ArticleScientific reports2020
Golden Gate assembly with a bi-directional promoter (GBid): A simple, scalable method for phage display Fab library creation.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 16 citations in OpenAlex.
- Facile, high-throughput protein purification enabled by high-efficiency protease elution.Protein science : a publication of the Protein Society · 2026Article
- Integrating fab phage and yeast display through seamless transfer of paired heavy- and light-chain libraries for efficient antibody discovery.Antibody therapeutics · 2026Article
- Dual-mode ClfA-targeting DARPin biologics protect against diverse methicillin-resistantbioRxiv : the preprint server for biology · 2026Article
- Construction of Naïve and Immune Human Fab Phage Display Library.Methods in molecular biology (Clifton, N.J.) · 2026Article
- An integrated technology for quantitative wide mutational scanning of human antibody Fab libraries.Nature communications · 2024Article
- An integrated technology for quantitative wide mutational scanning of human antibody Fab libraries.bioRxiv : the preprint server for biology · 2024Article
- Standardizing cassette-based deep mutagenesis by Golden Gate assembly.Biotechnology and bioengineering · 2024Article
- Methods for Engineering Binders to Multi-Pass Membrane Proteins.Bioengineering (Basel, Switzerland) · 2023Review
- An efficient cloning method to expand vector and restriction site compatibility of Golden Gate Assembly.Cell reports methods · 2023Article
- Construction of Naïve and Immune Human Fab Phage Display Library.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Chicken-derived CD20 antibodies with potent B-cell depletion activity.British journal of haematology · 2022Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
Abstract
Fabs offer an attractive platform for monoclonal antibody discovery/engineering, but library construction can be cumbersome. We report a simple method - Golden Gate assembly with a bi-directional promoter (GBid) - for constructing phage display Fab libraries. In GBid, the constant domains of the Fabs are located in the backbone of the phagemid vector and the library insert comprises only the variable regions of the antibodies and a central bi-directional promoter. This vector design reduces the process of Fab library construction to "scFv-like" simplicity and the double promoter ensures robust expression of both constituent chains. To maximize the library size, the 3 fragments comprising the insert - two variable chains and one bi-directional promoter - are assembled via a 3-fragment overlap extension PCR and the insert is incorporated into the vector via a high-efficiency one-fragment, one-pot Golden Gate assembly. The reaction setup requires minimal preparatory work and enzyme quantities, making GBid highly scalable. Using GBid, we constructed a chimeric chicken-human Fab phage display library comprising 10
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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.