Label-Free Microscale Thermophoresis Discriminates Sites and Affinity of Protein–Ligand Binding

Look, no label! Microscale thermophoresis makes use of the intrinsic fluorescence of proteins to quantify the binding affinities of ligands and discriminate between binding sites. This method is suitable for studying binding interactions of very small amounts of protein in solution. The binding of ligands to iGluR membrane receptors, small-molecule inhibitorss to kinase p38, aptamers to thrombin, and Ca(2+) ions to synaptotagmin was quantified.

• Publication year

  2012

• Venue

  Angewandte Chemie

• Publication date

  2012-09-24

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1002/anie.201204268PMID 23001866PMCID 3588113
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Direct detection of antibody concentration and affinity in human serum using microscale thermophoresis.

  2012cited by this paper
• Studies of the binding mechanism between aptamers and thrombin by circular dichroism, surface plasmon resonance and isothermal titration calorimetry.

  2011cited by this paper
• Label-free quantification of membrane-ligand interactions using backscattering interferometry

  2011cited by this paper
• Kinetic Capillary Electrophoresis with Mass‐Spectrometry Detection (KCE‐MS) Facilitates Label‐Free Solution‐Based Kinetic Analysis of Protein–Small Molecule Binding

  2011cited by this paper
• Survey of the year 2009: applications of isothermal titration calorimetry

  2011cited by this paper
• Protein-binding assays in biological liquids using microscale thermophoresis.

  2010cited by this paper
• Pharmacology of ionotropic glutamate receptors: A structural perspective.

  2010cited by this paper
• Optical thermophoresis for quantifying the buffer dependence of aptamer binding.

  2010cited by this paper
• Quantifizierung der Puffer‐Abhängigkeit von Aptamer‐Bindungsreaktionen mit optischer Thermophorese

  2010cited by this paper
• The Ca2+ Affinity of Synaptotagmin 1 Is Markedly Increased by a Specific Interaction of Its C2B Domain with Phosphatidylinositol 4,5-Bisphosphate

  2009cited by this paper
• Dependence of aptamer activity on opposed terminal extensions: improvement of light-regulation efficiency

  2009cited by this paper
• I. Photocontrol of ionotropic glutamate receptors and II. Total synthesis of exiguamine A and B

  2008cited by this paper
• Reversibly caged glutamate: a photochromic agonist of ionotropic glutamate receptors.

  2007cited by this paper
• Free-Solution, Label-Free Molecular Interactions Studied by Back-Scattering Interferometry

  2007cited by this paper
• Probing protein folding and conformational transitions with fluorescence.

  2006cited by this paper
• Glutamate receptors at atomic resolution

  2006cited by this paper
• Why molecules move along a temperature gradient

  2006cited by this paper
• Kinetic studies of small molecule interactions with protein kinases using biosensor technology.

  2005cited by this paper
• p38 MAP kinase inhibitors: many are made, but few are chosen.

  2005cited by this paper
• Tuning activation of the AMPA-sensitive GluR2 ion channel by genetic adjustment of agonist-induced conformational changes

  2003cited by this paper
• Thermophoresis in protein solutions

  2003cited by this paper
• Unfolding and conformational studies on bovine adrenodoxin probed by engineered intrinsic tryptophan fluorescence.

  2002cited by this paper
• SB 239063, a second-generation p38 mitogen-activated protein kinase inhibitor, reduces brain injury and neurological deficits in cerebral focal ischemia.

  2001cited by this paper
• Direct measurement of protein binding energetics by isothermal titration calorimetry.

  2001cited by this paper
• Probing ligand protein binding equilibria with fluorescence fluctuation spectroscopy.

  2000cited by this paper
• Mechanisms for activation and antagonism of an AMPA-sensitive glutamate receptor: crystal structures of the GluR2 ligand binding core.

  2000cited by this paper
• Isothermal titration calorimetry and differential scanning calorimetry as complementary tools to investigate the energetics of biomolecular recognition

  1999cited by this paper
• The glutamate receptor ion channels.

  1999cited by this paper
• The activation state of p38 mitogen-activated protein kinase determines the efficiency of ATP competition for pyridinylimidazole inhibitor binding.

  1998cited by this paper
• Kinetic analysis of macromolecular interactions using surface plasmon resonance biosensors.

  1997cited by this paper
• Kinetic analysis of macromolecular interactions using surface plasmon resonance biosensors.

