Recombinant Protein Expression: Eukaryotic hosts
- 1st Edition, Volume 660 - November 4, 2021
- Imprint: Academic Press
- Editors: Zvi Kelman, William B. O'Dell
- Language: English
- Hardback ISBN:9 7 8 - 0 - 3 2 3 - 9 0 7 3 7 - 8
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 9 0 7 3 8 - 5
Recombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters… Read more
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Request a sales quoteRecombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more.
Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae.
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in the Methods in Enzymology serial
- Updated release includes the latest information on Recombinant Protein Expression
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
- Cover
- Title page
- Table of Contents
- Copyright
- Contributors
- Preface
- Yeast hosts
- Baculovirus–insect cell systems
- Plant hosts
- Mammalian hosts
- Section I: Yeast hosts
- Chapter One: Expression of recombinant multi-protein complexes in Saccharomyces cerevisiae
- Abstract
- 1: Introduction
- 2: Yeast strain
- 3: Gal 1/10 promotor and plasmid series
- 4: Protein affinity tags
- 5: Induction of recombinant protein/complexes from an episomal plasmid(s)
- 6: Expression of genes that are integrated into the yeast genome
- 7: Overview and conclusion
- Acknowledgments
- References
- Chapter Two: Recombinant production of membrane proteins in yeast
- Abstract
- 1: Introduction
- 2: The Saccharomyces cerevisiae expression platform
- 3: Initial screening for optimal expression and protein purification conditions
- 4: Generation of clean expression plasmids by homologous recombination
- 5: Application of yeast for in vivo measurement of recombinant membrane protein activity
- 6: Key resources table
- 7: Materials and equipment
- 8: Step-by-step method details
- 9: Expected outcomes
- 10: Advantages
- 11: Limitations
- 12: Optimization and troubleshooting
- 13: Safety considerations and standards
- 14: Alternative methods/procedures
- References
- Chapter Three: Expression of proteins in Pichia pastoris
- Abstract
- 1: Introduction
- 2: Before you begin
- 3: Key resources table
- 4: Materials and equipment
- 5: Step-by-step method details
- 6: Expected outcomes
- 7: Quantification and statistical analysis
- 8: Advantages
- 9: Limitations
- 10: Optimization and troubleshooting
- 11: Safety considerations and standards
- 12: Alternative methods/procedures
- 13: Notes
- References
- Chapter Four: Hybrid-architectured promoter design to engineer expression in yeast
- Abstract
- 1: Introduction
- 2: Hybrid-architectured promoter design to engineer recombinant protein expression
- 3: Method for hybrid-architectured EPV design
- 4: Protocol
- 5: Summary
- Acknowledgments
- References
- Chapter Five: Hybrid-architectured promoter design to deregulate expression in yeast
- Abstract
- 1: Introduction
- 2: Hybrid-architectured engineered promoter design for deregulated expression in yeast
- 3: Method for hybrid-architectured EPV design to deregulate expression
- 4: Protocol
- 5: Summary
- Acknowledgment
- References
- Section II: Baculovirus–insect cell systems
- Chapter Six: The MultiBac BEVS: Basics, applications, performance and recent developments
- Abstract
- 1: Introduction
- 2: MultiBac development
- 3: Baculovirus-vectored heterologous protein expression protocol
- 4: Key resources table
- 5: Materials and equipment
- 6: Basic step-by-step method for baculovirus-vectored heterologous protein expression
- 7: Expected outcomes
- 8: Advantages
- 9: Limitations
- 10: Safety considerations and standards
- 11: Alternative methods/procedures
- 12: Expanding the scope
- 13: Use of MultiBac for SARS-CoV-2 research
- 14: Conclusions and outlook
- References
- Chapter Seven: Applications of Golden Gate cloning to protein production using the baculovirus expression vector system
- Abstract
- 1: Introduction
- 2: Vector design
- 3: Protocol
- 4: Advantages
- 5: Limitations
- 6: Optimization and troubleshooting
- Acknowledgments
- References
- Chapter Eight: A fast-track protocol for protein expression using the BEV system
- Abstract
- 1: Introduction
- 2: Equipment and materials
- 3: Protocols
- 4: Notes
- 5: Advantages
- 6: Limitations
- References
- Section III: Plant hosts
- Chapter Nine: Transient expression of recombinant proteins in plants
- Abstract
- 1: Introduction
- 2: Vectors for transient protein expression in plants
- 3: Codon optimization
- 4: Transient expression
- 5: Extraction of soluble proteins
- 6: Application of high concentration of ascorbate via a foliar spray
