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This third edition expands on the previous editions with updated and new chapters on protein chromatography. Chapters detail protein stability and storage, avoiding proteolysis, protein quantitation methods, generation and purification of recombinant proteins, recombinant antibody production, and the tagging of proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Protein Chromatography: Methods and Protocols, Third Edition aims to provide commonly used methods and new approaches to help both new researchers and experts expand their knowledge.
List of contents
A Synopsis of Proteins and Their Purification.- Mixed-mode chromatography and its role in monoclonal antibody purification.- Application of Continuous Counter-current Chromatography in Protein Purification.- Chromatographic Purification of Viral Vectors for Gene Therapy Applications.- Scale-up of Protein Purification -Downstream Processing Issues.- Approaches to avoid Proteolysis During Protein Expression and Purification.- Tagging Recombinant Proteins to Enhance Solubility and Aid Purification.- Magnetic nanoparticles for protein separation and purification.- Ion-Exchange Chromatography: Basic Principles and Application.- Hydroxyapatite Chromatography.- Poly-Histidine-Tagged Protein Purification using IMAC.- Lectin Affinity Chromatography.- Single-step non-chromatographic purification of recombinant mammalian proteins using a split intein ELP tag system.- Clinical Proteomics: Liquid Chromatography-Mass Spectrometry (LC/MS) Purification Systems.- Immunoprecipitation: Variations, Considerations, and Applications.- Protein Quantitation and Analysis of Purity.- Protein Extraction and Purification by Differential Solubilisation.- Protein Stability: Enhancement and Measurement.- Storage and Lyophilisation of Pure Proteins.- Strategies for the Purification of Membrane Proteins.
