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179 results on '"Niko Hildebrandt"'

1. A sensitive homogeneous enzyme assay for euchromatic histone-lysine-N-methyltransferase 2 (G9a) based on terbium-to-quantum dot time-resolved FRET

Author

Mohammad Amjadi, Tooba Hallaj, and Niko Hildebrandt

Subjects
enzyme assay, histone-modifying enzymes, epigenetic enzymes, time-resolved fret, inhibitor screening, Medicine (General), R5-920, Biology (General), QH301-705.5
Abstract

Introduction: Histone modifying enzymes include several classes of enzymes that are responsible for various post-translational modifications of histones such as methylation and acetylation. They are important epigenetic factors, which may involve several diseases and so their assay, as well as screening of their inhibitors, are of great importance. Herein, a bioassay based on terbium-to-quantum dot (Tb-to-QD) time-resolved Förster resonance energy transfer (TR-FRET) was developed for monitoring the activity of G9a, the euchromatic histone-lysine N-methyltransferase 2. Overexpression of G9a has been reported in some cancers such as ovarian carcinoma, lung cancer, multiple myeloma and brain cancer. Thus, inhibition of this enzyme is important for therapeutic purposes. Methods: In this assay, a biotinylated peptide was used as a G9a substrate in conjugation with streptavidin-coated ZnS/CdSe QD as FRET acceptor, and an anti-mark antibody labeled with Tb as a donor. Time-resolved fluorescence was used for measuring FRET ratios. Results: We examined three QDs, with emission wavelengths of 605, 655 and 705 nm, as FRET acceptors and investigated FRET efficiency between the Tb complex and each of them. Since the maximum FRET efficiency was obtained for Tb to QD705 (more than 50%), this pair was exploited for designing the enzyme assay. We showed that the method has excellent sensitivity and selectivity for the determination of G9a at concentrations as low as 20 pM. Furthermore, the designed assay was applied for screening of an enzyme inhibitor, S-(5’-Adenosyl)-L-homocysteine (SAH). Conclusion: It was shown that Tb-to-QD FRET is an outstanding platform for developing a homogenous assay for the G9a enzyme and its inhibitors. The obtained results confirmed that this assay was quite sensitive and could be used in the field of inhibitor screening.

Published
2021
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3. Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET

Author

Shashi Bhuckory, K. David Wegner, Xue Qiu, Yu-Tang Wu, Travis L. Jennings, Anne Incamps, and Niko Hildebrandt

Subjects
lanthanides, nanoparticles, fluorescence, biosensing, multiplexing, PSA, Organic chemistry, QD241-441
Abstract

Time-gated Förster resonance energy transfer (TG-FRET) between Tb complexes and luminescent semiconductor quantum dots (QDs) provides highly advantageous photophysical properties for multiplexed biosensing. Multiplexed Tb-to-QD FRET immunoassays possess a large potential for in vitro diagnostics, but their performance is often insufficient for their application under clinical conditions. Here, we developed a homogeneous TG-FRET immunoassay for the quantification of carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and prostate-specific antigen (PSA) from a single serum sample by multiplexed Tb-to-QD FRET. Tb–IgG antibody donor conjugates were combined with compact QD-F(ab’)2 antibody acceptor conjugates with three different QDs emitting at 605, 650, and 705 nm. Upon antibody–antigen–antibody sandwich complex formation, the QD acceptors were sensitized via FRET from Tb, and the FRET ratios of QD and Tb TG luminescence intensities increased specifically with increasing antigen concentrations. Although limits of detection (LoDs: 3.6 ng/mL CEA, 3.5 ng/mL NSE, and 0.3 ng/mL PSA) for the triplexed assay were slightly higher compared to the single-antigen assays, they were still in a clinically relevant concentration range and could be quantified in 50 µL serum samples on a B·R·A·H·M·S KRYPTOR Compact PLUS clinical immunoassay plate reader. The simultaneous quantification of CEA, NSE, and PSA at different concentrations from the same serum sample demonstrated actual multiplexing Tb-to-QD FRET immunoassays and the potential of this technology for translation into clinical diagnostics.

Published
2020
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4. Terbium to Quantum Dot FRET Bioconjugates for Clinical Diagnostics: Influence of Human Plasma on Optical and Assembly Properties

Author

Niko Hildebrandt, Juan B. Blanco-Canosa, Philip E. Dawson, Michael H. Stewart, Kimihiro Susumu, W. Russ Algar, Frank Morgner, Igor L. Medintz, Daniel Geißler, and Stefan Stufler

Subjects
FRET, quantum dots, terbium, luminescence lifetime, blood, plasma, clinical diagnostics, biotin, streptavidin, histidin, immunoassay, Chemical technology, TP1-1185
Abstract

Förster resonance energy transfer (FRET) from luminescent terbium complexes (LTC) as donors to semiconductor quantum dots (QDs) as acceptors allows extraordinary large FRET efficiencies due to the long Förster distances afforded. Moreover, time-gated detection permits an efficient suppression of autofluorescent background leading to sub-picomolar detection limits even within multiplexed detection formats. These characteristics make FRET-systems with LTC and QDs excellent candidates for clinical diagnostics. So far, such proofs of principle for highly sensitive multiplexed biosensing have only been performed under optimized buffer conditions and interactions between real-life clinical media such as human serum or plasma and LTC-QD-FRET-systems have not yet been taken into account. Here we present an extensive spectroscopic analysis of absorption, excitation and emission spectra along with the luminescence decay times of both the single components as well as the assembled FRET-systems in TRIS-buffer, TRIS-buffer with 2% bovine serum albumin, and fresh human plasma. Moreover, we evaluated homogeneous LTC-QD FRET assays in QD conjugates assembled with either the well-known, specific biotin-streptavidin biological interaction or, alternatively, the metal-affinity coordination of histidine to zinc. In the case of conjugates assembled with biotin-streptavidin no significant interference with the optical and binding properties occurs whereas the histidine-zinc system appears to be affected by human plasma.

Published
2011
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5. Evaluating Quantum Dot Performance in Homogeneous FRET Immunoassays for Prostate Specific Antigen

Author

Shashi Bhuckory, Olivier Lefebvre, Xue Qiu, Karl David Wegner, and Niko Hildebrandt

Subjects
Qdots, antibodies, bioconjugation, terbium, FRET, time-gating, homogeneous, immunoassay, serum, TPSA, clinical diagnostics, Chemical technology, TP1-1185
Abstract

The integration of semiconductor quantum dots (QDs) into homogeneous Förster resonance energy transfer (FRET) immunoassay kits for clinical diagnostics can provide significant advantages concerning multiplexing and sensitivity. Here we present a facile and functional QD-antibody conjugation method using three commercially available QDs with different photoluminescence (PL) maxima (605 nm, 655 nm, and 705 nm). The QD-antibody conjugates were successfully applied for FRET immunoassays against prostate specific antigen (PSA) in 50 µL serum samples using Lumi4-Tb (Tb) antibody conjugates as FRET donors and time-gated PL detection on a KRYPTOR clinical plate reader. Förster distance and Tb donor background PL were directly related to the analytical sensitivity for PSA, ...which resulted in the lowest limits of detection for Tb-QD705 (2 ng/mL), followed by Tb-QD655 (4 ng/mL), and Tb-QD605 (23 ng/mL). Duplexed PSA detection using the Tb-QD655 and Tb-QD705 FRET-pairs demonstrated the multiplexing ability of our immunoassays. Our results show that FRET based on QD acceptors is suitable for multiplexed and sensitive biomarker detection in clinical diagnostics.

Published
2016
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8. Plasmonic quenching and enhancement: metal–quantum dot nanohybrids for fluorescence biosensing

Author

Niko Hildebrandt, Mihye Lim, Namjun Kim, Da Yeon Choi, and Jwa-Min Nam

Subjects
Materials Chemistry, Metals and Alloys, Ceramics and Composites, General Chemistry, Catalysis, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials
Abstract

Plasmonic “Quenchancement”: in this feature article, we discuss the interplay of different quenching and enhancement mechanisms that influence the development of metal–quantum dot nanohybrid biosensors.

