ArticleMolecular biotechnology2013
Directed evolution of a fluorogen-activating single chain antibody for function and enhanced brightness in the cytoplasm.
Article in Molecular biotechnology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 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
11 citing papers in PubMed, 22 citations in OpenAlex.
- Genetically Encoded Activators of Small Molecules for Imaging and Drug Delivery.Angewandte Chemie (International ed. in English) · 2020Review
- Next-Generation Fluorogen-Based Reporters and Biosensors for Advanced Bioimaging.International journal of molecular sciences · 2019Review
- Fluorogen-activating proteins: beyond classical fluorescent proteins.Acta pharmaceutica Sinica. B · 2018Review
- Development of cyanine probes with dinitrobenzene quencher for rapid fluorogenic protein labelling.Philosophical transactions. Series A, Mathematical, physical, and engineering sciences · 2017Article
- Breaking the color barrier - a multi-selective antibody reporter offers innovative strategies of fluorescence detection.Journal of cell science · 2017Article
- Fluorogenic Labeling Strategies for Biological Imaging.International journal of molecular sciences · 2017Review
- A Critical and Comparative Review of Fluorescent Tools for Live-Cell Imaging.Annual review of physiology · 2017Review
- Shedding light on biology of bacterial cells.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2016Review
- Engineering tandem single-chain Fv as cell surface reporters with enhanced properties of fluorescence detection.Protein engineering, design & selection : PEDS · 2015Article
- Rapid, specific, no-wash, far-red fluorogen activation in subcellular compartments by targeted fluorogen activating proteins.ACS chemical biology · 2015Article
- Localization microscopy using noncovalent fluorogen activation by genetically encoded fluorogen-activating proteins.Chemphyschem : a European journal of chemical physics and physical chemistry · 2014Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
Directed evolution is an exceptionally powerful tool that uses random mutant library generation and screening techniques to engineer or optimize functions of proteins. One class of proteins for which this process is particularly effective is antibodies, where properties such as antigen specificity and affinity can be selected to yield molecules with improved efficacy as molecular labels or in potential therapeutics. Typical antibody structure includes disulfide bonds that are required for stability and proper folding of the domains. However, these bonds are unable to form in the reducing environment of the cytoplasm, stymieing the effectiveness of optimized antibodies in many research applications. We have removed disulfide-forming cysteine residues in a single chain antibody fluorogen-activating protein (FAP), HL4, and employed directed evolution to select a derivative that is capable of activity in the cytoplasm. A subsequent round of directed evolution was targeted at increasing the overall brightness of the fluoromodule (FAP-fluorogen complex). Ultimately, this approach produced a novel FAP that exhibits strong activation of its cognate fluorogen in the reducing environment of the cytoplasm, significantly expanding the range of applications for which fluoromodule technology can be utilized.
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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.