ArticlePloS one2015
Optimizing Production of Antigens and Fabs in the Context of Generating Recombinant Antibodies to Human Proteins.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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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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Who cites it
13 citing papers in PubMed, 28 citations in OpenAlex.
- RP3Net: a deep learning model for predicting recombinant protein production in Escherichia coli.Bioinformatics (Oxford, England) · 2026Article
- Impact of ions, pH and the nature of substrate on the structure and activity of a robust single-stranded DNA binding (SSB)-like protein from Phi11.Archives of microbiology · 2025Article
- A potent synthetic nanobody with broad-spectrum activity neutralizes SARS-CoV-2 virus and the Omicron variant BA.1 through a unique binding mode.Journal of nanobiotechnology · 2022Article
- Article
- Multifaceted antibodies development against synthetic α-dystroglycan mucin glycopeptide as promising tools for dystroglycanopathies diagnostic.Glycoconjugate journal · 2020Article
- An efficient ORF selection system for DNA fragment libraries based on split beta-lactamase complementation.PloS one · 2020Article
- Circulating Levels of Interferon Regulatory Factor-5 Associates With Subgroups of Systemic Lupus Erythematosus Patients.Frontiers in immunology · 2019Article
- Biotin-tagged proteins: Reagents for efficient ELISA-based serodiagnosis and phage display-based affinity selection.PloS one · 2018Article
- A synthetic intrabody-based selective and generic inhibitor of GPCR endocytosis.Nature nanotechnology · 2017Article
- Structural basis for arginine methylation-independent recognition of PIWIL1 by TDRD2.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Effect of DNA sequence of Fab fragment on yield characteristics and cell growth of E. coli.Scientific reports · 2017Article
- Generating Recombinant Antibodies to Membrane Proteins through Phage Display.Antibodies (Basel, Switzerland) · 2016Review
- Systematic comparison of monoclonal versus polyclonal antibodies for mapping histone modifications by ChIP-seq.Epigenetics & chromatin · 2016Article
Corrections and comments
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Authors and funding
23 authors at 4 institutions in 2 countries.
Funding
Abstract
We developed and optimized a high-throughput project workflow to generate renewable recombinant antibodies to human proteins involved in epigenetic signalling. Three different strategies to produce phage display compatible protein antigens in bacterial systems were compared, and we found that in vivo biotinylation through the use of an Avi tag was the most productive method. Phage display selections were performed on 265 in vivo biotinylated antigen domains. High-affinity Fabs (<20nM) were obtained for 196. We constructed and optimized a new expression vector to produce in vivo biotinylated Fabs in E. coli. This increased average yields up to 10-fold, with an average yield of 4 mg/L. For 118 antigens, we identified Fabs that could immunoprecipitate their full-length endogenous targets from mammalian cell lysates. One Fab for each antigen was converted to a recombinant IgG and produced in mammalian cells, with an average yield of 15 mg/L. In summary, we have optimized each step of the pipeline to produce recombinant antibodies, significantly increasing both efficiency and yield, and also showed that these Fabs and IgGs can be generally useful for chromatin immunoprecipitation (ChIP) protocols.
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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.