ArticleProceedings of the National Academy of Sciences of the United States of America2012
Construction of a rationally designed antibody platform for sequencing-assisted selection.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Isolation of feline-derived scFvs against the VP1-CDE region of feline calicivirus from a phage display library and characterization of their antigen-binding and antiviral potential in vitro.BMC veterinary research · 2026Article
- Integrated scFv identification and CAR T cell generation for AML targeting in vivo.International journal of cancer · 2026Article
- A versatile platform for combinatorial antibody library cloning and NGS-based quality control with high accuracy.Nucleic acids research · 2025Article
- The rise of big data: deep sequencing-driven computational methods are transforming the landscape of synthetic antibody design.Journal of biomedical science · 2024Review
- Accurate profiling of full-length Fv in highly homologous antibody libraries using UMI tagged short reads.Nucleic acids research · 2023Article
- Mutational scan inferred binding energetics and structure in intrinsically disordered protein CcdA.Protein science : a publication of the Protein Society · 2023Article
- AI/ML combined with next-generation sequencing of VHH immune repertoires enables the rapid identification ofFrontiers in molecular biosciences · 2023Article
- Next-generation sequencing-guided identification and reconstruction of antibody CDR combinations from phage selection outputs.Nucleic acids research · 2019Article
- Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation.International journal of molecular sciences · 2019Review
- Integrating high-throughput screening and sequencing for monoclonal antibody discovery and engineering.Immunology · 2018Review
- Next-Generation Sequencing of Antibody Display Repertoires.Frontiers in immunology · 2018Review
- IG and TR single chain fragment variable (scFv) sequence analysis: a new advanced functionality of IMGT/V-QUEST and IMGT/HighV-QUEST.BMC immunology · 2017Article
- Pacific Biosciences Sequencing and IMGT/HighV-QUEST Analysis of Full-Length Single Chain Fragment Variable from anFrontiers in immunology · 2017Article
- Platform for high-throughput antibody selection using synthetically-designed antibody libraries.New biotechnology · 2016Article
- Express photolithographic DNA microarray synthesis with optimized chemistry and high-efficiency photolabile groups.Journal of nanobiotechnology · 2016Article
- High-Throughput Sequencing-Based Immune Repertoire Study during Infectious Disease.Frontiers in immunology · 2016Review
- Conceptual and methodological advances in cell-free directed evolution.Current opinion in structural biology · 2015Review
- Predominant structural configuration of natural antibody repertoires enables potent antibody responses against protein antigens.Scientific reports · 2015Article
- Conformation-dependent epitopes recognized by prion protein antibodies probed using mutational scanning and deep sequencing.Journal of molecular biology · 2015Article
- Effective Optimization of Antibody Affinity by Phage Display Integrated with High-Throughput DNA Synthesis and Sequencing Technologies.PloS one · 2015Article
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4 authors.
Funding
Abstract
Antibody discovery platforms have become an important source of both therapeutic biomolecules and research reagents. Massively parallel DNA sequencing can be used to assist antibody selection by comprehensively monitoring libraries during selection, thus greatly expanding the power of these systems. We have therefore constructed a rationally designed, fully defined single-chain variable fragment (scFv) library and analysis platform optimized for analysis with short-read deep sequencing. Sequence-defined oligonucleotide libraries encoding three complementarity-determining regions (L3 from the light chain, H2 and H3 from the heavy chain) were synthesized on a programmable microarray and combinatorially cloned into a single scFv framework for molecular display. Our unique complementarity-determining region sequence design optimizes for protein binding by utilizing a hidden Markov model that was trained on all antibody-antigen cocrystal structures in the Protein Data Bank. The resultant ~10(12)-member library was produced in ribosome-display format, and comprehensively analyzed over four rounds of antigen selections by multiplex paired-end Illumina sequencing. The hidden Markov model scFv library generated multiple binders against an emerging cancer antigen and is the basis for a next-generation antibody production platform.
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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.