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HTRF: A Technology Tailored for Drug Discovery - A Review of Theoretical Aspects and Recent Applications
Open accessAbstract
HTRF (Homogeneous Time Resolved Fluorescence) is the most frequently used generic assay technology to measure analytes in a homogenous format, which is the ideal platform used for drug target studies in high-throughput screening (HTS). This technology combines fluorescence resonance energy transfer technology (FRET) with time-resolved measurement (TR). In TR-FRET assays, a signal is generated through fluorescent resonance energy transfer between a donor and an acceptor molecule when in close proximity to each other. Buffer and media interference is dramatically reduced by dual-wavelength detection, and the final signal is proportional to the extent of product formation. The HTRF assay is usually sensitive and robust that can be miniaturized into the 384 and 1536-well plate formats. This assay technology has been applied to many antibody-based assays including GPCR signaling (cAMP and IP-One), kinases, cytokines and biomarkers, bioprocess (antibody and protein production), as well as the assays for protein-protein, proteinpeptide, and protein-DNA/RNA interactions.Since its introduction to the drug-screening world over ten years ago, researchers have used HTRF to expedite the study of GPCRs, kinases, new biomarkers, protein-protein interactions, and other targets of interest. HTRF has also been utilized as an alternative method for bioprocess monitoring. The first-generation HTRF technology, which uses Europium cryptate as a fluorescence donor to monitor reactions between biomolecules, was extended in 2008 through the introduction of a second-generation donor, Terbium cryptate (Tb), enhancing screening performance. Terbium cryptate possesses different photophysical properties compared to Europium, including increased quantum yield and a higher molar extinction coefficient. In addition to being compatible with the same acceptor fluorophors used with Europium, it can serve as a donor fluorophore to green-emitting fluors because it has multiple emission peaks including one at 490 nm. Moreover, all Terbium HTRF assays can be read on the same HTRF-compatible instruments as Europium HTRF assays.Overall, HTRF is a highly sensitive, robust technology for the detection of molecular interactions in vitro and is widely used for primary and secondary screening phases of drug development. This review addresses the general principles of HTRF and its current applications in drug discovery.
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Citations
Cites
Blume-Jensen, P., & Hunter, T. (2001) · Nature · doi:10.1038/35077225 · Cited by 3,738
PROTEIN SERINE/THREONINE KINASES
Edelman, A. M., Blumenthal, D., & Krebs, E. G. (1987) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.56.070187.003031 · Cited by 1,435
SIGNALLING BY RECEPTOR TYROSINE KINASES
Fantl, W. J., Johnson, D. E., & Williams, L. T. (1993) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.62.1.453 · Cited by 1,118
Ciruela, F., Casadó, V., Rodrigues, R. J., Luján, R., Burgueño, J., Canals, M., Borycz, J., Rebola, N., Goldberg, S. R., Mallol, J., et al. (2006) · Journal of Neuroscience · doi:10.1523/jneurosci.3574-05.2006 · Cited by 591
Principles and Biophysical Applications of Lanthanide-Based Probes
Selvin, P. R. (2002) · Annual Review of Biophysics and Biomolecular Structure · doi:10.1146/annurev.biophys.31.101101.140927 · Cited by 549
Maurel, D., Comps‐Agrar, L., Brock, C., Rives, M., Bourrier, E., Ayoub, M. A., Bazin, H., Tinel, N., Durroux, T., Prézeau, L., et al. (2008) · Nature Methods · doi:10.1038/nmeth.1213 · Cited by 500
Rare earth cryptates and homogeneous fluoroimmunoassays with human sera
Mathis, G. (1993) · Clinical Chemistry · doi:10.1093/clinchem/39.9.1953 · Cited by 393
Mathis, G. (1995) · Clinical Chemistry · doi:10.1093/clinchem/41.9.1391 · Cited by 339
