Inclusion Bodies: Methods and Protocols

ISBN-13: 9781071629291 / Angielski / Twarda / 2023 / 272 str.

Julian Kopp; Oliver Spadiut

Opis

This detailed volume presents a series of protocols dealing with different aspects of inclusion body (IB) processing, from cloning procedures to purification of refolded product. Commencing with chapters on upstream processing, looking into different expression strategies for IB production, the book continues with downstream applications, highlighting early protein purification and subsequent analytics, as well as success stories of IB-based processes. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, Inclusion Bodies: Methods and Protocols serves as an ideal resource for facilitating diverse aspects of IB processing.

Spis treści

1. Inclusion Bodies: Status Quo and Perspectives

            Julian Kopp and Oliver Spadiut

 

Part I: Upstream Processing

 

2. Thermoinducible E. coli for Recombinant Protein Production in Inclusion Bodies

            Norma A. Valdez-Cruz and Mauricio A. Trujillo-Roldán

 

3. High-Throughput Expression of Inclusion Bodies on an Automated Platform

            Annina Kemmer, Linda Cai, M. Nicolas Cruz-Bournazou, and Peter Neubauer

 

4. Design, Production, and Characterization of Catalytically-Active Inclusion Bodies

            Gizem Ölçücü, Karl-Erich Jaeger, and Ulrich Krauss

 

5. Bidirectional Promoter Libraries Enable the Balanced Co-Expression of Two Target Genes in E. coli

            Julia Jodlbauer, Lukas Rieder, Anton Glieder, and Birgit Wiltschi

 

6. Inclusion Body Production in Fed-Batch and Continuous Cultivation

            Stefan Kittler

 

7. Avoiding the All-or-None Response in Gene Expression during E. coli Continuous Cultivation Based on the On-Line Monitoring of Cell Phenotypic Switching Dynamics

            Frank Delvigne, Lucas Henrion, Vincent Vandenbroucke, and Juan Andres Martinez

 

8. Optimization of Inclusion Body Formation and Purification in Multi-Well Plates

            Sarah Kamel, Julia Schollmeyer, Anke Kurreck, and Peter Neubauer

 

Part II: Downstream Processing

 

9. High Pressure Homogenization for Inclusion Body Isolation

            Julian Ebner, Viktor Sedlmayr, and Robert Klausser

 

10. Development of Solubilization and Refolding Buffers

            Mohammad Roufarshbaf and Vajihe Akbari

 

11. Unit Operation-Spanning Investigation of the Redox System

            Julian Ebner, Diana Humer, and Viktor Sedlmayr

 

12. Using High Pressure and Alkaline pH for Refolding

            Ligia Morganti and Rosa Maria Chura-Chambi

 

13. Intensification of Inclusion Body Processing via Temperature-Based Refolding

            Ru Shen Wong, Nurul Nadia Mohamad Alias, Eugene Boon Beng Ong, and Mervyn W.O. Liew

 

14. Refolding of Proteins Expressed as Inclusion Bodies in E. coli

            Rashmi Sharma, Anupa Pahuja, and Anurag S. Rathore

 

15. Infrared Spectroscopy for Structure Analysis of Protein Inclusion Bodies

            Andreas Schwaighofer and Bernhard Lendl

 

Part III: Success Stories

 

16. The Purification of Heme Peroxidases from Escherichia coli Inclusion Bodies: A Success Story Shown by the Example of Horseradish Peroxidase

            Diana Humer and Julian Ebner

 

17. Production of α-Synuclein Fibrillar Specific scFv from Inclusion Bodies

            Vijay Gupta, Issam Hmila, Nishant N. Vaikath, Indulekha P. Sudhakaran, and Omar M.A. El-Agnaf

 

18. Method for Inclusion Bodies Production via E. coli Host System: rGCSF as Model Biotherapeutic Protein

            Abhilasha K. Rani, Venkateswara R. Naira, and Anurag S. Rathore

 

19. Antimicrobial Applications of Inclusion Bodies

            Ramon Roca-Pinilla, Anna Arís, and Elena Garcia-Fruitós