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Protein Chromatography

  • Kartonierter Einband
  • 540 Seiten
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This book examines vital topics in protein biochemistry and presents methods involving the generation and purification of recombin... Weiterlesen
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This book examines vital topics in protein biochemistry and presents methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, as well as variations on classic techniques.

A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Protocols, leading researchers present clear protocol-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular Biology(TM) series format, protocols chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Protein Chromatography: Methods and Protocols will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.


Part I: Overviews

1. A Digest of Protein Purification
Dermot Walls, Robert McGrath, and Sinéad T. Loughran

2. Gel-Filtration Chromatography
Ciarán Ó'Fágáin, Philip M. Cummins, and Brendan F. O'Connor

3. Immunoaffinity Chromatography
Jenny Fitzgerald, Paul Leonard, Elaine Darcy, and Richard O'Kennedy

4. Avoiding Proteolysis During Protein Chromatography
Barry J. Ryan

5. Scale-Up of Protein Purification: Downstream Processing Issues
John Joseph Milne

6. Phage Display: A Powerful Technology for the Generation of High Specificity Affinity Reagents from Alternative Immune Sources
William J.J. Finlay, Laird Bloom, and Orla Cunningham

7. Engineering Protein Stability
Ciarán Ó'Fágáin

8. Microfluidics in Protein Chromatography
Frank A. Gomez

9. Tagging Recombinant Proteins to Enhance Solubility and Aid Purification
Dermot Walls and Sinéad T. Loughran

Part II: Protocols

10. Storage and Lyophilization of Pure Proteins
Ciarán Ó'Fágáin

11. Differential Precipitation and Solubilization of Proteins
Barry J. Ryan

12. Ion-Exchange Chromatography: Basic Principles and Application to the Partial Purification of Soluble Mammalian Prolyl Oligopeptidase
Philip M. Cumminsa, Oonagh Dowling, and Brendan F. O'Connor

13. Protein Quantitation and Analysis of Purity
Eva M. Campion, Sinéad T. Loughran, and Dermot Walls

14. Purification of Proteins Fused to Glutathione S-Tranferase
Sandra Harper and David W. Speicher

15. Purification of Proteins Fused to Maltose-Binding Protein
Mario Lebendiker and Tsafi Danieli

16. Purification of Proteins from Baculovirus-Infected Insect Cells
Luke O'Shaughnessy and Sean Doyle

17. Purification of Poly-Histidine-Tagged Proteins
Sinéad T. Loughran and Dermot Walls

18. Immobilized Metal Affinity Chromatography/Reversed-Phase Enrichment of Phosphopeptides and Analysis by CID/ETD Tandem Mass Spectrometry
Rosana Navajas, Alberto Paradela, and Juan Pablo Albar

19. Tag Removal by Site-Specific Cleavage of Recombinant Fusion Proteins
Adam Charlton and Michael Zachariou

20. Purification of Antibodies Using Affinity Chromatography
Elaine Darcy, Paul Leonard, Jenny Fitzgerald, Martin Danaher, and Richard O' Kennedy

21. Optimized Generation of High-Affinity, High-Specificity Single-Chain Fv Antibodies from Multi-Antigen Immunized Chickens
William J.J. Finlay, Laird Bloom, and Orla Cunningham

22. Measuring Protein-Protein Interactions Using Biacore
Paul Leonard, Stephen Hearty, and Richard O'Kennedy

23. Ultra-Performance Liquid Chromatography-Mass Spectrometry of Proteins
Timothy R. Croley

24. Hydrophobic Interaction Chromatography
Philip Cummins and Brendan O'Connor

25. Fast Protein Liquid Chromatography
Ashkan Madadlou, Siobhan O'Sullivan, and David Sheehan

26. Shotgun Proteomics: A Qualitative Approach Applying Isoelectric Focusing on Immobilized pH Gradient and LC-MS/MS
Laurent Geiser, Ali R. Vaezzadeh, Jacques M. P. Deshusses, and Denis F. Hochstrasser

27. Shotgun Proteomics: A Relative Quantitative Approach Using Off-Gel Electrophoresis and LC-MS/MS
Laurent Geiser, Loïc Dayon, Ali R. Vaezzadeh, and Denis F. Hochstrasser

28. Clinical Proteomics: Liquid Chromatography-Mass Spectrometry (LC/MS) Purification Systems
Michael Henry and Paula Meleady

29. Strategies for the Purification of Membrane Proteins
Sinead Marian Smith

30. A Multi-Step Chromatographic Strategy to Purify Three Fungal Endo-ß-Glucanases
Tracey McCarthy and Maria G. Tuohy


Titel: Protein Chromatography
Untertitel: Methods and Protocols
EAN: 9781493961290
ISBN: 1493961292
Format: Kartonierter Einband
Herausgeber: Humana Press
Anzahl Seiten: 540
Gewicht: 1003g
Größe: H254mm x B178mm x T28mm
Jahr: 2016
Auflage: Softcover reprint of the original 1st ed. 2011

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