Functionalized Conjugated Polymers for Signal Amplifying Biosensors and Sensor Arrays

289 Pages·2008·4.61 MB·English

Functionalized Conjugated Polymers for Signal Amplifying Biosensors
and Sensor Arrays

by

Kangwon Lee

A dissertation submitted in partial fulfillment
of the requirements for the degree of
Doctor of Philosophy
(Materials Science and Engineering)
in The University of Michigan
2008

Doctoral Committee:

Assistant Professor Jinsang Kim, Chair
Professor Erdogan Gulari
Professor David C. Martin
Associate Professor Megan S. Lim
KangwonL ee
© 2008
All Rights Reserved
To my family for their support and love

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ACKNOWLEDGMENTS

It is really difficult to list all the people who have helped me along the way during
the long journey toward this point of my academic career. First of all, I would like to give
my sincere acknowledgements to my supervisor, Professor Jinsang Kim for his guidance,
support, and especially patience during my whole doctoral study. He led me to this
fabulous world of photophysics and conjugated polymers, which I hardly knew anything
about before I joined his group. His enthusiasm in science always encouraged me to keep
on pursuing many interesting research topics. I also would like to appreciate my
committee members; Professor Erdogan Gulari, Professor David Martin, and Professor
Megan, S. Lim for their valuable suggestions and comments on my research. I thank Ilju
foundation, Korea for Ilju fellowship for last four years.
I really want to thank microarray team including Dr. Jean-Marie Rouillard, Trinn
Pham and Dr. Suparna Mandal. Especially, Jean-Marie greatly helped me study DNA
microarray technique with valuable suggestions. I also appreciate the invaluable help of
Kojo Elenitoba-Johnson, Charles Seiler and Adam Kronk in pathology department
regarding cell-staining test. I have enjoyed collaborating with Professor Darrin Pochan
and Tuna Yucel in polymer characterization. I also would like to thank current and past
group members. Kim group is a great group to work in. All of them always gave me their
support and advice. Korean graduate students of MSE, Macro and ChemE departments at
Michigan have made my time profitable.
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I owe a great deal to my Lord, Jesus Christ. I also thank all of my friends of
Korean Bible Church of Ann Arbor for their love and support. Especially, I will not
forget enjoyable time with Seunghyun, Ilju, and Taeil family eternally.
Finally, I would love to thank all my family members including my parents, my
sister, my parents in law and two sisters in law. Without unconditional love, support, and
prayer from all of my family members, I could never have achieved this goal. Finally, I
deeply thank my lovely wife, Eunjoo, my son, Minseo, and my cute daughter, Yeseo for
their love, patience and support. I specially dedicate this work to Eunjoo with love from
the bottom of my heart.

