Therapeutic Proteins shares innovative and proven techniques for the production of therapeuic proteins downstream of the discovery stage.

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.

Contents

1. Biopharmaceutical Proteins: Opportunities and Challenges
2. Expression of Antibody Fragments by Periplasmic Secretion in Escherichia coli
3. Secretory Production of Therapeutic Proteins in Escherichia coli
4. Expression of Recombinant LTB Protein in Marine vibrio VSP60
5. Heterologous Gene Expression in Yeast
6. Pharmaceutical Proteins From Methylotrophic Yeasts
7. Expression of Human Papillomavirus Type 16 L1 in Baculovirus Expression Systems: A Case Study
8. Large-Scale Transient Expression of Therapeutic Proteins in Mammalian Cells
9. Site-Specific Integration for High-Level Protein Production in Mammalian Cells
10. Production of Recombinant Therapeutic Proteins by Mammalian Cells in Suspension Culture
11. Transgene Control Engineering in Mammalian Cells
12. Fusion to Albumin as a Means to Slow the Clearance of Small Therapeutic Proteins Using the Pichia pastoris Expression System: A Case Study
13. High-Throughput Recovery of Therapeutic Proteins from the Inclusion Bodies of Escherichia coli: An Overview
14. Isolation, Solubilization, Refolding, and Chromatographic Purification of Human Growth Hormone from Inclusion Bodies of Escherichia coli Cells: A Case Study
15. Large-Scale Preparation of Factor ViIa from Human Plasma: A Case Study
16. Purification of Clinical-Grade Monoclonal Antibodies by Chromatographic Methods
17. Virus Elimination and Validation
18. Virus Removal by Nanofiltration
19. Determining Residual Host Cell Antigen Levels in Purified Recombinant Proteins by Slot Blot and Scanning Laser Densitometry
20. Principles of Biopharmaceutical Protein Formulation: An Overview
21. Solid-State Protein Formulation: Methodologies, Stability, and Excipient Effects
22. Stabilization of Therapeutic Proteins by Chemical and Physical Methods
23. Extraction and Characterization of Vaccine Antigens from Water-in-Oil Adjuvant Formulations
24. Probing Reversible Self-Association of Therapeutic Proteins by Sedimentation Velocity in the Analytical Ultracentrifuge
25. Biological Characterization of Pegylated Interferons: A Case Study
26. Characterization of Interferon a2B Pegylated via Carboxyalkylation: A Case Study
27. Quantifying Recombinant Proteins and Their Degradation Products Using SDS-PAGE and Scanning Laser Densitometry
28. Extraction and Denaturing Gel Electrophoretic Methodology for the Analysis of Yeast Proteins
29. Characterization of Therapeutic Proteins by Membrane and In-Gel Tryptic Digestion
30. Oligosaccharide Release and MALDI-TOF MS Analysis of N-Linked Carbohydrate Structures from Glycoproteins
31. Capillary Electrophoresis of Carbohydrates Derivatized With Fluorophoric Compounds
32. High-Throughput LC/MS Methodology for a(1Æ3)Gal Determination of Recombinant Monoclonal Antibodies: A Case Study
33. Carbohydrate Structural Characterization of Fas Ligand Inhibitory Protein
34. Top-Down Characterization of Protein Pharmaceuticals by Liquid Chromatography/Mass Spectrometry: Application to Recombinant Factor IX Comparability– A Case Study
35. Sample Preparation Procedures for High-Resolution Nuclear Magnetic Resonance Studies of Aqueous and Stabilized Solutions of Therapeutic Peptides

Index

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