Development of spider silk protein particles for pharmaceutical applications:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Volltext http://d-nb.info/1047543559/34 kostenfrei Inhaltsverzeichnis |
| Beschreibung: | VII, 179 S. Ill., graph. Darst. |
| Format: | Langzeitarchivierung gewährleistet, LZA |
Internformat
MARC
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| 245 | 1 | 0 | |a Development of spider silk protein particles for pharmaceutical applications |c Markus Hofer |
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| 502 | |a München, Univ., Diss., 2013 | ||
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Datensatz im Suchindex
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| adam_text | TABLE OF CONTENTS
CHAPTER 1: GENERAL INTRODUCTION
1.1 I
NTRODUCTION
1
1.2 D
RUG DELIVERY SYSTEMS FOR THERAPEUTIC
PROTEINS
3
1.3 P
ROTEIN-BASED DRUG DELIVERY SYSTEMS
6
1.3.1 PREPARATION TECHNIQUES FOR PROTEIN
PARTICLES AS DRUG CARRIERS 6
1.3.1.1 EMULSION / SOLVENT EXTRACTION 6
1.3.1.2 COACERVATION - NON-SOLVENT 7
1.3.1.3 COACERVATION - SALT PRECIPITATION 7
1.3.2
ALBUMIN
8
1.3.3 GELATIN 8
1.3.4
SILK
9
1.3.4.1 STRUCTURE AND PROPERTIES OF SILK PROTEINS 10
1.3.4.2 NATURAL ASSEMBLY PROCESS 12
1.3.4.3 IMMUNOGENICITY AND BIODEGRADATION OF SILK PROTEINS 13
1.3.4.4 SILKWORM SILK FOR DRUG DELIVERY 14
1.4 S
PIDER SILK PROTEINS
19
1.4.1 DEVELOPMENT OF RECOMBINANT SPIDER SILK PROTEINS 19
1.4.1.1 TYPES OF SPIDER SILK PROTEINS 19
1.4.1.2 ENGINEERED SPIDER SILK PROTEINS 20
1.4.2
SPIDER SILK PROTEIN
EADF4(C16) 21
1.4.2.1 CHARACTERISTICS OF EADF4(C16) 21
1.4.2.2 PROCESSING OF EADF4(C16) 22
1.4.2.3 EADF4(C16) FOR DRUG DELIVERY 26
1.5
O
BJECTIVES OF THE
THESIS 28
1.6 R
EFERENCES
30
CHAPTER 2:
MATERIALS AND METHODS
2.1 M
ATERIALS
41
2.1.1 PROTEINS 41
2.1.1.1 SPIDER SILK PROTEIN EADF4(C16) 41
2.1.1.2 LYSOZYME 42
2.1.1.3 NERVE GROWTH FACTOR 42
I
HTTP://D-NB.INFO/1049218957
TABLE OF CONTENTS
2.1.2 REAGENTS, CHEMICALS AND EXCIPIENTS 43
2.2 M
ETHODS APPLIED
FOR SPIDER SILK PROTEIN OR SPIDER SILK PARTICLES
45
2.2.1 PREPARATION OF
LIQUID
EADF4
(CL6)
FORMULATIONS
45
2.2.2
CHARACTERIZATION OF
LIQUID
EADF4
(CL6)
FORMULATIONS
45
2.2.2.1 ASYMMETRICAL FLOW FIELD-FLOW FRACTIONATION 45
2.2.2.2 INCUBATION AT ELEVATED TEMPERATURE 46
2.2.2.3 SIZE EXCLUSION CHROMATOGRAPHY 46
2.2.2.4 SODIUM DODECYL SULPHATE POLYACRYLAMIDE GEL ELECTROPHORESIS 46
2.2.2.5 NEPHELOMETRY 47
