Synopses & Reviews
Synopsis
This second edition expands on the previous edition with new chapters that are suitable for newcomers, as well as more detailed chapters that cover protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the challenges that scale-up of production poses to the investigator. Many of the chapters also discuss generation and purification of recombinant proteins, recombinant antibody production, and the tagging of proteins as a means to enhance their solubility and simplify their purification on an individual scale or in high-throughput systems. This book also provides readers with chapters that describe not just the more commonly used methods, but also recently developed approaches such as proteomic/mass spectrometric techniques and Lectin-based affinity chromatography. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Protein Chromatography: Methods and Protocols, Second Edition is a valuable resource for anyone who is interested in the field of protein chromatography.
Synopsis
Part I Overview Chapters
1. A Synopsis of Protein and their Purification
Dermot Walls, Gary Cooney, and Sin ad T. Loughran
2. Gel-Filtration Chromatography
Ciar n 'F g in, Philip M. Cummins, and Brendan F. O'Connor
3. Immunoaffinity Chromatography: Concepts and Applications
Jenny Fitzgerald, Paul Leonard, Elaine Darcy, Shikha Sharma, and Richard O'Kennedy
4. Avoiding Proteolysis During Protein PurificationBarry J. Ryan and Gary T. Henehan
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, Joanne Grant, Edward Franklin, Deirdre N Sh illeabh in, and Orla Cunningham
7. Protein Stability: Enhancement and Measurement
Ciar n 'F g in
8. Tagging Recombinant Proteins to Enhance Solubility and Aid Purification
Sin ad T. Loughran and Dermot Walls
Part II Protocols Chapters
9. Storage and Lyophilisation of Pure Proteins
Ciar n 'F g in and Keith Colliton
10. Differential Precipitation and Solubilization of Proteins
Barry J. Ryan and Gemma K. Kinsella
11. Ion-Exchange Chromatography: Basic Principles and Application
Philip M. Cummins, Keith D. Rochfort, and Brendan F. O'Connor
12. Protein Quantitation and Analysis of Purity
Eva M. Campion, Sin ad T. Loughran, and Dermot Walls
13. Purification of Proteins Fused to Maltose-Binding Protein
Mario Lebendiker and Tsafi Danieli
14. Purification of Poly-Histidine-Tagged Proteins
Sin ad T. Loughran, Ronan T. Bree, and Dermot Walls
15. Purification of Antibodies Using Affinity Chromatography
Elaine Darcy, Paul Leonard, Jenny Fitzgerald, Martin Danaher, Hui Ma, and Richard O' Kennedy
16. Optimized Generation of High-Affinity, High