ArticleBiotechnology and bioengineering2016
Geometry and expression enhance enrichment of functional yeast-displayed ligands via cell panning.
Article in Biotechnology and bioengineering, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Engineering Murine Cross-Reactivity Into an Affibody to Human Death Receptor 5.Biotechnology and bioengineering · 2026Article
- A tethered yeast surface display system facilitates enriching epitope-specific binding interactions.Protein engineering, design & selection : PEDS · 2026Article
- An engineering strategy to target activated EGFR with CAR T cells.Cell reports methods · 2024Article
- Hyperstable Synthetic Mini-Proteins as Effective Ligand Scaffolds.ACS synthetic biology · 2023Article
- Discovery of antibodies targeting multipass transmembrane proteins using a suspension cell-based evolutionary approach.Cell reports methods · 2023Article
- Single-Cell B-Cell Sequencing to Generate Natively Paired scFab Yeast Surface Display Libraries.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Identification of lamprey variable lymphocyte receptors that target the brain vasculature.Scientific reports · 2022Article
- Design and Evaluation of Engineered Extracellular Vesicle (EV)-Based Targeting for EGFR-Overexpressing Tumor Cells Using Monobody Display.Bioengineering (Basel, Switzerland) · 2022Article
- A Hybrid Adherent/Suspension Cell-Based Selection Strategy for Discovery of Antibodies Targeting Membrane Proteins.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Antibody Library Screening Using Yeast Biopanning and Fluorescence-Activated Cell Sorting.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Extended yeast surface display linkers enhance the enrichment of ligands in direct mammalian cell selections.Protein engineering, design & selection : PEDS · 2021Article
- Display of Microbial Glucose Dehydrogenase and Cholesterol Oxidase on the Yeast Cell Surface for the Detection of Blood Biochemical Parameters.Biosensors · 2020Article
- CRISPR-Mediated Isogenic Cell-SELEX Approach for Generating Highly Specific Aptamers Against Native Membrane Proteins.Cellular and molecular bioengineering · 2020Article
- Magnetic Bead-Immobilized Mammalian Cells Are Effective Targets to Enrich Ligand-Displaying Yeast.ACS combinatorial science · 2020Article
- Ligand Engineering via Yeast Surface Display and Adherent Cell Panning.Methods in molecular biology (Clifton, N.J.) · 2020Article
- A yeast display immunoprecipitation screen for targeted discovery of antibodies against membrane protein complexes.Protein engineering, design & selection : PEDS · 2019Article
- Nature-inspired design and evolution of anti-amyloid antibodies.The Journal of biological chemistry · 2019Article
- Validation and Stabilization of a Prophage Lysin of Clostridium perfringens by Using Yeast Surface Display and Coevolutionary Models.Applied and environmental microbiology · 2019Article
- Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276.ACS combinatorial science · 2019Article
- Engineered Charge Redistribution of Gp2 Proteins through Guided Diversity for Improved PET Imaging of Epidermal Growth Factor Receptor.Bioconjugate chemistry · 2018Article
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Authors and funding
8 authors.
Funding
Abstract
Yeast surface display has proven to be an effective tool in the discovery and evolution of ligands with new or improved binding activity. Selections for binding activity are generally carried out using immobilized or fluorescently labeled soluble domains of target molecules such as recombinant ectodomain fragments. While this method typically provides ligands with high affinity and specificity for the soluble molecular target, translation to binding true membrane-bound cellular target is commonly problematic. Direct selections against mammalian cell surfaces can be carried out either exclusively or in combination with soluble target-based selections to further direct towards ligands for genuine cellular target. Using a series of fibronectin domain, affibody, and Gp2 ligands and human cell lines expressing a range of their targets, epidermal growth factor receptor and carcinoembryonic antigen, this study quantitatively identifies the elements that dictate ligand enrichment and yield. Most notably, extended flexible linkers between ligand and yeast enhance enrichment ratios from 1.4 ± 0.8 to 62 ± 57 for a low-affinity (>600 nM) binder on cells with high target expression and from 14 ± 13 to 74 ± 25 for a high-affinity binder (2 nM) on cells with medium valency. Inversion of the yeast display fusion from C-terminal display to N-terminal display still enables enrichment albeit with 40-97% reduced efficacy. Collectively, this study further enlightens the conditions-while highlighting new approaches-that yield successful enrichment of yeast-displayed binding ligands via panning on mammalian cells. Biotechnol. Bioeng. 2016;113: 2328-2341. © 2016 Wiley Periodicals, Inc.
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