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This volume aims at providing state-of-the-art protocols detailing ribosome display, cDNA display, phage display, yeast surface display, and mammalian display. Chapters guide readers through methods and protocols on in vitro methods over prokaryotic display systems, lower eukaryotes, and mammalian cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of all necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Genotype Phenotype Coupling aims to provide an overview of current technologies in this exciting and continuously evolving field.
List of contents
Screening, Post-SELEX Optimization and Application of DNA Aptamers Specific for Tobramycin.- Affitins: Ribosome Display for Selection of Aho7c-based Affinity Proteins.- cDNA Display: A Stable and Simple Genotype-phenotype Coupling Using a Cell-free Translation System.- cDNA Display of Disulfide-containing Peptide Library and in vitro Evolution.- Rapid Antigen and Antibody-like Molecule Discovery by Staphylococcal Surface Display.- Restriction-free Construction of a Phage-presented Very Short Macrocyclic Peptide Library.- In Vitro Maturation of a Humanized Shark VNAR Domain to Improve Its Biophysical Properties.- Antibody Phage Display: Antibody Selection in Solution Using Biotinylated Antigens.- Assessing Antibody Specificity in Human Serum Using Deep Sequence-coupled Biopanning.- Isolation of Antigen-specific VHH Single Domain Antibodies by Combining Animal Immunization with Yeast Surface Display.- Selection and Characterization of Anti-idiotypic Shark Antibody Domains.-Simplifying the Detection of Surface Presentation Levels in Yeast Surface Display by Intracellular tGFP Expression.- Methods for Construction of Yeast Display Libraries of Four-Domain T-cell Receptors.- Isolation of Tailor-made Antibody Fragments from Yeast-displayed B-Cell Receptor Repertoires by Multiparameter Fluorescence-Activated Cell Sorting.- Isolation of Anti-Hapten Antibodies by Fluorescence-Activated Cell Sorting of Yeast-Displayed B-Cell Receptor Gene Repertoires.- Rapid Generation of Chicken Immune Libraries for Yeast Surface Display.- Ligand Engineering via Yeast Surface Display and Adherent Cell Panning.- Simultaneous Soluble Secretion and Surface Display of Proteins in Saccharomyces Cerevisiae Using Inefficient Ribosomal Skipping.- Chemical Modification of the Yeast Cell Surface Allows the Switch between Display and Soluble Secretion of Full-length Antibodies.- Advanced Establishment of Stable Recombinant Human Suspension Cell Lines Using Geno-Phenotype Coupling Transposon Vectors.- Mammalian Surface Display Screening of Diverse Cystine-Dense Peptide Libraries for Difficult to Drug Targets.- Engineering Antibodies on the Surface of CHO Cells.- Single B Cell Cloning and Production of Rabbit Monoclonal Antibodies.
