Fundamentals of Recombinant Protein Production, Purification and Characterization
- 1st Edition - September 10, 2024
- Editors: Deepti Yadav, Abhishek Guldhe, Tukayi Kudanga
- Language: English
- Paperback ISBN:9 7 8 - 0 - 3 2 3 - 9 8 3 8 8 - 4
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 9 8 5 8 4 - 0
Fundamentals of Recombinant Protein Production, Purification and Characterization is organized into nine chapters in a logical fashion that cover an introduction to recombina… Read more
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Request a sales quoteFundamentals of Recombinant Protein Production, Purification and Characterization is organized into nine chapters in a logical fashion that cover an introduction to recombinant proteins and expression in different host expression systems, extraction, purification and analysis of proteins. This important reference features protocols, along with the advantages and disadvantage of each expression hosts and characterization technique (presented in tabular format) and offers detailed coverage of all aspects of protein production and processing (upstream and downstream processing) in one place. Finally, the book ends with different characterization techniques. Production of recombinant proteins for biotechnological and therapeutic applications at a large scale is an essential need of mankind. With the huge application potential of therapeutic and industrial proteins, there has been increasing demand for effective and efficient bioprocessing strategies. Recent progress around recombinant DNA technologies and bioprocessing strategies has paved the way for efficient production of recombinant proteins. Important factors such as insolubility and cost of production need to be considered for large scale production of these recombinant proteins.
- Includes step-by-step reproducible protocols while also providing updated information on the rationale and latest developments in expression systems
- Can also be used as a handbook for protein expression and purification as expression systems and chromatographic methods are explained in detail
- Consists of notes on troubleshooting from the eminent researchers in the field
- Provides comprehensive information on protein production, purification and characterization in a single volume
- Describes different purification methods for comparatively difficult to obtain proteins
- Brings the topics of recombinant protein expression, purification and characterization together, thereby making it the first resource on how to solve problems with respect to upstream and downstream processing of heterologous proteins
- Cover image
- Title page
- Table of Contents
- Copyright
- Dedication
- List of contributors
- Preface
- Section A: General background
- Chapter 1. Introduction to recombinant protein expression and purification
- Abstract
- 1.1 Introduction
- 1.2 The significance of recombinant proteins in biotechnology, medicine, and industry
- 1.3 Historical perspective
- 1.4 Fundamentals of protein structure and function
- 1.5 Protein functions and classes
- 1.6 Recombinant DNA technology
- 1.7 Recombinant protein expression systems
- 1.8 Protein expression strategies
- 1.9 Protein purification techniques
- 1.10 Case studies and applications
- 1.11 Impact of recombinant protein production on drug development and manufacturing
- 1.12 Biomedical and research applications
- 1.13 Emerging trends and future directions in protein expression and purification research
- 1.14 Opportunities and areas for further research and innovation
- 1.15 Conclusion
- References
- Section B: Recombinant protein expression in different expression hosts
- Chapter 2. Recombinant protein expression in Escherichia coli
- Abstract
- 2.1 Heterologous protein expression in Escherichia coli
- 2.2 Why Escherichia coli as a host?
- 2.3 Expression vectors for Escherichia coli
- 2.4 Strong promoters drive efficient expression of the genes
- 2.5 Selection markers
- 2.6 Different Escherichia coli strains used in heterologous protein production
- 2.7 Culture growth conditions and monitoring of the host cells
- 2.8 Osmotic shock protocol assists in appropriate protein folding
- 2.9 Escherichia coli has preferential codon utilization (codon bias)
- 2.10 Improper folding of heterologous protein may lead to the formation of inclusion bodies in the host cells
- 2.11 Molecular chaperones promote better protein folding and activity
- 2.12 List of chaperones frequently used in Escherichia coli expression host systems
- 2.13 Affinity tags promote the solubility of the desired protein and make purification easy
- 2.14 Removal of tags to obtain the desired product
- 2.15 Expression of industrial and therapeutics protein in Escherichia coli
- 2.16 Lower yield of protein after purification and the strategies to overcome them
- References
- Chapter 3. Recombinant protein expression in yeast
- Abstract
- 3.1 Introduction
- 3.2 Host yeast strain for recombinant protein expression
- 3.3 Yeast expression vectors
- 3.4 Optimization of the yeast expression plasmid
- 3.5 Secretion signals
- 3.6 Advantages and limitations for yeast recombinant expression
- 3.7 Engineering strategies for improved expression
- 3.8 High-cell-density fermentation for enhanced protein production
- 3.9 Down streaming process and strategy
- 3.10 Engineering yeast strain for enhanced protein production
- 3.11 Case studies and application of recombinant protein expression in yeast
- 3.12 Challenges and future perspective
- 3.13 Conclusion
- References
- Chapter 4. Recombinant protein expression in mammalian cells
- Abstract
- 4.1 Introduction
- 4.2 Mammalian cell biology basics
- 4.3 Expression vectors
- 4.4 Host cell lines for recombinant protein expression
