Ultrasensitive bioanalytical assays using time-resolved fluorescence detection

Dickson, Eva F. Gudgin; Pollak, Alfred; Diamandis, Eleftherios P.

doi:10.1016/0163-7258(94)00078-h

Cited by 123 publications

(12 citation statements)

References 165 publications

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“…Like fluorescence resonance energy transfer (FRET), distance-dependent changes in intramolecular LRET efficiency can be employed to detect molecular interactions or conformational states 20 , 22 , 36 . To explore this capability in the context of trLRET imaging, we tested whether we could visualize the binding of two macromolecules in a live animal.…”

Section: Resultsmentioning

confidence: 99%

“…Lanthanide imaging systems have not yet achieved the signal intensities and detection sensitivities required for routine applications, and they do not surpass the capabilities of conventional fluorescence microscopy. This is paradoxical, given the predominance of lanthanide probes in ultrasensitive solution- and cell-based photometric assays 20 – 22 . Here we identify and solve three problems that have limited current lanthanide imaging systems.…”

Section: Introductionmentioning

confidence: 99%

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Ultrasensitive optical imaging with lanthanide lumiphores

et al. 2017

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In principle, the millisecond emission lifetimes of lanthanide chelates should enable their ultrasensitive detection in biological systems by time-resolved optical microscopy. In practice, however, lanthanide imaging techniques have provided no better sensitivity than conventional fluorescence microscopy. Here, we identify three fundamental problems that have impeded lanthanide microscopy: low photon flux, inefficient excitation, and optics-derived background luminescence. We overcome these limitations with a new lanthanide imaging modality, trans-reflected illumination with luminescence resonance energy transfer (trLRET), which increases the time-integrated signal intensities of lanthanide lumiphores by 170-fold and the signal-to-background ratios by 75-fold. We demonstrate that trLRET provides at least an order-of-magnitude increase in detection sensitivity over conventional epifluorescence microscopy when used to visualize endogenous protein expression in zebrafish embryos. We also show that trLRET can be used to optically detect molecular interactions in vivo. trLRET promises to unlock the full potential of lanthanide lumiphores for ultrasensitive, autofluorescence-free biological imaging.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Ultrasensitive optical imaging with lanthanide lumiphores

et al. 2017

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“…A major step forward in the high throughput screening of biological entities came with the advent of homogeneous time resolved FRET assays such as LANCE ® and HTRF ® [8,9] coupled with a tool box of reagents and labelling chemistries, Many of the heterogeneous assay formats used for studying ligand-receptor interactions were easily adapted to simple mix and measure homogeneous assay formats which could also be miniaturised to 384 well format, allowing an increase in throughput. These assays were also tolerant of crude bacterial supernatants from E. coli , crucial for the screening of antibodies from phage display libraries.…”

Section: Homogeneous Biochemical Assay Formatsmentioning

confidence: 99%

Evolution of Biologics Screening Technologies

Cariuk¹,

Gardener²,

Vaughan³

2013

Pharmaceuticals

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Screening for biologics, in particular antibody drugs, has evolved significantly over the last 20 years. Initially, the screening processes and technologies from many years experience with small molecules were adopted and modified to suit the needs of biologics discovery. Since then, antibody drug discovery has matured significantly and is today investing earlier in new technologies that commercial suppliers are now developing specifically to meet the growing needs of large molecule screening. Here, we review the evolution of screening and automation technologies employed in antibody discovery and highlight the benefits that these changes have brought.

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“…Using europium (Eu) chelates as the labels, Time-resolved fluoroimmunoassay (TRFIA) was considered as a successful non-isotopic detection method since it was first reported by Lovgren et al . 18 , and had been noticed as an excellent performance method and employed in sorts of biomedical sciences fields 19 – 26 . Time-resolved fluoroimmunoassay gets excellent precision, higher sensitivity and extremely wider detection range relative to traditional ELISA 27 .…”

Section: Introductionmentioning

confidence: 99%

A time-resolved fluoroimmunoassay to assay the rabies virus glycoprotein: application for estimation of human rabies vaccine potency

Lin

Chen

Zhao

et al. 2017

Sci Rep

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Replacement of the in vivo rabies vaccine potency test (NIH test) by in vitro methods had been discussed by several researcher including WHO expert working groups. In this paper, a time-resolved fluoroimmunoassay (TRFIA) for the assay of rabies virus glycoprotein in rabies vaccine was first established to estimate the rabies vaccine potency by using specific monoclonal antibody that only recognized the native, trimeric and immunogenic form of rabies virus glycoprotein. Potency of the rabies virus glycoprotein was assayed with satisfactory performance under optimal conditions, and the method demonstrated satisfactory results when applied in practical samples. The correlation coefficient of potency values obtained from the present TRFIA and ELISA was 0.912, and 0.903 for those from the present TRFIA and NIH test. These preliminary results confirmed that this TRFIA can replace ELISA with higher performance, and could be a promising replacement of the NIH test. Based upon these results, the present TRFIA seemed to be a convenient tool for evaluating rabies vaccine potency and its products at different stages accordingly.

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Ultrasensitive bioanalytical assays using time-resolved fluorescence detection

Cited by 123 publications

References 165 publications

Ultrasensitive optical imaging with lanthanide lumiphores

Ultrasensitive optical imaging with lanthanide lumiphores

Evolution of Biologics Screening Technologies

A time-resolved fluoroimmunoassay to assay the rabies virus glycoprotein: application for estimation of human rabies vaccine potency

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