Affinity Chromatography - Methods and Protocols (2nd edition)

Hardcover  348 pp  ISBN 9781588296597      £64.00

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.

This book 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.

Written for geneticists, molecular biologists, biotechnologists, pharmaceutical scientists

Contents

• Affinity Chromatography: History, Perspectives, Limitations and Prospects - Ana Cecília A. Roque and Christopher R. Lowe

VARIOUS MODES OF AFFINITY CHROMATOGRAPHY

• Immobilized Metal Ion Affinity Chromatography of Native Proteins - Adam Charlton and Michael Zachariou
• Affinity Precipitation of Proteins Using Metal Chelates - Ashok Kumar, Igor Yu. Galaev and Bo Mattiasson
• Immunoaffinity Chromatography - Stuart R. Gallant, Vish Koppaka and Nick Zecherle
• Dye-ligand Chromatography - Stuart R. Gallant, Vish Koppaka and Nick Zecherle
• Purification of Proteins Using Displacement Chromatography - Nihal Tugcu

AFFINITY CHROMATOGRAPHY USING PURIFICATION TAGS

• Rationally Designed Ligands for use in Affinity Chromatography: an Artificial Protein L - Ana Cecília A. Roque and Christopher Lowe
• Phage display of Peptides in Ligand Selection for use in Affinity Chromatography - Joanne L Casey, Andrew M. Coley and Michael Foley
• Preparation, Analysis and Use of an Affinity Adsorbent for the Purification of GST Fusion Protein - Gareth M. Forde
• Immobilized Metal Ion Affinity Chromatography of Histidine-tagged fusion proteins - Adam Charlton and Michael Zachariou
• Methods for the Purification of HQ-Tagged Proteins - Beckky Godat, Laurie Engel, Natalie Betz and Tonny M. Johnson
• Amylose Affinity Chromatography of Maltose-Binding Protein: Purification by both Native and Novel Matrix-Assisted Dialysis-Refolding Methods. - Leonard K. Pattenden and Walter G. Thomas
• Methods for the Detection of Protein-Protein and Protein-DNA Interactions using Halo-Tag ^TM - Marjeta Urh, Danette Hartzell, Jacqui Mendez and Keith Wood
• Site-specific Cleavage of Fusion Proteins - Adam Charlton
• The Use of TAGzyme for the efficient Removal of N-terminal His-tags - José Arnau, Conni Lauritzen, Gitte E. Petersen and John Pedersen

VARIOUS APPLICATIONS OF AFFINITY CHROMATOGRAPHY.

• Affinity Processing of Cell-containing Feeds Using Monolithic Macroporous Hydrogels, Cryogels - Igor Yu. Galaev and Bo Mattiasson
• Monolithic Bioreactors for Macromolecules - Mojca Ben_ina, Katja Ben_ina, Ale_ Podgornik and Ale_ _trancar
• Plasmid DNA Purification via the Use of a Dual Affinity Protein - Gareth M. Forde
• Affinity Chromatography of Phosphorylated Proteins - Grigoriy S. Tchaga
• Protein Separation Using Phospholipid Chromatography - Tzong-Hsien Lee and Marie-Isabel Aguilar 21 Analysis of Proteins in Solution Using Affinity Capillary Electrophoresis - Neils H.H. Heegaard, Christian Schou and Jesper Ostergaard