Levoye, A., Dam, J., Ayoub, M. A., Guillaume, J., Couturier, C., Delagrange, P., & Jockers, R. (2006) · The EMBO Journal · doi:10.1038/sj.emboj.7601193 · Cited by 335
Dopamine D2 receptors form higher order oligomers at physiological expression levels
Guo, W. X., Urizar, E., Králíková, M., Mobarec, J. C., Shi, L., Filizola, M., & Javitch, J. A. (2008) · The EMBO Journal · doi:10.1038/emboj.2008.153 · Cited by 334
Flagged first, then by citations
Research works printed but not linked: 1
- [23]Amoravain M Lyotard S Jaga D Lebreton ML Servent F Bomer U Introduction of a new HTRF acceptor, d2: Development of a complete GPCR platform for a Gs, Gi and Gq screening SBS 2005 11th Annual Conference Geneva
Cited by
Sorted by citations
Items 401–500 of 522
Jaengwang, K., Pommayon, S., Sonklin, C., Choowongkomon, K., Waraho‐Zhmayev, D., & Tabtimmai, L. (2025) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/j.bbagen.2025.130854 · Cited by 1
Rêgo, M., Silva, L., Medeiros, J., Figueiredo, R., Alves, S., & Beltrão, E. (2013) · Biotechnic & Histochemistry · doi:10.3109/10520295.2013.846479 · Cited by 1
Hyper sensitive protein detection by Tandem-HTRF reveals Cyclin D1 dynamics in adult mouse
Zampieri, A., Champagne, J., Auzemery, B., Fuentes, I., Maurel, B., & Bienvenu, F. (2015) · Scientific Reports · doi:10.1038/srep15739 · Cited by 2
Schrade, K., & Klußmann, E. (2017) · Cardiac and vascular biology · doi:10.1007/978-3-319-54579-0_4 · Cited by 1
Fu, Y., Xie, Z., Zhao, P., Lv, S., & Chen, S. (2020) · Planta Medica · doi:10.1055/a-1272-3997 · Cited by 1
Evaluation of Homogeneous Proximity Immunoassays for Preclinical Bioanalysis
Eangoor, P., Das, S., & Pucci, V. (2020) · Bioanalysis · doi:10.4155/bio-2020-0258 · Cited by 1
Drug discovery processes: When and where the rubber meets the road
Miller, S. M., Moos, W. H., Munk, B. H., Munk, S. A., Hart, C. P., & Spellmeyer, D. C. (2023) · Elsevier eBooks · doi:10.1016/b978-0-12-824304-6.00025-0 · Cited by 1
Garnovskaya, M. N., Feshuk, M., Stewart, W. M., Friedman, K. P., Thomas, R. S., & Deisenroth, C. (2023) · Toxicology in Vitro · doi:10.1016/j.tiv.2023.105659 · Cited by 1
Photophysical Properties and Applications of Lanthanoid Helicates
Bünzli, J. G. (2013) · Wiley eBooks · doi:10.1002/9781118517413.ch6 · Cited by 1
Novel Modulation of Adenylyl Cyclase Type 2
Conley, J. M. (2013) · Purdue e-Pubs (Purdue University System) · Cited by 1
Biochemical assays and biophysical tools for inhibitor identification and validation
Fowkes, A., Bertrand, H. C., & Wells, G. (2013) · doi:10.4155/ebo.13.164 · Cited by 1
Branon, T. C. (2025) · Wiley eBooks · doi:10.1002/9781394236145.ch16 · Cited by 1
High‐Throughput Screening of Marine Resources
Hochard, A., Reininger, L., Ruchaud, S., & Bach, S. (2014) · Wiley eBooks · doi:10.1002/9783527681501.ch23
Baldo, B. (2011) · edoc (University of Basel) · doi:10.5451/unibas-005705769
High‐Throughput and ‐Content Screening/Screening for New Pharmaceutics
Tannert, A., & Schaefer, M. (2012) · Wiley eBooks · doi:10.1002/9783527643981.bphot090
The function of TCF4 in Pitt-Hopkins syndrome and schizophrenia
Forrest, M. P. (2013) · ORCA Online Research @Cardiff (Cardiff University)
Rau, S. (2014) · Publications of the UdS (Saarland University) · doi:10.22028/d291-22988
Luminescence-based assay methods in drug discovery
Huttunen, R. J. (2011) · UTUPub (University of Turku)
Zhou, H. (2014) · Digital Repository Service (Northeastern University) · doi:10.17760/d20004824
Herlitz, J. D. (1991) · PubMed
Jeppsson, F. (2016) · KTH Publication Database DiVA (KTH Royal Institute of Technology)
Seneci, P. (2018) · Elsevier eBooks · doi:10.1016/b978-0-08-099420-8.00003-1
Saha, B. B., Thu, K., & Wijayanta, A. T. (2018) · Evergreen · doi:10.5109/1957499
Nomura, S., Taichi, M., & Tanaka, K. (2021) · Springer eBooks · doi:10.1007/978-981-16-2458-2_4
Furger, C. (2016) · Wiley eBooks · doi:10.1002/9781119330141.biblio
Faculty Opinions recommendation of Crystal structure of the human oxytocin receptor.
