Table Of ContentTopics in
Fluorescence
Spectroscopy
Volume 10
Advanced Concepts in
Fluorescence Sensing
Part B: Macromolecular Sensing
Topics in Fluorescence Spectroscopy
Edited by JOSEPH R. LAKOWICZ and CHRIS D. GEDDES
Volume ]: Techniques
Volume 2: Principles
Volume 3: Biochemical Applications
Volume 4: Probe Design and Chemical Sensing
Volume 5: Nonlinear and Two-Photon-Induced Fluorescence
Volume 6: Protein Fluorescence
Volume 7: DNA Technology
Volume 8: Radiative Decay Engineering
Volume 9: Advanced Concepts in Fluorescence Sensing
PartA: Sn1all Molecule Sensing
Volume 10: Advanced Concepts in Fluorescence Sensing
Part B: Macromolecular Sensing
Topics in
Fluorescence
Spectroscopy
Volume 10
Advanced Concepts in
Fluorescence Sensing
Part B: Macromolecular Sensing
Edited by
CHRIS D. GEDDES
The Institute ofFluorescence
Medical Biotechnology Center
University ofMaryland Biotechnology Institute
BaltiJnore, Maryland
and
R.
JOSEPH LAKOWICZ
Centerfor Fluorescence Spectroscopy and
Department ofBiochelnistry and Molecular Biology
University ofMaryland School ofMedicine
Baltimore, Maryland
~
Springer
The Library ofCongresscatalogedthe first volume ofthis title as follows:
Topics in fluorescence spectroscopy/edited by Chris D. Geddes andJoseph R. Lakowicz.
p. cm.
Includes bibliographical references and index.
Contents: v.I. Techniques
I. Fluorescence spectroscopy. 1. Geddes.Chris D. II. Lakowicz, Joseph R.
QD96.F56T66 1991 91-32671
543'.0858-dc20 CIP
ISSN: 1574-1036
ISBN 0-387-23644-9 Printedon acid-freepaper
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CONTRIBUTORS
Caleb Behrend. Department of Chemistry, University of Michigan, Ann Arbor,
Michigan, 48109-1055.
Murphy Brasuel. DepartmentofEnvironmentalHealthSciences, UniversityofMichigan,
AnnArbor, Michigan, 48109-1055.
Sarah M Buck. Raoul Kopelman. Department of Chemistry, University of Michigan,
AnnArbor, Michigan, 48109-1055.
Eun Jeong Cho. Department ofChemistry and Biochemistry, Institute for Cellular and
MolecularBiology,TheUniversityofTexas atAustin, Austin, TX78712.
Sonja Draxler. Institut fur Experimentalphysik, Karl-Franzens-Universitat Graz, A-8010
GrazAustria.
Andrew D. Ellington. Department ofChemistry and Biochemistry, Institute for Cellular
andMolecularBiology,TheUniversityofTexas atAustin, Austin, TX78712.
Peter M Haggie. Departments ofMedicine and Physiology, Cardiovascular Research
Institute,UniversityofCalifornia, SanFrancisco, CA, 94143-0521.
TonyDJames. DepartmentofChemistry, UniversityofBath, BathBA2 7AYUK.
HuiJiang. BostonUniversity, ChemistryDepartmentand Photonics Center, Boston, MA
02215.
Guilford Jones. II. Boston University, Chemistry Department and Photonics Center,
Boston, MA02215.
Yong-Eun Lee Koo. Department of Chemistry, University of Michigan, Ann Arbor,
Michigan, 48109-1055.
RobertMasse. Applied Research and Development, MDS Pharma Services, 2350 Cohen
Street, Montreal, QC, Canada.
Eric Monson. Department ofChemistry, University ofMichigan, Ann Arbor, Michigan,
48109-1055.