  1997cited by this paper
• Kinetics of folding of the IgG binding domain of peptostreptococcal protein L.

  1997cited by this paper
• Oligonucleotide inhibitors of human thrombin that bind distinct epitopes.

  1997cited by this paper
• Stable expression of a functional GluR6 homomeric glutamate receptor channel in mammalian cells.

  1994cited by this paper
• Selection of single-stranded DNA molecules that bind and inhibit human thrombin

  1992cited by this paper
• Detection of antigen—antibody interactions by surface plasmon resonance. Application to Epitope Mapping

  1990cited by this paper
• Measurement of protein‐DNA interaction parameters by electrophoresis mobility shift assay

  1989cited by this paper
• Piezoelectric Sorption Detector.

  1964cited by this paper

• Label-Free and Immobilization-Free Protein-Binding Assays by Ultraviolet Transient Absorption Microscopy.

  2026cites this paper
• Functional consequences of genetic variations in DgoR, a GntR/FadR family transcriptional repressor of D-galactonate metabolism in Escherichia coli

  2025cites this paper
• Unprecedented ent-kaurane heterodimers from Icacina oliviformis: Potent anti-Helicobacter pylori leads targeting enoyl-ACP reductase FabI

  2025cites this paper
• Influence of α‑Helical Content on the Thermodiffusion of Apomyoglobin

  2025cites this paper
• Molecular mechanisms underlying allosteric behavior of Escherichia coli DgoR, a GntR/FadR family transcriptional regulator

  2025cites this paper
• Hirudin-Like Noninvasive Polyurethane Solid Anticoagulant Fabricated by One-Pot In Situ Solution Polymerization and Phase Conversion Methods.

  2025cites this paper
• Multivalent 2D- and 3D-nanogels as carbohydrate-lectin binders

  2025cites this paper
• Understanding the physiological role and cross-interaction network of VapBC35 toxin-antitoxin system from Mycobacterium tuberculosis

  2025cites this paper
• Drug discovery through biophysical techniques: Methods and applications.

  2025cites this paper
• Residence time in drug discovery: current insights and future perspectives

  2025cites this paper
• A single chain variable fragment antibody (Tn 64) cognate to fibronectin type III repeats promotes corneal wound healing by inhibiting fibrosis.

  2024cites this paper
• Microscale Thermophoresis Analysis of Chromatin Interactions.

  2024cites this paper
• Biophysical and Biochemical Characterization of Structurally Diverse Small Molecule Hits for KRAS Inhibition

  2024cites this paper
• AI-guided pipeline for protein–protein interaction drug discovery identifies a SARS-CoV-2 inhibitor

  2024cites this paper
• Analyzing aptamer structure and interactions: in silico modelling and instrumental methods

  2024cites this paper
• Analytical Methods for Protein Kinase and Inhibitor Screening including Kinetic Evaluation

  2024cites this paper
• Microscale thermophoresis (MST) and Spectral Shift (SpS) in drug discovery

  2024cites this paper
• A review of biophysical strategies to investigate protein-ligand binding: What have we employed?

  2024cites this paper
• Detection of small molecule compounds bound to proteins

  2024cites this paper
• Label-Free Quantification of DNA Hybridization Kinetics and Affinity Enabled by an Electro-Optical Modulation on Single Nanoparticles.

  2023cites this paper
• Measuring the Interaction of Sterols and Steroids with Proteins by Microscale Thermophoresis.

  2023cites this paper
• Accumulation and Stretching of DNA Molecules in Temperature-Induced Concentration Gradients.

  2023cites this paper
• Discovery of berberine analogs as potent and highly selective p300/CBP HAT inhibitors.

  2023cites this paper
• Biosensor-based active ingredient recognition system for screening TNF-α inhibitors from lotus leaves

  2023cites this paper
• New Insights into the Cooperativity and Dynamics of Dimeric Enzymes.

  2023cites this paper
• AI-guided pipeline for protein-protein interaction drug discovery identifies a SARS-CoV-2 inhibitor

  2023cites this paper
• Thermo-osmotic slip flows around a thermophoretic microparticle characterized by optical trapping of tracers

  2023cites this paper
• Overcoming the Solubility Problem in E. coli: Available Approaches for Recombinant Protein Production.

