Therapeutic Proteins (2^nd Ed., Softcover reprint of the original 2nd ed. 2012)
Methods and Protocols
Methods in Molecular Biology Series, Vol. 899

After the identification of a potential protein drug, the next critical step is the production of sufficient authentic material for testing, characterization, and clinical trials, which, when successful, leads to the need for robust methodologies for large-scale production, purification, characterization, viral inactivation, and continued testing of the final protein product.  Building on the valuable first edition, Therapeutic Proteins: Methods and Protocols, Second Edition aims to cover each of these key aspects of protein drug production through the contributions of authors from highly esteemed industrial and academic institutions around the world.  Emphasizing the newest developments in the field, this second edition also includes additional emphasis on discovery, including new display and screening methods as well as the design and engineering of new types of therapeutic proteins.  There is also discussion of computational and bioinformatics methods, and chapters on safety aspects of therapeutic protein development.  Written in the highly successful Methods in Molecular Biology? format, protocol chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Therapeutic Proteins.- Synthetic Antibody Libraries.- The Construction of ‘Phylomer’ Peptide Libraries as a Rich Source of Potent Inhibitors of Protein/Protein Interactions.- Ribosome Display and Screening for Protein Therapeutics.- Yeast Display of Engineered Antibody Domains.- Design, Expression, Purification, and Characterization of Engineered Antibody CH2 and VH Domains.- Engineering of Affibody Molecules for Therapy and Diagnostics.- Protein Design for Diversity of Sequences and Conformations Using Dead-End Elimination.- Design and Generation of DVD-Ig^TM Molecules for Dual-Specific Targeting.- Engineering and Expression of Bibody and Tribody Constructs in Mammalian Cells and in the Yeast Pichia pastoris.- Use of E. coli for the Production of a Single Protein.- Folding Engineering Strategies for Efficient Membrane Protein Production in E. coli.- Transient Expression Technologies: Past, Present, and Future.- Stable Transfection Pools for Large Quantity of Protein Production.- Mammalian Stable Expression of Biotherapeutics.- Transgenic Expression of Therapeutic Proteins in Arabidopsis thaliana Seed.- Methods for Chromatographic Removal of Endotoxin.- Effectiveness of Various Processing Steps for Viral Clearance of Therapeutic Proteins: Database Analyses of Commonly-Used Steps.- High-Throughput Quantitative N-Glycan Analysis of Glycoproteins.- High-Throughput Multimodal Strong Anion Exchange Purification and N-Glycan Characterization of Endogenous Glycoprotein Expressed in Glycoengineered Pichia pastoris.- Databases and Tools in Glycobiology.- Characterization of PEGylated Biopharmaceutical Products by LC/MS and LC/MS/MS.- Identification of Asp Isomerization in Proteins by ¹⁸O Labeling and Tandem Mass Spectrometry.- Monitoring of Subvisible Particles in Therapeutic Proteins.- Size-Exclusion Chromatography with Multi-Angle Light Scattering (SEC-MALS) for Elucidating Protein Aggregation Mechanisms.-Computational Methods to Predict Therapeutic Protein Aggregation.- Coarse-Grained Simulations of Protein Aggregation.- Chitosan-Based Nanoparticles as Delivery Systems of Therapeutic Proteins.- Challenges in the Development and Manufacturing of Antibody-Drug Conjugates.