ArticleAnalytical biochemistry1995
Fluorescence energy transfer immunoassay based on a long-lifetime luminescent metal-ligand complex.
Article in Analytical biochemistry, 1995. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 87 citations in OpenAlex.
- Applications of nanoparticles in plant disease identification and control for sustainable crop production.Discover nano · 2026Review
- Long-distance tmFRET using bipyridyl- and phenanthroline-based ligands.Biophysical journal · 2024Article
- Long-distance tmFRET using bipyridyl- and phenanthroline-based ligands.bioRxiv : the preprint server for biology · 2024Article
- Ru(II)-diimine functionalized metalloproteins: From electron transfer studies to light-driven biocatalysis.Biochimica et biophysica acta · 2016Review
- Photophysical studies of bioconjugated ruthenium metal-ligand complexes incorporated in phospholipid membrane bilayers.Inorganic chemistry · 2013Article
- Nonfluorescent quenchers to correlate single-molecule conformational and compositional dynamics.Journal of the American Chemical Society · 2012Article
- A high throughput fluorescence polarization assay for inhibitors of the GoLoco motif/G-alpha interaction.Combinatorial chemistry & high throughput screening · 2008Article
- Comparing the performance of the optical glucose assay based on glucose binding protein with high-performance anion-exchange chromatography with pulsed electrochemical detection: efforts to design a low-cost point-of-care glucose sensor.Journal of diabetes science and technology · 2007Article
- Real-time background suppression during frequency domain lifetime measurements.Analytical biochemistry · 2002Article
- High-molecular-weight protein hydrodynamics studied with a long-lifetime metal-ligand complex.Biochimica et biophysica acta · 2002Article
- Novel fluorescence sensing methods for high throughput screening.Journal of biomolecular screening · 2000Article
- Resonance energy transfer study using a rhenium metal-ligand lipid conjugate as the donor in a model membrane.Chemistry and physics of lipids · 1999Article
- Polarization sensing with visual detection.Analytical chemistry · 1999Article
- Temporally and spectrally resolved imaging microscopy of lanthanide chelates.Biophysical journal · 1998Article
- Creation of Metal-to-Ligand Charge Transfer Excited States with Two-Photon Excitation.Inorganic chemistry · 1997Article
- Synthesis and spectral characterization of a thiol-reactive long-lifetime Ru(II) complex.Analytical biochemistry · 1997Article
- Lifetime-based sensing of glucose using energy transfer with a long lifetime donor.Analytical biochemistry · 1997Article
- Long-lifetime metal-ligand pH probe.Analytical biochemistry · 1997Article
- Synthesis and evaluation of Ru-complexes as anisotropy probes for protein hydrodynamics and immunoassays of high-molecular-weight antigens.Biophysical chemistry · 1996Article
- Synthesis and characterization of a sulfhydryl-reactive rhenium metal-ligand complex.Bioconjugate chemistryArticle
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
We describe an immunoassay based on fluorescence resonance energy transfer (FRET). The antigen was human serum albumin (HSA), which was labeled with a ruthenium-ligand complex, [Ru(bpy)2(phen-ITC)]2+. The antibody (IgG) to HSA was labeled with a nonfluorescent absorber, Reactive Blue 4. Association of the Ru-labeled HSA with the antibody was detected by three spectral parameters, a decreased quantum yield of Ru-HSA, a decrease in its fluorescence lifetime, and an increase in its fluorescence anisotropy. The steady-state anisotropy of Ru-HSA increased approximately eightfold upon binding to the antibody. These spectral effects were observed both in the direct association of the Ru-HSA with Reactive Blue 4-labeled antibody, and in a competitive assay format wherein unlabeled HSA competed with Ru-HSA for the binding sites on the antibody. Some nonspecific interactions of HSA may have occurred with Reactive Blue 4-labeled AHA, a difficulty which can be avoided with a different acceptor. The use of FRET provides a reliable means to alter the spectral properties upon antigen-antibody binding. The advantages of a ruthenium-ligand fluorophore include its long-wavelength absorption and emission, long fluorescence lifetime, and high photo-stability. Long wavelengths minimize problems of autofluorescence from biological samples, and long life-times allow off-gating of the prompt autofluorescence.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.