Protein crystallization in stirred-tank reactors:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20130207-1115472-0-9 Inhaltsverzeichnis |
| Beschreibung: | IV, 153 S. Ill., graph. Darst. |
Internformat
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| 100 | 1 | |a Hebel, Dirk Peter |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Protein crystallization in stirred-tank reactors |c Dirk Peter Hebel |
| 264 | 1 | |c 2013 | |
| 300 | |a IV, 153 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
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| 502 | |a München, Techn. Univ., Diss., 2013 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 856 | 4 | 1 | |u http://mediatum.ub.tum.de/node?id=1115472 |x Verlag |z kostenfrei |3 Volltext |
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Datensatz im Suchindex
| _version_ | 1804150252616286208 |
|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS
1 INTRODUCTION 1
2 THESIS MOTIVATION AND OBJECTIVES 3
3 THEORETICAL BACKGROUND 5
3.1 PROTEINS 5
3.1.1 FUNDAMENTALS 5
3.1.2 PURIFICATION TECHNIQUES 6
3.1.3 FORMULATION AND STORAGE 1 1
3.2 PROTEIN CRYSTALLIZATION 12
3.2.1 MECHANISMS 12
3.2.2 EFFECTS OF CRYSTALLIZATION AGENTS, ADDITIVES AND OTHER VARIABLES
18
3.2.3 CRYSTALLIZATION METHODS 22
3.2.4 TECHNICAL-SCALE PROTEIN CRYSTALLIZATION 2 6
3.2.5 PROPERTIES OF PROTEIN CRYSTALS 26
3.2.6 PROTEINS UNDER STUDY 28
3.3 IONIC LIQUIDS AND PROTEINS 35
4 MATERIAL AND METHODS 38
4.1 PROTEIN ANALYTICS 3 8
4.1.1 PROTEIN CONCENTRATION DETERMINATION 3 8
4.1.2 SODIUM DODECYL SULFATE POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE).. 39
4.1.3 SIZE EXCLUSION CHROMATOGRAPHY (SEC] 39
4.1.4 FUNCTIONAL LIPASE ACTIVITY ASSAY 3 9
4.2 PREPARATION OF THE PROTEIN SOLUTIONS 40
4.2.1 DIALYSIS 4 1
4.2.2 CROSS-FLOW DIA- AND ULTRAFILTRATION 4 1
4.2.3 PAPAIN DIGESTION 4 2
4.2.4 CATION EXCHANGE CHROMATOGRAPHY (CEC) 4 2
4.3 PROTEIN CRYSTALLIZATION 4 3
4.3.1 VAPOR DIFFUSION CRYSTALLIZATION 4 3
4.3.2 MICROBATCH CRYSTALLIZATION 4 3
4.3.3 AGITATED BATCH CRYSTALLIZATION AND OVERVIEW OF THE CRYSTALLIZATION
SYSTEMS44
4.3.4 MICROSCOPY AND IMAGE ANALYSIS 4 7
HTTP://D-NB.INFO/1036694445
IMAGE 2
4.3.5 WASHING OF PROTEIN CRYSTALS AND ANALYSIS OF THE REMOVED SUBSTANCES
4 8
5 RESULTS AND DISCUSSION 49
5.1 PHASE DIAGRAMS AND PROTEIN CRYSTALLIZATION ON A 5 ML-SCALE 49
5.1.1 LYSOZYME FROM GALLUSGALLUS (LYS) 4 9
5.1.1.1 MICROBATCH-SCALE VALIDATION OF CRYSTALLIZATION CONDITIONS 4 9
5.1.1.2 CRYSTALLIZATION ON A 5 ML-SCALE 50
5.1.1.3 DISCUSSION 53
5.1.2 LIPASE FROM THERMOMYCES LANUGINOSUS (LIP) 54
5.1.3 XYLANASE II FROM TRICHODERMA REESEI (XYL) 57
5.1.3.1 MICROBATCH-SCALE SCREENING OF CRYSTALLIZATION CONDITIONS 57
5.1.3.2 CRYSTALLIZATION ON A 5 ML-SCALE 59
5.1.3.3 DISCUSSION 62
5.1.4 GLUCOSE ISOMERASE FROM STREPTOMYCES RUBIGINOSUS (GLU) 63
5.1.4.1 MICROBATCH-SCALE SCREENING OF CRYSTALLIZATION CONDITIONS 6 3
5.1.4.2 CRYSTALLIZATION ON A 5 ML-SCALE 65
5.1.4.3 DISCUSSION 67
5.1.5 ANTIGEN-BINDING FRAGMENT OF THE ANTIBODY CETUXIMAB (FAB) 68
5.1.5.1 MICROBATCH-SCALE SCREENING OF CRYSTALLIZATION CONDITIONS 68
5.1.5.2 CRYSTALLIZATION ON A 5 ML-SCALE 70
5.1.5.3 CRYSTALLIZATION IN THE PRESENCE OF PROTEIN CONTAMINANTS 72
5.1.5.4 DISCUSSION 75
5.1.6 CONCLUDING REMARKS 76
5.2 PROTEIN CRYSTALLIZATION USING ADDITIVES 7 8
5.2.1 LYSOZYME FROM GALLUS GALLUS (LYS) 78
5.2.1.1 IDENTIFICATION OF CRYSTALLIZATION CONDITIONS ON A