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TABLE OF CONTENTS
DEDICATION................................................................................................................... ii
ACKNOWLEDGMENTS ............................................................................................... iii
LIST OF FIGURES……………..………………………………………….....................x
LIST OF SCHEMES………………………………….………………………..…….xviii
LIST OF TABLES……………………………………………………………………...xx
ABSTRACT…………………………………………………………………………….xxi
CHAPTER 1. Introduction and Background.……...…………………………………..1
1.1. Conjugated Polymers…………...…….…….………………………………...2
1.2. Optoelectronic Property of CPs………………………………...………..…...4
1.3. Sensory Signal Amplification of CPs…………………...……………………7
1.4. Energy and Electron Transfer of CPs……………………………………...…8
1.5. Signal Transduction Modes of CPs………………………………...………..14
1.5.1. Turn-on Mode…………………………………………...…………….15
1.5.2. Turn-off Mode……………………………………………………...…19
1.5.3. Fluorescence Color Change Mode………………………………….…22
1.6. Objectives…………………………………………………………….……..29
1.7. References………...…………………………………………………………31
CHAPTER 2. Modulating the Side-chain Design of Poly(p-phenyleneethynylene)
Derivatives to Make Water-Soluble and Highly Emissive Conjugated
Polyelectrolytes………………………………………………………………………....35
2.1. Abstract………...………………………………………………..…………..36
2.2. Introduction…………...……………………………………………………..37
2.3. Experimental Section..……………………………………………..………..39
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2.3.1. General Methods………………………...…………………………….39
2.3.2. Synthesis and Characterization………...………..…………………… 41
2.4. Results and Discussion………………....……...……………………………49
2.5. Conclusion.……………...…………………………………………………..60
2.6. References………………...………………………………………………....61
CHAPTER 3. Synthesis and Functionalization of a Highly Fluorescent and
Completely Water Soluble Poly(para-phenyleneethynylene) Copolymer for
Bioconjugation………..………………………………………………………………...66
3.1. Abstract…………………………...………………………………………....67
3.2. Introduction…………………………………...……………………………..68
3.3. Experimental Section…………………………………………..……………69
3.3.1. Materials and Methods………………………………………………..69
3.3.2. Synthesis of 2,5-Diiodo-1,4-dimethoxybenzene……………………...70
3.3.3. Synthesis of 1,4-Diiodo-2,5-hydroquinone……………………………71
3.3.4. Synthesis of 1.3-Bis(3,6,9-trioxadecanyl)glycerol-2-toluenesulfonic
ester……………………………………………..……………………..71
3.3.5. Synthesis of 1,4-Bis(1,3-bis(3,6,9-trioxadecyl)-2-glyceryl)2,5-
diiodobenzene……...…………………………………………………..72
3.3.6. Synthesis of (2,5-Bis(2,5,8,11,15,18,21,24-octaoxapentacosan-13-
yloxy)-1,4-phenylene)bis(ethyne-2,1-diyl)bis(trimethylsilane)……....73
3.3.7. Synthesis of Monomer………………………...……………………...73
3.3.8. Synthesis of 1,4-Disodiumpropanesulfonoxy-2,5-diiodobenzene….....74
3.3.9. Synthesis for PPE-R and PPE-R -COOH…………………………….75
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3.4. Results and Discussion……………...………………………………………75
3.5. Conclusions…………………………………………...…………….……….82
3.6. References…………………………………………………………………...82
CHAPTER 4. Label-Free and Self-Signal Amplifying Molecular DNA Sensors
Based on Bioconjugated Polyelectrolytes ………………….……………………..…...85
4.1. Abstract………………………………………………….…………………..86
4.2. Introduction……………………………………………………...……..……87
4.3. Experimental Section......................................................................................91
4.4. Results and Discussion……………...……………………...….…………....94
4.5. Conclusion.………………………………………...………..……….….…107
4.6. References……………………………………………...……..……………107
CHAPTER 5. Conjugated Polyelectrolyte-Antibody Hybrid Molecules for Live Cell-
Imaging………………………………………………………………………………...112
5.1. Abstract……………………..…………………...……………..…………..113
5.2. Introduction………………………...……...……………………...………..114
5.3. Experimental Section….…..…………………………...………………..…116
5.4. Results and Discussion…………………………………………………….126
5.5. Conclusion...…………………………………...…………………………..142
5.6. References………………………………………...………………………..142
CHAPTER 6. Chemically and Photochemically Stable Conjugated Poly(oxadiazole)
Derivatives: A Comparison with Polythiophenes and Poly(p-
phenyleneethynylenes)………………………………………………………………...145
6.1. Abstract…………………………………………...………………………..146
6.2 Introduction………………………………………………………………....147
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6.3. Experimental Section...…………………………………………...……..…149
6.4. Results and discussion………………………………………....…………..153
6.5. Conclusion.………………………………...….…………………………...165
6.6. References…………………………………...………………………….….166
CHAPTER 7. Signal-Amplifying Conjugated Polymer-DNA Hybrid Chips…......170
7.1. Abstract………………………………………………………..…………...171
7.2. Introduction…………………………………...…………………...……….172
7.3. Experimental Section………………………………………...…………….173
7.3.1. P1 Synthesis…………….…………………………...…………….…173
7.3.2. DNA Chip Fabrication………………………………………...……..179
7.3.3. Hybridization Test……………………………………………...…....181
7.4. Results and Discussion………………………………………………..…...184
7.5. Conclusion…...………………………………...……………………….….191
7.6. References………………………………………...………………………..192
CHAPTER 8. Sensitive and Selective Label-free DNA Detection by Conjugated
Polymer-based Microarray and Intercalating Dye…………………………….........195
8.1. Abstract…………………………………….……...…………..…………...196
8.2. Introduction…………………………………...…………………...……….197
8.3. Experimental Section………………………………………...…………….199
8.4. Results and Discussion……………………………………...………...…...202
8.5. Conclusion…...………………………………...……………………….….211
8.6. References……………………………………...…………………………..211
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CHAPTER 9. Label-Free and Self-Signal Amplifying “Molecular Beacon”
Biosensors using Conjugated Polymers……………………………………..….........213
9.1. Abstract………………...………………….…………………..…………...214
9.2. Introduction……………...………………………………………...……….215
9.3. Experimental Section…………………...………………………………….217
9.4. Results and Discussion………………...……………………………...…...224
9.5. Conclusion…...…………...…………………………………………….….238
9.6. References…………………...……………………………………………..240
CHAPTER 10. Label-Free Identification of Prostate-Specific Antigen using
Conjugated Polymer-Peptide Hybrid Chips……………………………..…….........242
10.1. Abstract…………………………...…….………..…………..…………...243
10.2. Introduction…………………………...…………..……………...……….244
10.3. Experimental Section…………………………...……..………………….246
10.4. Results and Discussion………………………...……..……………...…...250
10.5. Conclusion…...………………………...……………..……………….….257
10.6. References……………………...……………………..…………………..258
CHAPTER 11. Summary and Future Prospective….…...……………...…….........260
11.1. Research Summary….………………….………..…………..…………...261
11.2. Future Consideration………………………….…..……………...……….265

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