Protein Chromatography

1. Front Matter

   Pages i-x

   PDF

2. Overviews

   1. Front Matter

      Pages 1-1

      PDF

   2. A Digest of Protein Purification

      • Dermot Walls, Robert McGrath, Sinéad T. Loughran

      Pages 3-23

   3. Gel-Filtration Chromatography

      • Ciarán Ó’Fágáin, Philip M. Cummins, Brendan F. O’Connor

      Pages 25-33

   4. Immunoaffinity Chromatography

      • Jenny Fitzgerald, Paul Leonard, Elaine Darcy, Richard O’Kennedy

      Pages 35-59

   5. Avoiding Proteolysis During Protein Chromatography

      • Barry J. Ryan

      Pages 61-71

   6. Scale-Up of Protein Purification: Downstream Processing Issues

      • John Joseph Milne

      Pages 73-85

   7. Phage Display: A Powerful Technology for the Generation of High Specificity Affinity Reagents from Alternative Immune Sources

      • William J. J. Finlay, Laird Bloom, Orla Cunningham

      Pages 87-101

   8. Engineering Protein Stability

      • Ciarán Ó’Fágáin

      Pages 103-136

   9. Microfluidics in Protein Chromatography

      • Frank A. Gomez

      Pages 137-150

   10. Tagging Recombinant Proteins to Enhance Solubility and Aid Purification

       • Dermot Walls, Sinéad T. Loughran

       Pages 151-175

3. Protocols

   1. Front Matter

      Pages 177-177

      PDF

   2. Storage and Lyophilisation of Pure Proteins

      • Ciarán Ó’Fágáin

      Pages 179-202

   3. Differential Precipitation and Solubilization of Proteins

      • Barry J. Ryan

      Pages 203-213

   4. Ion-Exchange Chromatography: Basic Principles and Application to the Partial Purification of Soluble Mammalian Prolyl Oligopeptidase

      • Philip M. Cummins, Oonagh Dowling, Brendan F. O’Connor

      Pages 215-228

   5. Protein Quantitation and Analysis of Purity

      • Eva M. Campion, Sinéad T. Loughran, Dermot Walls

      Pages 229-258

   6. Purification of Proteins Fused to Glutathione S-Transferase

      • Sandra Harper, David W. Speicher

      Pages 259-280

   7. Purification of Proteins Fused to Maltose-Binding Protein

      • Mario Lebendiker, Tsafi Danieli

      Pages 281-293

   8. Purification of Proteins from Baculovirus-Infected Insect Cells

      • Luke O’Shaughnessy, Sean Doyle

      Pages 295-309

   9. Purification of Poly-Histidine-Tagged Proteins

      • Sinéad T. Loughran, Dermot Walls

      Pages 311-335

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™ 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.

• Chromatographic methods
• Microfluidics
• Native protein isolation
• Protein biochemistry
• Protein quantitation
• Purification schemes
• Recombinant antibody production
• chromatography

• National Centre for Sensor Research, School of Biotechnology, Dublin City University, Dublin, Ireland

  Dermot Walls, Sinéad T. Loughran

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