ArticleExperimental & molecular medicine2017
Next-generation sequencing enables the discovery of more diverse positive clones from a phage-displayed antibody library.
Article in Experimental & molecular medicine, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
- Article
- The Evolution of Artificial Intelligence in Antibody Design: From Structure-Based Engineering to Generative Models.Antibodies (Basel, Switzerland) · 2026Review
- Phage Display Technology: Design, Library Construction, and Panning Strategies for Antibody Development.Biomolecules & therapeutics · 2026Review
- The characterization of variable new antigen receptors targeting FAP isolated from a novel immunized library.Communications biology · 2025Article
- Development of FAP-targeted theranostics discovered by next-generation sequencing-augmented mining of a novel immunized VNAR library.bioRxiv : the preprint server for biology · 2025Article
- Single-Stranded Variable Fragment Gene Libraries Built for Phage Display: An Updated Review of Design, Selection and Application.Journal of microbiology and biotechnology · 2024Review
- In vivo phage display identifies novel peptides for cardiac targeting.Scientific reports · 2024Article
- Development of a new affinity maturation protocol for the construction of an internalizing anti-nucleolin antibody library.Scientific reports · 2024Article
- The rise of big data: deep sequencing-driven computational methods are transforming the landscape of synthetic antibody design.Journal of biomedical science · 2024Review
- ExpoSeq: simplified analysis of high-throughput sequencing data from antibody discovery campaigns.Bioinformatics advances · 2024Article
- Development of a novel sandwich immunoassay based on targeting recombinant Francisella outer membrane protein A for the diagnosis of tularemia.Frontiers in cellular and infection microbiology · 2024Article
- Accurate profiling of full-length Fv in highly homologous antibody libraries using UMI tagged short reads.Nucleic acids research · 2023Article
- Concerted Antibody and Antigen Discovery by Differential Whole-cell Phage Display Selections and Multi-omic Target Deconvolution.Journal of molecular biology · 2023Article
- Towards better antivenoms: navigating the road to new types of snakebite envenoming therapies.The journal of venomous animals and toxins including tropical diseases · 2023Article
- Variability among the Isolates of Broad Bean Mottle Virus and Encapsidation of Host RNAs.Pathogens (Basel, Switzerland) · 2022Article
- Potential of Phage Display Antibody Technology for Cardiovascular Disease Immunotherapy.Journal of cardiovascular translational research · 2022Review
- Review
- Generation of synthetic antibody fragments with optimal complementarity determining region lengths for Notch-1 recognition.Frontiers in microbiology · 2022Article
- Identification of New Antibodies Targeting Malignant Plasma Cells for Immunotherapy by Next-Generation Sequencing-Assisted Phage Display.Frontiers in immunology · 2022Article
- Reconstruction of full antibody sequences in NGS datasets and accurate VComputational and structural biotechnology journal · 2022Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Phage display technology provides a powerful tool to screen a library for a binding molecule via an enrichment process. It has been adopted as a critical technology in the development of therapeutic antibodies. However, a major drawback of phage display technology is that because the degree of the enrichment cannot be controlled during the bio-panning process, it frequently results in a limited number of clones. In this study, we applied next-generation sequencing (NGS) to screen clones from a library and determine whether a greater number of clones can be identified using NGS than using conventional methods. Three chicken immune single-chain variable fragment (scFv) libraries were subjected to bio-panning on prostate-specific antigen (PSA). Phagemid DNA prepared from the original libraries as well as from the Escherichia coli pool after each round of bio-panning was analyzed using NGS, and the heavy chain complementarity-determining region 3 (HCDR3) sequences of the scFv clones were determined. Subsequently, through two-step linker PCR and cloning, the entire scFv gene was retrieved and analyzed for its reactivity to PSA in a phage enzyme immunoassay. After four rounds of bio-panning, the conventional colony screening method was performed for comparison. The scFv clones retrieved from NGS analysis included all clones identified by the conventional colony screening method as well as many additional clones. The enrichment of the HCDR3 sequence throughout the bio-panning process was a positive predictive factor for the selection of PSA-reactive scFv clones.
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