Martin A. Philbert. Department of Environmental Health Sciences, University of
Michigan, AnnArbor, Michigan, 48109-1055.
v
vi CONTRIBUTORS
William S. Powell. Meakins-Christie Laboratories, McGill University, 3626 St. Urbain
Street, Montreal, QC, Canada.
Manjula Rajendran. Department of Chemistry and Biochemistry, Institute for Cellular
andMolecularBiology,TheUniversityofTexasatAustin, Austin,TX78712.
AlnawazRehemtulla.MolecularTherapeuticsInc., AnnArbor, Michigan,48109.
BrianRoss. MolecularTherapeuticsInc., AnnArbor, Michigan,48109
Seiji Shinkai. Department of Chemistry and Biochemistry, Graduate School of
Engineering,KyushuUniversity, Fukuoka812-8581 JAPAN.
Richard B. Thompson. Department ofBiochemistry and Molecular Biology, School of
Medicine and Center for Fluorescence Spectroscopy, UniversityofMaryland, Baltimore,
Maryland21201.
Petra Turkewitsch. Applied Research and Development, MDS Pharma Services, 2350
CohenStreet,Montreal, QC, Canada.
A.S. Verkman. Departments of Medicine and Physiology, Cardiovascular Research
Institute,UniversityofCalifornia, SanFrancisco, CA, 94143-0521.
Valentine 1. Vullev. Boston University, Chemistry Department and Photonics Center,
Boston,MA02215.
Hao Xu. Department of Chemistry, University of Michigan, Ann Arbor, Michigan,
48109-1055.
PREFACE
Over the last decade fluorescence has become the dominant tool in
biotechnology and medical imaging. These exciting advances have beenunderpinned by
the advances in time-resolved techniques and instrumentation, probe design, chemical /
biochemicalsensing, coupledwithourfurtheredknowledge inbiology.
Ten years ago Volume 4 of the Topics in Fluorescence Spectroscopy series
outlined the emerging trends in time resolved fluorescence in analytical and clinical
chemistry. These emerging applications of fluorescence were the result of continued
advances inboth laser and computer technology and a drive to develop red/near-infrared
fluorophores. Based on the advancements in these technologies, it was envisaged that
small portable devices would find future common place in a doctor's office or for home
healthcare.
Today, these past emerging trends in fluorescence sensing are now widely used
as eitherstandardpractices inclinicalassessmentorcommercializedhealthcareproducts.
Miniature lasers in the form oflaser diodes and even light emitting diodes are widely
used in applications oftime-resolved fluorescence. Computer clock-speed is now not
considered a hurdle in data analysis. Even our choice of fluorophores has changed
dramatically inthe last decade, the traditional fluorophore finding continued competition
byfluorescentproteins andsemi-conductorquantumdots, tonamebutjustafew.
This volume "Advanced Concepts in Fluorescence Sensing: Macromolecular
Sensing" aims to summarize the current state ofthe art in fluorescence sensing. For this
reason we have invited chapters, encompassing a board range of macromolecular
fluorescence sensing techniques. Chapters in this volume deal with macromolecular
sensing, suchas using GFP, Aptamers and fluorescent pebble nano-sensors. This volume
directlycompliments volume 9ofthe Topics inFluorescence Spectroscopyseries, which
deals withadvancedconcepts insmallmolecule fluorescence sensing.
While many ofthe changes in recent fluorescence have been well received, its
continued growth inthe world has created achallenge intrying to archive and document
its use. Subsequently Chris D. Geddes has now become co-series editor ofthe Topics in
Fluorescence Spectroscopy series. We have also recently launched the Reviews in
Fluorescence series, which co-edited also by Dr's Geddes and Lakowicz and published
annually, is meant to directly compliment the Topics in Fluorescence Spectroscopy
series, withsmallchapters summarizingthe yearlyprogress influorescence.
Finally we would like to thank all the authors for their excellent contributions,
Mary Rosenfeld for administrative support and Kadir AsIan for help in typesettingboth
volumes 9and 10.
