Long-Lifetime Luminescent Europium(III) Complex as an Acceptor in an Upconversion Resonance Energy Transfer Based Homogeneous Assay

2015 ◽  
Vol 88 (1) ◽  
pp. 653-658 ◽  
Author(s):  
Satu Lahtinen ◽  
Qi Wang ◽  
Tero Soukka
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Homogeneous Assay ◽  
Long Lifetime

scholarly journals Enzyme Inhibitor Screening Using a Homogeneous Proximity-Based Immunoassay for Estradiol

2005 ◽  
Vol 10 (4) ◽  
pp. 348-354 ◽  
Author(s):  
Leena Kokko ◽  
Nina Johansson ◽  
Timo Lövgren ◽  
Tero Soukka
Keyword(s):  
Energy Transfer ◽  
High Performance ◽  
Near Infrared ◽  
Enzyme Inhibitor ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Enzyme Reaction ◽  
Hplc Method ◽  
Time Resolved ◽  
Homogeneous Assay

The authors have previously reported a homogeneous time-resolved fluorescence proximity immunoassay for estradiol. The assay was based on luminescence resonance energy transfer between a long lifetime fluorescent europium(III) chelate-dyed nanoparticle donor and a short lifetime, near-infrared fluorescent acceptor. The energy transfer prolonged the lifetime of the sensitized acceptor emission, and the fluorescence of the acceptor was measured using a time-resolved detection. The developed immunoassay was employed to screen inhibitors for enzyme 17β-hydroxysteroid dehydrogenase type 1. The enzyme overexpressed in MCF-7 cells catalyzed a reversible conversion of estroneto17β-estradiol. The inhibition efficiency of the tested molecule was obtained by comparing the final concentration of converted estradiol after 60 min of conversion reaction in a sample and in a conversion control not containing an inhibitor. The Zβ factor calculated using the E2 concentrations of the homogeneous assay was 0.64, demonstrating a relatively good performance of the assay. The results from the homogeneous assay were comparable with the results obtained using radioactively labeled estrone as a substrate and high-performance liquid chromatography (HPLC) separation of estrone and converted estradiol after the enzyme reaction. Thus, this homogeneous assay can simplify the primary screening of potential new drug molecules by replacing a tedious radiometric HPLC method.

Luminescence Energy Transfer–Based Screening and Target Engagement Approaches for Chemical Biology and Drug Discovery

2021 ◽  
pp. 247255522110360
Author(s):  
Eun Jeong Cho ◽  
Kevin N. Dalby
Keyword(s):  
Energy Transfer ◽  
Drug Discovery ◽  
Early Stage ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Live Cells ◽  
Molecular Binding ◽  
Compound Screening ◽  
Homogeneous Assay

Luminescence is characterized by the spontaneous emission of light resulting from either chemical or biological reactions. Because of their high sensitivity, reduced background interference, and applicability to numerous situations, luminescence-based assay strategies play an essential role in early-stage drug discovery. Newer developments in luminescence-based technologies have dramatically affected the ability of researchers to investigate molecular binding events. At the forefront of these developments are the nano bioluminescence resonance energy transfer (NanoBRET) and amplified luminescent proximity homogeneous assay (Alpha) technologies. These technologies have opened up numerous possibilities for analyzing the molecular biophysical properties of complexes in environments such as cell lysates. Moreover, NanoBRET enables the validation and quantitation of the interactions between therapeutic targets and small molecules in live cells, representing an essential benchmark for preclinical drug discovery. Both techniques involve proximity-based luminescence energy transfer, in which excited-state energy is transferred from a donor to an acceptor, where the efficiency of transfer depends on proximity. Both approaches can be applied to high-throughput compound screening in biological samples, with the NanoBRET assay providing opportunities for live-cell screening. Representative applications of both technologies for assessing physical interactions and associated challenges are discussed.

Homogeneous assay for biotin based on Aequorea victoria bioluminescence resonance energy transfer system

2003 ◽  
Vol 313 (1) ◽  
pp. 68-75 ◽  
Author(s):  
Andrey Yu Gorokhovatsky ◽  
Natalia V Rudenko ◽  
Victor V Marchenkov ◽  
Vitaly S Skosyrev ◽  
Maxim A Arzhanov ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Transfer System ◽  
Bioluminescence Resonance Energy Transfer ◽  
Aequorea Victoria ◽  
Homogeneous Assay

Homogeneous assay of target molecules based on chemiluminescence resonance energy transfer (CRET) using DNAzyme-linked aptamers

2014 ◽  
Vol 58 ◽  
pp. 308-313 ◽  
Author(s):  
Hyoyoung Mun ◽  
Eun-Jung Jo ◽  
Taihua Li ◽  
Hyou-Arm Joung ◽  
Dong-Gu Hong ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Homogeneous Assay ◽  
Target Molecules

scholarly journals A 15-min non-competitive homogeneous assay for microcystin and nodularin based on time-resolved Förster resonance energy transfer (TR-FRET)

2021 ◽  
Author(s):  
Sultana Akter ◽  
Urpo Lamminmäki
Keyword(s):  
Energy Transfer ◽  
Water Samples ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
World Health ◽  
Förster Resonance Energy Transfer ◽  
Time Resolved ◽  
Homogeneous Assay ◽  
Förster Resonance

AbstractSimple and rapid methods are required for screening and analysis of water samples to detect cyanobacterial cyclic peptide hepatotoxins: microcystin/nodularin. Previously, we reported a highly sensitive non-competitive heterogeneous assay for microcystin/nodularin utilizing a generic anti-immunocomplex (anti-IC) single-chain fragment of antibody variable domains (scFv) isolated from a synthetic antibody library together with a generic adda ((2S,3S,4E,6E,8S,9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid)-specific monoclonal antibody (Mab) recognizing the common adda part of the microcystin/nodularin. Using the same antibody pair, here we report a homogeneous non-competitive assay for microcystin/nodularin based on TR-FRET (time-resolved Förster resonance energy transfer) measurement. The anti-IC scFv labeled with Alexa Fluor 680 and the Mab labeled with europium enabled the FRET process to occur in the presence of microcystin/nodularin. The TR-FRET signal is proportional to the toxin concentration in the sample. The rapid (15 min) homogeneous assay without requiring any washing step detected all the tested nine toxin variants (microcystin-LR, -dmLR, -RR, -dmRR, -YR, -LY, -LF -LW, and nodularin-R). Very good signal to blank ratio (~13) was achieved using microcystin-LR and the sample detection limit (blank+3SD of blank) for microcystin-LR was ~0.3 μg/L (~0.08 μg/L in 80-μL reaction well). The practical application of the TR-FRET assay was demonstrated with water samples spiked with microcystin-LR as well as with environmental water. The average recoveries of microcystin-LR from spiked water ranged from 65 to 123%. Good correlation (r2 = 0.73 to 0.99) with other methods (liquid chromatography-mass spectrometry and previously reported heterogeneous assay) was found when environmental samples were analyzed. The developed wash-free assay has the potential to play as a quick screening tool to detect microcystin/nodularin from water below the World Health Organization’s guideline limit (1 μg/L of microcystin-LR). Graphical abstract

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