Transcription of Size Exclusion Chromatography
1 Size Exclusion Chromatography Principles and Methods18-1022-18 AL 11/2014GE and GE monogram are trademarks of General Electric Company. KTA, Biacore, BioProcess, HiLoad, HiPrep, HiScale, HiTrap, MabSelect , MidiTrap, MiniTrap, MultiTrap, Sephacryl, Sephadex, Sepharose, SpinTrap, Superdex, Superose, Tricorn, and UNICORN are trademarks of GE Healthcare Company or one of its subsidiaries. BCA is a trademark of Thermo Fisher Scientific LLC. Coomassie is a trademark of Imperial Chemical Industries Limited. Microsoft and Excel are registered trademarks of the Microsoft Corporation.
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3 A copy of these terms and conditions is available on request . Contact your local GE Healthcare representative for the most current Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire, HP7 9NA UKGE Healthcare Europe, GmbH Munzinger Strasse 5 D-79111 Freiburg GermanyGE Healthcare Bio-Sciences Corp. 800 Centennial Avenue, Box 1327 Piscataway, NJ 08855-1327 USAGE Healthcare Bio-Sciences KK Sanken Bldg., 3-25-1, Hyakunincho Shinjuku-ku, Tokyo 169-0073 JapanFor local office contact information,please visit GE Healthcare Bio-Sciences ABBj rkgatan 30751 84 UppsalaSwedenSize Exclusion ChromatographyPrinciples and MethodsGE HealthcareLife Sciencesimagination at workimagination at workHandbooks from GE Healthcare Life SciencesFor more information refer to ElectrophoresisPrinciples and MethodsGE HealthcareLife Sciences2-D Electrophoresis using Immobilized pH Gradients Principles and Methods 80-6429-60 Affinity ChromatographyPrinciples and MethodsGE HealthcareLife SciencesAffinity Chromatography Principles and Methods 18-1022-29GE HealthcareLife Sciences KTA
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7 7 Symbols ..8 Common acronyms and abbreviations ..9 Chromatography terminology ..10 Chapter 1 Size Exclusion Chromatography (SEC) in practice ..15 Introduction ..15 Purification by SEC ..15 Group separation ..16 High-resolution fractionation ..17 Rapid purity check and screening ..18 Resolution in SEC ..18 Sample volume and column dimensions ..18 Media selection ..20 Sample and buffer preparation ..24 Sample buffer composition ..24 Sample concentration and viscosity ..24 Sample volume ..26 Buffer composition ..26 Denaturing (chaotropic) agents and detergents ..26 Column and media preparation.
8 27 Setting column pressure application ..30 Elution and flow rates ..30 Controlling flow rates ..32 Method development for high-resolution fractionation ..32 Maintenance of SEC columns ..33 Equipment selection ..33 Scaling up ..33 BioProcess media for large-scale production ..34 Troubleshooting ..35 Chapter 2 Superdex: for high resolution, short run times, and high recovery ..39 Superdex Increase ..40 Superdex ..42 Superdex prep grade ..43 Separation options ..45 Separation examples ..46 Performing a separation ..48 First-time use or after long-term storage ..48 Cleaning.
9 49 Removing severe contamination ..49 Media characteristics ..50 Chemical stability ..50 Storage ..5018-1022-18 AL 3 Chapter 3 Superose: broad fractionation range for laboratory scale ..51 Superose Increase ..51 Superose ..53 Superose prep grade ..53 Separation options ..54 Separation examples ..55 Performing a separation ..56 First-time use or after long-term storage ..56 Cleaning ..57 Media characteristics ..57 Chemical stability ..57 Storage ..57 Chapter 4 Sephacryl: fast , high-recovery separations at laboratory and industrial scale ..59 Separation options ..62 Separation examples.
10 63 Performing a separation ..64 First-time use or after long-term storage ..64 Cleaning ..64To remove severe contamination ..65 Media characteristics ..65 Chemical stability ..65 Storage ..66 Chapter 5 Sephadex: desalting, buffer exchange, and sample clean up ..67 Separation options ..68 Separation examples ..72 Performing a separation ..73 General considerations ..73 Small-scale desalting of samples ..73 Desalting larger sample volumes using HiTrap and HiPrep columns ..73 Buffer preparation ..74 Sample preparation ..74 Buffer exchange ..74 HiTrap Desalting purification with a syringe.
