AFFINITY CHROMATOGRAPHY Edited by Sameh Magdeldin Affinity Chromatography Edited by Sameh Magdeldin Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2012 InTech All chapters are Open Access distributed under the Creative Commons Attribution 3.0 license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. As for readers, this license allows users to download, copy and build upon published chapters even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. Publishing Process Manager Bojan Rafaj Technical Editor Teodora Smiljanic Cover Designer InTech Design Team First published March, 2012 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from orders@intechopen.com Affinity Chromatography, Edited by Sameh Magdeldin p. cm. ISBN 978-953-51-0325-7 Contents Preface IX Part 1 Principles and Applications of Affinity Chromatography 1 Chapter 1 Affinity Chromatography: Principles and Applications 3 Sameh Magdeldin and Annette Moser Chapter 2 Affinity Interactions as a Tool for Protein Immobilization 29 Eva Benešová and Blanka Králová Part 2 Lectin Affinity Chromatography 47 Chapter 3 Affinity Chromatography of Lectins 49 Jure Pohleven, Borut Štrukelj and Janko Kos Chapter 4 The Difference of Lectin Recovery by Sugar-Bound Resin 75 Mitsuru Jimbo, Shin Satoh, Hirofumi Hasegawa, Hiroshi Miyake, Takao Yoshida, Tadashi Maruyama and Hisao Kamiya Part 3 Immunoaffinity Purification 89 Chapter 5 Immunoaffinity Chromatography: A Review 91 Daad A. Abi-Ghanem and Luc R. Berghman Chapter 6 Affinity Chromatography for Purification of IgG from Human Plasma 107 Lucia Hofbauer, Leopold Bruckschwaiger, Harald Arno Butterweck and Wolfgang Teschner Part 4 GST – Tagged Affinity Chromatography 125 Chapter 7 A Study of the Glutathione Transferase Proteome of Drosophila melanogaster: Use of S- Substituted Glutathiones as Affinity Ligands 127 Ramavati Pal, Milana Blakemore, Michelle Ding and Alan G. Clark VI Contents Part 5 Nucleic Acids Affinity Purification 143 Chapter 8 RNA Affinity Chromatography 145 Nehal Thakor and Martin Holcik Part 6 Immobilized Metallic Ion Affinity Chromatography (IMAC) 165 Chapter 9 Ion Metallic Affinity Chromatography and Purification of Bacterial Toxin 167 Luciano Moura Martins and Tomomasa Yano Part 7 Usage of Affinity Chromatography in Purification of Proteases, Protease Inhibitors, and Other Enzymes 175 Chapter 10 Polyhistidine Affinity Chromatography for Purification and Biochemical Analysis of Fungal Cell Wall-Degrading Enzymes 177 Takumi Takeda Chapter 11 Novel Detection Methods Used in Conjunction with Affinity Chromatography for the Identification and Purification of Hydrolytic Enzymes or Enzyme Inhibitors from Insects and Plants 187 Alexander V. Konarev and Alison Lovegrove Chapter 12 Affinity Chromatography as a Key Tool to Purify Protein Protease Inhibitors from Plants 211 Elizeu Antunes dos Santos, Adeliana Silva de Oliveira, Luciana Maria Araújo Rabêlo, Adriana Ferreira Uchôa and Ana Heloneida Araújo Morais Chapter 13 Identification of cGMP-Kinase Complexes by Affinity Chromatography 255 Salb Katharina, Schinner Elisabeth and Schlossmann Jens Part 8 Affinity Chromatography as a Quantitative Tool 273 Chapter 14 Affinity Chromatography as a Tool for Quantification of Interactions Between Drug Molecules and Their Protein Targets 275 Piotr Draczkowski, Dariusz Matosiuk and Krzysztof Jozwiak Contents VII Part 9 Practical Application of Affinity Chromatography in Research Field 305 Chapter 15 The Value of Fungal Protease Inhibitors in Affinity Chromatography 307 Jerica Sabotič, Katarina Koruza, Boštjan Gabor, Matjaž Peterka, Miloš Barut, Janko Kos and Jože Brzin Chapter 16 Affinity-Based Methods for the Separation of Parasite Proteins 333 C.R. Alves, F.S. Silva, F.O. Oliveira Jr, B.A.S. Pereira, F.A. Pires and M.C.S. Pereira Preface With the huge number of books and publications that utilizes different aspects of affinity chromatography, it is still hard to find a freely accessed book that gathers a solid and concise understanding of affinity principles together with its applications in a single volume. The vision of this book is not more than an attempt to provide an open source single volume demonstrating the concept of affinity chromatography with some of its applications that meets the current throughput screening demands of scientists and researchers. This book “Affinity chromatography” starts with introductory chapter that passes through the basic principles and snapshot applications of this analytical technique, followed by a wide ranged practical research chapters utilizing affinity chromatography, written by leading experts worldwide. It is worthy to say that the scope of the information contained in this book is still limited to be covered in a single volume. Affinity chromatography is aimed mainly at those interested in different analytical and separation techniques, particularly, biochemists, biologists, pharmacists, advanced graduate students and post graduate researchers. Finally, I am grateful to the all experts who showed positive and specific “affinity“to participate and share in this book with their valuable experience. Indeed, without their participation, this book won’t come to light. Sameh Magdeldin, Ph.D Senior Post Doc Researcher and Proteomics Team Leader Medical School, Niigata University, Japan Ass. Prof. (Lecturer), Physiology Dept. Suez Canal University, Egypt [...]