Protein Chromatography

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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains keynotes and implementation advice from the experts

Inhalt

A Synopsis of Protein and their Purification.- Gel-Filtration Chromatography.- Immunoaffinity Chromatography: Concepts and Applications.- Avoiding Proteolysis During Protein Purification.- Scale-Up of Protein Purification: Downstream Processing Issues.- Phage Display: A Powerful Technology for the Generation of High Specificity Affinity Reagents from Alternative Immune Sources.- Protein Stability: Enhancement and Measurement.- Tagging Recombinant Proteins to Enhance Solubility and Aid Purification.- Storage and Lyophilisation of Pure Proteins.- Differential Precipitation and Solubilization of Proteins.- Ion-Exchange Chromatography: Basic Principles and Application.- Protein Quantitation and Analysis of Purity.- Purification of Proteins Fused to Maltose-Binding Protein.- Purification of Poly-Histidine-Tagged Proteins.- Purification of Antibodies Using Affinity Chromatography.- Optimized Generationof High-Affinity, High-Specificity Single-Chain Fv Antibodies from Multi-Antigen Immunized Chickens.- Measuring Protein-Protein Interactions Using Biacore.- Hydrophobic Interaction Chromatography.