ArticlemAbs
Combining phage display with SMRTbell next-generation sequencing for the rapid discovery of functional scFv fragments.
Article in mAbs. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 23 citations in OpenAlex.
- Antibody format matters: A comparative analysis of VHH and scFv domains reveals superior in vivo CAR T cell function with VHH domains.Molecular therapy : the journal of the American Society of Gene Therapy · 2026Article
- Integrated scFv identification and CAR T cell generation for AML targeting in vivo.International journal of cancer · 2026Article
- Whole-genome sequencing ofMicrobiology resource announcements · 2025Article
- The characterization of variable new antigen receptors targeting FAP isolated from a novel immunized library.Communications biology · 2025Article
- Preclinical development of anti-CD21 chimeric antigen receptor T cells to treat T cell acute lymphoblastic leukemia.Science translational medicine · 2025Article
- Discovery of Antibodies Against Endemic Coronaviruses with NGS-Based Human Fab Phage Display Platform.Antibodies (Basel, Switzerland) · 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
- Selection of Affibody Affinity Proteins from Phagemid Libraries.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Antibody Isolation from Human Synthetic Libraries of Single-Chain Antibodies and Analysis Using NGS.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Review
- Reconstruction of full antibody sequences in NGS datasets and accurate VComputational and structural biotechnology journal · 2022Article
- Application of recombinant antibodies for treatment ofFrontiers in immunology · 2022Review
- High-Throughput Sequencing of Phage Display Libraries Reveals Parasitic Enrichment of Indel Mutants Caused by Amplification Bias.International journal of molecular sciences · 2021Article
- Article
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Authors and funding
13 authors at 3 institutions in 1 country.
Funding
Abstract
Phage display technology in combination with next-generation sequencing (NGS) currently is a state-of-the-art method for the enrichment and isolation of monoclonal antibodies from diverse libraries. However, the current NGS methods employed for sequencing phage display libraries are limited by the short contiguous read lengths associated with second-generation sequencing platforms. Consequently, the identification of antibody sequences has conventionally been restricted to individual antibody domains or to the analysis of single domain binding moieties such as camelid VHH or cartilaginous fish IgNAR antibodies. In this study, we report the application of third-generation sequencing to address this limitation. We used single molecule real time (SMRT) sequencing coupled with hairpin adaptor loop ligation to facilitate the accurate interrogation of full-length single-chain Fv (scFv) libraries. Our method facilitated the rapid isolation and testing of scFv antibodies enriched from phage display libraries within days following panning. Two libraries against CD160 and CD123 were panned and monitored by NGS. Analysis of NGS antibody data sets led to the isolation of several functional scFv antibodies that were not identified by conventional panning and screening strategies. Our approach, which combines phage display selection of immune libraries with the full-length interrogation of scFv fragments, is an easy method to discover functional antibodies, with a range of affinities and biophysical characteristics.
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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 Socratic graph.