2.2.2.6 LIGHT OBSCURATION 47
2.2.3
PREPARATION OF
EADF4(CL
6)
PARTICLES
48
2.2.3.1 MICROMIXING SYSTEM 48
2.2.3.2 DIALYSIS 49
2.2.3.3 ULTRASONIC NOZZLE 49
2.2.3.4 ULTRASONIC TREATMENT AND PURIFICATION 50
2.2.3.5 GRAVIMETRICAL DETERMINATION OF PARTICLE CONCENTRATION 50
2.2.3.6 FREEZE-DRYING OF NANOSUSPENSIONS 51
2.2
.4
CHARACTERIZATION OF
EADF4
(CL6)
PARTICLES 51
2.2.4.1 DYNAMIC LIGHT SCATTERING 51
2.2.4.2 LASER DIFFRACTION 52
2.2.4.3 FOURIER TRANSFORMED INFRARED SPECTROSCOPY 53
2.2.4.4 SCANNING ELECTRON MICROSCOPY 53
2.2.4.5 COLLOIDAL STABILITY OF EADF4(C16) PARTICLES 53
2.2.4.6 DETERMINATION OF RESIDUAL MOISTURE 53
2.2.4.7 DIFFERENTIAL SCANNING CALORIMETRY 54
2.2.5 REMOTE LOADING
OF SPIDER SILK PARTICLES 55
2.2.5.1 LOADING PROCEDURE 55
2.2.5.2 DETERMINATION OF LOADING AND LOADING EFFICIENCY 55
2.2.5.3 CONFOCAL LASER SCANNING MICROSCOPY 56
2.2.6 ENCAPSULATION OF
DRUGS INTO SPIDER SILK PARTICLES 57
2.2.6.1 CO-PRECIPITATION OF RHODAMINE B 57
2.2.6.2 CO-PRECIPITATION OF LYSOZYME 57
2.2.7 IN VITRO RELEASE
FROM SPIDER SILK PARTICLES 59
2.2.7.1 EXPERIMENTAL SETUP 59
2.2.7.2 MICRO BCA PROTEIN ASSAY 61
2.2.7.3 SIZE EXCLUSION CHROMATOGRAPHY - LYSOZYME 61
2.2.7.4 ACTIVITY ASSAY - LYSOZYME 61
II
TABLE OF CONTENTS
2.3 METHODS APPLIED FOR
STUDIES WITH NERVE
GROWTH FACTOR 63
2.3.1 FORCED DEGRADATION OF
NERVE GROWTH FACTOR 63
2.3.1.1 EXPERIMENTAL SETUP 63
2.3.1.2 DESORPTION PROCEDURE 64
2.3.1.3 FREEZE-THAW 65
2.3.1.4 STIRRING 65
2.3.1.5 LIGHT EXPOSURE 65
2.3.1.6 INCUBATION AT ELEVATED TEMPERATURE 66
2.3.2 ANALYTICAL METHODS 66
2.3.2.1 SIZE EXCLUSION CHROMATOGRAPHY 66
2.3.2.2 REVERSED PHASE CHROMATOGRAPHY 66
2.3.2.3 NEPHELOMETRY 67
2.3.2.4 LIGHT OBSCURATION 67
2.3.2.5 SODIUM DODECYL SULPHATE POLYACRYLAMIDE GEL ELECTROPHORESIS 67
2.3.2.6 MICROCALORIMETRY 68
2.4 REFERENCES 69
CHAPTER 3: PREPARATION TECHNIQUES, CHARACTERIZATION
AND
FORMULATION DEVELOPMENT OF
SPIDER SILK PARTICLES
3.1 INTRODUCTION
71
3.2 PREPARATION OF EADF4(CL
6) PARTICLES 74
3.2.1 SPIDER SILK NANOPARTICLES 74
3.2.2 SPIDER SILK MICROPARTICLES 80
3.2.2.1 SIMULTANEOUS ATOMIZATION OF EADF4(C16) SOLUTION AND POTASSIUM
PHOSPHATE SOLUTION 80
3.2.2.2 ATOMIZATION OF EADF4(C16) INTO POTASSIUM PHOSPHATE SOLUTIONS 82
3.2.2.3 ATOMIZATION OF POTASSIUM PHOSPHATE INTO EADF4(C16) SOLUTIONS 84
3.3 CHARACTERIZATION OF LIQUID
EADF4(C16)
FORMULATIONS 88
3.3.1 CHARACTERIZATION OF
EADF4
(C16)
BY
AF4 88
3.3.2 STABILITY OF
EADF4
(CL6)
