Entwicklung LSPR-basierter optischer Biosensoren zum markierungsfreien Nachweis biomolekularer Wechselwirkungen in hochintegrierten Peptidbibliotheken [Elektronische Ressource] / vorgelegt von Fanny Caroline Liu

Liu Ren

Informations

• Publié par: ruprecht-karls-universitat_heidelberg
• Publié le: 01 janvier 2011
• Nombre de lectures: 121
• Langue: Deutsch
• Poids de l'ouvrage: 70 Mo

Extrait

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