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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.
Fully updated and practical, Therapeutic Proteins: Methods and Protocols, Second Edition provides an essential resource to all scientists working in the field of therapeutic proteins.
1. Therapeutic Proteins
Dimiter S. Dimitrov
2. Synthetic Antibody Libraries
Bryce Nelson and Sachdev S. Sidhu
3. The Construction of ‘Phylomer’ Peptide Libraries as a Rich Source of Potent Inhibitors of Protein/Protein Interactions
Nadia Milech and Paul Watt
4. Ribosome Display and Screening for Protein Therapeutics
Damjana Kastelic and Mingyue He
5. Yeast Display of Engineered Antibody Domains
Qi Zhao, Zhongyu Zhu, and Dimiter S. Dimitrov
6. Design, Expression, Purification, and Characterization of Engineered Antibody CH2 and VH Domains
Rui Gong, Weizao Chen, and Dimiter S. Dimitrov
7. Engineering of Affibody Molecules for Therapy and Diagnostics
Joachim Feldwisch and Vladimir Tolmachev
8. Protein Design for Diversity of Sequences and Conformations Using Dead-End Elimination
Karl J.M. Hanf
9. Design and Generation of DVD-IgTM Molecules for Dual-Specific Targeting
Enrico DiGiammarino, Tariq Ghayur, and Junjian Liu
10. Engineering and Expression of Bibody and Tribody Constructs in Mammalian Cells and in the Yeast Pichia pastoris
Steve Schoonooghe
11. Use of E. coli for the Production of a Single Protein
Lili Mao and Masayori Inouye
12. Folding Engineering Strategies for Efficient Membrane Protein Production in E. coli
Brent L. Nannenga and François Baneyx
13. Transient Expression Technologies: Past, Present, and Future
Sabine Geisse and Bernd Voedisch
14. Stable Transfection Pools for Large Quantity of Protein Production
Jianxin Ye
15. Mammalian Stable Expression of Biotherapeutics
Thomas Jostock and Hans-Peter Knopf
16. Transgenic Expression of Therapeutic Proteins in Arabidopsis thaliana Seed
Cory L. Nykiforuk and Joseph G. Boothe
17. Methods for Chromatographic Removal of Endotoxin
Adam J. Lowe, Cameron L. Bardliving, and Carl A. Batt
18. Effectiveness of Various Processing Steps for Viral Clearance of Therapeutic Proteins: Database Analyses of Commonly-Used Steps
Dana Cipriano, Michael Burnham, and Joseph V. Hughes
19. High-Throughput Quantitative N-Glycan Analysis of Glycoproteins
Margaret Doherty, Ciara A. McManus, Rebecca Duke, and Pauline M. Rudd
20. High-Throughput Multimodal Strong Anion Exchange Purification and N-Glycan Characterization of Endogenous Glycoprotein Expressed in Glycoengineered Pichia pastoris
Sujatha Gomathinayagam, Erik Hoyt, Alissa M. Thompson, Eric Brown, Khanita Karaveg, Stephen Hamilton, and Huijuan Li
21. Databases and Tools in Glycobiology
Natalia V. Artemenko, Andrew G. McDonald, Gavin P. Davey, and Pauline M. Rudd
22. Characterization of PEGylated Biopharmaceutical Products by LC/MS and LC/MS/MS
Lihua Huang and P. Clayton Gough
23. Identification of Asp Isomerization in Proteins by 18O Labeling and Tandem Mass Spectrometry
Jennifer Zhang and Viswanatham Katta
24. Monitoring of Subvisible Particles in Therapeutic Proteins
Satish K. Singh and Maria R. Toler
25. Size-Exclusion Chromatography with Multi-Angle Light Scattering (SEC-MALS) for Elucidating Protein Aggregation Mechanisms
Erinc Sahin and Christopher J. Roberts
26. Computational Methods to Predict Therapeutic Protein Aggregation
Patrick M. Buck, Sandeep Kumar, Xiaoling Wang, Neeraj J. Agrawal, Bernhardt L. Trout, and Satish K. Singh
27. Coarse-Grained Simulations of Protein Aggregation
Troy Cellmer and Nicolas L. Fawzi
28. Chitosan-Based Nanoparticles as Delivery Systems of Therapeutic Proteins
Pedro Fonte, José Carlos Andrade, Vítor Seabra, and Bruno Sarmento
29. Challenges in the Development and Manufacturing of Antibody-Drug Conjugates
Laurent Ducry
Lihua Huang and P. Clayton Gough
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