ArticleeLife2022
A general approach for stabilizing nanobodies for intracellular expression.
Article in eLife, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed.
- The Promise of Single-Domain Antibodies as Ocular Therapeutics: A Narrative Review.International journal of molecular sciences · 2026Review
- Design principles for optogenetic-based targeted protein degradation.Synthetic and systems biotechnology · 2026Review
- Hypervariable loop profiling decodes sequence determinants of antibody stability.Nature structural & molecular biology · 2026Article
- High-Affinity Nanobody Against the LEDGF PWWP Domain Inhibits Chromatin Binding In Vitro.Biomolecules · 2026Article
- Proteolysis activity mapping and substrate discovery platform for identifying tumor-activated biosensors.Nature chemical biology · 2026Article
- Article
- Tiny tools closing the gap: nanobodies in research and therapy.Function (Oxford, England) · 2026Review
- Functionally-derivatized Nanoantibodies as novel tools for Giardia lamblia cyst detection.PLoS neglected tropical diseases · 2026Article
- Programmable nanobody circuits for cell selection.bioRxiv : the preprint server for biology · 2026Article
- Single-Molecule Imaging of Endogenous Proteins.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Nanobody-based gene therapy targeting complement component C3 reduces choroidal neovascularization in mice.Molecular therapy. Methods & clinical development · 2025Article
- Nanobodies targeting hnRNPA2/B1 and tau.bioRxiv : the preprint server for biology · 2025Article
- Modulating Subcellular Localization to Preserve the Stability and Functionality of Intracellular Nanobodies.Antibodies (Basel, Switzerland) · 2025Article
- Incidental vs. Engineered Nanoparticles in Alzheimer's and Parkinson's Disease: Pathological Pathways and Therapeutic Interventions.Nano research · 2025Article
- Progress toward new function and design of extracellular G protein-coupled receptor nanobodies.Molecular pharmacology · 2025Review
- Protein-based Radiopharmaceuticals that target fibroblast activation protein alpha: a review of current progress.EJNMMI radiopharmacy and chemistry · 2025Review
- Protease-Containing Nanobodies for Detecting and Manipulating Intracellular Antigens Using Antiviral Drugs.ACS chemical biology · 2025Article
- Nanobodies: From Discovery to AI-Driven Design.Biology · 2025Review
- A chimeric viral platform for directed evolution in mammalian cells.Nature communications · 2025Article
- Unique mechanisms to increase structural stability and enhance antigen binding in nanobodies.Structure (London, England : 1993) · 2025Article
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5 authors.
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Abstract
Conventional antibodies and their derived fragments are difficult to deploy against intracellular targets in live cells, due to their bulk and structural complexity. Nanobodies provide an alternative modality, with well-documented examples of intracellular expression. Despite their promise as intracellular reagents, there has not been a systematic study of nanobody intracellular expression. Here, we examined intracellular expression of 75 nanobodies from the Protein Data Bank. Surprisingly, a majority of these nanobodies were unstable in cells, illustrated by aggregation and clearance. Using comparative analysis and framework mutagenesis, we developed a general approach that stabilized a great majority of nanobodies that were originally unstable intracellularly, without significantly compromising target binding. This approach led to the identification of distinct sequence features that impacted the intracellular stability of tested nanobodies. Mutationally stabilized nanobody expression was found to extend to in vivo contexts, in the murine retina and in
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