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Affinity Chromatography

Methods and Protocols

Series: Methods in Molecular Biology

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by Michael Zachariou

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Main description:

Forty years after the term “affinity chromatography” was introduced, this mode of chromatography remains a key tool in the armory of separation techniques that are available to separation and interaction scientists. Affinity chromat- raphy is favored because of its high selectivity, speed, and ease of use. The rapid and selective isolation of molecules using affinity chromatography has allowed a better understanding of biological processes, accelerated the identi- cation of target molecules, and spawned new process areas such as immobilized enzyme reactors. It has had ubiquitous application in most areas of science ranging from small molecule isolation to biopolymers such as DNA, proteins, polysaccharides, and even whole cells. The number of applications of affinity chromatography continues to expand at a rapid rate. For example, more than 60% of purification protocols include some sort of affinity chromatography step, while a database search of PubMed reveals more than 36,000 publications making use of the term “affinity chromatography,” more than 3000 of which refer to it in their title. The US patent office reports more than 16,000 ref- ences to the term “affinity chromatography”, while there are more than 270 references to the same term in the patent title. The aim of this edition of Methods in Molecular Biology, Affinity Chromatography: Methods and Protocols, Second Edition is to provide the beginner with the practical knowledge to develop affinity separations suitable for various applications relevant to the post-genomic era.

Feature:

Contains state-of-the-art, brand new protocols developed for this second edition

Provides step-by-step laboratory instructions for readily reproducible results

Back cover:

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.

Contents:

PROPOSED TABLE OF CONTENTS 1 Affinity Chromatography: History, Perspectives, Limitations and Prospects Ana Cecília A. Roque and Christopher R. Lowe VARIOUS MODES OF AFFINITY CHROMATOGRAPHY 2 Immobilized Metal Ion Affinity Chromatography of Native Proteins Adam Charlton and Michael Zachariou 3 Affinity Precipitation of Proteins Using Metal Chelates Ashok Kumar, Igor Yu. Galaev and Bo Mattiasson 4 Immunoaffinity Chromatography Stuart R. Gallant, Vish Koppaka and Nick Zecherle 5 Dye-ligand Chromatography Stuart R. Gallant, Vish Koppaka and Nick Zecherle 6 Purification of Proteins Using Displacement Chromatography Nihal Tugcu
AFFINITY CHROMATOGRAPHY USING PURIFICATION TAGS 7 Rationally Designed Ligands for use in Affinity Chromatography: an Artificial Protein L Ana Cecília A. Roque and Christopher Lowe 8 Phage display of Peptides in Ligand Selection for use in Affinity Chromatography Joanne L Casey, Andrew M. Coley and Michael Foley 9 Preparation, Analysis and Use of an Affinity Adsorbent for the Purification of GST Fusion Protein Gareth M. Forde 10 Immobilized Metal Ion Affinity Chromatography of Histidine-tagged fusion proteins Adam Charlton and Michael Zachariou 11 Methods for the Purification of HQ-Tagged Proteins Beckky Godat, Laurie Engel, Natalie Betz and Tonny M. Johnson 12 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 13 Methods for the Detection of Protein-Protein and Protein-DNA Interactions using Halo-Tag™ Marjeta Urh, Danette Hartzell, Jacqui Mendez and Keith Wood 14 Site-specific Cleavage of Fusion Proteins Adam Charlton 15 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. 16 Affinity Processing of Cell-containing Feeds Using Monolithic Macroporous Hydrogels, Cryogels Igor Yu. Galaev and Bo Mattiasson 17 Monolithic Bioreactors for Macromolecules Mojca Ben_ina, Katja Ben_ina, Ale_ Podgornik and Ale_ _trancar 18 Plasmid DNA Purification via the Use of a Dual Affinity Protein Gareth M. Forde 19 Affinity Chromatography of Phosphorylated Proteins Grigoriy S. Tchaga 20 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 Østergaard

PRODUCT DETAILS

ISBN-13: 9781588296597
Publisher: Springer (Humana Press)
Publication date: November, 2007
Pages: 348
Weight: 695g
Availability: Not available (reason unspecified)
Subcategories: Genetics, Pharmacology

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Average Rating

From the reviews of the second edition:

"The book consists of three parts including 21 chapters that have been written by a large group of authors from many countries. … recommends this book as a manual for many specialists involved in purification of biomolecules in various fields of biochemistry, molecular biology, biotechnology, and bioorganic chemistry. The book can also be recommended as a handbook for university students and their teachers as well as for other schools preparing specialists in the above-mentioned fields." (G. Ya. Wiederschain, Biochemistry, November, 2008)