Entwicklung LSPR-basierter optischer Biosensoren zum markierungsfreien Nachweis biomolekularer Wechselwirkungen in hochintegrierten Peptidbibliotheken [Elektronische Ressource] / vorgelegt von Fanny Caroline Liu
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2011
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154
pages
Deutsch
Ebook
2011
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Publié par
Publié le
01 janvier 2011
Nombre de lectures
121
Langue
Deutsch
Poids de l'ouvrage
70 Mo
Publié par
Publié le
01 janvier 2011
Nombre de lectures
121
Langue
Deutsch
Poids de l'ouvrage
70 Mo
INAUGURAL-DISSERTATION
Zur Erlangung der Doktorwürde
der Naturwissenschaftlich-Mathematischen Gesamtfakultät
der
Ruprecht-Karls-Universität
Heidelberg
vorgelegt von
Dipl. –Chem. Fanny Caroline Liu
aus Yogyakarta/Indonesien
Tag der mündlichen Prüfung: 11. Februar 2011
Entwicklung LSPR-basierter optischer Biosensoren zum
markierungsfreien Nachweis biomolekularer Wechselwirkungen in
hochintegrierten Peptidbibliotheken
Gutachter:
Prof. Dr. rer. nat. Reiner Dahint
Prof. Dr. rer. nat. Joachim Spatz
Development of LSPR-based Optical Biosensors
for the Label-Free Detection of Biomolecular Interactions
in High-Density Peptide Arrays
Referees:
Prof. Dr. rer. nat. Reiner Dahint
Prof. Dr. rer. nat. Joachim Spatz
CONTENTS 1
CONTENTS
CONTENTS .............................................................................................................................. 1
ABSTRACT .............................................................................................................................. 5
LIST OF PUBLICATIONS ..................................................................................................... 8
ABBREVIATIONS .................................................................................................................. 9
1. INTRODUCTION .............................................................................................................. 11
2. THEORY ............................................................................................................................. 14
2.1. In situ peptide synthesis ............................................................................................................................... 14
2.1.1 Photolithographic synthesis ..................................................................................................................... 15
2.1.2 SPOT synthesis ........................................................................................................................................ 16
2.1.3. Particlebased synthesis .......................................................................................................................... 16
2.2. Surface plasmon resonance as a tool in molecular biosensing ................................................................. 19
2.2.1. Surface plasmon resonance ..................................................................................................................... 19
2.2.1.1. Theory of propagating surface plasmon .......................................................................................... 19
2.2.1.2. Theory of localized surface plasmon ............................................................................................... 21
2.2.2. Surface plasmons in optical sensing ....................................................................................................... 21
2.2.2.1. Excitation of surface plasmons ........................................................................................................ 21
2.2.2.2. Quantitative interpretation of SPR response .................................................................................... 23
2.2.2.3. Methods of measurements ............................................................................................................... 25
2.2.2.4. Application of surface plasmons in biosensing ............................................................................... 26
2.3. Protein/resistant films .................................................................................................................................. 26
2.3.1. Poly(ethylene glycol) (PEG) ................................................................................................................... 27
2.3.2. Immobilizing PEG onto surfaces ............................................................................................................ 29
2.3.2.1. Chemisorption ................................................................................................................................. 29
2.3.2.2. Covalent grafting ............................................................................................................................. 29
2.3.2.3. Physisorption ................................................................................................................................... 30
2.3.2.4. Atom Transfer Radical Polymerization ........................................................................................... 31
2.3.2. Zwitterionic polymers ............................................................................................................................. 33
2.4. Surface analysis methods ............................................................................................................................. 34
2.4.1. Excitation and detection of surface plasmons ......................................................................................... 34
2.4.2. Xray Photoelectron Spectroscopy (XPS) ............................................................................................... 35
2.4.3. Scanning Electron Microscopy (SEM) ................................................................................................... 37
2.4.3.1. Basic Principle ................................................................................................................................. 38
2.4.3.2. SampleBeam Interactions ............................................................................................................... 38
3. MATERIALS AND METHODS ....................................................................................... 40
3.1. Fabrication of LSPR biosensors based on optical responsive materials ................................................. 40 CONTENTS 2
3.1.1. Materials ................................................................................................................................................. 40
3.1.2. Deposition of the bottom gold layer ....................................................................................................... 40
3.1.3. Deposition of the dielectric layer ............................................................................................................ 41
3.1.3.1. By simple incubation ....................................................................................................................... 41
3.1.3.2. By spin coating ................................................................................................................................ 41
3.1.3.3. By floating ....................................................................................................................................... 42
3.1.3.4. By thermal evaporation ................................................................................................................... 43
3.1.4. Deposition of the upper gold layer .......................................................................................................... 43
3.1.4.1. By metallization .............................................................................................................................. 43
3.1.4.2. By Physical Vapor Deposition ........................................................................................................ 43
3.2. Synthesis of protein/resistant coatings on the LSPR biosensor via Atom Transfer Radical
Polymerization (ATRP) ...................................................................................................................................... 44
3.2.1. Materials ................................................................................................................................................. 44
3.2.2. Direct synthesis of a proteinresistant coating on the LSPR biosensor ................................................... 45
3.2.2.1. Direct attachment of ATRP initiators on the LSPR biosensor ........................................................ 45
3.2.2.2. Synthesis of a graft polymer film on the LSPR biosensor via ATRP .............................................. 45
3.2.3. Indirect synthesis of a proteinresistant coating on the LSPR biosensor ................................................ 45
3.2.3.1. Synthesis of a silica gel on the LSPR biosensor .............................................................................. 45
3.2.3.2. Synthesis of a graft polymer film on the silica gelcoated LSPR biosensor .................................... 45
3.2.3.2.1. Attachment of ATRP initiators on the silicagel coated LSPR biosensor ................................ 45
3.2.3.2.2. Synthesis of a graft polymer film on the silicagel coated LSPR biosensor via ATRP
