Fluorophore Multimerization as an Efficient Approach towards Bright Protein Labels

Reiber, Thorge; Zavoiura, Oleksandr; Dose, Christian

doi:10.1002/ejoc.202100117

Cited by 8 publications

(7 citation statements)

References 136 publications

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“…For example, incorporation of multiple methylene blue redox markers into a C-reactive protein sensor enhances signal output and enables a LoD of 1 pM, which is considerably lower than that of previous electrochemical and optical approaches using single reporter molecules 49 . Alternatively, secondary reporter complexes in the form of oligonucleotide systems or reporter-labelled nanomaterials (for example, dendrimers) may be used to produce amplified responses 50 . However, many of these multimerization amplification strategies are designed for use in more traditional immunoassays with externally added reporter molecules, and thus may not be compatible with body-based biomolecular sensing.…”

Section: Strategies For Signal Amplificationmentioning

confidence: 99%

Biomolecular sensors for advanced physiological monitoring

et al. 2023

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Body-based biomolecular sensing systems, including wearable, implantable and consumable sensors allow comprehensive health-related monitoring. Glucose sensors have long dominated wearable bioanalysis applications owing to their robust continuous detection of glucose, which has not yet been achieved for other biomarkers. However, access to diverse biological fluids and the development of reagentless sensing approaches may enable the design of body-based sensing systems for various analytes. Importantly, enhancing the selectivity and sensitivity of biomolecular sensors is essential for biomarker detection in complex physiological conditions. In this Review, we discuss approaches for the signal amplification of biomolecular sensors, including techniques to overcome Debye and mass transport limitations, and selectivity improvement, such as the integration of artificial affinity recognition elements. We highlight reagentless sensing approaches that can enable sequential real-time measurements, for example, the implementation of thin-film transistors in wearable devices. In addition to sensor construction, careful consideration of physical, psychological and security concerns related to body-based sensor integration is required to ensure that the transition from the laboratory to the human body is as seamless as possible.

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Section: Strategies For Signal Amplificationmentioning

confidence: 99%

Biomolecular sensors for advanced physiological monitoring

et al. 2023

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“…[13] To achieve high fluorescent brightness, multimerization strategy by increasing the number of fluorophores per label is an efficient approach. [14] Unfortunately, the fluorescence intensity of certain CEs in the membrane reduces as dye concentration increases due to the aggregation-caused quenching (ACQ) effect, limiting the overall brightness. [12] In this study, we designed and synthesized three new narrow-band gap CE molecules named EPV, EBen, and EFBT, which emit in the NIR-II region and possess an enlarged Stokes shift.…”

Section: Introductionmentioning

confidence: 99%

NIR‐II Conjugated Electrolytes as Biomimetics of Lipid Bilayers for In Vivo Liposome Tracking

Meng,

Gao,

Zhou

et al. 2024

Angewandte Chemie

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Liposomes serve as promising and versatile vehicles for drug delivery. Tracking these nanosized vesicles, particularly in vivo, is crucial for understanding their pharmacokinetics. This study introduces the design and synthesis of three new conjugated electrolyte (CE) molecules, which emit in the second near‐infrared window (NIR‐II), facilitating deeper tissue penetration. Additionally, these CEs, acting as biomimetics of lipid bilayers, demonstrate superior compatibility with lipid membranes compared to commonly used carbocyanine dyes like DiR. To counteract the aggregation‐caused quenching effect, CEs employ a twisted backbone, as such their fluorescence intensities can effectively enhance after a fluorophore multimerization strategy. Notably, a “passive” method was employed to integrate CEs into liposomes during the liposome formation, and membrane incorporation efficiency was significantly promoted to nearly 100%. To validate the in vivo tracking capability, the CE‐containing liposomes were functionalized with cyclic arginine‐glycine‐aspartic acid (cRGD) peptides, serving as tumor‐targeting ligands. Clear fluorescent images visualizing tumor site in living mice were captured by collecting the NIR‐II emission. Uniquely, these CEs exhibit additional emission peak in visible region, enabling in vitro subcellular analysis using routine confocal microscopy. These results underscore the potential of CEs as a new‐generation of membrane‐targeting probes to facilitate the liposome‐based medicine research.

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“…Therefore, recent strategies focused mainly on multimerization of small-molecule dyes. [18][19][20] In the last decades, π-conjugated polymers have been developed and were introduced for various applications reaching from organic solar cells over light emitting diodes and organic field-effect transistors to different utilizations for life science applications. [21][22][23][24][25] Compared to organic smallmolecule dyes, the polymeric structure allows for delocalization of the π-conjugation over many repeating units, leading to high extinction coefficients which can be controlled by synthetic design.…”

Section: Introductionmentioning

confidence: 99%

“…by rigidifying structures, or by blocking non‐fluorescent excited state relaxation mechanisms, [13–17] but the extinction coefficient is an inherent property of the π‐conjugated system and thus cannot be altered easily. Therefore, recent strategies focused mainly on multimerization of small‐molecule dyes [18–20] …”

Section: Introductionmentioning

confidence: 99%

Water‐Soluble Fluorescent Polymer Dyes with Tunable Emission Spectra for Flow Cytometry Applications

Berndt,

Glaap,

Jennings

et al. 2024

Angewandte Chemie

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The application of spectrally unique, bright, and water‐soluble fluorescent dyes is indispensable for the analysis of biological systems. Multiparameter flow cytometry is a powerful tool for characterization of mixed cell populations. To discriminate the different cell populations, they are typically stained by a set of fluorescent reagents, e.g., antibody‐fluorophore conjugates. The number of parameters which can be studied simultaneously strongly depends on the availability of reagents which can be differentiated by their spectral properties. In this study a series of fluorescent polymer dyes was developed, that can be excited with a single violet laser (405 nm) but distinguished by their unique emission spectra. The polyfluorene‐based polymers can be used on their own, or in combination with covalently bound small‐molecule dyes to generate energy transfer constructs to red‐shift the emission wavelength based on Förster resonance energy transfer (FRET). The polymer dyes were utilized in a biological flow cytometry assay by conjugating several of them to antibodies, demonstrating their effectiveness as reagents. This report represents the first systematic investigation of structure‐property relationships for this type of fluorescent dyes.

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Fluorophore Multimerization as an Efficient Approach towards Bright Protein Labels

Cited by 8 publications

References 136 publications

Biomolecular sensors for advanced physiological monitoring

Biomolecular sensors for advanced physiological monitoring

NIR‐II Conjugated Electrolytes as Biomimetics of Lipid Bilayers for In Vivo Liposome Tracking

Water‐Soluble Fluorescent Polymer Dyes with Tunable Emission Spectra for Flow Cytometry Applications

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