Published
2023
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9. Nucleic Acid Hybridization Enhanced Luminescence for Rapid and Sensitive RNA and DNA Based Diagnostics

Author

Cui Liu, Xiaoyuan Wei, Huimin Zhang, Mingzhen Zhang, Xue-Feng Yu, Niko Hildebrandt, Qing-Ying Luo, and Zongwen Jin

Subjects
Nucleic Acid Probes, Luminescence, Nucleic Acids, RNA, Nucleic Acid Hybridization, DNA, Phosphates, Analytical Chemistry
Abstract

Long-lived emissive nucleic acid probes are widely used in biochemical analysis due to their programmable structures, high signal-to-background ratio, and high sensitivity. Homogeneous detection based on long-lived emissive nucleic acid probes is often achieved through Förster resonance energy transfer (FRET), which suffers from the limitation of a narrow effective distance range. Herein, a new strategy of accessing nucleic acid hybridization-responsive luminescent probes is presented. The photoluminescence (PL) of a Lumi4-Tb complex internally modified with DNA is switched on by nucleic acid hybridization, after which the PL is increased up to 20 times. PL lifetime analysis revealed a possible mechanism of luminescence enhancement. Due to the flexibility of single-stranded nucleic acid chains, the bases and phosphate groups can coordinate with the Tb(III), which reduces the stability of the Tb complex and results in weak PL. After hybridization, the rigid double helix structure suppresses the coordination between Tb(III) and the bases or phosphate groups, causing luminescence enhancement. As the DNA sequence can be freely designed, an array of probes for different DNA or RNA targets can be created with the same Tb complex. Moreover, the novel probe design can afford pM detection limits of DNA or RNA without any nucleic acid amplification and exhibits great potential for nucleic acid detection in clinical diagnosis.

Published
2022
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12. Guest Editorial Introduction to JSTQE Issue on Nanobiophotonics

Author

James W. Tunnell, Wei-Chuan Shih, Andrea M. Armani, Ilko K. Ilev, Andrew M. Fales, Niko Hildebrandt, and Beop Min Kim

Subjects
Noninvasive imaging, Computer science, Research areas, Tissue imaging, business.industry, Nanotechnology, Technology development, Atomic and Molecular Physics, and Optics, Molecular level, Ultrahigh resolution, Special section, Electrical and Electronic Engineering, Photonics, business
Abstract

The papers in this special section focus on nanobiophotonics. This is an emerging field of modern science and biomedical nanotechnology. This new field continues to vastly expand with state-of-the-art developments across the entire spectrum of biomedical applications ranging from fundamental studies of light-nanobiomaterial interactions to clinical diagnostics and therapeutics. In nanobiophotonics research areas, there has been a great impetus recently for the development of noninvasive imaging and sensing of intracellular structures and cellular functions as well as obtaining quantitative information for light-tissue interactions at the cellular, intracellular, and molecular level. Existing photonics technologies provide unique capabilities for investigating these scientific questions in terms of using non-ionizing optical radiation for providing noninvasive intracellular, cellular, and tissue imaging, and in-vivo sensing with ultrahigh resolution in the subwavelength nanoscale range (smaller than 100 nm). Furthermore, recent technology development efforts leverage the advanced features of photonics to move beyond the diffraction limit, providing sub-nanometer resolution into cellular processes.

Published
2021
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14. When Nanoworlds Collide: Implementing DNA Amplification, Nanoparticles, Molecules, and FRET into a Single MicroRNA Biosensor

Author

Niko Hildebrandt, Xue Qiu, Jingyue Xu, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE12-0030,PhenX,Base moléculaire de l'activation de la recombinaison Xer par les phages(2016)

Subjects
Materials science, [SDV]Life Sciences [q-bio], Nanoparticle, Bioengineering, Nanotechnology, Biosensing Techniques, 02 engineering and technology, Multiplexing, Quantum Dots, Fluorescence Resonance Energy Transfer, [CHIM]Chemical Sciences, Molecule, General Materials Science, ComputingMilieux_MISCELLANEOUS, [PHYS]Physics [physics], Mechanical Engineering, DNA, General Chemistry, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Dna amplification, MicroRNAs, Förster resonance energy transfer, Quantum dot, Rolling circle replication, 0210 nano-technology, Biosensor
Abstract

Isothermal nucleic acid amplification strategies have been combined with nanotechnology for advanced biosensing, material design, and biomedical applications. However, merging phenomena and materials of different nanoscales with the aim of exploiting all their benefits at once has remained a challenging endeavor. Here, we exemplify the various problems one can encounter when combining the nanodimensions of lanthanide complexes (∼2 nm), Förster resonance energy transfer (FRET, ∼5 nm), quantum dots (QDs, ∼20 nm), and rolling circle amplification (RCA, ∼250 nm) into a single microRNA biosensor and how these challenges can be overcome. Six different approaches, including simple FRET-RCA, enzyme-digesting FRET-RCA, and FRET-hyperbranched-RCA were investigated. We demonstrated specific miR-21 detection with 80 fM limit of detection and multiplexing capability with FRET from a Tb complex to different QDs. The detailed view on the various complex multi-nanodimensional assay systems elucidated the limited clinical translation of such sophisticated multicomponent nanobiosensors.

Published
2021
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17. A sensitive homogeneous enzyme assay for euchromatic histone-lysine-N-methyltransferase 2 (G9a) based on terbium-to-quantum dot time-resolved FRET

Author

Niko Hildebrandt, Tooba Hallaj, and Mohammad Amjadi

Subjects
Medicine (General), Epigenetic enzymes, QH301-705.5, Inhibitor screening, Pharmaceutical Science, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, R5-920, Biology (General), Original Research, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, biology, Substrate (chemistry), General Medicine, Enzyme assay, Histone-modifying enzymes, 0104 chemical sciences, 010404 medicinal & biomolecular chemistry, Förster resonance energy transfer, Enzyme, Histone, chemistry, Biochemistry, Time-resolved FRET, Enzyme inhibitor, Acetylation, Biotinylation, biology.protein
Abstract

Introduction: Histone modifying enzymes include several classes of enzymes that are responsible for various post-translational modifications of histones such as methylation and acetylation. They are important epigenetic factors, which may involve several diseases and so their assay, as well as screening of their inhibitors, are of great importance. Herein, a bioassay based on terbium-to-quantum dot (Tb-to-QD) time-resolved Förster resonance energy transfer (TR-FRET) was developed for monitoring the activity of G9a, the euchromatic histone-lysine N-methyltransferase 2. Overexpression of G9a has been reported in some cancers such as ovarian carcinoma, lung cancer, multiple myeloma and brain cancer. Thus, inhibition of this enzyme is important for therapeutic purposes. Methods: In this assay, a biotinylated peptide was used as a G9a substrate in conjugation with streptavidin-coated ZnS/CdSe QD as FRET acceptor, and an anti-mark antibody labeled with Tb as a donor. Time-resolved fluorescence was used for measuring FRET ratios. Results: We examined three QDs, with emission wavelengths of 605, 655 and 705 nm, as FRET acceptors and investigated FRET efficiency between the Tb complex and each of them. Since the maximum FRET efficiency was obtained for Tb to QD705 (more than 50%), this pair was exploited for designing the enzyme assay. We showed that the method has excellent sensitivity and selectivity for the determination of G9a at concentrations as low as 20 pM. Furthermore, the designed assay was applied for screening of an enzyme inhibitor, S-(5’-Adenosyl)-L-homocysteine (SAH). Conclusion: It was shown that Tb-to-QD FRET is an outstanding platform for developing a homogenous assay for the G9a enzyme and its inhibitors. The obtained results confirmed that this assay was quite sensitive and could be used in the field of inhibitor screening.

Published
2020
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18. Rapid and Wash-Free Time-Gated FRET Histamine Assays Using Antibodies and Aptamers

Author

Hui-Jun Fu, Ruifang Su, Lin Luo, Zi-Jian Chen, Thomas Just Sørensen, Niko Hildebrandt, and Zhen-Lin Xu

Subjects
Fluid Flow and Transfer Processes, Immunoassay, food testing, Process Chemistry and Technology, Oligonucleotides, Reproducibility of Results, aptasensors, Bioengineering, Biosensing Techniques, Antibodies, immunoassays, Foodborne Diseases, Förster resonance energy transfer, Fluorescence Resonance Energy Transfer, Animals, fluorescence, Instrumentation, Histamine
Abstract

Histamine (HA) is an indicator of food freshness and quality. However, high concentrations of HA can cause food poisoning. Simple, rapid, sensitive, and specific quantification can enable efficient screening of HA in food and beverages. However, conventional assays are complicated and time-consuming, as they require multiple incubation, washing, and separation steps. Here, we demonstrate that time-gated Förster resonance energy transfer (TG-FRET) between terbium (Tb) complexes and organic dyes can be implemented in both immunosensors and aptasensors for simple HA quantification using a rapid, single-step, mix-and-measure assay format. Both biosensors could quantify HA at concentrations relevant in food poisoning with limits of detection of 0.19 μg/mL and 0.03 μg/mL, respectively. Excellent specificity was documented against the structurally similar food components tryptamine and l-histidine. Direct applicability of the TG-FRET assays was demonstrated by quantifying HA in spiked fish and wine samples with both excellent concentration recovery and agreement with conventional multistep enzyme-linked immunosorbent assays (ELISAs). Our results show that the simplicity and rapidity of TG-FRET assays do not compromise sensitivity, specificity, and reliability, and both immunosensors and aptasensors have a strong potential for their implementation in advanced food safety screening.

