Protein Downstream Processing
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Protein Downstream Processing : Design, Development and Application of High and Low-Resolution Methods

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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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Product details

  • Paperback | 555 pages
  • 178 x 254 x 29.46mm | 979.76g
  • Totowa, NJ, United States
  • English
  • Softcover reprint of the original 1st ed. 2014
  • 23 Tables, black and white; 62 Illustrations, color; 59 Illustrations, black and white; XV, 555 p. 121 illus., 62 illus. in color.
  • 1493960407
  • 9781493960408
  • 1,483,941

Back cover copy

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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Table of contents

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 Orti, 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, Iris L. Batalha, and Ana Cecilia 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 Astrand, and Sophia Hober



19. Phage Display of Engineered Binding Proteins

Mark Levisson, Ruud B. Spruijt, Ingrid Nolla Winkel, Serve W.M. Kengen, and John van der Oost



20. Biomimetic Affinity Ligands for Protein Purification

Isabel T. Sousa and M. Angela 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. Martinez-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 Schluter





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 Draber, Vadym Sulimenko, Tetyana Sulimenko, and Eduarda Draberova



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, Serve 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 Muller, and Heiner Boettinger



38. One-Step Purification of Glutamate decarboxylase from E. coli Using Aqueous Two Phase System

W. Yao, J. Zhu, and B. Sun
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