Protein Downstream Processing (2nd Ed., 2nd ed. 2021)
Design, Development, and Application of High and Low-Resolution Methods

Methods in Molecular Biology Series, Vol. 2178

Coordinator: Labrou Nikolaos E.

Language: English

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Protein Downstream Processing
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Protein Downstream Processing
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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. 


Preface…
Table of Contents…
Contributing Authors…

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

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts