Six-Color Time-Resolved Förster Resonance Energy Transfer for Ultrasensitive Multiplexed Biosensing

Geißler, Daniel; Stufler, S.; Löhmannsröben, Hans‐Gerd; Hildebrandt, Niko

doi:10.1021/ja310317n

Cited by 172 publications

(186 citation statements)

References 41 publications

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“…RET processes include Förster resonance energy transfer (FRET) and bioluminescence or chemiluminescence (BRET and CRET) and can involve a variety of materials, including organic dyes, lanthanide complexes, fluorescent proteins, graphene, chemi-and bioluminescent enzymes, and inorganic materials like AuNPs, which may function as acceptors or donors and can be arranged in relays (63). The use of long-lifetime luminescent nanoparticles and RET agents allows for time-gated detection, which will be an important area of development for commercial nanoparticle biosensors (64). Minimizing donoracceptor separation is paramount, and nanoparticle biosensing components must be carefully chosen and constructed to achieve maximum sensitivity.…”

Section: Biosensing Mechanismsmentioning

confidence: 99%

Colloidal nanoparticles as advanced biological sensors

Howes

Chandrawati

Stevens

2014

Science

893

686

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Colloidal nanoparticle biosensors have received intense scientific attention and offer promising applications in both research and medicine. We review the state of the art in nanoparticle development, surface chemistry, and biosensing mechanisms, discussing how a range of technologies are contributing toward commercial and clinical translation. Recent examples of success include the ultrasensitive detection of cancer biomarkers in human serum and in vivo sensing of methyl mercury. We identify five key materials challenges, including the development of robust mass-scale nanoparticle synthesis methods, and five broader challenges, including the use of simulations and bioinformatics-driven experimental approaches for predictive modeling of biosensor performance. The resultant generation of nanoparticle biosensors will form the basis of high-performance analytical assays, effective multiplexed intracellular sensors, and sophisticated in vivo probes.Evolution has given rise to organisms of staggering complexity. Our now extensive knowledge of biological systems pales in comparison to the remaining mysteries. Unraveling these requires tools that probe the molecular machinery of life and provide detailed feedback on complex networks of subtle interactions. Such tools are cornerstones of biomedical research and practice, and improvements in these lead directly to a better understanding of fundamental biology, monitoring of health, and diagnosis of disease. Colloidal nanoparticle biosensors are a class of biological probe that will not only yield improved biological sensing but also provide a step change in our ability to probe the biomolecular realm. Nanoparticles can act as high-performance sensors because nanomaterials exhibit unique and useful behaviors not present in their bulk form: for example, bright tunable fluorescence from semiconductor nanoparticles and localized surface plasmon resonance (LSPR) phenomena in metallic nanoparticles. These particles exhibit intense responses to incident light (or other stimuli), and the ability to modulate this response by interaction with target analytes makes them excellent biosensor signal transducers. Outputs can be quantitative or qualitative depending on the functionality of

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Section: Biosensing Mechanismsmentioning

confidence: 99%

Colloidal nanoparticles as advanced biological sensors

Howes

Chandrawati

Stevens

2014

Science

893

686

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“…Algar et al, has also demonstrated the multiplexed tracking of protease activity using a single color of QD vector and a time-gated FRET mechanisms [21]. The steady state fluorescence based diagnostic methods have lately been complemented by their time resolved counterparts as Geißler and co-workers have shown a. Six-color time-resolved FRET for ultrasensitive multiplexed biosensing [22]. To establish a strong theoretical basis to this kind of platforms, Claussenet et al, have constructed complex logic functions implemented with QD bionanophotonic circuits [23].…”

Section: Quantum Dotsmentioning

confidence: 99%

Nano-Technological Approaches to Improve the Efficiency of Bio-Assays

Roy¹

2016

Glob J Biotechnol Biomater Sci

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One of the biggest issues in today's healthcare industry is to find a very fast yet effective diagnostic platform, which is suitable for our busy lifestyle without compromising the detection efficiency. The major requirements of an efficient diagnostic platform can be summarized as minimum sample requirement, least reagent requirement, having options of multiple tests in a single platform, high through put analysis and last but never the least a non-expensive sample run. The largest contributor of the bio-assay industry is protein based chromogenic bio-assays, which depends on strong antigen-antibody interaction and high emissive properties of reporter dye molecule attached to the antigen. In reality, this apparently simple looking reaction system has to face several difficulties before an appreciable signal is received by the photo-detector to give an analysable dataset. With the rapid emergence of nanotechnology during last few decades, some of the critical problems have been solved [1][2][3][4][5]. However, a large number of unresolved issues still remain there. This editorial will briefly address some of those critical issues and how they have been tackled by nanotechnology. The two-most tunable variables of a bio-assay platform are the reporter molecules and the sensing platform as described by the Figure 1. We will describe both the issues separately in the next sections. Optical efficiency of the reporter moleculeThe optical efficiency of a reporter molecule suffers from two major issues, firstly the biological auto-fluorescence and secondly the fluorescence quenching of the reporter dye by the aqueous environment. The biological autofluorescence originates from the

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“…The letters A and D denote whether the signal was collected in the donor or acceptor channel. The DCM has already proven its potential in multiplexed lung cancer assays 33 and protein concentration assays. 6 The method allows enhancing signal-to-background ratio.…”

Section: Pccp Papermentioning

confidence: 99%

Photophysical evaluation of a new functional terbium complex in FRET-based time-resolved homogenous fluoroassays

Cywiński

Nono²,

Charbonnière³

et al. 2014

Phys. Chem. Chem. Phys.

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A new functional luminescent lanthanide complex (LLC) has been synthesized with terbium as a central lanthanide ion and biotin as a functional moiety. Unlike in typical lanthanide complexes assembled via carboxylic moieties, in the presented complex, four phosphate groups are chelating the central lanthanide ion. This special chemical assembly enhances the complex stability in phosphate buffers conventionally used in biochemistry. The complex synthesis strategy and photophysical properties are described as well as the performance in time-resolved Förster Resonance Energy Transfer (FRET) assays. In those assays, this biotin-LLC transferred energy either to acceptor organic dyes (Cy5 or AF680) labelled on streptavidin or to quantum dots (QD655 or QD705) surfacefunctionalised with streptavidins. The permanent spatial donor-acceptor proximity is assured through strong and stable biotin-streptavidin binding. The energy transfer is evidenced from the quenching observed in donor emission and from a decrease in donor luminescence decay, both associated with simultaneous increase in acceptor intensity and in the decay time. The dye-based assays are realised in TRIS and in PBS, whereas QD-based systems are studied in borate buffer. The delayed emission analysis allows for quantifying the recognition process and for auto-fluorescence-free detection, which is particularly relevant for application in bioanalysis. In accordance with Förster theory, Förster-radii (R 0 ) were found to be around 60 Å for organic dyes and around 105 Å for QDs. The FRET efficiency (Z) reached 80% and 25% for dye and QD acceptors, respectively. Physical donor-acceptor distances (r) have been determined in the range 45-60 Å for organic dye acceptors, while for acceptor QDs between 120 Å and 145 Å. This newly synthesised biotin-LLC extends the class of highly sensitive analytical tools to be applied in the bioanalytical methods such as time-resolved fluoroimmunoassays (TR-FIA), luminescent imaging and biosensing.

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Six-Color Time-Resolved Förster Resonance Energy Transfer for Ultrasensitive Multiplexed Biosensing

Cited by 172 publications

References 41 publications

Colloidal nanoparticles as advanced biological sensors

Colloidal nanoparticles as advanced biological sensors

Nano-Technological Approaches to Improve the Efficiency of Bio-Assays

Photophysical evaluation of a new functional terbium complex in FRET-based time-resolved homogenous fluoroassays

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