- 4.5 Strategies for recombinant protein expression
- 4.6 Cell culture format and media
- 4.7 Current strategy for recombinant cell-line development
- 4.8 Downstream processing and purification
- 4.9 Improving productivity
- 4.10 Challenges and future directions
- 4.11 Case studies and applications
- 4.12 Conclusion
- References
- Chapter 5. Chlamydomonas reinhardtii for the production of recombinant proteins: current knowledge and perspectives
- Abstract
- 5.1 Introduction
- 5.2 Transformation methods
- 5.3 Vector construction strategies
- 5.4 Promoters
- 5.5 Other strategies
- 5.6 Current claims of heterologous proteins expressed in Chlamydomonas reinhardtii
- 5.7 Antibodies
- 5.8 Antigens vaccines
- 5.9 Enzymes
- 5.10 Immunotoxins
- 5.11 Other therapeutic proteins
- 5.12 Future challenges
- 5.13 Conclusion
- References
- Chapter 6. Filamentous fungi as cell factories for heterogeneous protein production
- Abstract
- 6.1 Heterologous proteins
- 6.2 Common filamentous fungi as host
- 6.3 General fungal metabolism and protein secretion
- 6.4 Transformation methods
- 6.5 Fermentation
- 6.6 Applications of filamentous fungi as cell factories for heterologous protein production
- 6.7 Recent advances for utilizing fungi as cell factories
- References
- Further reading
- Section C: Purification strategies and characterization of recombinant proteins
- Chapter 7. Current scenario of recombinant proteins: extraction, purification, concentration, and storage
- Abstract
- 7.1 Introduction
- 7.2 Methods of extraction of recombinant proteins
- 7.3 Purification of the extracted protein
- 7.4 Concentration of the extracted protein
- 7.5 Storage of purified recombinant proteins
- 7.6 Conclusion
- References
- Chapter 8. Protein purification: chromatographic and nonchromatographic techniques for purifying recombinant proteins
- Abstract
- 8.1 Introduction
- 8.2 Chromatographic methods for protein purification
- 8.3 Chromatography types
- 8.4 Chromatographic resins
- 8.5 Binding and elution of proteins
- 8.6 Storage of chromatographic columns
- 8.7 Application of chromatography to purification of biologics
- 8.8 Nonchromatographic methods for protein purification
- 8.9 Important points to consider for strategizing protein purification
- 8.10 Conclusion and future perspectives
- References
- Chapter 9. An overview of the contamination sources in recombinant proteins and their characterization
- Abstract
- Abbreviations
- 9.1 Introduction
- 9.2 Sources of contamination
- 9.3 Characterization methods
- 9.4 Conclusion
- References
- Chapter 10. Analytical methods for evaluating and characterizing recombinant proteins
- Abstract
- 10.1 Introduction
- 10.2 Applications of recombinant protein analysis
- 10.3 Protein quantification
- 10.4 Enzyme activity
- 10.5 Electrophoretic analyses of protein
- 10.6 Mass spectrometry
- 10.7 Conclusion and future direction
- References
- Index
- No. of pages: 320
- Language: English
- Edition: 1
- Published: September 10, 2024
- Imprint: Academic Press
- Paperback ISBN: 9780323983884
- eBook ISBN: 9780323985840
DY
Deepti Yadav
Dr. Deepti Yadav grew up in Lucknow, Uttar Pradesh, India. After getting a master’s degree in microbiology from Delhi University, she joined the Durban University of Technology in South Africa to pursue her PhD in biotechnology with Prof. Tukayi Kudanga’s research group within the Department of Biotechnology and Food Science.
Dr. Yadav has worked in a number of research labs, where she gained hands-on experience in techniques such as molecular biology, tissue culture, cloning, protein expression in recombinant hosts and protein purification. She is particularly interested in the fields of enzyme biotechnology, enzyme kinetics, and enzyme applications.
After obtaining her PhD degree, Dr. Yadav was hired as a Research Associate by the School of Energy Materials at Mahatma Gandhi University, Kottayam, where she is working on synthesis of polyhydroxybutyrate from a locally available water hyacinth plant. She has written several scholarly papers and has been invited to speak at conferences. She has also coached and encouraged numerous young students to seek professions in biotechnology.
AG
Abhishek Guldhe
Dr. Abhishek Guldhe is an Associate Professor at Amity Institute of Biotechnology, Amity University, Maharashtra, India. He was awarded with the prestigious Ramalingaswami Re-entry Fellowship of the Department of Biotechnology, Government of India. His research focus areas are microalgal biotechnology, biofuels, bioactive compounds, environmental biotechnology, and waste beneficiation. He teaches various subjects of biotechnology and environmental sciences. He has over 45 publications which include papers and book chapters in reputed international journals and books.
TK
Tukayi Kudanga
Prof. Tukayi Kudanga holds a PhD in biotechnology and is a professor in the Department of Biotechnology and Food Science at Durban University of Technology, South Africa. He is a recognized authority in laccase applications, particularly in biocatalysis and surface functionalization of lignocellulose materials. His areas of expertise include enzyme biotechnology, biocatalysis, and biomaterials. His current research focuses on enzymatic synthesis of bioactive compounds, enzymatic modification of food biopolymers, and development of green technologies for functionalization of lignocellulose materials. He is also interested in the search for novel biocatalysts, developing new applications and modifying enzymes for relevant biotechnological applications. Prof. Kudanga has extensive experience in teaching and research and has published his works widely. He has been invited to be plenary speaker at international conferences and reviews for several scientific journals.