ArticleBMC biotechnology2007
A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm.
Article in BMC biotechnology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed.
- Egyptian rousette bat humoral immunity to H9 influenza hemagglutinin.bioRxiv : the preprint server for biology · 2026Article
- AI-assisted protein design to rapidly convert antibody sequences to intrabodies targeting diverse peptides and histone modifications.Science advances · 2026Article
- Generation Using Phage-Display of pH-Dependent Antibodies Against the Tumor-Associated Antigen AXL.Antibodies (Basel, Switzerland) · 2025Article
- AI-assisted protein design to rapidly convert antibody sequences to intrabodies targeting diverse peptides and histone modifications.bioRxiv : the preprint server for biology · 2025Article
- Comprehensive characterization and database construction of immune repertoire in the largest Chinese glioma cohort.iScience · 2024Article
- Stealth-Engineered Albumin-Coated Nanoparticles for Targeted Therapy: Effective Drug Delivery and Tumor Suppression in Xenograft-Zebrafish Model.International journal of nanomedicine · 2024Article
- Antibody-mediated neutralization of galectin-3 as a strategy for the treatment of systemic sclerosis.Nature communications · 2023Article
- Designed Ankyrin Repeat Proteins as a tool box for analyzing p63.Cell death and differentiation · 2022Article
- Antibody-mediated enzyme formation: Its legacy at age fifty-four.Journal of molecular recognition : JMR · 2021Review
- Anti-Müllerian hormone concentration regulates activin receptor-like kinase-2/3 expression levels with opposing effects on ovarian cancer cell survival.International journal of oncology · 2021Article
- Anti-Müllerian hormone (AMH) autocrine signaling promotes survival and proliferation of ovarian cancer cells.Scientific reports · 2021Article
- An Inside Job: Applications of Intracellular Single Domain Antibodies.Biomolecules · 2020Review
- Immunotherapy of triple-negative breast cancer with cathepsin D-targeting antibodies.Journal for immunotherapy of cancer · 2019Article
- Tuning recombinant protein expression to match secretion capacity.Microbial cell factories · 2018Article
- A Two-Step Approach for the Design and Generation of Nanobodies.International journal of molecular sciences · 2018Article
- Review
- Protein engineering strategies for the development of viral vaccines and immunotherapeutics.FEBS letters · 2014Review
- In-cell intrabody selection from a diverse human library identifies C12orf4 protein as a new player in rodent mast cell degranulation.PloS one · 2014Article
- Expression of recombinant antibodies.Frontiers in immunology · 2013Article
- Article
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Authors and funding
10 authors.
Funding
Abstract
backgroundIntrabodies are defined as antibody molecules which are ectopically expressed inside the cell. Such intrabodies can be used to visualize or inhibit the targeted antigen in living cells. However, most antibody fragments cannot be used as intrabodies because they do not fold under the reducing conditions of the cell cytosol and nucleus.
resultsWe describe the construction and validation of a large synthetic human single chain antibody fragment library based on a unique framework and optimized for cytoplasmic expression. Focusing the library by mimicking the natural diversity of CDR3 loops ensured that the scFvs were fully human and functional. We show that the library is highly diverse and functional since it has been possible to isolate by phage-display several strong binders against the five proteins tested in this study, the Syk and Aurora-A protein kinases, the alphabeta tubulin dimer, the papillomavirus E6 protein and the core histones. Some of the selected scFvs are expressed at an exceptional high level in the bacterial cytoplasm, allowing the purification of 1 mg of active scFv from only 20 ml of culture. Finally, we show that after three rounds of selection against core histones, more than half of the selected scFvs were active when expressed in vivo in human cells since they were essentially localized in the nucleus.
conclusionThis new library is a promising tool not only for an easy and large-scale selection of functional intrabodies but also for the isolation of highly expressed scFvs that could be used in numerous biotechnological and therapeutic applications.
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