Protein Purification (3^rd Ed.) Principles, High Resolution Methods, and Applications Methods of Biochemical Analysis Series

The authoritative guide on protein purification?now completely updated and revised

Since the Second Edition of Protein Purification was published in 1998, the sequencing of the human genome and other developments in bioscience have dramatically changed the landscape of protein research. This new edition addresses these developments, featuring a wealth of new topics and several chapters rewritten from scratch.

Leading experts in the field cover all major biochemical separation methods for proteins in use today, providing professionals in biochemistry, organic chemistry, and analytical chemistry with quick access to the latest techniques. Entirely new or thoroughly revised content includes:

• High-resolution reversed-phase liquid chromatography

• Electrophoresis in gels

• Conventional isoelectric focusing in gel slabs and capillaries and immobilized pH gradients

• Affinity ligands from chemical and biological combinatorial libraries

• Membrane separations

• Refolding of inclusion body proteins from E. coli

• Purification of PEGylated proteins

• High throughput screening techniques in protein purification

• The history of protein chromatography

Preface to The Third Edition vii

Preface to The Second Edition ix

Preface to The First Edition xi

Contributors xiii

Part I Introduction 1

1 Introduction to Protein Purification 3
Bo Ersson, Lars Rydén, and Jan-Christer Janson

Part II Chromatography 23

2 Introduction to Chromatography 25
Jan-Christer Janson and Jan Åke Jönsson

3 Gel Filtration: Size Exclusion Chromatography 51
Lars Hagel

4 Ion Exchange Chromatography 93
Evert Karlsson and Irwin Hirsh

5 High-Resolution Reversed-Phase Chromatography of Proteins 135
Sylvia Winkel Pettersson

6 Hydrophobic Interaction Chromatography 165
Kjell-Ove Eriksson and Makonnen Belew

7 Immobilized Metal Ion Affinity Chromatography 183
Lennart Kågedal

8 Covalent Chromatography 203
Francisco Batista-Viera, Lars Rydén, and Jan Carlsson

9 Affinity Chromatography 221
Francisco Batista-Viera, Jan-Christer Janson, and Jan Carlsson

10 Affinity Ligands from Chemical Combinatorial Libraries 259
Enrique Carredano and Herbert Baumann

11 Affinity Ligands from Biological Combinatorial Libraries 269
Per- Åke Nygren

Part III Other Separation Methods and Related Techniques 279

12 Membrane Separations 281
Joachim K. Walter, Zuwei Jin, Maik W. Jornitz, and Uwe Gottschalk

13 Refolding of Inclusion Body Proteins from E. coli 319
Zhiguo Su, Diannan Lu, and Zheng Liu

14 Purification of PEGylated Proteins 339
Conan J. Fee and James M. Van Alstine

Part IV Electrophoresis 363

15 Electrophoresis in Gels 365
Reiner Westermeier

16 Conventional Isoelectric Focusing in Gel Slabs and Capillaries and Immobilized pH Gradients 379
Pier Giorgio Righetti, Elisa Fasoli, and Sabina Carla Righetti

17 Two-Dimensional Electrophoresis in Proteomics 411
Reiner Westermeier and Angelika Görg

18 Protein Elution and Blotting Techniques 441
Reiner Westermeier

19 Capillary Electrophoretic Separations 451
Wolfgang Thormann

Part V Separation Method Optimization 487

20 High Throughput Screening Techniques in Protein Purification 489
Karol M. Lacki and Eggert Brekkan

Index 507

JAN-CHRISTER JANSON is Professor Emeritus at the Department of Physical and Analytical Chemistry, Uppsala University, Uppsala, Sweden.