Freidinger, R. M. (2020) · Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature · doi:10.3410/f.738337689.793579103
Analytical Measurement Systems
Thurow, K., & Junginger, S. (2022) · Wiley eBooks · doi:10.1002/9783527829446.ch9
Old tricks and new trends to address tumors: a medicinal chemist’s perspective
Seneci, P. (2023) · Istituto Lombardo - Accademia di Scienze e Lettere - Rendiconti di Scienze · doi:10.4081/scienze.2022.826
Nachweismethoden zur Protein-Interaktion
Pusterla, T., & Volz, A. (2023) · Springer eBooks · doi:10.1007/978-3-662-62671-9_14
Facen, E., Assoni, G., Donati, G., Paladino, D., Carreira, Á. S. A., Pietra, V. L., Lotti, R., Marinelli, L., Arosio, D., Seneci, P., et al. (2023) · SSRN Electronic Journal · doi:10.2139/ssrn.4597592
Rational drug design targeting G-protein-coupled receptors: ligand search and screening (review)
Luginina, A. P., Khnykin, A. N., Khorn, P. A., Moiseeva, O. V., Safronova, N. A., Поспелов, В. А., Dashevskii, D. E., Belousov, A. S., Borshchevskiy, I. V., & Mishin, A. (2024) · Biochemistry (Moscow) · doi:10.31857/s0320972524050158
Clerigué, J., Pascual-Guerra, J., Leon, R. M., & Menéndez, J. C. (2024) · Neuromethods · doi:10.1007/978-1-0716-4232-0_5
Patent highlights April–May 2024
Mucke, H. A. (2024) · Pharmaceutical Patent Analyst · doi:10.1080/20468954.2025.2459593
Advanced Protein Modeling Method: Benchmarking and Applications in Computer-Aided Drug Discovery
Phatak, S. S. (2012) · Digital Commons-TMC (Texas Medical Center)
Kapur, B., Martin, E., Bondar, A., Schuler, M. J., Schmidt, E., Nar, H., & Schnapp, G. (2025) · ACS Pharmacology & Translational Science · doi:10.1021/acsptsci.5c00516
Wier, S. P. V., Goddard, Z. R., Lei, X., Hyde, E., Picard, D. J., Hobson, E., Cominetti, M. M. D., O'Connell, M. A., Searcey, M., Remke, M., et al. (2025) · ChemRxiv · doi:10.26434/chemrxiv-2024-lf2kt-v3
Accelerating the discovery of MET inhibitors powered by high-throughput hit identification
Zhang, H., Zhang, L., & Liu, C. (2025) · Bioanalysis · doi:10.1080/17576180.2025.2601855
Ben-Yehuda, H., Arad, M., Ramos, J. M. P., Sharon, E., Castellani, G., Ferrera, S., Cahalon, L., Colaiuta, S. P., Salame, T.-M., & Schwartz, M. (2021) · Figshare · doi:10.6084/m9.figshare.14850601
Ben-Yehuda, H., Arad, M., Ramos, J. M. P., Sharon, E., Castellani, G., Ferrera, S., Cahalon, L., Colaiuta, S. P., Salame, T.-M., & Schwartz, M. (2021) · Figshare · doi:10.6084/m9.figshare.14850595
Ben-Yehuda, H., Arad, M., Ramos, J. M. P., Sharon, E., Castellani, G., Ferrera, S., Cahalon, L., Colaiuta, S. P., Salame, T.-M., & Schwartz, M. (2021) · Figshare · doi:10.6084/m9.figshare.14850598
Ben-Yehuda, H., Arad, M., Ramos, J. M. P., Sharon, E., Castellani, G., Ferrera, S., Cahalon, L., Colaiuta, S. P., Salame, T.-M., & Schwartz, M. (2021) · Figshare · doi:10.6084/m9.figshare.14850592
Ben-Yehuda, H., Arad, M., Ramos, J. M. P., Sharon, E., Castellani, G., Ferrera, S., Cahalon, L., Colaiuta, S. P., Salame, T.-M., & Schwartz, M. (2021) · Figshare · doi:10.6084/m9.figshare.14850589
Ben-Yehuda, H., Arad, M., Ramos, J. M. P., Sharon, E., Castellani, G., Ferrera, S., Cahalon, L., Colaiuta, S. P., Salame, T.-M., & Schwartz, M. (2021) · Figshare · doi:10.6084/m9.figshare.14850604
