ArticleScientific reports2020
A novel epitope tagging system to visualize and monitor antigens in live cells with chromobodies.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Antigen-Detected NMR for Minimal Epitope Engineering and Structure-Guided Selection of a NaAdvanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Intracellular protein binders for imaging, control and future therapeutics.Nature biomedical engineering · 2026Review
- A nanobody-based degron system for targeted protein knockdown in Dictyostelium discoideum.BMC molecular and cell biology · 2026Article
- IPTG- and estradiol-inducible gene expression systems in the unicellular red alga Cyanidioschyzon merolae.Plant physiology · 2026Article
- Broad application of the ALFA tagging technology for in planta nanobody-based imaging and biochemical characterization of plant proteins.Plant communications · 2025Article
- Evaluation of AlphaFold modeling for elucidation of nanobody-peptide epitope interactions.The Journal of biological chemistry · 2025Article
- Fluorescent labeling of proteins in vitro and in vivo using encoded peptide tags.The Journal of biological chemistry · 2025Review
- Evaluation of Alphafold modeling for elucidation of nanobody-peptide epitope interactions.bioRxiv : the preprint server for biology · 2025Article
- Nanobodies as novel tools to monitor the mitochondrial fission factor Drp1.Life science alliance · 2024Article
- A nanobody-based strategy for rapid and scalable purification of human protein complexes.Nature protocols · 2024Review
- Making the effect visible - OX40 targeting nanobodies forFrontiers in immunology · 2024Article
- Protein expression/secretion boost by a novel unique 21-mer cis-regulatory motif (Exin21) via mRNA stabilization.Molecular therapy : the journal of the American Society of Gene Therapy · 2023Article
- Single-molecule visualization of mRNA circularization during translation.Experimental & molecular medicine · 2023Review
- Characterization of a Nanobody-Epitope Tag Interaction and Its Application for Receptor Engineering.ACS chemical biology · 2022Article
- Affimers and nanobodies as molecular probes and their applications in imaging.Journal of cell science · 2022Article
- A Light-Activatable Photocaged Variant of the Ultra-High Affinity ALFA-Tag Nanobody.Chembiochem : a European journal of chemical biology · 2022Article
- Single-domain near-infrared protein provides a scaffold for antigen-dependent fluorescent nanobodies.Nature methods · 2022Article
- Biparatopic nanobodies protect mice from lethal challenge with SARS-CoV-2 variants of concern.EMBO reports · 2022Article
- Tripartite split-GFP assay to identify selective intracellular nanobody that suppresses GTPase RHOA subfamily downstream signaling.Frontiers in immunology · 2022Article
- A Nanobody-Based Toolset to Monitor and Modify the Mitochondrial GTPase Miro1.Frontiers in molecular biosciences · 2022Article
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Epitope tagging is a versatile approach to study different proteins using a well-defined and established methodology. To date, most epitope tags such as myc, HA, V5 and FLAG tags are recognized by antibodies, which limits their use to fixed cells, tissues or protein samples. Here we introduce a broadly applicable tagging strategy utilizing a short peptide tag (PepTag) which is specifically recognized by a nanobody (PepNB). We demonstrated that the PepNB can be easily functionalized for immunoprecipitation or direct immunofluorescence staining of Pep-tagged proteins in vitro. For in cellulo studies we converted the PepNB into a fluorescently labeled Pep-chromobody (PepCB) which is functionally expressed in living cells. The addition of the small PepTag does not interfere with the examined structures in different cellular compartments and its detection with the PepCB enables optical antigen tracing in real time. By employing the phenomenon of antigen-mediated chromobody stabilization (AMCBS) using a turnover-accelerated PepCB we demonstrated that the system is suitable to visualize and quantify changes in Pep-tagged antigen concentration by quantitative live-cell imaging. We expect that this novel tagging strategy offers new opportunities to study the dynamic regulation of proteins, e.g. during cellular signaling, cell differentiation, or upon drug action.
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