... been used as well Figure 2 shows a wide variety of molecules that can be purified by affinity chromatography based on their polarity and volatility Fig 2 Illustration showing different molecules that can be purified using affinity chromatography 6 Affinity Chromatography 3.2 Components of affinity medium When affinity chromatography is used for the purification and separation of large biomolecules from... longer occurs Since the beginning of this technique, the term affinity chromatography has raised many controversies among researchers Some say it would be more accurate if termed bioaffinity chromatography (O'Carra et al, 1974) or hydrophobic affinity (Shaltiel, 1974) Nonetheless, the term affinity chromatography has been expanded to describe a potential method of separating biomolecule mixtures on the... advantages of using peptides as affinity ligands are their low cost and stability Other ligands can be used in affinity chromatography for biomolecule purification For more information on all types of affinity ligands see references (Clonis, 2006; Hage, 2006) 3.2.4 Immobilization of affinity ligands Immobilization of the affinity ligand is also very important when designing an affinity chromatography method... previously Finally, the coupling of reversed phase affinity chromatography with mass spectrometry has ultimately aided in discovery of protein biomarkers 3 Fundamental principles of affinity chromatography Separation of a desired protein using affinity chromatography relies on the reversible interactions between the protein to be purified and the affinity ligand coupled to chromatographic matrix As... tightly bind the target These candidates can then be cloned, sequenced and used for affinity chromatography Affinity Chromatography: Principles and Applications 11 A similar process can be used to develop RNA affinity ligands Once a potential aptamer sequence is identified, it can synthesized in vitro and used as the affinity ligand on a chromatographic support An example of aptamers usage as in purification... collected in a concentrated form 3.1 Biomolecules purified by affinity chromatography Antibodies were first purified using affinity chromatography in 1951 when Campbell et al used affinity chromatography to isolate rabbit anti-bovine serum albumin antibodies (Campbell et al, 1951) For their purification, bovine serum albumin was used as the affinity ligand on a cellulose support Two years later, this... 14 Affinity Chromatography top 14 high-abundance proteins has been shown to result in a 25% increase in identified proteins (Tu et al, 2010) Moreover, affinity chromatography is widely used in many ‘omics’ studies (e.g proteomics, metabolomics and genomics) and is currently used in tandem with other methods to develop high-throughput screening methods for potential drugs 5 Biokinetics of affinity chromatography. .. Webb, 1978) The most commonly used dye is Cibacron blue F-3-GA which can be immobilized onto 22 Affinity Chromatography sepharose to create an affinity column See Figure 6 This dye is capable of removing over 90% of albumin in the sample (Travis et al, 1976) 6.6 Lectin affinity chromatography Lectin affinity chromatography is one of the most powerful techniques for studying glycosylation as a protein... 36-40 Kim, HS, Hage, DS, Immobilization methods for affinity chromatography, In: Handbook of Affinity Chromatography, Hage, DS, pp 35-78, Taylor & Francis, Boca Raton, FL Labrou NE (2000) Dye-ligand affinity chromatography for protein separation and purification Methods Mol Biol 147: 129-139 Labrou NE, Karagouni A, Clonis YD (1995) Biomimetic-dye affinity adsorbents for enzyme purification: application... within the cross-linked silica gel 4 Current techniques involving affinity chromatography Affinity chromatography is currently being used for a wide variety of applications ranging from the study of drug-protein binding interactions to the depletion of high abundance proteins to enhance the detection/quantification of dilute proteins Affinity chromatography can be used to study drug-protein binding interactions . different molecules that can be purified using affinity chromatography. Affinity Chromatography 6 3.2 Components of affinity medium When affinity chromatography is used for the purification. 3.1 Biomolecules purified by affinity chromatography Antibodies were first purified using affinity chromatography in 1951 when Campbell et al. used affinity chromatography to isolate rabbit. Complexes by Affinity Chromatography 255 Salb Katharina, Schinner Elisabeth and Schlossmann Jens Part 8 Affinity Chromatography as a Quantitative Tool 273 Chapter 14 Affinity Chromatography