Therapeutic Proteins
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Therapeutic Proteins : Methods and Protocols

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With the recent completion of the sequencing of the human genome, it is widely anticipated that the number of potential new protein drugs and targets will escalate at an even greater rate than that observed in recent years. However, identification of a potential target is only part of the process in developing these new next generation protein-based "drugs" that are increasingly being used to treat human disease. Once a potential protein drug has been identified, the next rate-limiting step on the road to development is the production of sufficient authentic material for testing, charact- ization, clinical trials, and so on. If a protein drug does actually make it through this lengthy and costly process, methodology that allows the production of the protein on a scale large enough to meet demand must be implemented. Furthermore, large-scale production must not compromise the authenticity of the final product. It is also nec- sary to have robust methods for the purification, characterization, viral inactivation and continued testing of the authenticity of the final protein product and to be able to formulate it in a manner that retains both its biological activity and lends itself to easy administration. Therapeutic Proteins: Methods and Protocols covers all aspects of protein drug production downstream of the discovery stage. This volume contains contributions from leaders in the field of therapeutic protein expression, purification, characterization, f- mulation, and viral inactivation.show more

Product details

  • Hardback | 482 pages
  • 157.5 x 231.1 x 30.5mm | 952.56g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • 2005 ed.
  • biography
  • 1588293904
  • 9781588293909

Review quote

"...scientists in the area will find the material of value." - Doody's Health Sciences Book Review Journalshow more

Back cover copy

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(TM) 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.show more

Table of contents

Biopharmaceutical Proteins: Opportunities and Challenges John R. Birch and Yemi Onakunle Expression of Antibody Fragments by Periplasmic Secretion in Escherichia coli Andrew G. Popplewell, Mukesh Sehdev, Mariangela Spitali, and A. Neil C. Weir Secretory Production of Therapeutic Proteins in Escherichia coli Sang Yup Lee, Jong Hyun Choi, and Sang Jun Lee Expression of Recombinant LTB Protein in Marine Vibrio VSP60 Wang Yili and Si Lusheng Heterologous Gene Expression in Yeast Lee J. Byrne, Kenneth J. O'Callaghan, and Mick F. Tuite Pharmaceutical Proteins from Methylotrophic Yeasts Eric C. de Bruin, Erwin H. Duitman, Arjo L. de Boer, Marten Veenhuis, Ineke G. A. Bos, and C. Erik Hack Expression of Human Papillomavirus Type 16 L1 in Baculovirus Expression Systems: A Case Study Zheng Jin, Si Lusheng, and Wang Yili Large-Scale Transient Expression of Therapeutic Proteins in Mammalian Cells Sabine Geisse, Martin Jordan, and Florian M. Wurm Site-Specific Integration for High-Level Protein Production in Mammalian Cells Bhaskar Thyagarajan and Michele P. Calos Production of Recombinant Therapeutic Proteins by Mammalian Cells in Suspension Culture Lily Chu, Ilse Blumentals, and Gargi Maheshwari Transgene Control Engineering in Mammalian Cells Beat P. Kramer and Martin Fussenegger Fusion to Albumin as a Means to Slow the Clearance of Small Therapeutic Proteins Using the Pichia pastoris Expression System: A Case Study William P. Sheffield, Teresa R. McCurdy, and Varsha Bhakta High Throughput Recovery of Therapeutic Proteins from the Inclusion Bodies of Escherichia coli: An Overview Amulya K. Panda Isolation, Solubilization, Refolding, and Chromatographic Inclusion Bodies of Escherichia coli cells: A Case Study Surinder M. Singh, A. N. S. Eshwari, Lalit C. Garg, and Amulya K. Panda Large-Scale Preparation of Factor VIIa from Human Plasma: A Case Study Teruhisa Nakashima and Kazuhiko Tomokiyo Purification of Clinical-Grade Monoclonal Antibodies by Chromatographic Methods Alberto L. Horenstein, Ilaria Durelli, and Fabio Malavasi Virus Elimination and Validation Nicola Boschetti and Anna Johnston Virus Removal by Nanofiltration Marina Korneyeva and Scott Rosenthal Determining Residual Host Cell Antigen Levels in Purified Recombinant Proteins by Slot Blot and Scanning Laser Densitometry Aaron P. Miles, Daming Zhu, and Allan Saul Principles of Biopharmaceutical Protein Formulation: An Overview Scott P. Sellers and Yuh-Fun Maa Solid-State Protein Formulation: Methodologies, Stability, and Excipient Effects Yuh-Fun Maa and Scott P. Sellers Stabilization of Therapeutic Proteins by Chemical and Physical Methods Ken-ichi Izutsu Extraction and Characterization of Vaccine Antigens from Water-In-Oil Adjuvant Formulations Aaron P. Miles and Allan Saul Probing Reversible Self-Association of Therapeutic Proteins by Sedimentation Velocity in the Analytical Ultracentrifuge Bernardo Perez-Ramirez and John J. Steckert Biological Characterization of Pegylated Interferons: A Case Study Lei Xie, Constance Cullen, Sheri Bradshaw, Marc DeLorenzo, and Michael J. Grace Characterization of Interferon a2B Pegylated Via Carboxyalkylation: A Case Study David C. Wylie, Marcio Voloch, Seoju Lee, Yan-Hui Liu, Collette Cutler, Brittany Larkin, and Susan Cannon-Carlson Quantifying Recombinant Proteins and Their Degradation Products Using SDS-PAGE and Scanning Laser Densitometry Aaron P. Miles and Allan Saul Extraction and Denaturing Gel Electrophoretic Methodology for the Analysis of Yeast Proteins Kenneth J. O'Callaghan, Lee J. Byrne, and Mick Fshow more