Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Göttingen
Cuvillier
2012
|
| Ausgabe: | 1. Aufl. |
| Schriftenreihe: | ibvt-Schriftenreihe
63 |
| Schlagworte: | |
| Online-Zugang: | Inhaltstext Inhaltsverzeichnis |
| Beschreibung: | VI, 143 S. Ill., graph. Darst. 21 cm |
| ISBN: | 9783954041152 |
Internformat
MARC
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| 264 | 1 | |a Göttingen |b Cuvillier |c 2012 | |
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| 490 | 1 | |a ibvt-Schriftenreihe |v 63 | |
| 502 | |a Zugl.: Braunschweig, Techn. Univ., Diss., 2011 | ||
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| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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Datensatz im Suchindex
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IMAGE 1
TABLE OF CONTENTS
1 INTRODUCTION 1
1.1 ANTIBODIES 2
1.1.1 BENCHMARK ANALYSIS 2
1.1.2 ANTIBODY FORMATS 3
1.1.3 ABF - KEY BENEFITS 6
1.1.4 SPECIFIC EXAMPLES 6
1.1.5 ANTIBODY PRODUCTION SYSTEMS 8
1.2 BACILLUS MEGATERIUM AS PRODUCTION HOST 12
1.3 OBJECTIVES 14
2 THEORY 15
2.1 "OMICS" APPROACH 15
2.2 FLOW CYTOMETRY AND CELL HETEROGENEITY 17
2.2.1 FLOW CYTOMETRY - HISTORY AND FUNCTION 17
2.2.2 ASSESSING CELL STATES 18
2.2.3 CELL HETEROGENEITIES 20
2.3 PROCESS DEVELOPMENT 22
3 MATERIAL A N D METHODS 25
3.1 STRAIN AND PLASMID 25
3.2 SAMPLING OF CULTIVATION SUPERNATANT 26
3.3 HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY ANALYSIS 27
3.4 BIOMASS MEASUREMENTS 27
3.5 QUANTIFICATION OF THE ANTIBODY FRAGMENT D1.3 SCFV CONCENTRATION VIA
ELISA 27
3.6 CELL BANK - BACILLUS MEGATERIUM 29
3.7 SCREENING IN DEEP WELL PLATES 29
3.8 CULTIVATION CONDITIONS - DEEP WELL PLATES 30
3.9 CULTIVATION CONDITIONS - BATCH 31
HTTP://D-NB.INFO/102301159X
IMAGE 2
3.10 CULTIVATION CONDITIONS - FED-BATCH 32
3.11 GAS ANALYSIS 32
3.12 OSCILLATING FED-BATCH STRATEGY 32
3.13 FLOW CYTOMETRY 34
3.14 DYE SCREENING FOR MEMBRANE POTENTIAL ESTIMATION 34
3.15 DIOC 2(3) RATIO ANALYSIS 35
3.16 MEMBRANE POTENTIAL CALIBRATION 35
3.17 VIABILITY ASSAY 36
3.18 SINGLE CELL PRODUCTION INTENSITY ASSAY 36
3.19 IMMUNO FIELD EMISSION SCANNING ELECTRON MICROSCOPY 36
3.20 CONFOCAL LASER SCANNING MICROSCOPY 37
3.21 POPULATION CLUSTER ANALYSIS 37
3.21.1 DIOC 2 MEMBRANE POTENTIAL ASSAY (SS, EV) (FL1, FL3) 38
3.21.2 PRODUCTION INTENSITY ASSAY (FL1, FL3) (FL1, FL3, SS, EV) 43
3.22 CENTRAL COMPOSITE DESIGN 44
3.23 GENETIC ALGORITHM 44
3.24 PREDICTION OF METAL-DEPENDENT ENZYMES O F BACILLUS MEGATERIUM 45
3.25 DNA MICRO ARRAYS 46
3.25.1 RNA-PREPARATION 46
3.25.2 ONE COLOUR MICROARRAY 46
3.26 CHEMICALS 46
4 RESULTS A N D DISCUSSION 47
4.1 AIMS O F THE WORK 47
4.2 MEDIUM DESIGN 48
4.2.1 CARBON SOURCE SCREENING 48
4.2.2 INDUCER SCREENING 49
4.2.3 GENETIC ALGORITHM APPROACH - OPTIMIZATION OF METAL ION
CONCENTRATIONS 50 4.2.4 STATISTICAL DOE - OPTIMIZATION OF MGS0 4 AND
(NH4)2S04 CONCENTRATIONS 58
IMAGE 3
4.3 SINGLE CELL ANALYSIS
59
4.3.1 MEMBRANE POTENTIAL 59
4.3.2 CELL INTEGRITY 63
4.3.3 PRODUCTION INTENSITY 64
4.4 PROCESS DEVELOPMENT 67
4.4.1 BATCH CULTURE 67
4.4.2 FED-BATCH CULTURE 73
4.4.3 UP-SCALE APPROACH 81
4.5 TRANSCRIPTOME ANALYSIS 83
4.5.1 DIFFERENTIAL GENE EXPRESSION DEPENDENT ON RECOMBINANT ABF D1.3
SCFV PRODUCTION/SECRETION IN BACILLUS MEGATERIUM 85
4.5.2 GROWTH PHASE DEPENDENT GENE EXPRESSION 88
4.5.3 GROWTH PHASE DEPENDENT GENE EXPRESSION RELATED TO RECOMBINANT ABF
D1.3 SCFV PRODUCTION 92
4.5.4 SUMMARY 94
4.6 CULTURE HETEROGENEITY 98
4.6.1 LATE INDUCTION OF ABF D1.3 SCFV PRODUCTION 98
4.6.2 BATCH PROCESS WITH 50 G/L INITIAL CARBON SOURCE 106
4.6.3 PROCESS RELEVANCE OF HETEROGENEITIES 108
5 CONCLUSIONS A N D OUTLOOK 110
5.1 MEDIUM DESIGN 110
5.2 ADVANCED PROCESS CONTROL 111
5.3 PROCESS DEVELOPMENT AND LIMITATIONS 113
5.4 ERA O F CYTOMICS AND DATA MODELING 114
5.5 ANTIBODIES - A FUTURE PERSPECTIVE 115
5.5.1 POTENTIAL OF MICROBIAL SYSTEMS 115
5.5.2 MOST PROMISING ANTIBODY FORMATS 117