NN-01-195, a Novel Conjugate of HSP90 and AURKA Inhibitors, Effectively Targets Solid Tumors
Nguyen, T. T., Nandwana, N. K., Srikanth, Y. V. V., Bagnyukova, T., Chkhalo, O., Cai, K. Q., Lamperelli, J., Pirestani, S., Mehra, M. K., Burtness, B. A., et al. (2026) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-25-0857
G protein-coupled receptor pharmacology: Tools, techniques and assays for drug discovery
Gonzalez, R. D. L. F., Ranasinghe, I., & Langmead, C. J. (2026) · Elsevier eBooks · doi:10.1016/b978-0-443-29808-0.00029-7
Kamel, E. M., Allam, A. A., Rudayni, H. A., Alkhedhairi, S., Ahmed, N. A., Alkhayl, F. F. A., & Lamsabhi, A. M. (2026) · Epigenetics & Chromatin · doi:10.1186/s13072-026-00663-4
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Mach, L., & Nazaré, M. (2026) · CHIMIA International Journal for Chemistry · doi:10.2533/chimia.2026.165
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Hajj, M. (2012) · theses.fr (ABES) · doi:10.70675/e75ab04fzf6ddz4c25zb55dz5eefb493d85c
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Milhas, S. (2016) · theses.fr (ABES) · doi:10.70675/7dcf6f45z231cz483fzb8f5z5cd27c8b0499
Martin, B. (2017) · theses.fr (ABES) · doi:10.70675/bdbbffa0za5a7z46e6z9494z59bdacd89ba5
Les méthyltransférases de la coiffe du MERS-CoV : étude fonctionnelle et recherches d'inhibiteurs
Aouadi, W. (2017) · theses.fr (ABES) · doi:10.70675/8e1435e3zc63az4f17z8e10zc76c0a7a533b
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Heuninck, J. (2019) · theses.fr (ABES) · doi:10.70675/e4fae036z07a1z4ab5z9730z70b3c09c88d4
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Patel, K. B., Akinwumi, I. A., Weber, T. M., MacDonald, M. R., Collins, S. J., Chitnis, S. P., Zack, C. L., Ogboo, B. C., Nasief, N. N., Ouellette, E. A., et al. (2026) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.6c02714
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Galbiati, A., Colasurdo, E., Patsilinakos, A., Lanfredini, S., Bonifati, S., Massignan, T., Varricchio, C., Bencheva, L., Boldrini, A., Terruzzi, L., et al. (2026) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.64898/2026.10.22.760876
Metadata
- Authors
- Degorce, F.
- Venue
- Current Chemical Genomics
- Publisher
- Bentham Science Publishers Ltd.
- Published
- 2009-03-17
- Volume
- 3
- Issue
- 1
- Pages
- 22-32
- ISSN
- 1875-3973 (ISSN-L)
- PMC
- PMC2802762
- OpenAlex
- W2073113196
- Cited by
- 501
- References
- 74
- Citations by year
- 2026: 29 · 2025: 31 · 2024: 35 · 2023: 37 · 2022: 31 · 2021: 53 · 2020: 35 · 2019: 29 · 2018: 34 · 2017: 28 · 2016: 23 · 2015: 25 · 2014: 39 · 2013: 28 · 2012: 26
- FWCI
- 5.90
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- cc-by
- Locations
- Current Chemical Genomics (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Keywords
- homogeneous time-resolved fluorescence, time-resolved fluorescence resonance energy transfer, drug discovery, high-throughput screening, fluorescence resonance energy transfer, europium cryptate, G protein-coupled receptor signaling, kinase activity assay, protein-protein interaction, bioprocess monitoring, antibody-based assays, biomarkers
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