Published
2022
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19. Spatial and Temporal Resolution of Luminescence Quenching in Small Upconversion Nanocrystals

Author

Federico Pini, Laura Francés-Soriano, Nicola Peruffo, Antonio Barbon, Niko Hildebrandt, and Marta Maria Natile

Subjects
upconversion, Kinetics, Luminescence, Models, Chemical, energy migration, kinetic modeling, luminescence, nanoparticles, Luminescent Measurements, Solvents, Nanoparticles, General Materials Science, Particle Size, Nanostructures
Abstract

Luminescent upconversion nanocrystals (UCNCs) have become one of the most promising nanomaterials for biosensing, imaging, and theranostics. However, their ultimate translation into robust luminescent probes for daily use in biological and medical laboratories requires comprehension and control of the many possible deactivation pathways that cause upconversion luminescence (UCL) quenching. Here, we demonstrate that thorough modeling of UCL rise and decay kinetics using a freely accessible software can identify the UCL quenching mechanisms in small (40 nm) UCNCs with spatial and temporal resolution. Applied to the most relevant β-NaYF

Published
2022

20. Multiplexed Biosensing and Bioimaging Using Lanthanide-Based Time-Gated Förster Resonance Energy Transfer

Author

Xue Qiu, Jingyue Xu, Marcelina Cardoso Dos Santos, and Niko Hildebrandt

Subjects
Quantum Dots, Fluorescence Resonance Energy Transfer, Metal Nanoparticles, General Medicine, General Chemistry, Biosensing Techniques, Lanthanoid Series Elements, Fluorescent Dyes
Abstract

The necessity to scrutinize more and more biological molecules and interactions both in solution and on the cellular level has led to an increasing demand for sensitive and specific multiplexed diagnostic analysis. Photoluminescence (PL) detection is ideally suited for multiplexed biosensing and bioimaging because it is rapid and sensitive and there is an almost unlimited choice of fluorophores that provide a large versatility of photophysical properties, including PL intensities, spectra, and lifetimes.The most frequently used technique to detect multiple parameters from a single sample is spectral (or color) multiplexing with different fluorophores, such as organic dyes, fluorescent proteins, quantum dots, or lanthanide nanoparticles and complexes. In conventional PL biosensing approaches, each fluorophore requires a distinct detection channel and excitation wavelength. This drawback can be overcome by Förster resonance energy transfer (FRET) from lanthanide donors to other fluorophore acceptors. The lanthanides' multiple and spectrally narrow emission bands over a broad spectral range can overlap with several different acceptors at once, thereby allowing FRET from one donor to multiple acceptors. The lanthanides' extremely long PL lifetimes provide two important features. First, time-gated (TG) detection allows for efficient suppression of background fluorescence from the biological environment or directly excited acceptors. Second, temporal multiplexing, for which the PL lifetimes are adjusted by the interaction with the FRET acceptor, can be used to determine specific biomolecules and/or their conformation

Published
2022

22. Cooperative Luminescence and Cooperative Sensitisation Upconversion of Lanthanide Complexes in Solution

Author

Richard C. Knighton, Niko Hildebrandt, Guillaume Pilet, Carlos Platas-Iglesias, Lohona K. Soro, Loïc J. Charbonnière, Aurora Rodríguez-Rodríguez, Marc Lenertz, and Laura Francés-Soriano

Subjects
Lanthanide, Quenching (fluorescence), Materials science, Quantum yield, General Chemistry, 02 engineering and technology, General Medicine, Photochemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 7. Clean energy, Catalysis, Photon upconversion, 0104 chemical sciences, Upconversion nanoparticles, Excited state, Solar energy conversion, Luminescence, 0210 nano-technology
Abstract

Upconversion nanoparticles have led to various breakthrough applications in solar energy conversion, imaging, and biomedicine. One key impediment is the facilitation of such processes at the molecular scale in solution where quenching effects are much more pronounced. In this work, molecular solution-state cooperative luminescence (CL) upconversion arising from a Yb excited state is explored and the mechanistic origin behind cooperative sensitisation (CS) upconversion in Yb/ Tb systems is investigated. Counterintuitively, the best UC performances were obtained for Yb/Tb ratios close to parity, resulting in the brightest molecular upconversion complexes with a quantum yield of 2.8 × 10-6 at a low laser power density of 2.86 W/cm2.

Published
2021

23. Elucidating Cooperative Luminescence and Cooperative Sensitisation Upconversion of Nonanuclear Lanthanide Complexes in Solution

Author

Richard C. Knighton, Niko Hildebrandt, Aurora Rodríguez-Rodríguez, Laura Francés-Soriano, Loïc J. Charbonnière, Lohona K. Soro, Guillaume Pilet, Carlos Platas-Iglesias, and Marc Lenertz

Subjects
Lanthanide, Laser power density, Materials science, Quenching (fluorescence), Excited state, Quantum yield, Luminescence, Photochemistry, Spectroscopy, Photon upconversion
Abstract

Upconversion nanoparticles have led to various breakthrough applications in solar energy conversion, imaging, and biomedicine. One key impediment is the facilitation of such processes at the molecular scale in solution where quenching effects are much more pronounced. In this work we explore, for the first time, molecular solution-state cooperative luminescence (CL) upconversion arising from a Yb excited state and investigate the mechanistic origin behind cooperative sensitisation (CS) upconversion in Yb/ Tb systems. Counterintuitively, the best UC performances were obtained for Yb/Tb ratios close to parity, resulting in the brightest molecular upconversion complexes with an unprecedented quantum yield of 2.8 × 10-6 at a low laser power density of 2.86 W/cm2.

Published
2021
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24. pH dependence of water anomaly temperature investigated by Eu(III) cryptate luminescence

Author

Niko Hildebrandt, Jurriaan M. Zwier, Patricia Haro-González, Daniel Jaque, Carlos Mingoes, Lucía Labrador-Páez, Herve Bazin, F. Jaque, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)

Subjects
Luminescence, Materials science, [SDV]Life Sciences [q-bio], Analytical chemistry, chemistry.chemical_element, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Biochemistry, Hydrogen bonds, Analytical Chemistry, Ph dependence, [CHIM]Chemical Sciences, Molecule, [PHYS]Physics [physics], Hydrogen bond, Water, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Highly sensitive, chemistry, 13. Climate action, Lanthanide ions, Ph range, Anomaly (physics), 0210 nano-technology, Europium
Abstract

International audience; Although water has been extensively studied, not all of its unique properties have been fully understood. There is still controversy about the temperature at which hydrogen bonds are broken or weakened, producing the anomalous temperature dependence of many water properties. Different temperatures between 23 and 48 °C have been reported, but no study has scrutinized the reasons for this discrepancy. We suggest the determining role of pH in the alteration of the water anomaly temperature. We employed a luminescent europium trisbipyridine cryptate, which is highly sensitive to changes in the arrangement of water molecules and whose luminescence intensity and lifetime are not significantly influenced by variations over a broad pH range. Our results revealed an increase of the crossover temperature from circa 35 °C at pH 3.5 to circa 45 °C at pH 7 to 9, which explains the discrepancies of previous studies. The pH dependence of water anomaly temperature is an important property for a better understanding of water and water-based systems and applications.

