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High Throughput Protein Expression and Purification
Methods and Protocols
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Main description:

Despite exciting advances in genome sequencing, isolating a protein from its expression system in its native form still presents a complex challenge. In High Throughput Protein Expression and Purification: Methods and Protocols, leading scientists detail the most successful protocols currently in use, including various high throughput cloning schemes, protein expression analysis, and production protocols. This volume describes the use of E. coli, insect, and mammalian cells, as well as cell-free systems for the production of a wide variety of proteins, including glycoproteins and membrane proteins, in order to best represent strategies that create and exploit common features to enable simplified cloning, stable expression, and purification of proteins. Written in the highly successful Methods in Molecular Biology™ series format, the chapters present brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section for tips on troubleshooting and avoiding known pitfalls.


Cutting-edge and comprehensive, High Throughput Protein Expression and Purification: Methods and Protocols is an ideal reference for protein biochemists and all those who wish to apply these easy-to-use protocols to the many applicable fields.


Feature:

Provides step-by-step protocols for protein production that utilize a wide range of cutting-edge techniques


Covers cell-based and cell-free methodologies needed to produce bacterial proteins, eukaryotic proteins, soluble and membrane proteins


Examines a variety of cloning methodologies that enable high-throughput cloning for protein production


Describes the use of a variety of tags to facilitate parallel processing for high-throughput protein production


Includes several reviews to help readers choose which protocols best suits their needs


Provides protocols that utilize robotics for sample handling, as well as protocols designed small laboratories performing experiments by hand


Back cover:

Despite exciting advances in genome sequencing, isolating a protein from its expression system in its native form still presents a complex challenge. In High Throughput Protein Expression and Purification: Methods and Protocols, leading scientists detail the most successful protocols currently in use, including various high throughput cloning schemes, protein expression analysis, and production protocols. This volume describes the use of E. coli, insect, and mammalian cells, as well as cell-free systems for the production of a wide variety of proteins, including glycoproteins and membrane proteins, in order to best represent strategies that create and exploit common features to enable simplified cloning, stable expression, and purification of proteins. Written in the highly successful Methods in Molecular Biology™ series format, the chapters present brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section for tips on troubleshooting and avoiding known pitfalls.


Cutting-edge and comprehensive, High Throughput Protein Expression and Purification: Methods and Protocols is an ideal reference for protein biochemists and all those who wish to apply these easy-to-use protocols to the many applicable fields.


Contents:

Part I: Reviews

1. High Throughput Protein Production (HTPP): A Review of Enabling Technologies to Expedite Protein Production
Jim Koehn and Ian Hunt

2. Designing Experiments for High-Throughput Protein Expression
Stephen P. Chambers and Susanne E. Swalley


Part II: Protocols for High-throughput Cloning

3. Gateway Cloning for Protein Expression
Dominic Esposito, Leslie A. Garvey, and Chacko S. Chakiath

4. Flexi Vector Cloning
Paul G. Blommel, Peter A. Martin, Kory D. Seder, Russell L. Wrobel, and Brian G. Fox

5. The Precise Engineering of Expression Vectors Using High Throughput In-Fusion™ PCR Cloning
Nick S. Berrow, David Alderton, and Raymond J Owens

6. The Polymerase Incomplete Primer Extension (PIPE) Method Applied to High-Throughput Cloning and Site-Directed Mutagenesis
Heath E. Klock and Scott A. Lesley

7. A Family of LIC Vectors for High-Throughput Cloning and Purification of Proteins
William H. Eschenfeldt, Lucy Stols, Cynthia Sanville Millard, Andrzej Joachimiak, and Mark I. Donnelly

8. 'System 48' High Throughput Cloning and Protein Expression Analysis
James M. Abdullah, Andrzej Joachimiak, and Frank R. Collart

Part III: Protocols for High-throughput Protein Expression and Purification

9. Automated 96-Well Purification of Hexahistidine-Tagged Recombinant Proteins on MagneHis Ni2+-Particles
Chiann-Tso Lin, Priscilla A. Moore, and Vladimir Kery
10. E.coli and Insect Cell Expression, Automated Purification and Quantitative Analysis
Stephen P. Chambers, John R. Fulghum, Douglas A. Austen, Fan Lu, and Susanne E. Swalley

11. Hexahistidine-Tagged Maltose-Binding Protein as a FusionPartner for the Production of Soluble Recombinant Proteins in Escherichia coli
Brian P. Austin, Sreedevi Nallamsetty, and David S. Waugh

12. PHB-Intein-Mediated Protein Purification Strategy
Alison R. Gillies, Mahmoud Reza Banki, and David W. Wood

13. High-Throughput Biotinylation of Proteins
Brian K. Kay, Sang Thai, and Veronica V. Volgina

14. High-Throughput Insect Cell Protein Expression Applications
Mirjam Buchs, Ernie Kim, Yann Pouliquen, Michael Sachs, Sabine Geisse, Marion Mahnke, and Ian Hunt

15. High-Throughput Protein Expression Using Cell-free System
Kalavathy Sitaraman and Deb K. Chatterjee

16. The Production of Glycoproteins by Transient Expression in Mammalian Cells
Joanne E. Nettleship, Nahid Rahman-Huq, and Raymond J. Owens

17. High-Throughput Expression and Detergent Screening of Integral Membrane Proteins
Said Eshaghi

18. Cell-Free Expression for Nanolipoprotein Particles (NLPs): Building a High-Throughput Membrane Protein Solubility Platform
Jenny A. Cappuccio, Angela K. Hinz, Edward A. Kuhn, Julia Fletcher, Erin S. Arroyo, Paul T. Henderson, Craig D. Blanchette, Vicki L. Walsowrth, Michele H. Corzett, Richard J. Law, Joseph B. Pesavento, Brent W. Segelke, Todd A. Sulchek, Brett A. Chromy, Federico Katzen, Todd Peterson, Graham Bench, Wieslaw Kudlicki, Paul D. Hoeprich Jr., and Matthew A. Coleman

19. Expression and Purification of Soluble His6-Tagged TEV Protease
Joseph E. Tropea, Scott Cherry, and David S. Waugh

20. High-Throughput Protein Concentration and Buffer Exchange: Comparison of Ultrafiltration and Ammonium Sulfate Precipitation
Priscilla A. Moore and Vladimir Kery


PRODUCT DETAILS

ISBN-13: 9781588298799
Publisher: Springer (Humana Press)
Publication date: October, 2008
Pages: 322
Weight: 917g
Availability: Not available (reason unspecified)
Subcategories: Biochemistry
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CUSTOMER REVIEWS

Average Rating 

From the reviews:

"The book consists 20 chapters, which cover methods in protein expression and purification summarized by leading experts in this area from USA, UK, and Switzerland. … Numerous pictures in the text, a references list at the end of each chapter, and a subject index help better understand the presented materials. The volume is true handbook for protein biochemists, molecular biologists, biotechnologists, and other researchers and technicians who are involved in the many applicable fields." (G. Wiederschain, Biochemistry (Moscow), Vol. 74 (8), 2009)

“High Throughput Protein Expression and Purification: Methods and Protocols, edited by Sharon A. Doyle, is an interesting attempt showing what one currently has at one’s disposal if one wishes to make a first step toward this aim. … this book should be recommended especially to researchers who would like to organize a new laboratory focused on high throughput protein production, or to improve an existing one. It can also prove helpful to those who wish to optimize individual protein expression, isolation or purification.” (Marek Figlerowicz, Acta Biochimica Polonica, Vol. 56 (4), December, 2009)