Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20121212-1115861-0-8 Inhaltsverzeichnis |
| Beschreibung: | III, 189 S. Ill., graph. Darst. |
Internformat
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| 100 | 1 | |a Villa, Valentina |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions |c Valentina Villa |
| 264 | 1 | |c 2012 | |
| 300 | |a III, 189 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Techn. Univ., Diss., 2012 | ||
| 650 | 0 | 7 | |a Wechselwirkung |0 (DE-588)4064937-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Antikörper |0 (DE-588)4002290-0 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Protein p53 |0 (DE-588)4176550-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Antigen |0 (DE-588)4002272-9 |2 gnd |9 rswk-swf |
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| 689 | 0 | 0 | |a Antikörper |0 (DE-588)4002290-0 |D s |
| 689 | 0 | 1 | |a Antigen |0 (DE-588)4002272-9 |D s |
| 689 | 0 | 2 | |a Protein p53 |0 (DE-588)4176550-3 |D s |
| 689 | 0 | 3 | |a Wechselwirkung |0 (DE-588)4064937-4 |D s |
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| 912 | |a ebook | ||
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Datensatz im Suchindex
| _version_ | 1804150123538677760 |
|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS
1. ABSTRACT 1
2. ZUSAMMENFASSUNG 3
3. OBJECTIVES 5
4. MATERIALS & METHODS 7
4.1 MATERIALS 7
4.2 METHODS 1 1
4.2.1 BUFFER SOLUTIONS 1 1
4.2.2 GENERAL METHODS IN MOLECULAR BIOLOGY AND MICROBIOLOGY 12
4.2.3 EXPRESSION AND PURIFICATION OF FULL-LENGTH WILD TYPE HUMAN P53 1 3
4.2.4 METHODS IN ELECTROPHORESIS 15
4.2.5 METHODS FOR HCAL-DNA MONO CONJUGATE PROJECT 19
4.2.6 METHODS IN SPR (BIACORE* X-100) 2 4
5. INTRODUCTION TO SWITCHSENSE 27
5.1 PHYSICAL BASICS 2 8
5.1.1 DNA STRUCTURE 2 8
5.1.2 THE ELECTRICAL DOUBLE LAYER 29
5.1.3 ENERGY TRANSFER OF FLUORESCENCE TO METAL SURFACE 30
5.2 EXPERIMENTAL SETUP 3 1
5.2.1 CHIP DESIGN AND FLOW CHANNEL IN PROTOTYPEL AND PROTOTYPE2 3 1
5.2.2 CHIP DESIGN A N D FLOW CHANNEL SETUP IN SWA1 3 3
5.2.3 OPTICAL SETUP 3 4
5.3 ELECTRICAL MANIPULATION OF SURFACE-TETHERED DNA STRANDS 35
5.3.1 METHODS 35
5.3.2 FACTORS AFFECTING THE SWITCHING BEHAVIOR O F DNA STRANDS 4 1
6. SWITCHABLE HCAL-DNA LAYERS 4 3
6.1 INTRODUCTION 4 3
6.1.1 SOLULINK* CONJUGATION CHEMISTRY 4 3
I
HTTP://D-NB.INFO/1033344869
IMAGE 2
6.1.2 HUMAN CARBONIC ANHYDRASE 1 4 6
6.1.3 BINDING KINETICS 4 7
6.2 RESULTS, P A R T 1: HCAL-DNA CONJUGATE PREPARATION 5 0
6.2.1 ALDEHYDIC FUNCTIONALIZATION OF AMINATED OLIGONUCLEOTIDE 5 0
6.2.2 HYDRAZIDE FUNCTIONALIZATION OF HCAL 5 1
6.2.3 OPTIMIZATION OF CONJUGATION AND PURIFICATION STEPS 5 4
6.3. DISCUSSION, P A R T 1: OPTIMIZED PROTOCOL OF HCAL-DNA MONO
CONJUGATE 6 4
6.4 RESULTS A N D DISCUSSION, PART 2: HCAL-DNA CONJUGATES ON SWITCHSENSE
6 7
6.4.1 HYBRIDIZATION KINETICS OF HCAL-DNA CONJUGATES ON SURFACE 6 7
6.4.2 VOLTAGE RESPONSE OF HYBRIDIZED HCAL-DNA 70
6.4.3 TIME RESOLVED MEASUREMENT OF HYBRIDIZED HCAL-DNA 7 4
6.4.4 SIZING OF HCA1 7 9
6.4.5 ACETAZOLAMIDE INFLUENCE ON HCAL FIINCTIONALIZED LAYERS 8 2
6.5 CONCLUSION 8 3
7. ANTIBODY KINETICS ANALYSED IN SWITCHSENSE 8 5
7.1 INTRODUCTION 8 5
7.1.1 SURFACE-BASED SENSOR 8 5
7.1.2 SPR-BASED SENSOR 8 6
7.2 RESULTS 8 8
7.2.1 EMPLOYED MONOCLONAL ANTIBODIES AND EXPERIMENTAL DESIGN 8 8
7.2.2 KINETIC ANALYSIS ON THE SPR SYSTEM BIACORE* X100 9 0
7.2.3 KINETIC ANALYSIS ON SWITCHSENSE 9 5
7.2.4 TESTED CROSS-BINDING OF MONOCLONAL ANTIBODIES ON SWITCHSENSE 103
7.2.5 DISCUSSION 106
7.3 CONCLUSION 119
8. HUMAN P53 ANALYSED IN SWITCHSENSE 1 2 1
8.1 INTRODUCTION: P53 TUMOUR SUPPRESSOR 1 2 1
