This detailed volume explores a wide variety of applications of yeast surface display, an extensively used protein engineering technology. Beginning with detailed protocols for the construction and efficient selection/screening of yeast surface display libraries, as well as for the analysis of individual yeast-displayed protein variants, the book continues with protocols describing the selection of yeast surface display libraries for binding to mammalian cells or to extracellular matrix as well as protocols for a broad spectrum of specialized yeast surface display applications, demonstrating the versatility of this display platform. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible methodologies, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Yeast Surface Display serves as a comprehensive resource that enables the implementation of this powerful and versatile technique in virtually any molecular biology laboratory, even in the absence of any prior yeast surface display experience.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Klappentext

Yeast Surface Display: New Opportunities for a Time-Tested Protein Engineering System.- Yeast Surface Display for Protein Engineering: Library Generation, Screening, and Affinity Maturation.- Site-Wise Diversification of Combinatorial Libraries Using Insights from Structure-Guided Stability Calculations.- Ancestral Sequence Reconstruction and Alternate Amino Acid States Guide Protein Library Design for Directed Evolution.- Machine Learning-Driven Protein Library Design: A Path Toward Smarter Libraries.- Kinetic Competition Screening of Yeast-Displayed Libraries for the Generation of High Affinity Binders.- Engineering Proteins by Combining Deep Mutational Scanning and Yeast Display.- Engineering Binders with Exceptional Selectivity.- Affinity and Stability Analysis of Yeast Displayed Proteins.- Antibody Library Screening Using Yeast Biopanning and Fluorescence-Activated Cell Sorting.- A Hybrid Adherent/Suspension Cell-Based Selection Strategy for Discovery of Antibodies Targeting Membrane Proteins.- Ligand Selection by Combination of Recombinant and Cell Panning Selection Techniques.- Identification of Brain ECM Binding Variable Lymphocyte Receptors Using Yeast Surface Display.- Guidelines, Strategies, and Principles for the Directed Evolution of Cross-Reactive Antibodies Using Yeast Surface Display Technology.- Yeast Display for the Identification of Peptide-MHC Ligands of Immune Receptors.- Yeast Display Guided Selection of pH Dependent Binders.- Yeast Mating as a Tool for Highly Effective Discovery and Engineering of Antibodies via Display Methodologies.- Humanization of Chicken-Derived Antibodies by Yeast Surface Display.- Engineering Tissue Inhibitors of Metalloproteinases Using Yeast Surface Display.- Discovery of Cyclic Peptide Binders from Chemically Constrained Yeast Display Libraries.- Generation of Thermally Stable Affinity Pairs for Sensitive, Specific Immunoassays.- Isolating Anti-Amyloid Antibodies from Yeast-Displayed Libraries.- EngineeringProteins Containing Noncanonical Amino Acids on the Yeast Surface.- Construction of Yeast Display Libraries for Selection of Antigen-Binding Variants of Large Extracellular Loop of CD81, a Major Surface Marker Protein of Extracellular Vesicles.- Isolating and Engineering Fluorescence-Activating Proteins Using Yeast Surface Display.- Simultaneous Display of Multiple Kinds of Enzymes on the Yeast Cell Surface for Multistep Reactions.

Inhalt

Yeast Surface Display: New Opportunities for a Time-Tested Protein Engineering System.- Yeast Surface Display for Protein Engineering: Library Generation, Screening, and Affinity Maturation.- Site-Wise Diversification of Combinatorial Libraries Using Insights from Structure-Guided Stability Calculations.- Ancestral Sequence Reconstruction and Alternate Amino Acid States Guide Protein Library Design for Directed Evolution.- Machine Learning-Driven Protein Library Design: A Path Toward Smarter Libraries.- Kinetic Competition Screening of Yeast-Displayed Libraries for the Generation of High Affinity Binders.- Engineering Proteins by Combining Deep Mutational Scanning and Yeast Display.- Engineering Binders with Exceptional Selectivity.- Affinity and Stability Analysis of Yeast Displayed Proteins.- Antibody Library Screening Using Yeast Biopanning and Fluorescence-Activated Cell Sorting.- A Hybrid Adherent/Suspension Cell-Based Selection Strategy for Discovery of Antibodies Targeting Membrane Proteins.- Ligand Selection by Combination of Recombinant and Cell Panning Selection Techniques.- Identification of Brain ECM Binding Variable Lymphocyte Receptors Using Yeast Surface Display.- Guidelines, Strategies, and Principles for the Directed Evolution of Cross-Reactive Antibodies Using Yeast Surface Display Technology.- Yeast Display for the Identification of Peptide-MHC Ligands of Immune Receptors.- Yeast Display Guided Selection of pH Dependent Binders.- Yeast Mating as a Tool for Highly Effective Discovery and Engineering of Antibodies via Display Methodologies.- Humanization of Chicken-Derived Antibodies by Yeast Surface Display.- Engineering Tissue Inhibitors of Metalloproteinases Using Yeast Surface Display.- Discovery of Cyclic Peptide Binders from Chemically Constrained Yeast Display Libraries.- Generation of Thermally Stable Affinity Pairs for Sensitive, Specific Immunoassays.- Isolating Anti-Amyloid Antibodies from Yeast-Displayed Libraries.- EngineeringProteins Containing Noncanonical Amino Acids on the Yeast Surface.- Construction of Yeast Display Libraries for Selection of Antigen-Binding Variants of Large Extracellular Loop of CD81, a Major Surface Marker Protein of Extracellular Vesicles.- Isolating and Engineering Fluorescence-Activating Proteins Using Yeast Surface Display.- Simultaneous Display of Multiple Kinds of Enzymes on the Yeast Cell Surface for Multistep Reactions.