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.
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Product details

  • Hardback | 482 pages
  • 157.5 x 231.1 x 30.5mm | 952.56g
  • Totowa, NJ, United States
  • English
  • 2005 ed.
  • XIV, 482 p.
  • 1588293904
  • 9781588293909

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.
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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 F
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Review quote

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