Gel Filtration Principles and Methods Back to Collection ● Amersham 18-1022-18 Biosciences Edition Al
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Handbooks from Amersham Biosciences 18109746 Gel Filtration 7a2gicaiom3ngsmpePRictan The recombinant protein Handbook Protein Purification Percoll Reversed Phase Chromatography 113229 dMethods Ficoll-Paque Plus lon Exchange Chromatography Expanded Bed Adsorption 18-1152.690 nof lymphocytes nd Methods Methods GST Gene Fusion System Affinity Chromatography Chromatofocusing Metnods rand PE 18.71g7258 2-D Electrophoresis sing immo ed pH grac
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Gel filtration Principles and Methods
1 Gel filtration Principles and Methods
Contents lntroducetion n abbreviations Chapter 1 Gel filtration in practice 9 Introduction. Separation by gel filtration Resolution in gel filtration 13 da elecrto Buffer composition and Denaturing (chaotropic)agents and detergents. Column and media preparation . 33 Sample application . .23 Elution and flow rates. .24 Method development for high resolution fractionation. 26 Care of gel filtration media .27 Equipment selection 27 Scaling up BioProcess Media for large-scale production Troubleshooting. .29 Chapter 2 Gel filtration media Components of gel filtration media. Superdex:first choice for high resolution,short run times and high recovery. 6 Separation options. aration examples 43 Media characteristics Chemical stability 44 torage. Sephacryl:fast,high recovery separations at laboratory and industrial scale . eparation options Media characteristics 51 Chemical stability. 51 storage. Superose:broad fractionation range,but not suitable for industrial scale separations.52
2 Contents Introduction . 5 Symbols . 6 Common abbreviations . 7 Chapter 1 Gel filtration in practice . 9 Introduction . 9 Separation by gel filtration . 9 Resolution in gel filtration . 13 Media selection . 16 Sample preparation . 20 Buffer composition and preparation . 21 Denaturing (chaotropic) agents and detergents .22 Column and media preparation . 23 Sample application . 23 Elution and flow rates . 24 Method development for high resolution fractionation . 26 Care of gel filtration media . 27 Equipment selection . 27 Scaling up . 27 BioProcess Media for large-scale production . 29 Troubleshooting . 29 Chapter 2 Gel filtration media . 35 Components of gel filtration media . 35 Superdex: first choice for high resolution, short run times and high recovery. 36 Separation options . 38 Separation examples . 39 Performing a separation .42 Cleaning .43 Media characteristics .44 Chemical stability . 44 Storage . 44 Sephacryl: fast, high recovery separations at laboratory and industrial scale . 45 Separation options . 48 Separation examples . 48 Performing a separation .49 Cleaning .50 Media characteristics .51 Chemical stability . 51 Storage . 51 Superose: broad fractionation range, but not suitable for industrial scale separations . 52
Separation options mple ing a sep Media characteristics 56 Chemical stability. 56 Storage .56 mple 57 61 Scale up and processing large sample volumes .6 nn Packing cal stability 69 Chapter 3 Gel filtration in theory .71 Defining the process. Selectivity curves and media selection. .74 Resolution .75 Chapter 4 Molecular weight determination and molecular weight distribution analysis. .79 Chapter adex LH-20 81 Packing a column 83 Performing a separation. 84 Transferring Sephadex LH-20 between organicent 85 Chapter 6 Gel filtration in a Purification Strategy(CPP). 81 Applying CiPP. 8 Selection and combination of purification techniques .88 Gel filtration as a polishing step. .91
3 Separation options . 53 Separation examples . 54 Performing a separation .54 Cleaning .55 Media characteristics .56 Chemical stability . 56 Storage . 56 Sephadex: rapid group separation of high and low molecular weight substances, such as desalting, buffer exchange and sample clean up . 57 Separation options . 59 Separation examples . 61 Performing a separation .61 Scale up and processing large sample volumes .66 Media characteristics .68 Column Packing . 68 Cleaning .68 Chemical stability . 68 Storage . 69 Chapter 3 Gel filtration in theory . 71 Defining the process . 71 Selectivity curves and media selection . 74 Resolution . 75 Chapter 4 Molecular weight determination and molecular weight distribution analysis . 79 Chapter 5 Sephadex LH-20 . 81 Separation options . 82 Separation examples . 82 Packing a column . 83 Performing a separation .84 Cleaning .84 Medium characteristics .84 Chemical stability . 85 Storage . 85 Transferring Sephadex LH-20 between organic solvents .85 Chapter 6 Gel filtration in a Purification Strategy (CIPP) . 87 Applying CIPP . 87 Selection and combination of purification techniques . 88 Gel filtration as a polishing step .91