ArticleJournal of nucleic acids2012
In vitro selection of fab fragments by mRNA display and gene-linking emulsion PCR.
Article in Journal of nucleic acids, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 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
12 citing papers in PubMed, 28 citations in OpenAlex.
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Directing evolution of novel ligands by mRNA display.Chemical Society reviews · 2021Review
- Liquid drop of DNA libraries reveals total genome information.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Cell-Free Protein Synthesis: A Promising Option for Future Drug Development.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2020Review
- Review
- Synthetic Biology Goes Cell-Free.BMC biology · 2019Review
- Naïve Human Antibody Libraries for Infectious Diseases.Advances in experimental medicine and biology · 2017Review
- Directed evolution of anti-HER2 DARPins by SNAP display reveals stability/function trade-offs in the selection process.Protein engineering, design & selection : PEDS · 2015Article
- Conceptual and methodological advances in cell-free directed evolution.Current opinion in structural biology · 2015Review
- Advances in the directed evolution of proteins.Current opinion in chemical biology · 2014Review
- Towards personalized medicine mediated by in vitro virus-based interactome approaches.International journal of molecular sciences · 2014Review
- In vitro Fab display: a cell-free system for IgG discovery.Protein engineering, design & selection : PEDS · 2014Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In vitro selection by display methods has been an effective tool for engineering recombinant antibodies. mRNA display based on a cell-free translation system has the advantages of larger library sizes and quicker selection procedures compared with cell-based display methods such as phage display. However, mRNA display has been limited to select single-chain polypeptides such as scFvs due to its characteristic of linking a nascent polypeptide with its encoding mRNA on the ribosome. Here we demonstrated a new way of selecting heterodimeric Fab fragments by using mRNA display combined with emulsion PCR. We designed a pair of complementary 5' UTR sequences that can link the Fab heavy and light chain genes together by overlap-extension PCR in water-in-oil emulsions. We confirmed that two mRNA-displayed polypeptides for heavy and light chain of a model Fab fragment were associated into the active form and that a specific Fab fragment gene was enriched over 100-fold per round of a model affinity selection followed by the gene-linking emulsion PCR. We further performed directed evolution of Fab fragments with higher binding activity from a randomized Fab fragment library.
Identifiers
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.