Lanthanides and quantum dots [Elektronische Ressource] : time-resolved laser spectroscopy of biochemical Förster resonance energy transfer (FRET) systems / by Niko Hildebrandt

Niko Hildebrandt

Sujets

• Biologie

Informations

• Publié par: universitat_potsdam
• Publié le: 01 janvier 2006
• Nombre de lectures: 22
• Langue: English
• Poids de l'ouvrage: 7 Mo

Extrait

Lanthanides and Quantum Dots -
Time-Resolved Laser Spectroscopy of Biochemical
F¨orster Resonance Energy Transfer (FRET) Systems
by
Niko Hildebrandt
A Dissertation
submitted to the
Mathematics-Natural Science Faculty
of the University of Potsdam
in partial fulfillment of the
requirements for the degree of
Doctor of Natural Science
Potsdam, December 2006To Ella and Monique.
You make my life complete.Contents
List of publications III
Abstract V
Kurzzusammenfassung VII
1 Introduction 1
2 Theoretical and practical background 3
2.1 F¨orsterresonanceenergytransfer.......................... 3
2.1.1 Introduction.................................. 3
2.1.2 Theoreticaltreatment ............................ 3
2.1.3 FRETapplications.............................. 6
2.2 Luminescencequenching............................... 7
2.3 Lanthanidecomplexes ................................ 8
2.3.1 Lanthanideions 8
2.3.2 Lanthanideioncomplexingligands..................... 15
2.3.3 Biochemical applications of lanthanide complexes . . . . . . . . . . . . . 17
2.4 Quantum dot semiconductor nanocrystals . . . . . . . . . . . . . . . . . . . . . 19
2.4.1 Introduction.................................. 19
2.4.2 The quasiparticle approach in semiconductor crystals . . . . . . . . . . . 19
2.4.3 Quantumconfinement............................ 20
2.4.4 Core/shell semiconductor QD . . . . . . . . . . . . . . . . . . . . . . . . 23
2.4.5 BiocompatibleQD.............................. 25
2.5 BiochemicalFRETsystems............................. 27
2.5.1 Immunoassays................................. 27
2.5.2 Biotin-(strept)avidin 30
2.5.3 Conjugationmethods 32
3 Experimental section 35
3.1 Measuringsolutions.................................. 35
3.2 FRET donors and acceptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
3.3 Analyticalmethods 36
3.3.1 Stationary absorption and luminescence spectroscopy . . . . . . . . . . . 36
3.3.2 Time-resolved luminescence spectroscopy and fluoro immunoassays . . . 36
3.3.3 MALDI-TOF 38
3.4 Dataprocessing.................................... 38
III
4 Results and discussion 39
4.1 CharacterizationoftheFRETdonors........................ 39
4.1.1 Bioconjugation................................ 39
4.1.2 Photophysicalcharacterization 42
4.2 Characterization of the FRET acceptors . . . . . . . . . . . . . . . . . . . . . . 49
4.2.1 Bioconjugation 50
4.2.2 Photophysicalcharacterization 50
4.3 FRETexperiments.................................. 53
4.3.1 Streptavidin-biotin FRET system . . . . . . . . . . . . . . . . . . . . . . 53
4.3.2 HCGFRETimmunoasays ......................... 62
5 Summary and outlook 67
6 Appendix 71
6.1 Fundamental constants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
6.2 Abbreviations..................................... 71
6.3 Emisionspectra................................... 73
6.4 Luminescencedecaykinetics............................. 82
6.5 FRETexperiments.................................. 88
6.5.1 Biot-QD655 as acceptor . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
6.5.2 Biot-APC and Biot-DY633 as acceptors . . . . . . . . . . . . . . . . . . 92
References 97List of publications
Original
1. Hildebrandt N, Charbonni`ere LJ, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Homogeneous FRET
Immunoassay Based on Lanthanides to Quantum Dots Energy Transfer” In: Osinski M, Jovin
TM, Yamamoto K, editors. Proceedings of SPIE Vol. 6448 - Colloidal Quantum Dots for
Biomedical Applications II; 2007; 64480K (12 pages).
2. Niederkruger¨ M, Salb C, Marowsky G, Beck M, Hildebrandt N, L¨ ohmannsr¨ oben H-G;
”Improvement of a Fluorescence Immunoassay with a Compact Diode-Pumped Solid State
Laser at 315 nm” In: Proceedings of SPIE Vol. 6380 - Smart Medical and Biomedical Sensor
Technology IV; 2006; 63800M (9 pages).
3. Charbonni`ere LJ, Hildebrandt N, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Lanthanides to
Quantum Dots Resonance Energy Transfer in Time-Resolved FluoroImmunoAssays and Lu-
minescence Microscopy” Journal of the American Chemical Society2006;128(39): 12800-12809.
4. Hildebrandt N, Charbonni`ere LJ, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Quantum Dots
as Resonance Energy Transfer Acceptors for Monitoring Biological Interactions” In: Grzymala
R, Haeberle O, editors. Proceedings of SPIE Vol. 6191 - Biophotonics and New Therapy
Frontiers; 2006; 61910W (9 pages).
