Protein purification applications: a practical approach
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Oxford [u.a.]
Oxford Univ. Press
2001
|
| Edition: | 2. ed. |
| Series: | The practical approach series
245 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Hier auch später erschienene, unveränderte Nachdrucke |
| Physical Description: | XVI, 165 S. Ill., graph. Darst. |
| ISBN: | 0199636729 0199636710 |
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| 245 | 1 | 0 | |a Protein purification applications |b a practical approach |c ed. by Simon Roe |
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Record in the Search Index
| _version_ | 1804128493413335040 |
|---|---|
| adam_text | Protein purification
applications
Second Edition
h [Pira©ftil©al [^p)pi?®®©lhi
Edited by
Simon Roe
AEA Technology pic, Bioprocessing Facility,
Transport Way, Watlington Road,
Oxford 0X4 6LY, U K
OXFORD
UNIVERSITY PRESS
Contents
List of protocols page xiii
Abbreviations xv
1 Fusion protein purification methods 1
Stephen J Brewer, Charles E Glatz, and Charles Dickerson
1 Introduction 1
2 Protecting proteins from proteolysis 2
Purification of fused AP11I and release of protein using V8 protease
Isolation of EGF from tryp E-fusion using trypsin 5
3 Engineering to improve purification processes 7
Fluidized bed IMAC for polyHis-tagged proteins 8
Engineering metal affinity sites in somatotropins 10
Engineering proteins for polyelectrolyte precipitation 11
4 Fusions of generic application 12
PolyHistidine 13
FLAG 14
Recombinant phage antibody system 15
Fusion tag detection and purification: S-Tag system 15
T7-Tag affinity purification kit 25
Fusions based on maltose-binding protein 16
~ - Intein-mediated purification with an affinity chitin binding tag 16
5 Concluding remarks 16
References 17
2 Initial purification of inclusion bodies 19
Bernard N Violand
1 Introduction 19
2 Mechanism of inclusion body formation 19
3 Isolation of inclusion bodies from cell homogenates 20
4 Washing of inclusion bodies 22
References 26
CONTENTS
3 Purification for crystallography 29
S P Wood and A R Coker
1 Introduction 29
2 Methods for protein crystallization 33
Batch 33
Vapour diffusion 35
Dialysis 37
3 Influence of heterogeneity on crystallization 38
Effects of impurities 38
Origins of microheterogeneity 39
Detection of microheterogeneity 41
Additives and proteolysis 43
4 Concluding remarks 45
References 45
4 Protein purifications from mammalian cell culture 49
Terry Cartvvright
1 Introduction 49
2 Nature of recombinant proteins produced in mammalian cells 50
3 Transgenic animals 51
4 Overview of extraction and purification process 52
5 Process design—preventive measures 53
Cell culture system 53
Production cells 54
Culture medium 54
6 Cell separation 55
Purification without cell separation 56
7 Initial product recovery and fractionation 56
Recovery of recombinant protein from milk 57
Direct capture of product from feedstock 57
8 Main purification 57
Ion exchange chromatography 58
Hydrophobic interaction chromatography 58
Affinity chromatography systems 59
Small molecule affinity ligands 60
9 Particle-tolerant chromatographic supports 61
10 Purification of monoclonal antibodies 62
Separation using protein A 62
Monoclonal antibody purification by ion exchange 64
Other recent approaches to purification of monoclonal antibodies 65
11 The elimination of viral contamination risks 66
Spiking and the use of model viruses 66
Scale-down of unit operations 67
Calculation of virus clearance 68
A cautionary note 68
Virus removal by membrane filtration 69
12 Elimination of contaminating DNA 69
viii
CONTENTS
13 Other biologically active contaminants 70
14 Polishing steps 70
15 Problems 70
References 71
5 Protein purification from microbial cell culture 73
Peter R Levison
1 Introduction 73
2 Chromatographic processes 73
Positive or negative steps 74
Batch or column contactors 75
Gradient elution 75
3 Ion exchange protein purifications from microbial cell culture 76
Isolation of yeast enzymes 76
Isolation of amylase from Bacillus subtilis 79
Isolation of DNA-modifying enzymes 81
4 Ion exchange nucleic acid purification from microbial culture 83
References 85
6 Protein purification from milk 87
Richard Burr
1 Introduction 87
2 Milk proteins 87
Micelle structure 88
Major milk protein groups 88
3 Factors to consider in preparation and handling milk 93
Source 93
Proteolysis 93
Solubility 93
Protein modification 94