8.1.1 STRUCTURE AND FUNCTION OF INDIVIDUAL P53 DOMAINS 122
8.2 MOTIVATION 129
I I
IMAGE 3
8.3 EXPERIMENTAL DESIGN 130
8.3.1 THE PROTEIN CHARACTERIZATION 1 3 1
8.3.2 OLIGONUCLEOTIDE DESIGN 133
8.4 RESULTS 134
8.4.1 EMSA RESULTS 134
8.4.2 DETECTION OF P53-DNA BINDING ACTIVITY ON SWITCHSENSE 135
8.5 DISCUSSION 146
8.5.1 THE NONSPECIFIC BINDING OF P53 TO DNA 146
8.5.2 THE P53 DIMER-TETRAMER TRANSITION DETECTED ON SURFACE 148
8.5.3 THE DNA BENDING UPON P53 BINDING 150
8.5.4 THE ROLE OF THE HALF SITE RESPONSE ELEMENTS 152
8.6 CONCLUSION 153
9. CONCLUSION 155
10. REFERENCES 157
11. PUBLICATIONS 165
12. ACKNOWLEDGEMENTS 167
13. DECLARATION 169
14. APPENDIX 1 7 1
14.1 C-48-MER FOLDING 1 7 1
14.2 HYBRIDIZATION KINETICS 1 7 1
14.3 ANTIBODY KINETIC ANALYSIS IN BIACORE 173
14.4 ANTIBODY KINETIC ANALYSIS IN SW/TCFCSENSE 177
14.5 CROSS-BINDING EXPERIMENTS OF ANTIBODIES ON PROTOTYPE2 185
14.6 LFM DATA OF INITIAL DS-DNA LAYERS ON SWA1 AND ON PROTOTYPE2 187
14.7 P53 PROJECT 188
I I I
|
| any_adam_object | 1 |
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| ctrlnum | (OCoLC)835308846 (DE-599)BVBBV040791304 |
| discipline | Chemie |
| format | Thesis Book |
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| spelling | Villa, Valentina Verfasser aut Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions Valentina Villa 2012 III, 189 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2012 Wechselwirkung (DE-588)4064937-4 gnd rswk-swf Antikörper (DE-588)4002290-0 gnd rswk-swf Protein p53 (DE-588)4176550-3 gnd rswk-swf Antigen (DE-588)4002272-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Antikörper (DE-588)4002290-0 s Antigen (DE-588)4002272-9 s Protein p53 (DE-588)4176550-3 s Wechselwirkung (DE-588)4064937-4 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20121212-1115861-0-8 http://mediatum.ub.tum.de/node?id=1115861 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20121212-1115861-0-8 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025769428&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Villa, Valentina Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions Wechselwirkung (DE-588)4064937-4 gnd Antikörper (DE-588)4002290-0 gnd Protein p53 (DE-588)4176550-3 gnd Antigen (DE-588)4002272-9 gnd |
| subject_GND | (DE-588)4064937-4 (DE-588)4002290-0 (DE-588)4176550-3 (DE-588)4002272-9 (DE-588)4113937-9 |
| title | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions |
| title_auth | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions |
| title_exact_search | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions |
| title_full | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions Valentina Villa |
| title_fullStr | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions Valentina Villa |
| title_full_unstemmed | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions Valentina Villa |
| title_short | Electro-switchable DNA layers for the analysis of antibody-antigen and p53-DNA interactions |
| title_sort | electro switchable dna layers for the analysis of antibody antigen and p53 dna interactions |
| topic | Wechselwirkung (DE-588)4064937-4 gnd Antikörper (DE-588)4002290-0 gnd Protein p53 (DE-588)4176550-3 gnd Antigen (DE-588)4002272-9 gnd |
| topic_facet | Wechselwirkung Antikörper Protein p53 Antigen Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1115861 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20121212-1115861-0-8 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025769428&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT villavalentina electroswitchablednalayersfortheanalysisofantibodyantigenandp53dnainteractions |