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Affinity Chromatography Principles
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Affinity Chromatography Principles
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Serine proteases and zymogens with
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Appendix 4 ........................
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This handbook describes the role of
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Affinity chromatography is also use
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BioProcess Media for large-scale pr
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Chapter 2 Affinity chromatography i
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Avoid using magnetic stirrers as th
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pH elution A change in pH alters th
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0.6 0.4 0.2 0 A280 pH selected for
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Situation Cause Remedy Protein does
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Chapter 3 Purification of specific
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Table 2. Relative binding strengths
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Purification examples Figure 11 sho
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MAbTrap Kit Fig. 13. MAbTrap Kit, r
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Media characteristics Ligand Compos
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A 280 nm 2.0 1.5 1.0 0.5 Column: rP
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Desalt and/or transfer purified IgG
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Performing a separation Column: Rec
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M r 97 000 66 000 45 000 30 000 20
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in The Recombinant Protein Handbook
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Cleaning These procedures are appli
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Purification examples Figures 24 an
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Nickel solution: 0.1 M NiSO 4 Bindi
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Protein A fusion proteins IgG Sepha
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Purification or removal of serine p
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Sample: 2 ml clarified E. coli homo
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Serine proteases and zymogens with
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DNA binding proteins HiTrap Heparin
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Sample: Column: Binding buffer: Elu
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Media characteristics Ligand densit
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Sample: Pooled and frozen human pla
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The harsh conditions required to br
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Purification or removal of albumin
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IFN-act. A units 280 nm 250 0.12 20
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Purification options Binding capaci
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Sepharose NH (CH 2) n NH N N O N N
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If detergent or denaturing agents h
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Glycoproteins or polysaccharides Co
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For complex samples containing glyc
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For complex samples containing glyc
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Chemical stability Avoid exposure t
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Proteins and peptides with exposed
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Several methods can be used to dete
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Media characteristics Composition M
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- Page 99 and 100: Sepharose has been modified and dev
- Page 101 and 102: Ligand coupling Several methods are
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- Page 121 and 122: Media characteristics Product Compo
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- Page 127 and 128: Resolution is achieved by the selec
- Page 129 and 130: For any capture step, select the te
- Page 131 and 132: Appendix 1 Sample preparation Sampl
- Page 133 and 134: Remove salts from proteins with mol
- Page 135 and 136: 1. Filter (0.45 µm) or centrifuge
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- Page 139 and 140: Removal of lipoproteins Lipoprotein
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- Page 143 and 144: 4. Estimate the amount of slurry (r
- Page 145: Appendix 5 Conversion data: protein
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- Page 153 and 154: Appendix 8 Analytical assays during
- Page 155 and 156: Appendix 9 Storage of biological sa
- Page 157 and 158: Ordering information Product Quanti
- Page 159 and 160: STREAMLINE, Sepharose, HiTrap, ÄKT