ArticleChemistry & biology2015
A 45-Amino-Acid Scaffold Mined from the PDB for High-Affinity Ligand Engineering.
Article in Chemistry & biology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed.
- Engineered B7-H3 Binding in Modular Gp2 Miniproteins.Biotechnology and bioengineering · 2026Article
- Incorporation of azide groups into an engineered Fn3 scaffold protein for site-specific, bioorthogonal conjugation.Protein engineering, design & selection : PEDS · 2026Article
- Development of a Mesothelin-Binding Engineered Scaffold Protein as a Theranostic for Pleural Mesothelioma.Bioconjugate chemistry · 2025Article
- HOPS-Dependent Endosomal Escape Demands Protein Unfolding.ACS central science · 2024Article
- An engineering strategy to target activated EGFR with CAR T cells.Cell reports methods · 2024Article
- Extensive antibody search with whole spectrum black-box optimization.Scientific reports · 2024Article
- Hyperstable Synthetic Mini-Proteins as Effective Ligand Scaffolds.ACS synthetic biology · 2023Article
- Design rules for efficient endosomal escape.bioRxiv : the preprint server for biology · 2023Article
- The smallest functional antibody fragment: Ultralong CDR H3 antibody knob regions potently neutralize SARS-CoV-2.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Predicting and Interpreting Protein Developability Via Transfer of Convolutional Sequence Representation.ACS synthetic biology · 2023Article
- Titrating Avidity of Yeast-Displayed Proteins Using a Transcriptional Regulator.ACS synthetic biology · 2023Article
- Yeast biopanning against site-specific phosphorylations in tau.Protein engineering, design & selection : PEDS · 2023Article
- Protein scaffolds: antibody alternatives for cancer diagnosis and therapy.RSC chemical biology · 2022Review
- Machine Learning-driven Protein Library Design: A Path Toward Smarter Libraries.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Yeast Surface Display for Protein Engineering: Library Generation, Screening, and Affinity Maturation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Site-wise Diversification of Combinatorial Libraries Using Insights from Structure-guided Stability Calculations.Methods in molecular biology (Clifton, N.J.) · 2022Article
- A Protein-Engineered, Enhanced Yeast Display Platform for Rapid Evolution of Challenging Targets.ACS synthetic biology · 2021Article
- A Platform for Deep Sequence-Activity Mapping and Engineering Antimicrobial Peptides.ACS synthetic biology · 2021Article
- An engineered CD81-based combinatorial library for selecting recombinant binders to cell surface proteins: Laminin binding CD81 enhances cellular uptake of extracellular vesicles.Journal of extracellular vesicles · 2021Article
- High-throughput developability assays enable library-scale identification of producible protein scaffold variants.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
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Authors and funding
5 authors.
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Abstract
Small protein ligands can provide superior physiological distribution compared with antibodies, and improved stability, production, and specific conjugation. Systematic evaluation of the PDB identified a scaffold to push the limits of small size and robust evolution of stable, high-affinity ligands: 45-residue T7 phage gene 2 protein (Gp2) contains an α helix opposite a β sheet with two adjacent loops amenable to mutation. De novo ligand discovery from 10(8) mutants and directed evolution toward four targets yielded target-specific binders with affinities as strong as 200 ± 100 pM, Tms from 65 °C ± 3 °C to 80°C ± 1 °C, and retained activity after thermal denaturation. For cancer targeting, a Gp2 domain for epidermal growth factor receptor was evolved with 18 ± 8 nM affinity, receptor-specific binding, and high thermal stability with refolding. The efficiency of evolving new binding function and the size, affinity, specificity, and stability of evolved domains render Gp2 a uniquely effective ligand scaffold.
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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.