AT ELEVATED TEMPERATURE 93
3.4 COLLOIDAL STABILITY OF
SPIDER SILK NANOPARTICLES 96
3.5 FREEZE-DRYING OF EADF4(C16)
NANOSUSPENSIONS 99
3.5.1 FEASIBILITY STUDY 99
3.5.2 FORMULATION DEVELOPMENT 101
3.6 SUMMARY AND CONCLUSIONS :...
104
III
TABLE OF CONTENTS
3.7 R
EFERENCES
106
CHAPTER4: SPIDER SILK PARTICLES FOR
DRUG DELIVERY
4.1 I
NTRODUCTION 109
4.2 P
ROTEIN LOADED
EADF4(C16)
PARTICLES
VIA REMOTE LOADING
112
4.2.1 LYSOZYME 112
4.2.2 NERVE GROWTH
FACTOR 116
4.2.3 INVESTIGATION OF LOADING
MECHANISM 119
4.2.4
IN VITRO RELEASE
FROM PROTEIN LOADED SPIDER SILK PARTICLES
123
4.2.4.1 INFLUENCE OF PH, IONIC STRENGTH AND PAYLOAD 123
4.2.4.2 ANALYSIS OF DRUG RELEASE FROM REMOTE LOADED EADF4(C16)
PARTICLES. 125
4.2.4.3 IN VITRO RELEASE IN THE PRESENCE OF ALBUMIN 129
4.3 E
NCAPSULATION OF DRUGS
INTO
EADF4(C16)
PARTICLES
131
4.3.1
RHODAMINEB
132
4.3.2
LYSOZYME
133
4.3.2.1 LYSOZYME-LOADED NANOPARTICLES 134
4.3.2.2 LYSOZYME-LOADED MICROPARTICLES 137
4.4 S
UMMARY AND CONCLUSIONS
- 140
4.5 R
EFERENCES 142
CHAPTER 5: SPIDER SILK PARTICLES AS NOVEL
EXCIPIENT FOR THE
STABILIZATION OF
NERVE GROWTH FACTOR
5.1 I
NTRODUCTION
145
5.2 D
EVELOPMENT OF A STANDARD
DESORPTION PROCEDURE
148
5.3 F
ORCED DEGRADATION
STUDIES
151
5.3.1 FREEZE-THAW STUDY 151
5.3.2 STIRRING 155
5.3.3 LIGHT EXPOSURE 159
5.3.4
INCUBATION AT ELEVATED TEMPERATURE
164
5.4 S
UMMARY AND CONCLUSIONS
170
5.5 R
EFERENCES
172
IV
TABLE OF CONTENTS
CHAPTER 6:
FINAL SUMMARY AND CONCLUSION
F
INAL SUMMARY AND CONCLUSION 175
V
|
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| spelling | Hofer, Markus Verfasser aut Development of spider silk protein particles for pharmaceutical applications Markus Hofer 2013 VII, 179 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2013 Langzeitarchivierung gewährleistet, LZA (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-165970 https://nbn-resolving.org/urn:nbn:de:bvb:19-165970 Resolving-System kostenfrei Volltext http://d-nb.info/1047543559/34 Langzeitarchivierung Nationalbibliothek http://edoc.ub.uni-muenchen.de/16597/ Verlag kostenfrei DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027236879&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hofer, Markus Development of spider silk protein particles for pharmaceutical applications |
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| title | Development of spider silk protein particles for pharmaceutical applications |
| title_auth | Development of spider silk protein particles for pharmaceutical applications |
| title_exact_search | Development of spider silk protein particles for pharmaceutical applications |
| title_full | Development of spider silk protein particles for pharmaceutical applications Markus Hofer |
| title_fullStr | Development of spider silk protein particles for pharmaceutical applications Markus Hofer |
| title_full_unstemmed | Development of spider silk protein particles for pharmaceutical applications Markus Hofer |
| title_short | Development of spider silk protein particles for pharmaceutical applications |
| title_sort | development of spider silk protein particles for pharmaceutical applications |
| topic_facet | Hochschulschrift |
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