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High Throughput Protein Expression and Purification

Methods and Protocols

Editors:

  • 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

Part of the book series: Methods in Molecular Biology (MIMB, volume 498)

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Table of contents (20 protocols)

  1. Front Matter

    Pages i-xii
  2. Designing Experiments for High-Throughput Protein Expression

    • Stephen P. Chambers, Susanne E. Swalley
    Pages 19-29
  3. Gateway Cloning for Protein Expression

    • Dominic Esposito, Leslie A. Garvey, Chacko S. Chakiath
    Pages 31-54
  4. Flexi Vector Cloning

    • Paul G. Blommel, Peter A. Martin, Kory D. Seder, Russell L. Wrobel, Brian G. Fox
    Pages 55-73
  5. The Precise Engineering of Expression Vectors Using High-Throughput In-Fusion™ PCR Cloning

    • Nick S. Berrow, David Alderton, Raymond J. Owens
    Pages 75-90
  6. A Family of LIC Vectors for High-Throughput Cloning and Purification of Proteins

    • William H. Eschenfeldt, Stols Lucy, Cynthia Sanville Millard, Andrzej Joachimiak, I. Donnelly Mark
    Pages 105-115
  7. “System 48” High-Throughput Cloning and Protein Expression Analysis

    • James M. Abdullah, Andrzej Joachimiak, Frank R. Collart
    Pages 117-127
  8. E. coli and Insect Cell Expression, Automated Purification and Quantitative Analysis

    • Stephen P. Chambers, John R. Fulghum, Douglas A. Austen, Fan Lu, Susanne E. Swalley
    Pages 143-156
  9. PHB-Intein-Mediated Protein Purification Strategy

    • Alison R. Gillies, Reza Banki Mahmoud, David W. Wood
    Pages 173-183
  10. High-Throughput Biotinylation of Proteins

    • Brian K. Kay, Sang Thai, Veronica V. Volgina
    Pages 185-198
  11. High-Throughput Insect Cell Protein Expression Applications

    • Mirjam Buchs, Ernie Kim, Yann Pouliquen, Michael Sachs, Sabine Geisse, Marion Mahnke et al.
    Pages 199-227
  12. High-Throughput Protein Expression Using Cell-Free System

    • Kalavathy Sitaraman, Deb K. Chatterjee
    Pages 229-244
  13. The Production of Glycoproteins by Transient Expression in Mammalian Cells

    • Joanne E. Nettleship, Nahid Rahman-Huq, Raymond J. Owens
    Pages 245-263
  14. Cell-Free Expression for Nanolipoprotein Particles: Building a High-Throughput Membrane Protein Solubility Platform

    • Jenny A. Cappuccio, Angela K. Hinz, Edward A. Kuhn, Julia E. Fletcher, Erin S. Arroyo, Paul T. Henderson et al.
    Pages 273-295
  15. Expression and Purification of Soluble His6-Tagged TEV Protease

    • Joseph E. Tropea, Scott Cherry, David S. Waugh
    Pages 297-307

About this book

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.

Reviews

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)

Editors and Affiliations

  • Glaxo Smith Kline, Walnut Creek, USA

    Sharon A. Doyle

Bibliographic Information

  • Book Title: High Throughput Protein Expression and Purification

  • Book Subtitle: Methods and Protocols

  • Editors: Sharon A. Doyle

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-59745-196-3

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2009

  • Hardcover ISBN: 978-1-58829-879-9Published: 20 October 2008

  • Softcover ISBN: 978-1-61737-821-8Published: 19 November 2010

  • eBook ISBN: 978-1-59745-196-3Published: 06 November 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XII, 322

  • Number of Illustrations: 51 b/w illustrations, 6 illustrations in colour

  • Topics: Biochemistry, general, Protein Science, Protein Structure

  • Industry Sectors: Biotechnology, Consumer Packaged Goods, Pharma

Buy it now

Buying options

eBook USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Other ways to access