5.5.3 FUTURE - APPLICATION 118
6 REFERENCES 119
7 NOMENCLATURE 136
7.1 ABBREVIATIONS 136
7.2 SYMBOLS 139 |
| any_adam_object | 1 |
| author | David, Florian Bodo 1981- |
| author_GND | (DE-588)1024740676 |
| author_facet | David, Florian Bodo 1981- |
| author_role | aut |
| author_sort | David, Florian Bodo 1981- |
| author_variant | f b d fb fbd |
| building | Verbundindex |
| bvnumber | BV040537800 |
| ctrlnum | (OCoLC)811273122 (DE-599)DNB102301159X |
| dewey-full | 660.63 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 660 - Chemical engineering |
| dewey-raw | 660.63 |
| dewey-search | 660.63 |
| dewey-sort | 3660.63 |
| dewey-tens | 660 - Chemical engineering |
| discipline | Chemie / Pharmazie Biologie |
| edition | 1. Aufl. |
| format | Thesis Book |
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| id | DE-604.BV040537800 |
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| isbn | 9783954041152 |
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| physical | VI, 143 S. Ill., graph. Darst. 21 cm |
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| spelling | David, Florian Bodo 1981- Verfasser (DE-588)1024740676 aut Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium von Florian Bodo David 1. Aufl. Göttingen Cuvillier 2012 VI, 143 S. Ill., graph. Darst. 21 cm txt rdacontent n rdamedia nc rdacarrier ibvt-Schriftenreihe 63 Zugl.: Braunschweig, Techn. Univ., Diss., 2011 Proteingewinnung (DE-588)4175994-1 gnd rswk-swf Bacillus megaterium (DE-588)4278034-2 gnd rswk-swf Prozessoptimierung (DE-588)4176074-8 gnd rswk-swf Bioverfahrenstechnik (DE-588)4307166-1 gnd rswk-swf Antikörper (DE-588)4002290-0 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Antikörper (DE-588)4002290-0 s Proteingewinnung (DE-588)4175994-1 s Bacillus megaterium (DE-588)4278034-2 s Bioverfahrenstechnik (DE-588)4307166-1 s Prozessoptimierung (DE-588)4176074-8 s DE-604 ibvt-Schriftenreihe 63 (DE-604)BV011182135 63 X:MVB text/html http://deposit.dnb.de/cgi-bin/dokserv?id=4052017&prov=M&dok_var=1&dok_ext=htm Inhaltstext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025383767&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | David, Florian Bodo 1981- Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium ibvt-Schriftenreihe Proteingewinnung (DE-588)4175994-1 gnd Bacillus megaterium (DE-588)4278034-2 gnd Prozessoptimierung (DE-588)4176074-8 gnd Bioverfahrenstechnik (DE-588)4307166-1 gnd Antikörper (DE-588)4002290-0 gnd |
| subject_GND | (DE-588)4175994-1 (DE-588)4278034-2 (DE-588)4176074-8 (DE-588)4307166-1 (DE-588)4002290-0 (DE-588)4113937-9 |
| title | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium |
| title_auth | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium |
| title_exact_search | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium |
| title_full | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium von Florian Bodo David |
| title_fullStr | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium von Florian Bodo David |
| title_full_unstemmed | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium von Florian Bodo David |
| title_short | Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium |
| title_sort | holistic bioprocess engineering of antibody fragment secreting bacillus megaterium |
| topic | Proteingewinnung (DE-588)4175994-1 gnd Bacillus megaterium (DE-588)4278034-2 gnd Prozessoptimierung (DE-588)4176074-8 gnd Bioverfahrenstechnik (DE-588)4307166-1 gnd Antikörper (DE-588)4002290-0 gnd |
| topic_facet | Proteingewinnung Bacillus megaterium Prozessoptimierung Bioverfahrenstechnik Antikörper Hochschulschrift |
| url | http://deposit.dnb.de/cgi-bin/dokserv?id=4052017&prov=M&dok_var=1&dok_ext=htm http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025383767&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV011182135 |
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