Drying and storage 95
4 Fractionation of milk proteins 95
fractionation of casein and whey 95
Fractionation of milk proteins by differential solubility 100
5 Chromatography of milk proteins 101
Ion exchange 101
Size exclusion chromatography 103
Affinity chromatography 104
Reverse-phase chromatography 105
Chromatofocusing 105
6 Identification and analysis of purity 106
Polyacrylamide gel electrophoresis 106
Isoelectric focusing 109
Capillary electrophoresis 110
Analytical chromatography 110
Mass spectrometry 110
ix
CONTENTS
7 Developments 110
Preparative electrophoresis 110
Microfiltration and ultrafiltration 111
Aqueous two-phase partitioning 111
Acknowledgements 112
References 112
7 Protein purification from animal tissue 117
Nigel M Hooper
1 Introduction 117
2 Choice of tissue 117
3 Tissue disruption 118
4 Prevention of unwanted proteolysis 119
5 Subcellular fractionation 119
6 Solubilization of membrane proteins 124
7 Example purifications of animal proteins 126
Purification of angiotensin converting enzyme 126
Purification of aminopeptidase P 129
Purification of fructose-l,6-biphosphatase 129
References 132
Appendix 132
8 Protein purification from plant sources 135
G Paul Bolwefl
1 Introduction 135
2 Special considerations 136
Low protein concentration 136
Proteolysis 137
Inhibition and tanning 138
3 Source of material 138
- Leaf, 138
Stem 140 _
Roots 140
Seeds and storage organs 140
Fruits 140
Tissue cultures 141
Organelles 141
4 Direct and indirect strategies for identification of subsets of plant
proteins 147
5 Examples of successful purification protocols: enzymes of carbohydrate
metabolism 149
Calvin cycle 24 150
Polysaccharide synthases 150
6 Enzymes of secondary metabolism 152
Phenolic metabolism 152
X
CONTENTS
Terpenoids 154
Alkaloids 154
7 Preparation for Edman protein sequencing 155
References 156
A1 List of suppliers 159
Index 163
xi
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| series | The practical approach series |
| series2 | The practical approach series |
| spelling | Protein purification applications a practical approach ed. by Simon Roe 2. ed. Oxford [u.a.] Oxford Univ. Press 2001 XVI, 165 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 245 Hier auch später erschienene, unveränderte Nachdrucke Eiwitten gtt Protéines - Purification - Manuels de laboratoire ram Zuiveringstechnieken gtt Proteins Purification Laboratory manuals Proteins isolation & purification Methode (DE-588)4038971-6 gnd rswk-swf Proteine (DE-588)4076388-2 gnd rswk-swf Reinigung (DE-588)4049250-3 gnd rswk-swf Proteine (DE-588)4076388-2 s Reinigung (DE-588)4049250-3 s Methode (DE-588)4038971-6 s DE-604 Roe, Simon Sonstige oth The practical approach series 245 (DE-604)BV007921321 245 HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009347539&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Protein purification applications a practical approach The practical approach series Eiwitten gtt Protéines - Purification - Manuels de laboratoire ram Zuiveringstechnieken gtt Proteins Purification Laboratory manuals Proteins isolation & purification Methode (DE-588)4038971-6 gnd Proteine (DE-588)4076388-2 gnd Reinigung (DE-588)4049250-3 gnd |
| subject_GND | (DE-588)4038971-6 (DE-588)4076388-2 (DE-588)4049250-3 |
| title | Protein purification applications a practical approach |
| title_auth | Protein purification applications a practical approach |
| title_exact_search | Protein purification applications a practical approach |
| title_full | Protein purification applications a practical approach ed. by Simon Roe |
| title_fullStr | Protein purification applications a practical approach ed. by Simon Roe |
| title_full_unstemmed | Protein purification applications a practical approach ed. by Simon Roe |
| title_short | Protein purification applications |
| title_sort | protein purification applications a practical approach |
| title_sub | a practical approach |
| topic | Eiwitten gtt Protéines - Purification - Manuels de laboratoire ram Zuiveringstechnieken gtt Proteins Purification Laboratory manuals Proteins isolation & purification Methode (DE-588)4038971-6 gnd Proteine (DE-588)4076388-2 gnd Reinigung (DE-588)4049250-3 gnd |
| topic_facet | Eiwitten Protéines - Purification - Manuels de laboratoire Zuiveringstechnieken Proteins Purification Laboratory manuals Proteins isolation & purification Methode Proteine Reinigung |
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