- 7: Conclusion
- Acknowledgments
- References
- Chapter Ten: Transient expression of recombinant proteins in plants using potato virus X based vectors
- Abstract
- 1: Introduction
- 2: Recombinant expression vectors based on potato virus X genome
- 3: Materials and equipment
- 4: Step-by-step method details
- 5: Expected outcomes
- 6: Advantages and limitations
- 7: Current state, challenges, prospect of plants as biofactories
- 8: Summary
- Acknowledgments
- References
- Chapter Eleven: Design and expression of a bispecific antibody against dengue and chikungunya virus in plants
- Abstract
- 1: Introduction
- 2: Dengue and chikungunya infections
- 3: Antibodies against dengue and chikungunya
- 4: Dengue and chikungunya bispecific antibody design
- 5: Protocol
- 6: Results, discussion, and conclusions
- References
- Chapter Twelve: Manufacturing plant-made monoclonal antibodies for research or therapeutic applications
- Abstract
- 1: Introduction
- 2: Overview of the workflow
- 3: Upstream processing: Plant growth, Agrobacteria culture, infiltration, and biomass extraction
- 4: Upstream processing procedures
- 5: Downstream processing: Chromatography, filtration, and storage
- 6: Downstream processing procedures
- 7: Quality assurance, purity, and safety
- 8: Advantages and limitations
- 9: Scaling for research applications
- 10: Summary
- References
- Section IV: Mammalian hosts
- Chapter Thirteen: Creation of monoclonal antibody expressing CHO cell lines grown with sodium butyrate and characterization of resulting antibody glycosylation
- Abstract
- 1: Introduction
- 2: Background
- 3: Materials and equipment
- 4: Cell line development of mAb producing clones with varying productivities
- 5: Sodium butyrate (NaBu) concentration optimization and treatment of developed cell lines
- 6: Protein A chromatography
- 7: N-glycan analysis using mass spectrometry
- 8: Summary
- 9: Notes
- References
- Chapter Fourteen: Method to transfer Chinese hamster ovary (CHO) batch shake flask experiments to large-scale, computer-controlled fed-batch bioreactors
- Abstract
- 1: Introduction
- 2: Materials and equipment
- 3: Step-by-step method details
- 4: Advantages
- 5: Limitations
- 6: Safety considerations and standards
- Acknowledgments
- References
- Chapter Fifteen: Inducible protein expression in piggyBac transposase mediated stable HEK293 cell pools
- Abstract
- 1: Introduction
- 2: Equipment and materials
- 3: Protocols
- 4: Notes
- 5: Conclusions
- References
- Chapter Sixteen: Exploring the design space of AAV transient-transfection in suspension cell lines
- Abstract
- 1: Background
- 2: Study design
- 3: Materials
- 4: Equipment
- 5: Step-by-step method details
- 6: Conclusions
- Acknowledgment
- References
- Edition: 1
- Volume: 660
- Published: November 4, 2021
- Imprint: Academic Press
- No. of pages: 380
- Language: English
- Hardback ISBN: 9780323907378
- eBook ISBN: 9780323907385
ZK
Zvi Kelman
Zvi Kelman is the Director of the Biomolecular Labeling Laboratory (BL2), National Institute of Standards and Technology (NIST). He is also an Adjunct Professor in the Department of Cell Biology and Molecular Genetics at the University of Maryland, College Park and affiliated with the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine. Zvi earned a B.Sc. in Agriculture from the Hebrew University in Jerusalem and a M.Sc. in Cell Biology from the Weizmann Institute of Science. After receiving a Ph.D. in Molecular Biology from Cornell University, he was a Helen Hay Whitney Foundation Post-Doctoral Fellow at Johns Hopkins University and Memorial Sloan-Kettering Cancer Center. Upon completion of his postgraduate training, he became a Life Technologies Professor at the Center for Advanced Research in Biotechnology (CARB), University of Maryland Biotechnology Institute (UMBI). Zvi moved to the University of Maryland, College Park in 2010 as a Professor in the Department of Cell Biology and Molecular Genetics. In 2011 he was recruited to NIST to direct the Biomolecular Labeling Laboratory.
Affiliations and expertise
Director, Biomolecular Labeling Laboratory, NIST-IBBR, Rockville, MD, USAWO
William B. O'Dell
William Brad O'Dell is a PhD biologist experienced in in Molecular Biology, Protein Expression, Protein Purification, Crystallography, Lab-Scale Yeast Fermentation, Flow Cytometry and Small Angle Scattering of Molecules and Materials.
Affiliations and expertise
Biologist, National Institute of Standards and Technology (NIST), MD, USARead Recombinant Protein Expression: Eukaryotic hosts on ScienceDirect