Published
2019
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25. Discrimination of the V600E Mutation in BRAF by Rolling Circle Amplification and Förster Resonance Energy Transfer

Author

Niko Hildebrandt, Mariia O. Dekaliuk, Frédéric Troalen, Xue Qiu, Philippe Busson, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Signalisation, noyaux et innovations en cancérologie (UMR8126), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), and Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV]Life Sciences [q-bio], Bioengineering, 02 engineering and technology, medicine.disease_cause, 01 natural sciences, chemistry.chemical_compound, medicine, [CHIM]Chemical Sciences, Denaturation (biochemistry), Instrumentation, Gene, [PHYS]Physics [physics], Fluid Flow and Transfer Processes, Mutation, Process Chemistry and Technology, 010401 analytical chemistry, 021001 nanoscience & nanotechnology, Molecular biology, 3. Good health, 0104 chemical sciences, Förster resonance energy transfer, chemistry, Cell-free fetal DNA, Rolling circle replication, 0210 nano-technology, DNA, V600E
Abstract

International audience; The quantification of very low concentrations of circulating tumor DNA (ctDNA) biomarkers from liquid biopsies has become an important requirement for clinical diagnostics and personalized medicine. In particular, the simultaneous detection of wild-type (WT) dsDNA and their cancer-related counterparts presenting single-point mutations with simple, sensitive, specific, and reproducible technologies is paramount for ctDNA assays in clinical practice. Here, we present the development and evaluation of an amplified dsDNA assay based on a combination of isothermal rolling circle amplification (RCA) and time-gated Förster resonance energy transfer (TG-FRET) between a Tb donor and two dye (Cy3.5 and Cy5.5) acceptors. The RCA–FRET assay is free of washing and separation steps and can quantify both WT and mutated (MT) (V600E) dsDNA in the BRAF gene from a single sample in the 75 fM to 4.5 pM (4.5 × 105 to 2.7 × 107 copies) concentration range. This assay includes all steps from denaturation of the dsDNA targets to the final duplexed quantification of WT and MT targets. High assay performance at different dsDNA sequence lengths and high target specificity even in the presence of a large excess of nonspecific cell-free DNA from human plasma samples demonstrated the applicability to clinical samples. The RCA–FRET single-point mutation sensor has the potential to become an important complementary technique for analyzing liquid biopsies in advanced cancer diagnostics.

Published
2019
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26. FRET-Modulated Multihybrid Nanoparticles for Brightness-Equalized Single-Wavelength Barcoding

Author

Liang Huang, Niko Hildebrandt, Ben Corry, Chi Chen, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)

Subjects
Brightness, Photoluminescence, [SDV]Life Sciences [q-bio], chemistry.chemical_element, Nanoparticle, Terbium, 010402 general chemistry, 01 natural sciences, Biochemistry, Catalysis, Colloid and Surface Chemistry, Quantum Dots, Fluorescence Resonance Energy Transfer, Fluorescence microscope, Spectroscopy, Fluorescent Dyes, business.industry, General Chemistry, Carbocyanines, 0104 chemical sciences, Förster resonance energy transfer, chemistry, Nanoparticles, Optoelectronics, business, Excitation
Abstract

International audience; Semiconductor quantum dots (QDs) are the most versatile fluorophores for Förster resonance energy transfer (FRET) because they can function as both donors and acceptors for a multitude of fluorophores. However, a complete understanding of multidonor-multiacceptor FRET networks on QDs and their full employment into advanced fluorescence sensing and imaging have not been accomplished. Here, we provide a holistic photophysical analysis of such multidonor-QD-multiacceptor FRET systems using time-resolved and steady-state photoluminescence (PL) spectroscopy and Monte Carlo simulations. Multiple terbium complex (Tb) donors (1-191 units) and Cy5.5 dye acceptors (1-60 units) were attached to a central QD, and the entire range of combinations of FRET pathways was investigated by Tb, QD, and Cy5.5 PL. Experimental and simulation results were in excellent agreement and could disentangle the distinct contributions of hetero-FRET, homo-FRET, and dye dimerization. The FRET efficiency was independent of the number of Tb donors and dependent on the number of Cy5.5 acceptors, which could be used to independently adapt the PL intensity by the number of Tb donors and the PL lifetime by the number of Cy5.5 acceptors. We used this unique tuning capability to prepare Tb-QD-Cy5.5 conjugates with distinct QD PL lifetimes but similar QD PL intensities. These brightness-equalized multihybrid FRET nanoparticles were applied to optical barcoding via three time-gated PL intensity detection windows, which resulted in simple RGB ratios. Direct applicability was demonstrated by an efficient RGB distinction of different nanoparticle-encoded microbeads within the same field of view with both single-wavelength excitation and detection on a standard fluorescence microscope.

Published
2019
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29. Upconversion in molecular hetero-nonanuclear lanthanide complexes in solution

Author

Niko Hildebrandt, Richard C. Knighton, Lohona K. Soro, Alexandre Lecointre, Guillaume Pilet, Alexandra Fateeva, Loïc J. Charbonnière, Laura Francés-Soriano, Laurie Pontille, Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Multimatériaux et Interfaces (LMI), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), and Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Lanthanide, Materials science, Solution state, Metals and Alloys, 02 engineering and technology, General Chemistry, Molecular systems, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, Catalysis, Photon upconversion, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Materials Chemistry, Ceramics and Composites, Physical chemistry, Molecule, [CHIM.COOR]Chemical Sciences/Coordination chemistry, 0210 nano-technology, Luminescence, Excitation
Abstract

International audience; Here we show that nonanuclear lanthanide complexes respresent a new class of solution state upconversion (UC) molecules. For a composition of one Tb per eight Yb the nonanuclear complexes display a very efficient UC phenomenon with Tb luminescence in the visible region upon 980 nm NIR excitation of Yb. An unprecedented value of 1.0 × 10−7 was obtained for the UC efficiency at only 2.86 W cm−2, demonstrating these new molecular complexes to be up to 26 times more efficient than the best current molecular systems, the UC being observed down to a concentration of 10 nM.

Published
2021
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30. In situ rolling circle amplification Förster resonance energy transfer (RCA-FRET) for washing-free real-time single protein imaging

Author

Laura Francés-Soriano, Mattias Leino, Marcelina Cardoso Dos Santos, Daniel Kovacs, K. Eszter Borbas, Ola Söderberg, and Niko Hildebrandt

Subjects
RCA, time-gating, FRET, lanthanides, Imaging
Abstract

Fluorescence signal enhancementviaisothermal nucleic acid amplification is an important approach for sensitive imaging of intra- or extracellular nucleic acid or protein biomarkers. Rolling circle amplification (RCA) is frequently applied for fluorescencein situimaging but faces limitations concerning multiplexing, dynamic range, and the required multiple washing steps before imaging. Here, we show that Förster resonance energy transfer (FRET) between fluorescent dyes and between lanthanide (Ln) complexes and dyes that hybridize to β-actin-specific RCA products in HaCaT cells can afford washing-free imaging of single β-actin proteins. Proximity-dependent FRET could be monitored directly after or during (real-time monitoring) dye or Ln DNA probe incubation and could efficiently distinguish between photoluminescence from β-actin-specific RCA and DNA probes freely diffusing in solution or nonspecifically attached to cells. Moreover, time-gated FRET imaging with the Ln-dye FRET pairs efficiently suppressed sample autofluorescence and improved the signal-to-background ratio. Our results present an important proof of concept of RCA-FRET imaging with a strong potential to advancein situRCA toward easier sample preparation, higher-order multiplexing, autofluorescence-free detection, and increased dynamic range by real-time monitoring ofin situRCA.

Published
2020

31. Upconversion in Molecular Hetero-Nonanuclear Lanthanide Clusters in Solution

Author

Richard Knighton, Lohona K. Soro, Alexandre Lecointre, Guillaume Pilet, Alexandra Fateeva, laurie pontille, Laura Laura Francés-Soriano, Niko Hildebrandt, and Loïc J. Charbonnière

Abstract

Up until now restricted to solid state chemistry and nanoparticles, upconversion (UC) phenomena by which low energy photons are piled up to engender higher energy emission, are progressively entering the field of molecular probes with a handful of molecular/supramolecular discrete complexes. Here we show that nonanuclear lanthanide complexes respresent a new class of solution state UC materials, and are straightforwardly crystalized from LnCl3.6H2O salts, triethylamine and acetylacetone, retaining their structural integrity in solution. For a composition of one Tb per eight Yb the nonanuclear clusters display a very efficient upconversion phenomenon with Tb luminescence in the visible region upon 980 nm NIR excitation of Yb. An unprecedented value of 1.0×10-7 was obtained for the UC efficiency at only 2.86 W/cm2, demonstrating these new molecular clusters to be up to 26 times more efficient than the best current molecular systems, the UC being observed down to a concentration of 10 nM.

