ArticleProtein engineering, design & selection : PEDS2010
Physico-chemical determinants of soluble intrabody expression in mammalian cell cytoplasm.
Article in Protein engineering, design & selection : PEDS, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
What it found
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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
30 citing papers in PubMed, 63 citations in OpenAlex.
- Reliable repurposing of the antibody interactome inside the cell.Nature communications · 2026Article
- Modulating Subcellular Localization to Preserve the Stability and Functionality of Intracellular Nanobodies.Antibodies (Basel, Switzerland) · 2025Article
- A simeprevir-inducible molecular switch for the control of cell and gene therapies.Nature communications · 2023Article
- TauCell reports methods · 2022Article
- A Small Virus to Deliver Small Antibodies: New Targeted Therapies Based on AAV Delivery of Nanobodies.Microorganisms · 2021Review
- Nanobodies Right in the Middle: Intrabodies as Toolbox to Visualize and Modulate Antigens in the Living Cell.Biomolecules · 2020Review
- Applying Antibodies Inside Cells: Principles and Recent Advances in Neurobiology, Virology and Oncology.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2020Review
- Engineered Extracellular Vesicles/Exosomes as a New Tool against Neurodegenerative Diseases.Pharmaceutics · 2020Review
- Computational affinity maturation of camelid single-domain intrabodies against the nonamyloid component of alpha-synuclein.Scientific reports · 2018Article
- A comprehensive in silico characterization of bacterial signal peptides for the excretory production of3 Biotech · 2018Article
- Modifications of a signal sequence for antibody secretion from insect cells.Cytotechnology · 2018Article
- Elimination of TDP-43 inclusions linked to amyotrophic lateral sclerosis by a misfolding-specific intrabody with dual proteolytic signals.Scientific reports · 2018Article
- Exploiting Phage Display for Development of Novel Cellular Targeting Strategies.Methods in molecular biology (Clifton, N.J.) · 2018Article
- Targeted Intracellular Delivery of Antibodies: The State of the Art.Frontiers in pharmacology · 2018Review
- Engineered Autonomous Human Variable Domains.Current pharmaceutical design · 2016Review
- Bifunctional Anti-Non-Amyloid Component α-Synuclein Nanobodies Are Protective In Situ.PloS one · 2016Article
- Enhanced production of a single domain antibody with an engineered stabilizing extra disulfide bond.Microbial cell factories · 2015Article
- Visualizing posttranslational and epigenetic modifications of endogenous proteins in vivo.Histochemistry and cell biology · 2015Review
- Functional inhibition of β-catenin-mediated Wnt signaling by intracellular VHH antibodies.mAbs · 2015Article
- Lysine and arginine content of proteins: computational analysis suggests a new tool for solubility design.Molecular pharmaceutics · 2014Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 4 institutions in 1 country.
Funding
Abstract
Soluble antibody fragments are desirable not only as potential therapeutic and diagnostic agents for extracellular targets but also as 'intrabodies' for functional genomics, proteomics and gene therapy inside cells. However, antibody fragments are notoriously aggregation-prone when expressed intracellularly, due in part to unfavorable redox potential and macromolecular crowding in cell cytoplasm. Only a small proportion of intrabodies are soluble in cytoplasm and little is known about the sequence determinants that confer such stability. By comparing the cytoplasmic expression of several related human single-chain variable fragments and camelid V(HH)s in mammalian cells, we report that intrabody solubility is highly influenced by CDR content and is improved by an overall negative charge at cytoplasmic pH and reduced hydrophilicity. We hypothesize that ionic repulsion and weak hydrophobic interactions compensate, to different extents, for impaired disulfide bond formation in cytoplasm, thereby decreasing the risk for intrabody aggregation. As proof of principle, we demonstrate that the soluble expression of an aggregation-prone positively charged intrabody is modestly enhanced via cis or trans acidification using highly charged peptide tags (3XFLAG tag, SV40 NLS). These findings suggest that simple sequence analysis and electrostatic manipulation may aid in predicting and engineering solubility-enhanced intrabodies from antibody libraries for intracellular use.
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What OpenQuestion holds
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.