  2022cites this paper
• Complementary Experimental Methods to Obtain Thermodynamic Parameters of Protein Ligand Systems

  2022cites this paper
• An Optimized Microscale Thermophoresis Method for High-Throughput Screening of DNA Methyltransferase 2 Ligands.

  2022cites this paper
• Combining cysteine scanning with chemical labeling to map protein-protein interactions and infer bound structure in an intrinsically disordered region

  2022cites this paper
• KRAS Inhibitor that Simultaneously Inhibits Nucleotide Exchange Activity and Effector Engagement

  2022cites this paper
• New opportunities and challenges of natural products research: When target identification meets single-cell multiomics

  2022cites this paper
• Peptide adsorption on silica surfaces: Simulation and experimental insights.

  2022cites this paper
• Binding Affinity Quantifications of the Bacteriophage Mu DNA Modification Protein Mom Using Microscale Thermophoresis (MST).

  2022cites this paper
• Microscale thermophoresis as a powerful growing analytical technique for the investigation of biomolecular interaction and the determination of binding parameters

  2022cites this paper
• Thermomicrofluidic Biosensing Systems※

  2022cites this paper
• Functional and Biochemical Characterization of the MazEF6 Toxin-Antitoxin System of Mycobacterium tuberculosis

  2022cites this paper
• The current toolbox for APOBEC drug discovery.

  2022cites this paper
• PSnpBind: a database of mutated binding site protein–ligand complexes constructed using a multithreaded virtual screening workflow

  2022cites this paper
• Label-Free Physical Techniques and Methodologies for Proteins Detection in Microfluidic Biosensor Structures

  2022cites this paper
• Immunodiagnostic Technologies from Laboratory to Point-Of-Care Testing

  2021cites this paper
• Supramolecular Antagonists Promote Mitochondrial Dysfunction.

  2021cites this paper
• Thermomicrofluidics for biosensing applications

  2021cites this paper
• FoldAffinity: binding affinities from nDSF experiments

  2021cites this paper
• Opto-Thermophoretic Manipulation.

  2021cites this paper
• Kinetic Microscale Thermophoresis for Simultaneous Measurement of Binding Affinity and Kinetics

  2021cites this paper
• Drug Affinity Responsive Target Stability (DARTS) Accelerated Small Molecules Target Discovery: Principles and Application.

  2021cites this paper
• Temperature profile characterization with fluorescence lifetime imaging microscopy in a thermophoretic chip

  2021cites this paper
• Multi‐Arm PEG/Peptidomimetic Conjugate Inhibitors of DR6/APP Interaction Block Hematogenous Tumor Cell Extravasation

  2021cites this paper
• eSPC: an online data-analysis platform for molecular biophysics

  2021cites this paper
• Nanosecond Photochemical Reaction (nsPCR) for Enhanced Mass Spectrometric Identification, Quantification, and Visualization of Metabolites and Neuropeptides.

  2021cites this paper
• Assays to Estimate the Binding Affinity of Aptamers.

  2021cites this paper
• A feminizing switch in a hemimetabolous insect

  2021cites this paper
• Novel Method for Quantifying AhR-Ligand Binding Affinities Using Microscale Thermophoresis

  2021cites this paper
• Thermophoretic Micron-Scale Devices: Practical Approach and Review

  2020cites this paper
• Isothermal Analysis of ThermoFluor Data

  2020cites this paper
• Methods for Enhancing Aptamer Affinity for Antigen Detection and Its Characterization

  2020cites this paper
• Target-Directed Approaches for Screening Small Molecules against RNA Targets

  2020cites this paper
• DNA binds to a specific site of the adhesive blood-protein von Willebrand factor guided by electrostatic interactions

  2020cites this paper
• Real-time process quality control of ramulus cinnamomi by critical quality attribute using microscale thermophoresis and on-line NIR.

  2020cites this paper
• Homogeneous, Low-volume, Efficient and Sensitive Quantitation of Circulating Exosomal PD-L1 for Cancer Diagnosis and Immunotherapy Response Prediction.

  2020cites this paper
• In Vitro Analytical Approaches to Study Plant Ligand-Receptor Interactions1[OPEN]

  2020cites this paper
• Thermophoresis: The Case of Streptavidin and Biotin

  2020cites this paper
• Mitigating effects of osmolytes on the interactions between nanoparticles and supported lipid bilayer.

  2020cites this paper
• High-throughput screening for intrinsically disordered proteins by using biophysical methods

  2020cites this paper
• A modular RNA domain that confers differential ligand specificity.