Published
2020
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34. Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET

Author

Anne Incamps, Shashi Bhuckory, Xue Qiu, Niko Hildebrandt, K. David Wegner, Yu-Tang Wu, Travis L. Jennings, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), BAM Bundesanstalt für Materialforschung u. -prüfung, Ocean University of China (OUC), ThermoFisher, Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB), Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), ThermoFisher Scientific, Thermofisher Scientific, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Hildebrandt, Niko, Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)

Subjects
[SDV]Life Sciences [q-bio], Pharmaceutical Science, 02 engineering and technology, 01 natural sciences, 7. Clean energy, [PHYS] Physics [physics], Analytical Chemistry, PSA, Carcinoembryonic antigen, CEA, NSE, [CHIM] Chemical Sciences, Drug Discovery, Fluorescence Resonance Energy Transfer, [PHYS]Physics [physics], Immunoassay, multiplexing, biology, medicine.diagnostic_test, Chemistry, 021001 nanoscience & nanotechnology, Fluorescence, 3. Good health, [SDV] Life Sciences [q-bio], Chemistry (miscellaneous), Molecular Medicine, Kallikreins, fluorescence, 0210 nano-technology, Plate reader, 010402 general chemistry, GPI-Linked Proteins, Article, lcsh:QD241-441, lcsh:Organic chemistry, Quantum Dots, medicine, [CHIM]Chemical Sciences, Humans, lanthanides, Physical and Theoretical Chemistry, Terbium, Detection limit, Chromatography, Organic Chemistry, Prostate-Specific Antigen, 0104 chemical sciences, Carcinoembryonic Antigen, Förster resonance energy transfer, Immunoglobulin G, biology.protein, nanoparticles, biosensing, Luminescence, Biosensor
Abstract

Time-gated F&ouml, rster resonance energy transfer (TG-FRET) between Tb complexes and luminescent semiconductor quantum dots (QDs) provides highly advantageous photophysical properties for multiplexed biosensing. Multiplexed Tb-to-QD FRET immunoassays possess a large potential for in vitro diagnostics, but their performance is often insufficient for their application under clinical conditions. Here, we developed a homogeneous TG-FRET immunoassay for the quantification of carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and prostate-specific antigen (PSA) from a single serum sample by multiplexed Tb-to-QD FRET. Tb&ndash, IgG antibody donor conjugates were combined with compact QD-F(ab&rsquo, )2 antibody acceptor conjugates with three different QDs emitting at 605, 650, and 705 nm. Upon antibody&ndash, antigen&ndash, antibody sandwich complex formation, the QD acceptors were sensitized via FRET from Tb, and the FRET ratios of QD and Tb TG luminescence intensities increased specifically with increasing antigen concentrations. Although limits of detection (LoDs: 3.6 ng/mL CEA, 3.5 ng/mL NSE, and 0.3 ng/mL PSA) for the triplexed assay were slightly higher compared to the single-antigen assays, they were still in a clinically relevant concentration range and could be quantified in 50 &micro, L serum samples on a B&middot, R&middot, A&middot, H&middot, M&middot, S KRYPTOR Compact PLUS clinical immunoassay plate reader. The simultaneous quantification of CEA, NSE, and PSA at different concentrations from the same serum sample demonstrated actual multiplexing Tb-to-QD FRET immunoassays and the potential of this technology for translation into clinical diagnostics.

Published
2020
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35. Terbium–To–Quantum Dot Förster Resonance Energy Transfer for Homogeneous and Sensitive Detection of Histone Methyltransferase Activity

Author

Kimihiro Susumu, Xue Qiu, Tooba Hallaj, Igor L. Medintz, Niko Hildebrandt, Mohammad Amjadi, Urmia University, University of Tabriz [Tabriz], Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)

Subjects
Streptavidin, Histone methyltransferase activity, [SDV]Life Sciences [q-bio], Kinetics, 010402 general chemistry, 01 natural sciences, 03 medical and health sciences, chemistry.chemical_compound, Histone H3, Quantum Dots, Fluorescence Resonance Energy Transfer, [CHIM]Chemical Sciences, General Materials Science, Enzyme kinetics, Terbium, 030304 developmental biology, [PHYS]Physics [physics], 0303 health sciences, Combinatorial chemistry, 0104 chemical sciences, 3. Good health, Förster resonance energy transfer, chemistry, Histone methyltransferase, Histone Methyltransferases, Biosensor
Abstract

International audience; The development of rapid, simple, and versatile biosensors for monitoring the activity of histone modifying enzymes (HMEs) is needed for the improvement of diagnostic assays, screening of HME inhibitors, and a better understanding of HME kinetics in different environments. Nanoparticles can play an important role in this regard by improving or complementing currently available enzyme detection technologies. Here, we present the development and application of a homogeneous methyltransferase (SET7/9) assay based on time-gated Förster resonance energy transfer (TG-FRET) between terbium complexes (Tb) and luminescent semiconductor quantum dots (QDs). Specific binding of a Tb-antibody conjugate to a SET7/9-methylated Lys4 on a histone H3(1–21) peptide substrate attached to the QD surface resulted in efficient FRET and provided the mechanism for monitoring the SET7/9 activity. Two common peptide-QD attachment strategies (biotin–streptavidin and polyhistidine-mediated self-assembly), two different QD colors (625 and 705 nm), and enzyme sensing with post- or pre-assembled QD–peptide conjugates demonstrated the broad applicability of this assay design. Limits of detection in the low picomolar concentration range, high selectivity tested against non-specific antibodies, enzymes, and co-factors, determination of the inhibition constants of the SET7/9 inhibitors SAH and (R)-PFI-2, and analysis of the co-factor (SAM) concentration-dependent enzyme kinetics of SET7/9 which followed the Michaelis–Menten model highlighted the excellent performance of this TG-FRET HME activity assay.

Published
2020
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36. Single-Measurement Multiplexed Quantification of MicroRNAs from Human Tissue Using Catalytic Hairpin Assembly and Förster Resonance Energy Transfer

Author

Nicole Golob-Schwarzl, Jingyue Xu, Jiajia Guo, Xue Qiu, Niko Hildebrandt, Johannes Haybaeck, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Medical University Graz, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE12-0030,PhenX,Base moléculaire de l'activation de la recombinaison Xer par les phages(2016)

Subjects
Bioanalysis, [SDV]Life Sciences [q-bio], Bioengineering, 02 engineering and technology, 01 natural sciences, microRNA, Fluorescence Resonance Energy Transfer, Humans, cancer, [CHIM]Chemical Sciences, Terbium, Instrumentation, clinical diagnostics, Fluid Flow and Transfer Processes, Detection limit, [PHYS]Physics [physics], catalytic hairpin assembly, Chemistry, Process Chemistry and Technology, 010401 analytical chemistry, Translation (biology), tissue, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Autofluorescence, MicroRNAs, Förster resonance energy transfer, Biochemistry, Nucleic acid, FRET, miR-21, 0210 nano-technology, Biosensor, miR-20a
Abstract

International audience; Absolute quantification of microRNAs (miRNAs) or other nucleic acid biomarkers is an important requirement for molecular and clinical biosensing. Emerging technologies with beneficial features concerning simplicity and multiplexing present an attractive route for advancing diagnostic tools toward rapid and low-cost bioanalysis. However, the actual translation into the clinic by miRNA quantification in human samples is often missing. Here, we show that implementing time-gated Förster resonance energy transfer (TG-FRET) into a catalytic hairpin assembly (CHA) can be used for the simultaneous quantification of two miRNAs with a single measurement from total RNA extracts of human tissues. A single terbium–dye FRET pair was conjugated at two specific distances within target-specific CHA hairpin probes, such that each miRNA resulted in distinct amplified photoluminescence (PL) decays that could be distinguished and quantified by TG PL intensity detection. Enzyme-free amplification in a separation-free assay format and the absence of autofluorescence background allowed for simple, specific, and sensitive detection of miR-21 and miR-20a with limits of detection down to 1.8 pM (250 amol). Reliable duplexed quantification of both miRNAs at low picomolar concentrations was confirmed by analyzing total RNA extracts from different colon and rectum tissues with single- and dual-target CHA-TG-FRET and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for comparison. These simple and multiplexed nucleic acid biomarker assays present a capable method for clinical diagnostics and biomolecular research.

