ArticleCurrent pharmaceutical biotechnology2008
Single molecule studies of multiple-fluorophore labeled antibodies. Effect of homo-FRET on the number of photons available before photobleaching.
Article in Current pharmaceutical biotechnology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
18 citing papers in PubMed, 56 citations in OpenAlex.
- Improvement of the Stability and Optical Properties of CsPbBrNanomaterials (Basel, Switzerland) · 2023Article
- A method for parallel microscale protein labeling and precise control over the average degree of labeling (aDoL).Scientific reports · 2023Article
- Evaluating the Effect of Dye-Dye Interactions of Xanthene-Based Fluorophores in the Fluorosequencing of Peptides.Bioconjugate chemistry · 2022Article
- Exploiting Complex Fluorophore Interactions to Monitor Virus Capsid Disassembly.Molecules (Basel, Switzerland) · 2021Article
- Not so innocent: Impact of fluorophore chemistry on the in vivo properties of bioconjugates.Current opinion in chemical biology · 2021Review
- High-Performance Near-Infrared Fluorescent Secondary Antibodies for Immunofluorescence.Analytical chemistry · 2021Article
- In-gel fluorescence detection by DNA polymerase elongation.APL bioengineering · 2020Article
- The Effect of Fluorophore Conjugation on Antibody Affinity and the Photophysical Properties of Dyes.Biophysical journal · 2018Article
- Design and applications of a clamp for Green Fluorescent Protein with picomolar affinity.Scientific reports · 2017Article
- Strong signal increase in STED fluorescence microscopy by imaging regions of subdiffraction extent.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Superresolution Imaging Identifies That Conventional Trafficking Pathways Are Not Essential for Endoplasmic Reticulum to Outer Mitochondrial Membrane Protein Transport.Scientific reports · 2017Article
- Förster Resonance Energy Transfer between Quantum Dot Donors and Quantum Dot Acceptors.Sensors (Basel, Switzerland) · 2015Review
- FRET enhanced fluorescent nanodiamonds.Current pharmaceutical biotechnology · 2014Article
- Article
- Reversible fluorescence photoswitching in DNA.The journal of physical chemistry. B · 2012Article
- Quantitative characterization of fluorophores in multi-component nanoprobes by single-molecule fluorescence.Biomedical optics express · 2011Article
- Self-quenching of uranin: Instrument response function for color sensitive photo-detectors.Journal of luminescence · 2010Article
- Stoichiometry of a regulatory splicing complex revealed by single-molecule analyses.The EMBO journal · 2010Article
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 3 countries.
Funding
Abstract
Advancements in single molecule detection (SMD) continue to unfold powerful ways to study the behavior of individual and complex molecular systems in real time. SMD enables the characterization of complex molecular interactions and reveals basic physical phenomena underlying chemical and biological processes. We present here a systematic study of the quenching efficiency of Förster-type energy-transfer (FRET) for multiple fluorophores immobilized on a single antibody. We simultaneously monitor the fluorescence intensity, fluorescence lifetime, and the number of available photons before photobleaching as a function of the number of identical emitters bound to a single IgG antibody. The detailed studies of FRET between individual fluorophores reveal complex through-space interactions. In general, even for two or three fluorophores immobilized on a single protein, homo-FRET interactions lead to an overall non-linear intensity increase and shortening of fluorescence lifetime. Over-labeling of protein in solution (ensemble) results in the loss of fluorescence signal due to the self-quenching of fluorophores making it useless for assays applications. However, in the single molecule regime, over-labeling may bring significant benefits in regards to the number of available photons and the overall survival time. Our investigation reveals possibilities to significantly increase the observation time for a single macromolecule allowing studies of macromolecular interactions that are not obscured by ensemble averaging. Extending the observation time will be crucial for developing immunoassays based on single-antibody.
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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.