Protein Chromatography: Methods and Protocols (Methods in Molecular Biology)

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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.

Part I Overview Chapters1. A Synopsis of Protein and their PurificationDermot Walls, Gary Cooney, and Sinéad T. Loughran2. Gel-Filtration ChromatographyCiarán Ó'Fágáin, Philip M. Cummins, and Brendan F. O'Connor3. Immunoaffinity Chromatography: Concepts and ApplicationsJenny Fitzgerald, Paul Leonard, Elaine Darcy, Shikha Sharma, and Richard O'Kennedy4. Avoiding Proteolysis During Protein PurificationBarry J. Ryan and Gary T. Henehan5. Scale-Up of Protein Purification: Downstream Processing IssuesJohn Joseph Milne6. Phage Display: A Powerful Technology for the Generation of High Specificity Affinity Reagents from Alternative Immune SourcesWilliam J.J. Finlay, Laird Bloom, Joanne Grant, Edward Franklin, Deirdre Ní Shúilleabháin, and Orla Cunningham7. Protein Stability: Enhancement and Measurement Ciarán Ó'Fágáin8. Tagging Recombinant Proteins to Enhance Solubility and Aid Purification Sinéad T. Loughran and Dermot WallsPart II Protocols Chapters9. Storage and Lyophilisation of Pure ProteinsCiarán Ó'Fágáin and Keith Colliton