Published
2020
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37. Simple, Amplified, and Multiplexed Detection of MicroRNAs Using Time-Gated FRET and Hybridization Chain Reaction

Author

Carlos Mingoes, Xue Qiu, Niko Hildebrandt, Jiajia Guo, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), and ANR-16-CE12-0030,PhenX,Base moléculaire de l'activation de la recombinaison Xer par les phages(2016)

Subjects
construction, isothermal amplification, Time Factors, [SDV]Life Sciences [q-bio], Loop-mediated isothermal amplification, 02 engineering and technology, dna, 010402 general chemistry, 01 natural sciences, Analytical Chemistry, Nucleic acid thermodynamics, chemistry.chemical_compound, diagnostic assay, Fluorescence Resonance Energy Transfer, Detection limit, Chemistry, Multiple displacement amplification, Nucleic Acid Hybridization, 021001 nanoscience & nanotechnology, 0104 chemical sciences, signal amplification, MicroRNAs, Förster resonance energy transfer, in-situ, strand displacement amplification, Biophysics, Nucleic acid, sensitive detection, 0210 nano-technology, human activities, Biosensor, DNA
Abstract

International audience; The hybridization chain reaction (HCR) is a simple and sensitive method for quantifying nucleic acids. Current approaches cannot combine a washing-free sensing format with multiplexed target quantification at low concentrations, which would be highly desirable for detection both in solution and in situ. Here, we demonstrate the implementation of time-gated Förster resonance energy transfer (TG-FRET) between terbium donors and dye acceptors into HCR for multiplexed quantification of microRNAs (miR-20a and miR-21) and their DNA analogues. HCR-TG-FRET provided washing-free nucleic acid quantification with very low limits of detection down to 240 amol (1.7 pM) of microRNA and 123 amol (0.88 pM) of DNA. Efficient distinction from very homologous microRNAs demonstrated high target specificity. Multiplexing with a single measurement, a single excitation wavelength, and a single FRET pair allowed for a simultaneous quantification of miR-20a and miR-21 at concentrations between 30 and 300 pM from the same sample. HCR-TG-FRET showed similar performance for serum-free and serum-containing samples without the use of RNase inhibitors. Our results present a significant improvement in current HCR approaches regarding simplicity, sensitivity, and multiplexing. The versatile diagnostic performance of HCR-TG-FRET even in challenging biological environments presents an important advantage for advanced nucleic acid biosensing.

Published
2019
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38. Energy transfer with nanoparticles for in vitro diagnostics

Author

Niko Hildebrandt, Jiajia Guo, Jingyue Xu, Tooba Hallaj, Laura Francés-Soriano, Xue Qiu, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Urmia University, Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), and Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[PHYS]Physics [physics], Materials science, [SDV]Life Sciences [q-bio], Nanoparticle, Nanotechnology, 02 engineering and technology, Chromophore, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, Acceptor, Small molecule, Fluorescence, 0104 chemical sciences, Förster resonance energy transfer, Colloidal gold, [CHIM]Chemical Sciences, 0210 nano-technology, Luminescence, ComputingMilieux_MISCELLANEOUS
Abstract

In vitro diagnostics is an important application for nanoparticles in the life sciences and has attracted continuous research interest. While nanoparticles can be used as single luminescent probes, their unique photophysical properties provide many more advantages when they are combined with other nanoparticles or molecules to form energy transfer pairs. Energy transfer-based assays are homogeneous in nature and are often as simple as “mix and measure,” which has catalyzed their frequent use in clinical diagnostics. Most bioanalytical applications involve Forster resonance energy transfer (FRET), a distance-dependent nonradiative energy transfer from a donor to acceptor chromophore through dipole–dipole interaction. Nanosurface energy transfer (NSET) describes energy transfer to metal nanoparticles. Semiconductor quantum dots, upconversion nanoparticles, gold nanoparticles, and carbon dots are among the most applied nanoparticles in in vitro diagnostics and they have been largely explored in the analysis of disease-related nucleic acids, proteins, and small molecules. The utilizations of various nanoparticles with their intrinsic optical properties have changed the diagnostic horizon substantially by improving the assay simplicity, sensitivity, selectivity, robustness, and multiplexing capacity. In this chapter, we discuss the use of nanoparticles in energy transfer assays for in vitro diagnostics and show their strong potential for advancing the field beyond molecular fluorescent probes.

Published
2020
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39. Quantum dots for Förster Resonance Energy Transfer (FRET)

Author

Igor L. Medintz, Niko Hildebrandt, Marcelina Cardoso Dos Santos, W. Russ Algar, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), University of British Columbia (UBC), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), and Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
chemistry.chemical_classification, [PHYS]Physics [physics], Fluorescence-lifetime imaging microscopy, Materials science, Biomolecule, [SDV]Life Sciences [q-bio], 010401 analytical chemistry, technology, industry, and agriculture, Nanotechnology, [CHIM.MATE]Chemical Sciences/Material chemistry, 01 natural sciences, 0104 chemical sciences, Analytical Chemistry, Förster resonance energy transfer, chemistry, Quantum dot, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Surface modification, [CHIM]Chemical Sciences, Absorption (electromagnetic radiation), Spectroscopy
Abstract

International audience; The analysis of biomolecular interactions using quantum dots (QDs) as both FRET donors and acceptors has become an established technique in the life sciences. This development has been driven by the unique properties of QDs, which include large surfaces for the attachment of biomolecules, high brightness and photostability, strong and spectrally broad absorption, and color-tunability via QD size, shape, and material. Applications include molecular rulers for structural analysis, small-molecule sensors, immunoassays, enzyme assays, nucleic acid assays, fluorescence imaging in-vitro and in-vivo, and molecular logic gates. Here, we will explain the theory of QD-based FRET, review some aspects of QD surface functionalization that are important for FRET, and highlight and discuss the advantages and disadvantages of QDs in FRET-biosensing using both spectroscopy and imaging techniques.

Published
2020
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40. Picomolar Biosensing and Conformational Analysis Using Artificial Bidomain Proteins and Terbium-to-Quantum Dot Förster Resonance Energy Transfer

Author

Agathe Urvoas, Philippe Minard, Niko Hildebrandt, Igor L. Medintz, Corentin Leger, Kimihiro Susumu, Marie Valerio-Lepiniec, Akram Yahia-Ammar, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Modélisation et Ingénierie des Protéines (MIP), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Naval Research Laboratory (NRL), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Optical Sciences Division, Departments of Cell Biology and Chemistry, The Scripps Research Institute, Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), and Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Scaffold protein, Conformational change, [SDV]Life Sciences [q-bio], General Physics and Astronomy, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, molecular diagnostics, Protein structure, Quantum Dots, Fluorescence Resonance Energy Transfer, [CHIM]Chemical Sciences, lanthanides, General Materials Science, protein structure, Terbium, binding assay, [PHYS]Physics [physics], Chemistry, Ligand binding assay, General Engineering, molecular ruler, 021001 nanoscience & nanotechnology, Acceptor, 0104 chemical sciences, Förster resonance energy transfer, Semiconductors, Quantum dot, FRET, Biophysics, 0210 nano-technology, Biosensor
Abstract

Publisher: American Chemical Society; International audience; Although antibodies remain a primary recognition element in all forms of biosensing, functional limitations arising from their size, stability, and structure have motivated the development and production of many different artificial scaffold proteins for biological recognition. However, implementing such artificial binders into functional high-performance biosensors remains a challenging task. Here, we present the design and application of Förster resonance energy transfer (FRET) nanoprobes comprised of small artificial proteins (αRep bidomains) labeled with a Tb complex (Tb) donor on the C-terminus and a semiconductor quantum dot (QD) acceptor on the N-terminus. Specific binding of one or two protein targets to the αReps induced a conformational change that could be detected by time-resolved Tb-to-QD FRET. These single-probe FRET-switches were used in a separation-free solution-phase assay to quantify different protein targets at subnanomolar concentrations and measure the conformational changes with subnanometer resolution. Probing ligand-receptor binding under physiological conditions at very low concentrations in solution is a special feature of FRET that can be efficiently combined with other structural characterization methods to develop, understand, and optimize artificial biosensors. Our results suggest that the αRep FRET nanoprobes have a strong potential for their application in advanced diagnostics and intracellular live cell imaging of ligand-receptor interactions.

