Part I Screening and Design Purification Strategies
1. Protein Purification Technologies Nikolaos E. Labrou
2. High Throughput Process Development: I. Process Chromatography Anurag S. Rathore and R. Bhambure
3. High Throughput Process Development: II. Membrane Chromatography Anurag S. Rathore and S. Muthukumar
4. Media Selection in Ion Exchange Chromatography in a Single Microplate Charlotte Cabanne and Xavier Santarelli
5. High Throughput Screening of Dye-Ligands for Chromatography Sunil Kumar and Narayan S. Punekar
6. Technical Optimizations for the High Throughput Purification of Antibodies on Automated Liquid Handlers Peter M. Schmidt
Part II Low-Resolution Protein Purification Methods
7. Aqueous Two-Phase Systems for Clean-Up and Recovery of Enzymes from Plants and Plant-Derived Extracts Oscar Aguilar, Erick Heredia-Olea, Esther Perez-Carrillo, and Marco Rito-Palomares
8. Aqueous Two Phase Assisted Precipitation of Proteins: A Platform for Isolation of Process Related Impurities from Therapeutic Proteins Anurag S. Rathore and R. Bhambure
9. Recombinant Proteins Co-Expressed and Co-Purified in the Presence of Antibody Fragments Ario de Marco
Part III High-Resolution Protein Purification Methods
10. Affinity Tags in Protein Purification and Peptide Enrichment: An Overview Ana Sofia Pina, Íris L. Batalha, Ana M. G. C. Dias, and Ana Cecília A. Roque
11. Synthetic Ligand Affinity Chromatography Purification of Human Serum Albumin and Related Fusion Proteins Sharon Williams, Phil Morton, and Dev Baines
12. Zbasic - A Purification Tag for Selective Ion-Exchange Recovery My Hedhammar, Johan Nilvebrant, and Sophia Hober
13. An Orthogonal Fusion Tag for Efficient Protein Purification Johan Nilvebrant, Mikael Åstrand, and Sophia Hober
14. Biomimetic Affinity Ligands for Protein Purification Isabel T. Sousa and M. Ângela Taipa
15. Synthesis and Evaluation of Dye-Ligand Affinity Adsorbents for Protein Purification Evangelia G. Chronopoulou, Georgios Premetis, Christina Varotsou, Nikolaos Georgakis, Elisavet Ioannou and Nikolaos E. Labrou
16. Design of Affinity Chromatography Peptide Ligands through Combinatorial Peptide Libraries Screening G. R. Barredo, S. L. Saavedra, M. C. Martínez-Ceron, S. L. Giudicessi, M. M. Marani, F. Albericio, O. Cascone, and S. A. Camperi
17. ZCa: A Protein A-Derived Domain with Calcium-Dependent Affinity for Mild Antibody Purification Julia Scheffel, Sara Kanje, and Sophia Hober
18. Macroporous Polymer Monoliths for Affinity Chromatography and Solid Phase Enzyme Processing E.G. Korzhikova-Vlakha, G.A. Platonovaa, and T.B. Tennikova
19. Sample Displacement Batch Chromatography of Proteins Laura Heikaus, Siti Hidaya, Manasia Gaikwad, Marta Kotasinska, Verena Richter, Marcel Kwiatkowski, and Hartmut Schlüter
20. Lectin Affinity Chromatography: An Efficient Method to Purify Horse IgG3 Salvatore G. De-Simone and D. William Provance, Jr.
21. Heparin-Binding Affinity Tag: A Novel Affinity Tag for Simple and Efficient Purification of Recombinant Proteins Sanhita Maity, Musaab Al-ameer, Ravi Kumar Gundampati, Shilpi Agrawal, and Thallapuranam Krishnaswamy Suresh Kumar
22. Expression and Purification of Recombinant Proteins in Escherichia coli Tagged with the Metal-Binding Proteins SmbP and CusF3H+ Jessica J. Gomez-Lugo, Bryan D. Santos, David A. Perez-Perez, Jorge M. Montfort-Gardeazabal, Megan M. McEvoy, and Xristo Zarate
23. Antibody Aggregate Removal using a Mixed-Mode Chromatography Resin Tao Chen, Gaili Guo, Guoqing Tan, Ying Wang, and Yifeng Li
Part IV Assessing Protein Structural Integrity, Purity, and Stabilization
24. Proteomic Analysis of Food Allergens by MALDI TOF/TOF Mass Spectrometry Cosima D. Calvano, Mariachiara Bianco, Ilario Losito, and Tommaso R.I. Cataldi
25. Protein Structure Analysis and Validation with X-Ray Crystallography Anastassios C. Papageorgiou, Nirmal Poudel, and Jesse Mattsson
26. Measuring Binding Constants of His-Tagged Proteins using Affinity Chromatography and Ni-NTA Immobilized Enzymes Annette C. Moser, Benjamin White, and Frank A. Kovacs
27. Stabilization of Proteins 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
28. G Protein-Coupled Receptor Expression and Purification Karolina Corin, Lotta T. Tegler, and Sotirios Koutsopoulos
29. (Hyper)thermophilic Enzymes: Production and Purification Pierpaolo Falcicchio, Mark Levisson, Servé W.M. Kengen, Sotirios Koutsopoulos, and John van der Oost
30. Screening of Recombinant Lignocellulolytic Enzymes through Rapid Plate Assays Anthi Karnaouri, Anastasia Zerva, Paul Christakopoulos, and Evangelos Topakas
This second edition volume expands on the previous edition with updated research and techniques to help laboratory workers design and implement a successful purification strategy, emphasize critical aspects on practical problems, and answers questions encountered at the lab bench. The chapters in this book are divided into five parts: Part One discusses an overview of screening and design of purification strategies and covers initial aspects on high-throughput screening, methods development, and media selection; Parts Two and Three explore low- and high-resolution methods, with emphasis on affinity chromatography; Part Four describes analytical techniques of purified proteins; and Part Five presents selected examples and case studies to discuss the aforementioned. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and comprehensive, Protein Downstream Processing: Design, Development, and Application of High and Low-Resolution Methods, Second Edition is an ideal source of information to advanced students, junior researchers, and scientists involved in health sciences, cellular and molecular biology, biochemistry, biotechnology, and other related areas in both academia and industry.