Reaction-based phosphorescent nanosensor for ratiometric and time-resolved luminescence imaging of fluoride in live cells

2015 ◽  
Vol 51 (64) ◽  
pp. 12839-12842 ◽  
Author(s):  
Shujuan Liu ◽  
Jie Zhang ◽  
Danfeng Shen ◽  
Hua Liang ◽  
Xiangmei Liu ◽  
...  
Keyword(s):  
Live Cells ◽  
Time Resolved ◽  
Luminescence Imaging

A two-channel phosphorescent nanosensor has been designed for use in ratiometric and time-resolved luminescence imaging of intracellular F−.

scholarly journals KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements

2021 ◽  
Author(s):  
Diana Spiegelberg ◽  
Jonas Stenberg ◽  
Pascale Richalet ◽  
Marc Vanhove
Keyword(s):  
Flow Cytometry ◽  
Live Cells ◽  
Time Resolved ◽  
Surface Receptors ◽  
Full Characterization ◽  
End Point ◽  
Titration Curves ◽  
Kinetics Of

AbstractDesign of next-generation therapeutics comes with new challenges and emulates technology and methods to meet them. Characterizing the binding of either natural ligands or therapeutic proteins to cell-surface receptors, for which relevant recombinant versions may not exist, represents one of these challenges. Here we report the characterization of the interaction of five different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate target receptors using LigandTracer. The method offers the advantage of being performed on live cells, alleviating the need for a recombinant source of the receptor. Furthermore, time-resolved measurements, in addition to allowing the determination of the affinity of the studied drug to its target, give access to the binding kinetics thereby providing a full characterization of the system. In this study, we also compared time-resolved LigandTracer data with end-point KD determination from flow cytometry experiments and hypothesize that discrepancies between these two approaches, when they exist, generally come from flow cytometry titration curves being acquired prior to full equilibration of the system. Our data, however, show that knowledge of the kinetics of the interaction allows to reconcile the data obtained by flow cytometry and LigandTracer and demonstrate the complementarity of these two methods.

scholarly journals 3D Molecular Tracking in Live Cells with Simultaneous Time-Resolved Spectroscopy

2011 ◽  
Vol 100 (3) ◽  
pp. 475a
Author(s):  
James Werner ◽  
Peter Goodwin ◽  
Elizabeth Phipps ◽  
Patrick Cutler ◽  
Diane Lidke ◽  
...  
Keyword(s):  
Live Cells ◽  
Time Resolved ◽  
Time Resolved Spectroscopy ◽  
Molecular Tracking

Imaging the Redox States of Live Cells with the Time-Resolved Fluorescence of Genetically Encoded Biosensors

2019 ◽  
Vol 91 (6) ◽  
pp. 3869-3876 ◽  
Author(s):  
Lei Li ◽  
Changcheng Zhang ◽  
Peng Wang ◽  
Aoxue Wang ◽  
Jiasheng Zhou ◽  
...  
Keyword(s):  
Live Cells ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Redox States

Development of organelle-targetable europium complex probes for time-gated luminescence imaging of hypochlorous acid in live cells and animals

2017 ◽  
Vol 140 ◽  
pp. 407-416 ◽  
Author(s):  
Hua Ma ◽  
Bo Song ◽  
Yuanxiu Wang ◽  
Chaolong Liu ◽  
Xin Wang ◽  
...  
Keyword(s):  
Hypochlorous Acid ◽  
Europium Complex ◽  
Live Cells ◽  
Luminescence Imaging

scholarly journals Drug Derived Fluorescent Probes for the Specific Visualization of Cannabinoid Type 2 Receptor - A Toolbox Approach

2019 ◽  
Author(s):  
Gazzi, Thais ◽  
Benjamin Brennecke ◽  
Kenneth Atz ◽  
Claudia Korn ◽  
David Sykes ◽  
...  
Keyword(s):  
Fluorescent Probes ◽  
Tissue Injury ◽  
Expression Profiles ◽  
Tissue Expression ◽  
Live Cells ◽  
Time Resolved ◽  
Binding Data ◽  
Time Resolved Imaging ◽  
Human And Mouse

Cannabinoid type 2 receptor (CB<sub>2</sub>R) is a fundamental part of the endocannabinoid signaling system (eCB system), and is known to play an important role in tissue injury, inflammation, cancer and pain. In stark contrast to its significance, the underlying signaling mechanisms and tissue expression profiles are poorly understood. Due to its low expression in healthy tissue and lack of reliable chemical tools, CB<sub>2</sub>R visualization in live cells remains uncharted. Here we report the development of a drug derived toolbox of highly potent, CB<sub>2</sub>R-selective fluorescent probes based on reverse design. Extensive validation in several applications such as CB<sub>2</sub>R detection in flow cytometry and time-resolved imaging, and the development of a novel fluorescent-based TR-FRET assay to generate kinetic and equilibrium binding data demonstrate the high versatility of our toolbox. These probes are the first to preserve affinity and efficacy in both human and mouse CB<sub>2</sub>R, a crucial aspect for preclinical translatability, and to enable imaging of CB2R internalization in living cells using confocal microscopy.

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