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Protein Purification. the Strategy Gives Guidelines for How to Combine Purification Methods in the Best Way to Reach the Set Goals

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Protein Purification. the Strategy Gives Guidelines for How to Combine Purification Methods in the Best Way to Reach the Set Goals GE, imagination at work, and GE monogram are trademarks of General Electric Company. 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GE Healthcare UK Limited Amersham Place Strategies for Little Chalfont For local office contact information, Buckinghamshire, HP7 9NA please visit www.gelifesciences.com/contact UK GE Healthcare Europe, GmbH Munzinger Strasse 5 Protein Purif ication D-79111 Freiburg www.gelifesciences.com/protein-purification Germany GE Healthcare Bio-Sciences Corp. 800 Centennial Avenue, P.O. Box 1327 Handbook GE Healthcare Bio-Sciences AB Piscataway, NJ 08855-1327 USA Björkgatan 30 GE Healthcare Bio-Sciences KK 751 84 Uppsala Sanken Bldg., 3-25-1, Hyakunincho Shinjuku-ku, Tokyo 169-0073 Sweden Japan imagination at work imagination at work imagination at work 28-9833-31 AA 09/2010 Handbooks from GE Healthcare GST Gene Fusion System Gel Filtration Handbook Principles and Methods 18-1157-58 18-1022-18 Affinity Chromatography Recombinant Protein Purification Handbook Principles and Methods Principles and Methods 18-1022-29 18-1142-75 Antibody Purification Hydrophobic Interaction and Handbook Reversed Phase Chromatography 18-1037-46 Principles and Methods 11-0012-69 Ion Exchange Chromatography and Chromatofocusing 2-D Electrophoresis using Principles and Methods immobilized pH gradients 11-0004-21 Principles and Methods 80-6429-60 Cell Separation Media Methodology and Applications Microcarrier Cell Culture 18-1115-69 Principles and Methods 18-1140-62 Purifying Challenging Proteins Principles and Methods Nucleic Acid Sample Preparation for 28-9095-31 Downstream Analyses Principles and Methods Isolation of mononuclear cells 28-9624-00 Methodology and Applications 18-1152-69 Strategies for Protein Purification Handbook High-throughput Process Development 28-9833-31 with PreDictor Plates Principles and Methods 28-9403-58 Strategies for Protein Purification Handbook 28-9833-31 AA 1 Contents Introduction...........................................................................................................................................5 Common acronyms and abbreviations used in this handbook ..................................................................5 Chromatography terminology ....................................................................................................................................7 Symbols used in this handbook .............................................................................................................................. 11 Chapter.1. Brief.history.of.protein.purification................................................................................................13 Advances in methodology ......................................................................................................................................... 13 New chromatography media .................................................................................................................................. 13 Genomics and proteomics ......................................................................................................................................... 14 Where we are today .................................................................................................................................................... 14 Chapter.2 Protein.purification.methods...........................................................................................................17 Affinity chromatography (AC) .................................................................................................................................... 17 Immobilized metal ion affinity chromatography (IMAC) ............................................................................... 20 Gel filtration (GF) .............................................................................................................................................................. 21 Ion exchange chromatography (IEX) ..................................................................................................................... 23 Hydrophobic interaction chromatography (HIC) ............................................................................................. 25 Chromatofocusing ......................................................................................................................................................... 27 Reversed phase chromatography (RPC) .............................................................................................................. 28 Chapter.3 Desalting,.buffer.exchange,.and.protein.concentration.............................................................31 GF for desalting, removal of low-molecular-weight substances, and buffer exchange .............. 31 Other methods for desalting and buffer exchange ....................................................................................... 36 Protein concentration using ultrafiltration ........................................................................................................
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