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TitleTherapeutic Proteins [electronic resource] : Methods and Protocols
Author edited by C. Mark Smales, David C. James
ImprintTotowa, NJ : Humana Press, 2005
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Descript XIV, 482 p. online resource


With the completion of the human genome, the number of protein-based drug candidates has increased dramatically, along with the need to find methods for producing these proteins on a scale large enough to meet demand without compromising the authenticity of the final product. In Therapeutic Proteins: Methods and Protocols, world-class researchers share their innovative and proven techniques for the production of therapeutic proteins downstream of the discovery stage. The methods use a variety of sources for producing therapeutic proteins, including bacterial and yeast expression systems, and insect and mammalian cells, and then cover the purification of the resulting protein using both state-of-the-art and traditional methods, such as those sourced from plasma. Additional chapters offer methods for the characterization of therapeutic proteins throughout the production process, along with methods and strategies for viral inactivation and protein formulation. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Authoritative and highly practical, Therapeutic Proteins: Methods and Protocols offers both novice and experienced researchers the latest innovative, readily reproducible techniques, as well as helpful commentaries on the development and production of therapeutic proteins, ranging from their expression to actual formulation


Biopharmaceutical Proteins -- Expression of Antibody Fragments by Periplasmic Secretion in Escherichia coli -- Secretory Production of Therapeutic Proteins in Escherichia coli -- Expression of Recombinant LTB Protein in Marine vibrio VSP60 -- Heterologous Gene Expression in Yeast -- Pharmaceutical Proteins From Methylotrophic Yeasts -- Expression of Human Papillomavirus Type 16 L1 in Baculovirus Expression Systems -- Large-Scale Transient Expression of Therapeutic Proteins in Mammalian Cells -- Site-Specific Integration for High-Level Protein Production in Mammalian Cells -- Production of Recombinant Therapeutic Proteins by Mammalian Cells in Suspension Culture -- Transgene Control Engineering in Mammalian Cells -- Fusion to Albumin as a Means to Slow the Clearance of Small Therapeutic Proteins Using the Pichia pastoris Expression System -- High-Throughput Recovery of Therapeutic Proteins from the Inclusion Bodies of Escherichia coli -- Isolation, Solubilization, Refolding, and Chromatographic Purification of Human Growth Hormone from Inclusion Bodies of Escherichia coli Cells -- Large-Scale Preparation of Factor VIIa from Human Plasma -- Purification of Clinical-Grade Monoclonal Antibodies by Chromatographic Methods -- Virus Elimination and Validation -- Virus Removal by Nanofiltration -- Determining Residual Host Cell Antigen Levels in Purified Recombinant Proteins by Slot Blot and Scanning Laser Densitometry -- Principles of Biopharmaceutical Protein Formulation -- Solid-State Protein Formulation -- Stabilization of Therapeutic Proteins by Chemical and Physical Methods -- Extraction and Characterization of Vaccine Antigens from Water-in-Oil Adjuvant Formulations -- Probing Reversible Self-Association of Therapeutic Proteins by Sedimentation Velocity in the Analytical Ultracentrifuge -- Biological Characterization of Pegylated Interferons -- Characterization of Interferon ?2B Pegylated via Carboxyalkylation -- Quantifying Recombinant Proteins and Their Degradation Products Using SDS-PAGE and Scanning Laser Densitometry -- Extraction and Denaturing Gel Electrophoretic Methodology for the Analysis of Yeast Proteins -- Characterization of Therapeutic Proteins by Membrane and In-Gel Tryptic Digestion -- Oligosaccharide Release and MALDI-TOF MS Analysis of N-Linked Carbohydrate Structures from Glycoproteins -- Capillary Electrophoresis of Carbohydrates Derivatized With Fluorophoric Compounds -- High-Throughput LC/MS Methodology for ?(1?3)Gal Determination of Recombinant Monoclonal Antibodies -- Carbohydrate Structural Characterization of Fas Ligand Inhibitory Protein -- Top-Down Characterization of Protein Pharmaceuticals by Liquid Chromatography/Mass Spectrometry -- Sample Preparation Procedures for High-Resolution Nuclear Magnetic Resonance Studies of Aqueous and Stabilized Solutions of Therapeutic Peptides

Life sciences Biochemistry Life Sciences Biochemistry general


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