Published
2020
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41. Er

Author

Laura, Francés-Soriano, Nicola, Peruffo, Marta Maria, Natile, and Niko, Hildebrandt

Subjects
Ions, Neodymium, Europium, Infrared Rays, Fluorescence Resonance Energy Transfer, DNA, Single-Stranded, Metal Nanoparticles, Yttrium, Carbocyanines, Fluorescent Dyes
Abstract

The capability of upconverting nanoparticles (UCNPs) to convert near infrared (NIR) into visible light has become an important feature for biosensing, imaging, therapy, and their combination. While significant achievements have been accomplished during the last decade developing nanohybrids based on UCNPs as energy donors in Förster resonance energy transfer (FRET) systems, it is still challenging to understand and control FRET from UCNPs to dyes and to adapt the NIR excitation wavelength. Here, we describe the synthesis, characterization, and steady-state and time-resolved FRET analysis of UCNP-DNA nanohybrids, in which dye labelled single stranded (ss)DNA was attached to Yb-Er-co-doped core UCNPs (c-UCNPs) and c-UCNPs with a thin Nd-doped shell and a second thin undoped shell (css-UCNPs). Despite differences in sizes, compositions, donor-acceptor distances, brightness, and excitation wavelength (980 nm for Yb

Published
2020

42. Single‐Nanoparticle Cell Barcoding by Tunable FRET from Lanthanides to Quantum Dots

Author

David Parker, Emmanuel Bourrier, Chi Chen, Jurriaan M. Zwier, Yu-Tang Wu, Vjona Cifliku, Martina Delbianco, Liang Huang, Niko Hildebrandt, Lijiao Ao, Marcelina Cardoso Dos Santos, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Cisbio Bioassays, DEPARTMENT OF CHEMISTRY, Durham University, Institute of Molecular Chemistry, and Universiteit van Amsterdam (UvA)

Subjects
Optical Barcoding, Materials science, Photoluminescence, resonance energy-transfer, [SDV]Life Sciences [q-bio], Nanoparticle, 02 engineering and technology, bright, 010402 general chemistry, 01 natural sciences, biomolecules, Catalysis, nanocrystals, Europium, Live cell imaging, Microscopy, Quantum Dots, Fluorescence Resonance Energy Transfer, lanthanides, Terbium, lifetime, business.industry, Communication, imaging, General Chemistry, 021001 nanoscience & nanotechnology, biosensors, europium complexes, Fluorescence, photoluminescence lifetimes, Communications, 0104 chemical sciences, microspheres, Förster resonance energy transfer, Nanocrystal, Quantum dot, microscopy, FRET, Optoelectronics, Nanoparticles, 0210 nano-technology, business
Abstract

WOS:000446306600058; International audience; Fluorescence barcoding based on nanoparticles provides many advantages for multiparameter imaging. However, creating different concentration-independent codes without mixing various nanoparticles and by using single-wavelength excitation and emission for multiplexed cellular imaging is extremely challenging. Herein, we report the development of quantum dots (QDs) with two different SiO2 shell thicknesses (6 and 12nm) that are coated with two different lanthanide complexes (Tb and Eu). FRET from the Tb or Eu donors to the QD acceptors resulted in four distinct photoluminescence (PL) decays, which were encoded by simple time-gated (TG) PL intensity detection in three individual temporal detection windows. The well-defined single-nanoparticle codes were used for live cell imaging and a one-measurement distinction of four different cells in a single field of view. This single-color barcoding strategy opens new opportunities for multiplexed labeling and tracking of cells.

Published
2018
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43. Ultrabright Terbium Nanoparticles for FRET Biosensing and in Situ Imaging of Epidermal Growth Factor Receptors

Author

Laura Francés-Soriano, Vjona Cifliku, Cyrille Charpentier, Clémence Cheignon, Joan Goetz, Aline Nonat, Loïc J. Charbonnière, Niko Hildebrandt, Ka-Leung Wong, Marcelina Cardoso Dos Santos, Centre National de la Recherche Scientifique (CNRS), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), POLY-DTECH, Reconnaissance Ionique et Chimie de Coordination (RICC), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS), Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Hong Kong Baptist University (HKBU), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), ANR‐11‐IDEX‐0003‐02/ Saclay Plant Sciences, ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016), ANR-11-EQPX-0029,MORPHOSCOPE 2,Imagerie et reconstruction multiéchelles de la morphogenèse. (Plateforme d'innovation technologique et méthodologique pour l'imagerie in vivo et la reconstruction des dynamiques multiéchelles de la morphogenèse)(2011), and ANR-10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010)

Subjects
EGF Family of Proteins, [SDV]Life Sciences [q-bio], Biosensing Techniques, 010402 general chemistry, 7. Clean energy, 01 natural sciences, Catalysis, Mice, Epidermal growth factor, Fluorescence Resonance Energy Transfer, Animals, [CHIM]Chemical Sciences, Terbium, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, [PHYS]Physics [physics], Bioconjugation, 010405 organic chemistry, Chemistry, Biomolecule, Organic Chemistry, General Chemistry, 0104 chemical sciences, ErbB Receptors, Förster resonance energy transfer, Biotinylation, Biophysics, NIH 3T3 Cells, Nanoparticles, Biosensor, Linker, A431 cells
Abstract

International audience; Lanthanide‐doped nanoparticles (LnNPs) have become an important class of fluorophores for advanced biosensing and bioimaging. LnNPs that are photosensitized by surface‐attached antenna ligands can possess exceptional brightness. However, their functional bioconjugation remains an important challenge for their translation into bioanalytical applications. To solve this problem, we designed a ligand that can be simultaneously applied as efficient light harvesting antenna for Tb surface ions and strong linker of biomolecules to the LnNPs surfaces. To demonstrate generic applicability of the photosensitized TbNP‐bioconjugates, we applied them in two prototypical applications for biosensing and bioimaging. First, in‐solution biorecognition was shown by time‐resolved Förster resonance energy transfer (FRET) between streptavidin‐functionalized TbNPs to biotinylated dyes (ATTO 610). Second, in‐situ detection of ligand‐receptor binding on cells was accomplished with TbNP‐antibody (Matuzumab) conjugates that could specifically bind to transmembrane epidermal growth factor receptors (EGFR). High specificity and sensitivity were demonstrated by time‐gated imaging of EGFR on both strongly (A431) and weakly (HeLa and Cos7) EGFR‐expressing cell lines, whereas non‐expressing cell lines (NIH3T3) and EGFR‐passivated A431 cells did not show any signals. Despite the relatively large size of TbNP‐antibody conjugates, they could be internalized by A431 cells upon binding to extracellular EGFR, which showed their potential as bright and stable luminescence markers for intracellular signalling.

Published
2020
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44. Er3+-to-dye energy transfer in DNA-coated core and core/shell/shell upconverting nanoparticles with 980 nm and 808 nm excitation of Yb3+ and Nd3

Author

Marta Maria Natile, Niko Hildebrandt, Nicola Peruffo, Laura Francés-Soriano, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NanoBioPhotonics (NANO), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)

Subjects
Infrared devices, Materials science, Infrared Rays, [SDV]Life Sciences [q-bio], Kinetics, Metal Nanoparticles, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, 7. Clean energy, Biochemistry, Analytical Chemistry, Europium, Single-Stranded, Electrochemistry, Fluorescence Resonance Energy Transfer, Environmental Chemistry, Yttrium, Spectroscopy, Fluorescent Dyes, Ions, Neodymium, business.industry, Near-infrared spectroscopy, DNA, Orders of magnitude (numbers), Carbocyanines, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Förster resonance energy transfer, Energy transfer, DNA, Single-Stranded, FRET, Optoelectronics, Nanoparticles, 0210 nano-technology, Luminescence, business, Biosensor, Excitation, Upconversion, Visible spectrum
Abstract

The capability of upconverting nanoparticles (UCNPs) to convert near infrared (NIR) into visible light has become an important feature for biosensing, imaging, therapy, and their combination. While significant achievements have been accomplished during the last decade developing nanohybrids based on UCNPs as energy donors in Förster resonance energy transfer (FRET) systems, it is still challenging to understand and control FRET from UCNPs to dyes and to adapt the NIR excitation wavelength. Here, we describe the synthesis, characterization, and steady-state and time-resolved FRET analysis of UCNP–DNA nanohybrids, in which dye labelled single stranded (ss)DNA was attached to Yb–Er-co-doped core UCNPs (c-UCNPs) and c-UCNPs with a thin Nd-doped shell and a second thin undoped shell (css-UCNPs). Despite differences in sizes, compositions, donor–acceptor distances, brightness, and excitation wavelength (980 nm for Yb3+and 808 nm for Nd3+), all UCNP–DNA nanohybrids showed very similar concentration dependent FRET-quenching of UCNP luminescence with efficiencies between 0 and ∼20%. We analyzed luminescence intensities, decay times, and rise times and could show the entanglement of excitation and emission kinetics by simply changing the excitation wavelength from 980 nm to 808 nm for the same css-UCNPs. Time-gated FRET-sensitized dye luminescence showed dye-ssDNA concentration dependence over four orders of magnitude (1 nM to 10 μM), which suggested a possible application to nucleic acid biosensing for both 808 and 980 nm excitation.

