Affinity Chromatography

This expanded and updated volume aims to provide the beginner with the practical knowledge needed to develop affinity separations suitable for a variety of applications relevant to the post-genomic era. It contains state-of-the-art, brand new protocols.

Expanded and updated from the first edition, this second edition volume aims to provide the beginner with the practical knowledge needed to develop affinity separations suitable for a variety of applications relevant to the post-genomic era. It contains state-of-the-art, brand new protocols, and provides step-by-step laboratory instructions for readily reproducible results. Thirty-eight years after its introduction, affinity chromatography remains a key tool in the armory of separation techniques available to separation and interaction scientists.

Contains state-of-the-art, brand new protocols developed for this second edition Provides step-by-step laboratory instructions for readily reproducible results

Klappentext

Thirty-eight years after its introduction, affinity chromatography remains a key tool in the armory of separation techniques available to separation and interaction scientists. Expanded and updated from the first edition, Affinity Chromatography: Methods and Protocols, Second Edition, provides the beginner with the practical knowledge needed to develop affinity separations suitable for a variety of applications relevant to the post-genomic era. This second edition expands on the first edition by introducing more state-of-the-art protocols used in affinity chromatography. This new edition also describes protocols that demonstrate the concept of affinity chromatography being applied to meet the modern high throughput screening demands of researchers and development scientists whilst expanding on some more traditional affinity chromatography approaches that have become of greater interest to separation scientists. Chapters in this cutting-edge text expand on affinity chromatography techniques that currently enjoy frequent citation in the literature from those purifying biomolecules. Other chapters include protocols describing the use of a variety of fusion tags as well as how to cleave them, so as to allow the scientists to study the native phenotype of the protein. Renowned researchers also include protocols detailing diverse applications of affinity chromatography such as its use in catalytic reactions, DNA purification, whole cell separations and for the isolation of phosphorylated proteins. Affinity Chromatography: Methods and Protocols, Second Edition, is an essential reference for those interested in separation sciences, particularly in the pharmaceutical and biological research sectors, that have an interest in isolating macromolecules rapidly, quantitatively, and with high purity.

Inhalt
Preface.- Contributors.- Affinity Chromatography: History, Perspectives, Limitations and ProspectsAna Ceclia A. Roque and Christopher R. Lowe 1.- Various Modes of Affinity Chromatography.- Immobilized Metal Ion Affinity Chromatography of Native ProteinsAdam Charlton and Michael Zachariou.- Affinity Precipitation of Proteins Using Metal ChelatesAshok Kumar, Igor Yu. Galaev, and Bo Mattiasson.- Immunoaffinity ChromatographyStuart R. Gallant, Vish Koppaka, and Nick Zecherle.- Dye Ligand ChromatographyStuart R. Gallant, Vish Koppaka, and Nick Zecherle.- Purification of Proteins Using Displacement ChromatographyNihal Tugcu.- Affinity Chromatography Using Purification Tags.- Rationally Designed Ligands for Use in Affinity Chromatography: An Artificial Protein LAna Ceclia A. Roque and Christopher R. Lowe.- Phage Display of Peptides in Ligand Selection for Use in Affinity ChromatographyJoanne L. Casey, Andrew M. Coley, and Michael Foley.- Preparation, Analysis and Use of an Affinity Adsorbent for the Purification of GST Fusion ProteinGareth M. Forde.- Immobilized Metal Ion Affinity Chromatography of Histidine-Tagged Fusion ProteinsAdam Charlton and Michael Zachariou.- Methods for the Purification of HQ-Tagged ProteinsBecky Godat, Laurie Engel, Natalie A. Betz, and Tonny M. Johnson.- Amylose Affinity Chromatography of Maltose-Binding Protein: Purification by both Native and Novel Matrix-Assisted Dialysis Refolding MethodsLeonard K. Pattenden and Walter G. Thomas.- Methods for Detection of ProteinProtein and ProteinDNA Interactions Using HaloTagMarjeta Urh, Danette Hartzell, Jacqui Mendez, Dieter H. Klaubert, and Keith Wood.- Site-Specific Cleavage of Fusion ProteinsAdam Charlton.- The Use of TAGZyme for the Efficient Removal of N-Terminal His-TagsJose? Arnau, ConniLauritzen, Gitte Ebert Petersen, and John Pedersen.- Various Applications of Affinity Chromatography.- Affinity Processing of Cell-Containing Feeds Using Monolithic Macroporous Hydrogels, CryogelsIgor Yu. Galaev and Bo Mattiasson.- Monolithic Bioreactors for MacromoleculesMojca Ben?ina, Katja Ben?ina, Ale Podgornik, and Ale trancar.- Plasmid DNA Purification Via the Use of a Dual Affinity ProteinGareth M. Forde.- Affinity Chromatography of Phosphorylated ProteinsGrigoriy S. Tchaga.- Protein Separation Using Immobilized Phospholipid ChromatographyTzong-Hsien Lee and Marie-Isabel Aguilar.- Analysis of Proteins in Solution Using Affinity Capillary ElectrophoresisNiels H. H. Heegaard, Christian Schou, and Jesper Østergaard.