ArticleProtein engineering, design & selection : PEDS2014
Optimizing recombinant antibodies for intracellular function using hitchhiker-mediated survival selection.
Article in Protein engineering, design & selection : PEDS, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 29 citations in OpenAlex.
- Article
- Modified protein selection strategy based on Escherichia coli's Hitchhiker transport and validation through selection of nanobodies targeting bovine interferon gamma.Scientific reports · 2026Article
- Isolation of Antigen-Specific Nanobodies From Synthetic Libraries Using a Protein Selection Strategy That Combines MACS-Based Screening of YSD and FLI-TRAP.Bio-protocol · 2026Article
- Isolation of PCSK9-specific nanobodies from synthetic libraries using a combined protein selection strategy.Scientific reports · 2025Article
- Transcytosis-Driven Treatment of Neurodegenerative Disorders by mRNA-Expressed Antibody-Transferrin Conjugates.Biomedicines · 2024Review
- Efficient gene activation in plants by the MoonTag programmable transcriptional activator.Nucleic acids research · 2023Article
- Isolation of full-length IgG antibodies from combinatorial libraries expressed in the cytoplasm of Escherichia coli.Nature communications · 2023Article
- Efficient gene activation in plants by the MoonTag programmable transcriptional activator.bioRxiv : the preprint server for biology · 2023Article
- Disulfide-compatible phage-assisted continuous evolution in the periplasmic space.Nature communications · 2021Article
- A survival selection strategy for engineering synthetic binding proteins that specifically recognize post-translationally phosphorylated proteins.Nature communications · 2019Article
- Selection of Protein-Protein Interactions of Desired Affinities with a Bandpass Circuit.Journal of molecular biology · 2019Article
- Computational affinity maturation of camelid single-domain intrabodies against the nonamyloid component of alpha-synuclein.Scientific reports · 2018Article
- Continuous directed evolution of proteins with improved soluble expression.Nature chemical biology · 2018Article
- Directed evolution to improve protein folding in vivo.Current opinion in structural biology · 2018Review
- Recombinant Antibody Fragments for Neurodegenerative Diseases.Current neuropharmacology · 2017Review
- Review
- Article
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
Abstract
The 'hitchhiker' mechanism of the bacterial twin-arginine translocation pathway has previously been adapted as a genetic selection for detecting pairwise protein interactions in the cytoplasm of living Escherichia coli cells. Here, we extended this method, called FLI-TRAP, for rapid isolation of intracellular antibodies (intrabodies) in the single-chain Fv format that possess superior traits simply by demanding bacterial growth on high concentrations of antibiotic. Following just a single round of survival-based enrichment using FLI-TRAP, variants of an intrabody against the dimerization domain of the yeast Gcn4p transcription factor were isolated having significantly greater intracellular stability that translated to yield enhancements of >10-fold. Likewise, an intrabody specific for the non-amyloid component region of α-synuclein was isolated that has ~8-fold improved antigen-binding affinity. Collectively, our results illustrate the potential of the FLI-TRAP method for intracellular stabilization and affinity maturation of intrabodies, all without the need for purification or immobilization of the antigen.
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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.