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zhaoyongqiang

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[资源] Reversed Phase Chromatography(外文书籍)

下载地址http://www.namipan.com/d/Reverse ... 3e9f7057b6b0c7e0800

Contents
1. Introduction ..............................................................................................5
Theory of reversed phase chromatography ...............................................6
The matrix ................................................................................................9
The ligands ............................................................................................. 11
Resolution in reversed phase chromatography ........................................ 13
Resolution .............................................................................................. 13
Capacity factor ....................................................................................... 14
Efficiency ................................................................................................ 15
Selectivity ................................................................................................ 17
Binding capacity ..................................................................................... 18
Critical parameters in reversed phase chromatography........................... 19
Column length ...................................................................................19
Flow rate ............................................................................................19
Temperature ....................................................................................... 20
Mobile phase ..................................................................................... 20
Organic solvent .................................................................................. 20
Ion suppression .................................................................................. 21
Ion pairing agents .............................................................................. 22
Gradient elution ................................................................................. 23
Mode of use ............................................................................................24
Desalting ............................................................................................24
High resolution separations ............................................................... 25
Large scale preparative purification ................................................... 25
Stages in a purification scheme .......................................................... 26
Capture .............................................................................................. 26
Intermediate stages .............................................................................27
Polishing ............................................................................................27
2. Product Guide .........................................................................................29
SOURCE™ RPC ..................................................................................... 30
Product description ............................................................................ 30
High chemical stability ...................................................................... 32
Excellent flow/pressure characteristics ................................................ 34
High capacity ..................................................................................... 35
Availability ........................................................................................ 36
μRPC C2/C18 .........................................................................................37
Product description ............................................................................ 37
Chemical and physical stability .......................................................... 38
Flow/pressure characteristics ..............................................................38
Capacity ............................................................................................38
Availability ........................................................................................ 39
Sephasil™ Protein/Sephasil Peptide ......................................................... 39
Product description ............................................................................ 39
Chemical and physical stability .......................................................... 40
Flow/pressure characteristics ..............................................................40
Availability ........................................................................................ 40
3. Methods ................................................................................................. 41
Choice of separation medium .................................................................41
Unique requirements of the application .............................................. 41
Resolution .......................................................................................... 41
Scale of the purification ..................................................................... 42
Mobile phase conditions ....................................................................42
Throughput and scaleability ..............................................................42
Molecular weight of the sample components ......................................42
Hydrophobicity of the sample components ........................................ 43
Class of sample components ..............................................................43
Choice of mobile phase ........................................................................... 44
The organic solvent ............................................................................ 44
pH .....................................................................................................46
Ion pairing agents .............................................................................. 47
Sample preparation................................................................................. 49
Mobile phase preparation .......................................................................50
Storage of mobile phase ..................................................................... 50
Solvent disposal ................................................................................. 50
Detection ................................................................................................ 51
Ghosting ............................................................................................51
Mobile phase balancing ..................................................................... 51
Column conditioning .............................................................................. 52
Elution conditions ...................................................................................53
Column re-equilibration ......................................................................... 55
Column cleaning ..................................................................................... 55
Column storage ......................................................................................56
4. Applications............................................................................................57
Designing a biochemical purification ...................................................... 57
Naturally occurring peptides and proteins .............................................. 58
Purification of platelet-derived growth factor (PDGF) ........................ 59
Trace enrichment ............................................................................... 59
Purification of cholecystokinin-58 (CCK-58) from pig intestine ......... 60
Recombinant peptides and proteins ........................................................62
Process purification of inclusion bodies .............................................. 63
Purification of recombinant human epidermal growth factor ............. 63
Chemically synthesised peptides..............................................................65
Purification of a phosphorylated PDGF a-receptor derived peptide .... 65
Structural characterisation of a 165 kDa protein ............................... 66
Protein fragments from enzyme digests ................................................... 66
Protein characterisation at the micro-scale .........................................66
Protein identification by LC-MS ......................................................... 69
Chemically synthesised oligonucleotides ................................................. 70
5. Fault finding chart .................................................................................. 72
6. References ...............................................................................................81
7. Ordering information .............................................................................84

[ Last edited by zhaoyongqiang on 2008-3-17 at 12:07 ]

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