MICROBATCH-SCALE 78
5.2.1.2 CRYSTALLIZATION USING THE IONIC LIQUID 2-HEAF ON A 5 ML-SCALE 79
5.2.2 LIPASE FROM THERMOMYCES LANUGINOSUS (LIP) 8 1
5.2.2.1 IDENTIFICATION OF CRYSTALLIZATION CONDITIONS ON A
MICROBATCH-SCALE 8 1
5.2.2.2 CRYSTALLIZATION USING PHOSPHATE AND CHOLINE SALTS ON A 5
ML-SCALE 82
5.2.2.3 IDENTIFICATION OF A SUITABLE COMPLEMENTARY CRYSTALLIZATION AGENT
84
5.2.2.4 CRYSTALLIZATION USING PHOSPHATE AND PEG 10 000 ON A 5 ML-SCALE 8
5
5.2.3 GLUCOSE ISOMERASE FROM STREPTOMYCES RUBIGINOSUS (GLU) 86
IMAGE 3
5.2.4 ANTIGEN-BINDING FRAGMENT OF THE ANTIBODY CETUXIMAB (FAB] 8 7
5.2.5 SUMMARY AND DISCUSSION 90
5.3 PROTEIN CRYSTALLIZATION ON A 100 ML- AND A L-SCALE 92
5.3.1 LYSOZYME FROM GALLUSGALLUS (LYS] 92
5.3.2 LIPASE FROM THERMOMYCES LANUGINOSUS [LIP] 9 4
5.3.3 ANTIGEN-BINDING FRAGMENT OF THE ANTIBODY CETUXIMAB (FAB) 9 7
5.3.4 SUMMARY AND DISCUSSION 100
5.4 REMOVAL OF IMPURITIES FROM PROTEIN CRYSTALS 101
6 CONCLUSIONS AND OUTLOOK 103
7 REFERENCES 108
8 APPENDIX 122
8.1 ABBREVIATIONS 122
8.2 SYMBOLS AND VARIABLES 124
8.3 MATERIAL 126
8.3.1 EQUIPMENT 126
8.3.2 CHEMICALS 130
8.3.3 BUFFER COMPOSITIONS 133
8.4 PAPAIN DIGESTION OF CETUXIMAB 135
8.5 QUANTIFICATION OF CRYSTALLIZATION AGENTS 136
8.5.1 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) 136
8.5.2 PEG DETERMINATION USING A MODIFIED DRAGENDORFF REAGENT METHOD 136
8.5.3 SODIUM CHLORIDE DETERMINATION 137
8.6 PROTEIN SEQUENCES 138
8.7 TECHNICAL DRAWINGS OF THE 5 ML- AND 100 ML-CRYSTALLIZERS 140
8.7.1 TECHNICAL DRAWING OF THE 5 ML-SCALE CRYSTALLIZER 140
8.7.2 TECHNICAL DRAWING OF THE 100 ML-SCALE CRYSTALLIZER 141
8.8 LIST OF TABLES 143
8.9 LIST OF FIGURES 145
|
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| discipline | Chemie / Pharmazie Chemie Chemie-Ingenieurwesen |
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| physical | IV, 153 S. Ill., graph. Darst. |
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| spelling | Hebel, Dirk Peter Verfasser aut Protein crystallization in stirred-tank reactors Dirk Peter Hebel 2013 IV, 153 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20130207-1115472-0-9 http://mediatum.ub.tum.de/node?id=1115472 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20130207-1115472-0-9 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025926215&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hebel, Dirk Peter Protein crystallization in stirred-tank reactors |
| subject_GND | (DE-588)4113937-9 |
| title | Protein crystallization in stirred-tank reactors |
| title_auth | Protein crystallization in stirred-tank reactors |
| title_exact_search | Protein crystallization in stirred-tank reactors |
| title_full | Protein crystallization in stirred-tank reactors Dirk Peter Hebel |
| title_fullStr | Protein crystallization in stirred-tank reactors Dirk Peter Hebel |
| title_full_unstemmed | Protein crystallization in stirred-tank reactors Dirk Peter Hebel |
| title_short | Protein crystallization in stirred-tank reactors |
| title_sort | protein crystallization in stirred tank reactors |
| topic_facet | Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1115472 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20130207-1115472-0-9 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025926215&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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