  2020cites this paper
• Necator americanus Ancylostoma secreted protein-2 (Na-ASP-2) selectively binds an ascaroside (ascr#3)

  2020influential citation
• Necator americanus Ancylostoma Secreted Protein-2 (Na-ASP-2) Binds an Ascaroside (ascr#3) in Its Fatty Acid Binding Site

  2020influential citation
• Antibiotic Discovery with Synthetic Fermentation: Library Assembly, Phenotypic Screening, and Mechanism of Action of β-Peptides Targeting Penicillin-Binding Proteins.

  2019cites this paper
• Microscale thermophoresis: warming up to a new biomolecular interaction technique

  2019cites this paper
• Separation of Nano- and Microparticle Flows Using Thermophoresis in Branched Microfluidic Channels

  2019cites this paper
• Unveiling the druggable RNA targets and small molecule therapeutics

  2019cites this paper
• Modern Approaches to Analysis of Protein–Ligand Interactions

  2019cites this paper
• The plasmid‐borne quinolone resistance protein QnrB, a novel DnaA‐binding protein, increases the bacterial mutation rate by triggering DNA replication stress

  2019cites this paper
• Development and characterization of a new inhibitor of heme oxygenase activity for cancer treatment.

  2019cites this paper
• Do interactions between Protein and Phospholipids influence the release behavior from lipid-based exenatide depot systems?

  2019cites this paper
• Emerging strategies for the identification of protein–metabolite interactions

  2019cites this paper
• Synthesis and comparison of linear and hyperbranched multivalent glycosides for C-type lectin binding

  2019cites this paper
• The structural bases for agonist diversity in an Arabidopsis thaliana glutamate receptor-like channel

  2019cites this paper
• Towards time-resolved molecular interaction assays in living bacteria

  2019influential citation
• Nanosecond photochemically promoted click chemistry for enhanced neuropeptide visualization and rapid protein labeling

  2019cites this paper
• MCC1019, a selective inhibitor of the Polo-box domain of Polo-like kinase 1 as novel, potent anticancer candidate

  2019cites this paper
• Analysis of Molecular Interactions Using the Thermophoresis Method and its Applications in Neuroscience and Biological Processes

  2019cites this paper
• Thermophoresis of biological and biocompatible compounds in aqueous solution

  2019cites this paper
• Development and validation of new approaches for quantitative intermolecular interaction analysis by MicroScale Thermophoresis under near-native experimental conditions

  2019cites this paper
• Thermophoretic Manipulation of Micro- and Nanoparticle Flow through a Sudden Contraction in a Microchannel with Near-Infrared Laser Irradiation

  2018cites this paper
• Binding Specificities of Nanobody•Membrane Protein Complexes Obtained from Chemical Cross-Linking and High-Mass MALDI Mass Spectrometry.

  2018cites this paper
• Using two-site binding models to analyze microscale thermophoresis data.

  2018cites this paper
• A new strategy for hydrophobic drug delivery using a hydrophilic polymer equipped with stacking units.

  2018cites this paper
• Fusicoccane-Derived Diterpenoids from Alternaria brassicicola: Investigation of the Structure-Stability Relationship and Discovery of an IKKβ Inhibitor.

  2018cites this paper
• Thermophoretic forces on a mesoscopic scale.

  2018cites this paper
• A simple fluorescent aptamer based assay coupled with fluorescence scanning capillary array for aflatoxin B1.

  2018cites this paper
• On-line coupling between capillary electrophoresis and microscale thermophoresis (CE-MST); the proof-of-concept.

  2018cites this paper
• Small-molecule CaVα1⋅CaVβ antagonist suppresses neuronal voltage-gated calcium-channel trafficking

  2018influential citation
• Salvianolic acid A targeting the transgelin-actin complex to enhance vasoconstriction

  2018cites this paper
• nanoDSF as screening tool for enzyme libraries and biotechnology development

  2018cites this paper
• Prebiotic DNA replication and analyzing a genetic disorder with thermophoresis

  2018cites this paper
• The Toolbox to Study Protein–Protein Interactions in Plants

  2018influential citation
• Analyse der strukturchemischen Grundlagen der Wechselwirkungen zwischen der Phenylalaninhydroxylase und ihrem natürlichen Kofaktor Tetrahydrobiopterin

  2018cites this paper