer techniques have been developed and applied to HTS.…”

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confidence: 99%

A New Simple Cell-Based Homogeneous Time-Resolved Fluorescence QRET Technique for Receptor-Ligand Interaction Screening

Härmä¹,

Rozwandowicz-Jansen²,

Martikkala³

et al. 2009

SLAS Discovery

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In this article, a single-label separation-free fluorescence technique is presented as a potential screening method for cell-based receptor antagonists and agonists.The time-resolved fluorescence technique, quenching resonance energy transfer (QRET), relies on a single-labeled binding partner in combination with a soluble quencher. The quencher efficiently suppresses the luminescence of the unbound labeled ligand, whereas the luminescence of the bound fraction is not affected. This approach allows the development of cell-based screening assays in a simple and cost-effective manner. The authors have applied the technique to the screening of β 2 -adrenoreceptor (β 2 AR) antagonists and agonists in intact human embryonic kidney HEK293 i cells overexpressing human β 2 -adrenergic receptors. Two antagonists (propranolol, alprenolol) and 2 agonists (metaproterenol, terbutaline) for β 2 AR were investigated in a displacement assay using europium(III)-labeled pindolol ligand. The assay Z′ values ranged from 0.68 to 0.78, the coefficient of variation was less than 10%, and the K i values were 19 nM for propranolol and alprenolol and 14 and 5.9 µM for metaproterenol and terbutaline, respectively. The QRET technique with β 2 AR was also applied to LOPAC compound library screening, yielding nearly error-free recognition of known binders. This simple and cost-effective technique can be readily adapted to laboratory and industrial-scale screening. (Journal of Biomolecular Screening 2009:936-943)

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“…Time-resolved fluorescence (19): This involves a specialized pulsed laser and data management system to make use of a fluorochrome with a longer lifetime than that from the background particles. Up-conversion fluorescence (20): Using up-converting phosphors that provide an almost immediate visible response when excited by infrared light.…”

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confidence: 99%

Fluorescence resonance energy transfer (FRET)‐based specific labeling of Cryptosporidium oocysts for detection in environmental samples

Chung

Vesey²,

Gauci³

et al. 2004

Cytometry Pt A

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Background: Accurate detection and quantification of Cryptosporidium oocysts in water are a challenge to the water industry. This article demonstrates a way to fluorescently label Cryptosporidium oocysts, based on fluorescence resonance energy transfer (FRET). Labeled oocysts can then be applied to environmental waters and their movement followed by flow cytometric detection and enumeration of the FRET-labeled oocysts, as demonstrated here with environmental water samples. Methods: Cryptosporidium oocysts were labeled with three fluorochromes, FITC, Texas red, and Cy7, that through FRET yielded a Stokes shift of ϳ272 nm with excitation from a standard argon laser emitting at 488 nm. Defined flow cytometric settings and gatings were used to select FITC/green (530-nm), Texas red/red (650-nm), and Cy7/infrared (780-nm) fluorescing particles with light

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Time-Resolved Fluorescence of Lanthanide Probes and Applications in Biotechnology

Cited by 91 publications

References 121 publications

Simultaneous Quantification of Human Glandular Kallikrein 2 and Prostate-Specific Antigen mRNAs in Peripheral Blood from Prostate Cancer Patients

Simultaneous Quantification of Human Glandular Kallikrein 2 and Prostate-Specific Antigen mRNAs in Peripheral Blood from Prostate Cancer Patients

A New Simple Cell-Based Homogeneous Time-Resolved Fluorescence QRET Technique for Receptor-Ligand Interaction Screening

Fluorescence resonance energy transfer (FRET)‐based specific labeling of Cryptosporidium oocysts for detection in environmental samples

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