ChrisD. Geddes
JosephR. Lakowicz
Baltimore,Maryland, US.
August2004
vii
CONTENTS
1.PROTEIN-BASEDBIOSENSORS WITHPOLARIZATION
TRANSDUCTION .
RichardB. Thompson
1.1. INTRODUCTION 1
1.2.PRINCIPLES OF OPERATION 1
1.3.ADVANTAGES OFANISOTROPY-BASED SENSING 3
1.4.FLUORESCENCEPOLARIZATIONIMMUNOASSAy............................ 4
1.5.ANISOTROPY-BASEDMETALIONBIOSENSING 9
1.6. ANISOTROPY-BASED SENSINGOF OTHERANALYTES USING
PROTEINSAS TRANSDUCERS 15
1.7. CONCLUSIONS 16
1.8.ACKNOWLEDGMENTS 17
1.9.REFERENCES 17
2. GFP SENSORS 21
PeterM. Haggie andA.S. Verkman
2.1. INTRODUCTION 21
2.2. GENERALPRINCIPLES OFENGINEERINGFLUORESCENTPROTEIN
SENSORS 21
2.3. NEWGREENANDRELATEDFLUORESCENTPROTEINS 23
2.3.1. GFP Mutants 23
2.3.2.NovelFluorescentProteins 24
2.4. GFP-BASED SENSORS 24
2.4.1. pH Sensors ~............................................................ 24
2.4.2. Chloride/HalideSensors 26
2.4.3. SensorsofProteaseActivity 28
2.4.4. CalciumSensors 28
2.4.5. SensorsofCalcium-Calmodulin 30
2.4.6. SensorsofOtherSecondMessengers 30
ix
x CONTENTS
2.4.7. SensorsofProteinKinaseActivity....................................................... 31
2.4.8. SensorsofGproteins 32
2.4.9. MetaboliteSensors 33
2.4.10. SensorsofReduction-Oxidation(Redox)Potentia1............................. 33
2.4.11. NitrationSensors 34
2.4.12. Voltage Sensors 34
2.5. PERSPECTIVEANDFUTUREDIRECTIONS 35
2.6. ACKNOWLEDGMENTS 35
2.7. REFERENCES 36
3.FLUORESCENTSACCHARIDESENSORS 41
TonyDJamesandSeijiShinkai
3.1. INTRODUCTIONREFERENCES 41
3.2. READ-OUT 43
3.2.1. InternalChargeTransfer(ICT) 43
3.2.2. Photoinducedelectrontransfer(PET) 45
3.2.3. Others 46
3.3. INTERFACE 47
3.3.1. InternalChargeTransfer(ICT) 47
3.3.2.Photoinducedelectrontransfer(PET) 48
3.3.3. Others 54·
3.4. FLUORESCENTASSAY 56
3.5.POLYMERSUPPORTEDSENSORS 60
3.6. CONCLUSIONS 62
3.7.REFERENCES 62
4. FLUORESCENTPEBBLENANO-SENSORS AND NANOEXPLORERS
FORREAL-TIMEINTRACELLULARANDBIOMEDICAL
APPLICATIONS 69
HaoXu ,SarahM. Buck, RaoulKopelman, MartinA. Philbert,MurphyBrasuel,Eric
Monson, CalebBehrend,BrianRoss,AlnawazRehemtulla, andYong-EunLeeKoo
4.1. INTRODUCTION 69
4.1.1.BackgroundandHistory 69
4.2. PEBBLEMATRICES: DESIGN,PRODUCTION,AND QUALITY
CONTROL 72
4.2.1. PolyacrylamidePEBBLEs 73
4.2.2.Poly(decylmethacrylate)PEBBLEs 74
4.2.3. PolyethyleneGlycol-CoatedSol-GelSilicaPEBBLEs 76
4.3. CLASSIFICATIONAND CHARACTERIZATIONOFPEBBLE
SENSORS 79
4.3.1. IonSensors 79