5. Beck M, Hildebrandt N, L¨ ohmannsr¨ oben H-G; ”Quantum Dots as Acceptors in FRET
Assays Containing Serum” In: Grzymala R, Haeberle O, editors. Proceedings of SPIE Vol.
6191 - Biophotonics and New Therapy Frontiers; 2006; 61910X (8 pages).
6. L¨ ohmannsr¨ oben H-G, Beck M, Hildebrandt N, Schm¨ alzlin E, van Dongen JT; ”Laser-Based
Fluoroimmuno Analysis and In-Vivo Optical Oxygen Monitoring” In: Gries W, Pearsall TP,
editors. Proceedings of SPIE Vol. 6157 - Workshop on Laser Applications in Europe; 2006;
E1-E6.
7. Hildebrandt N, Charbonni`ere LJ, Beck M, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Quan-
tum Dots As Efficient Energy Acceptors in a Time-Resolved Fluoroimmunoassay” Angewandte
Chemie - International Edition 2005;44(46): 7612-7615.
8. Hildebrandt N, Charbonni`ere LJ, Beck M, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Quan-
tenpunkte als effiziente Energieacceptoren in einem zeitaufgel¨ osten Fluoroimmuntestsystem”
Angewandte Chemie 2005;117(46): 7784-7788.
Review
1. Hildebrandt N, L¨ ohmannsr¨ oben H-G; ”Quantum Dot Nanocrystals and Supramolecular Lan-
thanide Complexes - Energy Transfer Systems for Sensitive In Vitro Diagnostics and High
Throuput Screening in Chemical Biology” Current Chemical Biology 2007; accepted for pub-
lication.
IIIIV
Oral presentations at conferences and meetings
1. Hildebrandt N, Charbonni`ere LJ, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Homogeneous FRET
Immunoassay Based on Lanthanides to Quantum Dots Energy Transfer” BiOS 2007 -
Biomedical Optics - Colloidal Quantum Dots for Biomedical Applications II, San Jose, USA;
2007.
2. Hildebrandt N, Charbonni`ere LJ, Beck M, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Lan-
thanides and Quantum Dots as Efficient Donor-Acceptor Pairs for Sensitive Homogeneous
F¨ orster Resonance Energy Transfer Fluoroimmunoassays (FRET-FIA)” EOS Topical Meeting
on Biophotonics and Biomedical Optics 2006, Paris, France; 2006.
3. Hildebrandt N, Charbonni`ere LJ, Beck M, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Lan-
thanide Donors and Quantum Dot Acceptors for Homogeneous FRET Immunoassays” 105.
Bunsentagung 2006, Erlangen, Germany; 2006.
4. Hildebrandt N, Charbonni`ere LJ, Ziessel RF, L¨ ohmannsr¨ oben H-G; ”Quantum Dots
as Resonance Energy Transfer Acceptors for Monitoring Biological Interactions” Photonics
Europe 2006 - Biophotonics and New Therapy Frontiers, Strasbourg, France; 2006.
5. Hildebrandt N, Flehr R, Bois E, L¨ ohmannsr¨ oben H-G; ”Optimized Homogeneous Immunoas-
say Based on XeCl-Laser Excited F¨ orster Resonance Energy Transfer” CLEO/EUROPE 2005
- Conferences on Lasers and Electro-Optics/Europe - Photonic Manipulation and Bio-Sensing,
M¨ unchen, Germany; 2005.
6. Hildebrandt N,BoisE,NutiD,L¨ohmannsr¨ oben H-G; ”New lasers for better fluores-
cence immunoassays (FIA)” OPTO 2003, Paris - France; 2003.
Poster presentations at conferences and meetings
1. Hildebrandt N, Bois E, Nuti D, L¨ ohmannsr¨ oben H-G; ”Sensitivity Improvement of a Laser-
Based Homogeneous Fluorescence Immunoassay (FIA) Laboratory System” EPA summer
school: New Perspectives in Photochemistry, Egmond aan See - Netherlands; 2003.
2. Hildebrandt N, Bois E, Nuti D, L¨ ohmannsr¨ oben H-G; ”Miniaturized Laser-Based Ho-
mogeneous Fluorescence Immunoassay (FIA) Reader System” MINIT 2003, Potsdam,
Germany; 2003.Abstract
F¨ orster Resonance Energy Transfer (FRET) plays an important role for biochemical applica-
tions such as DNA sequencing, intracellular protein-protein interactions, molecular binding
studies, in vitro diagnostics and many others. For qualitative and quantitative analysis, FRET
systems are usually assembled through molecular recognition of biomolecules conjugated with
donor and acceptor luminophores. Lanthanide (Ln) complexes, as well as semiconductor
quantum dot nanocrystals (QD), possess unique photophysical properties that make them
especially suitable for applied FRET. In this work the possibility of using QD as very efficient
FRET acceptors in combination with Ln complexes as donors in biochemical systems is
demonstrated. The necessary theoretical and practical background of FRET, Ln complexes,
QD and the applied biochemical models is outlined. In addition, scientific as well as commercial
applications are presented.
FRET can be used to measure structural changes or dynamics at distances ranging from
approximately 1 to 10 nm. The very strong and well characterized binding process between
streptavidin (Strep) and biotin (Biot) is used as a biomolecular model system. A FRET
system is established by Strep conjugation with the Ln complexes and QD biotinylation. Three
+ +
3 3Ln complexes (one with Tb and two with Eu as central ion) are used as FRET donors.
Besides the QD two further acce