Published
2020
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45. Resonance energy transfer to gold nanoparticles: NSET defeats FRET

Author

Chi Chen, Niko Hildebrandt, National University of Defense Technology [China], Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)

Subjects
[PHYS]Physics [physics], Materials science, Energy transfer, [SDV]Life Sciences [q-bio], 010401 analytical chemistry, Nanotechnology, 01 natural sciences, 0104 chemical sciences, Analytical Chemistry, Formalism (philosophy of mathematics), Förster resonance energy transfer, Colloidal gold, [CHIM]Chemical Sciences, Luminescence, Biosensor, Spectroscopy, ComputingMilieux_MISCELLANEOUS
Abstract

Gold nanoparticles (AuNPs) have been widely applied as highly efficient luminescence quenchers in combination with a large variety of donor fluorophores. While the application of such systems in optical biosensing has resulted in significant advances of biological, biochemical, and biophysical research, the underlying energy transfer mechanism has been much less investigated. In this review, we discuss the differences between Forster resonance energy transfer (FRET) and nanosurface energy transfer (NSET), give an overview of the literature concerning the assignment of FRET or NSET to AuNP-based luminescence quenching, show why the development of improved NSET models and their application to experimental data provided strong arguments in favor of the NSET formalism, and discuss applications of NSET in biosensing, molecular rulers, and molecular tension probes. Taking into account the existing literature, the NSET formalism, including size and wavelength-dependent issues, is superior to FRET theory for a general description of energy transfer to AuNPs.

Published
2019
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46. Discrimination of the

Author

Mariia, Dekaliuk, Xue, Qiu, Frédéric, Troalen, Pierre, Busson, and Niko, Hildebrandt

Subjects
Proto-Oncogene Proteins B-raf, Base Sequence, Limit of Detection, Fluorescence Resonance Energy Transfer, Humans, Nucleic Acid Hybridization, Point Mutation, Biosensing Techniques, Carbocyanines, DNA Probes, Nucleic Acid Amplification Techniques, Circulating Tumor DNA, Fluorescent Dyes
Abstract

The quantification of very low concentrations of circulating tumor DNA (ctDNA) biomarkers from liquid biopsies has become an important requirement for clinical diagnostics and personalized medicine. In particular, the simultaneous detection of wild-type (WT) dsDNA and their cancer-related counterparts presenting single-point mutations with simple, sensitive, specific, and reproducible technologies is paramount for ctDNA assays in clinical practice. Here, we present the development and evaluation of an amplified dsDNA assay based on a combination of isothermal rolling circle amplification (RCA) and time-gated Förster resonance energy transfer (TG-FRET) between a Tb donor and two dye (Cy3.5 and Cy5.5) acceptors. The RCA-FRET assay is free of washing and separation steps and can quantify both WT and mutated (MT) (

Published
2019

47. FRET as a biomolecular research tool — understanding its potential while avoiding pitfalls

Author

Niko Hildebrandt, Igor L. Medintz, Steven S. Vogel, W. Russ Algar, Department of Chemistry, University of British Columbia (UBC), Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)

Subjects
[PHYS]Physics [physics], 0303 health sciences, Computer science, [SDV]Life Sciences [q-bio], Cell Biology, Biochemistry, 03 medical and health sciences, Förster resonance energy transfer, [CHIM]Chemical Sciences, Biochemical engineering, Instrumentation (computer programming), Molecular Biology, 030304 developmental biology, Biotechnology
Abstract

International audience; The applications of Förster resonance energy transfer (FRET) grow with each year. However, different FRET techniques are not applied consistently, nor are results uniformly presented, which makes implementing and reproducing FRET experiments challenging. We discuss important considerations for designing and evaluating ensemble FRET experiments. Alongside a primer on FRET basics, we provide guidelines for making experimental design choices such as the donor-acceptor pair, instrumentation and labeling chemistries; selecting control experiments to unambiguously demonstrate FRET and validate that the experiments provide meaningful data about the biomolecular process in question; analyzing raw data and assessing the results; and reporting data and experimental details in a manner that easily allows for reproducibility. Some considerations are also given for FRET assays and FRET imaging, especially with fluorescent proteins. Our goal is to motivate and empower all biologists to consider FRET for the powerful research tool it can be.

Published
2019
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48. A clinical role for Förster resonance energy transfer in molecular diagnostics of disease

Author

Xue Qiu, Niko Hildebrandt, Institut d'électronique fondamentale (IEF), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), and ANR-16-CE09-0015,NEUTRINOS,Suivi des interactions biologiques par détection optique ultrasensible à base de nanoparticules(2016)

Subjects
0301 basic medicine, Point-of-Care Systems, [SDV]Life Sciences [q-bio], Nanotechnology, Biosensing Techniques, Communicable Diseases, Pathology and Forensic Medicine, 03 medical and health sciences, 0302 clinical medicine, Neoplasms, Quantum Dots, Genetics, Fluorescence Resonance Energy Transfer, Humans, [CHIM]Chemical Sciences, Instrumentation (computer programming), Pathology, Molecular, Molecular Biology, ComputingMilieux_MISCELLANEOUS, Physics, [PHYS]Physics [physics], Molecular diagnostics, 3. Good health, 030104 developmental biology, Förster resonance energy transfer, 030220 oncology & carcinogenesis, Molecular Medicine, Reagent Kits, Diagnostic
Abstract

The large popularity of FRET-based applications started in the early 90s, driven by significant advances in new fluorophores, detection methods, and instrumentation [1]. Implementation of FRET tech...

Published
2019
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49. Aptamer and nanomaterial based FRET biosensors: a review on recent advances (2014–2019)

Author

Milad Torabfam, Zeki Semih Pehlivan, Niko Hildebrandt, Hasan Kurt, Cleva W. Ow-Yang, Meral Yüce, Institut de Biologie Intégrative de la Cellule (I2BC), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)

Subjects
Materials science, [SDV]Life Sciences [q-bio], Aptamer, Nanochemistry, Nanoprobe, Carbon Dots, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, 7. Clean energy, Metal-Organic Framework, Analytical Chemistry, law.invention, Nanomaterials, law, ssDNA, Quantum Dots, Gold Nanoparticles, [CHIM]Chemical Sciences, Reduced Graphene Oxide, ComputingMilieux_MISCELLANEOUS, Graphene Quantum Dots, [PHYS]Physics [physics], Up-Converting Nanoparticles, Graphene, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Biosensors, Förster resonance energy transfer, Quantum dot, FRET, 0210 nano-technology, Biosensor
Abstract

Fluorescence resonance energy transfer, one of the most powerful phenomena for elucidating molecular interactions, has been extensively utilized as a biosensing tool to provide accurate information at the nanoscale. Numerous aptamer- and nanomaterial-based FRET bioassays has been developed for detection of a large variety of molecules. Affinity probes are widely used in biosensors, in which aptamers have emerged as advantageous biorecognition elements, due to their chemical and structural stability. Similarly, optically active nanomaterials offer significant advantages over conventional organic dyes, such as superior photophysical properties, large surface-to-volume ratios, photostability, and longer shelf life. In this report (with 175 references), the use of aptamer-modified nanomaterials as FRET couples is reviewed: quantum dots, upconverting nanoparticles, graphene, reduced graphene oxide, gold nanoparticles, molybdenum disulfide, graphene quantum dots, carbon dots, and metal-organic frameworks. Tabulated summaries provide the reader with useful information on the current state of research in the field. Graphical abstract Schematic representation of a fluorescence resonance energy transfer-based aptamer nanoprobe in the absence and presence of a given target molecule (analyte). Structures are not drawn to their original scales.

Published
2019
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