Proteins are the most diverse group of biologically important substances. With the recent technological advances in the genomics area and the efforts in proteomics research, the rate of discovery for new proteins with unknown structure and function has increased. These proteins generated from genomic approaches present enormous opportunities for research and industrial application. Protein Downstream Processing: Design, Development and Application of High and Low-Resolution Methods is a compilation of chapters within the exciting area of protein purification designed to give the laboratory worker the information needed to design and implement a successful purification strategy. It presents reliable and robust protocols in a concise form, emphasizing the critical aspects on practical problems and questions encountered at the lab bench. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Protein Downstream Processing: Design, Development and Application of High and Low-Resolution Methods will be an ideal source of scientific information to advanced students, junior researchers, and scientists involved in health sciences, cellular and molecular biology, biochemistry, and biotechnology and other related areas in both academia and industry. ?
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Part I. Screening and Design Purification Strategies
1. Protein Purification: An Overview
Nikolaos Labrou
2. Establishment of a Design Space for Biopharmaceutical Purification Processes Using DoE
Ignacio Amadeo, Laura Mauro, Eduardo Ortí, and Guillermina Forno
3. High Throughput Process Development: I. Process Chromatography
A. S. Rathore and R. Bhambure
4. High Throughput Process Development: II. Membrane Chromatography
A. S. Rathore and S. Muthukumar
5. Media Selection in Ion Exchange Chromatography in a Single Microplate
Charlotte Cabanne and Xavier Santarelli
6. High Throughput Screening of Dye-Ligands for Chromatography
Sunil Kumar and Narayan S. Punekar
7. Measurement of Uptake Curves and Adsorption Isotherms by Automated Microscale Chromatography Pipette Tips
Sunil Chhatre and Daniel G. Bracewell
Part II. Low-Resolution Protein Purification Methods
8. Recovery of Recombinant Proteins from Plants Using Aqueous Two-Phase Partitioning Systems: An Outline
Zhengrong GU
9. Aqueous Two-Phase Systems Strategies for the Recovery of Proteins from Plants
Oscar Aguilar and Marco Rito-Palomares
10. Aqueous Two Phase Assisted Precipitation of Proteins: A Platform for Isolation of Process Related Impurities from Therapeutic Proteins
A. S. Rathore and R. Bhambure
11. Simultaneous Purification and Refolding of Proteins by Affinity Precipitation and Macro-(Affinity Ligand) Facilitated Three Phase Partitioning (MLFTPP)
Ipsita Roy, Kalyani Mondal, and Munishwar N. Gupta
12. Co-Expression and Co-Purification of Antigen-Antibody Complexes in Bacterial Cytoplasm and Periplasm
Ario de Marco
13. Immunoglobulin Purification by Caprylic Acid
Victor Morais, Patricia Berasain, and Hugo Massaldi
Part III: High-Resolution Protein Purification Methods
14. Affinity Tags in Protein Purification and Peptide Enrichment: An Overview
Ana Sofia Pina, Íris L. Batalha, and Ana Cecília A. Roque
15. Expression and Purification of Large Active GST Fusion Enzymes
Stefania Deceglie, Claudia Lionetti, Marina Roberti, Palmiro Cantatore, and Paola Loguercio Polosa
16. Synthetic Ligand Affinity Chromatography. Purification of Human Serum Albumin and Related Fusion Proteins
Sharon Williams, Phil Morton, and Dev Baines
17. Zbasic - A Purification Tag for Selective Ion-Exchange Recovery
My Hedhammar, Johan Nilvebrant, and Sophia Hober
18. An Orthogonal Fusion Tag for Efficient Protein Purification
Johan Nilvebrant, Mikael Åstrand, and Sophia Hober
19. Phage Display of Engineered Binding Proteins
Mark Levisson, Ruud B. Spruijt, Ingrid Nolla Winkel, Servé W.M. Kengen, and John van der Oost
20. Biomimetic Affinity Ligands for Protein Purification
Isabel T. Sousa and M. Ângela Taipa
21. Synthesis and Application of Dye-Ligand Affinity Adsorbents
Evangelia Chronopoulou and Nikolaos E. Labrou
22. Peptide Affinity Chromatography Based on Combinatorial Strategies for Protein Purification
S. A. Camperi, M. C. Martínez-Ceron, S. L. Giudicessi, M. M. Marani, F. Albericio, and O. Cascone
23. Affinity Chromatography of Proteins on Monolithic Columns
E.G. Vlakh, G.A. Platonova, and T.B. Tennikova
24. Sample Displacement Batch Chromatography of Proteins
Marta Kotasinska, Verena Richter, Marcel Kwiatkowski, and Hartmut Schlüter
Part IV. Assessing Protein Structural Integrity, Purity, and Stabilization
25. Analysis of Host-Cell Proteins in Biotherapeutic Proteins by LC/MS Approaches
Catalin E. Doneanu, Weibin Chen, and St John Skilton
26. Preparation of Monolithic Affinity Media for Nano-Liquid Chromatography Applications
Jens Sproß and Andrea Sinz
27. Proteomic Analysis of Complex Protein Samples by MALDI TOF Mass Spectrometry
Cosima Damiana Calvano, Cristina De Ceglie, and Carlo G. Zambonin
28. Modern Bioanalysis of Proteins by Electrophoretic Techniques
Sona Krizkova, Marketa Ryvolova, Michal Masarik, Ondrej Zitka, Vojtech Adam, Jaromir Hubalek, Tomas Eckschlager, and Rene Kizek
29. Protein Structure Validation and Analysis with X-Ray Crystallography
Anastassios C. Papageorgiou and Jesse Mattsson
30. Measuring Binding Constants of His-Tagged Proteins using Affinity Chromatography and Ni-NTA Immobilized Enzymes
Annette C. Moser, Benjamin White, and Frank A. Kovacs
31. Stabilization of Therapeutic Proteins in Aqueous Solutions and Freeze-Dried Solids: An Overview
Ken-ichi Izutsu
32. Stabilization of Protein by Freeze-Drying in the Presence of Trehalose: A Case Study of Tubulin
Pavel Dráber, Vadym Sulimenko, Tetyana Sulimenko, and Eduarda Dráberová
Part V. Applications/Case Studies
33. G Protein-Coupled Receptor Expression and Purification
Karolina Corin, Lotta T. Tegler, and Sotirios Koutsopoulos
34. (Hyper)thermophilic Enzymes: Production and Purification
Pierpaolo Falcicchio, Mark Levisson, Servé W.M. Kengen, Sotirios Koutsopoulos , and John van der Oost
35. Affinity Chromatography for Antibody Purification
Sushrut Arora, B. Vijayalakshmi Ayyar, and Richard O’Kennedy
36. Screening and Purification of Recombinant Lignocellulolytic Enzymes
Evangelos Topakas and Paul Christakopoulos
37. Purification of PEGylated Proteins, with the Example of PEGylated lysozyme and PEGylated scFv
Anna Moosmann, Egbert Müller, and Heiner Böttinger
38. One-Step Purification of Glutamate decarboxylase from E. coli Using Aqueous Two Phase System
W. Yao, J. Zhu, and B. Sun
Protein Downstream Processing: Design, Development And Application Of High And Low-Resolution Methods (Methods In Molecular Biology, Vol. 1129) by Labrou, Nikolaos, 9781627039765, Humana Press, 2014, Hardcover
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