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BiomaterialsLecture
17
Surfaces:InteractionsofProteins
withSurfaces
Unlessotherwisenoted,sourceInformationforthefollowingslides:
1) B.Ratner,A.Hoffman,F.Schoen,andJ.Lemons:BiomaterialsScience,2nd edition(SanDiego:ElsevierAcademicPress.2004).
2) Butt,HJ.;Graf,K.;Kappl PhysicsandChemistryofInterfaces,2nd Edition(WiletVCH:Weinheim 2006).
3) Modifiedfrom:Anne Mayes3.051J/20.340JMaterialsforBiomedical
Application,MIT,
http://ocw.mit.edu,
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ImportanceofProteinSurfaceInteractions
Modulatecelladhesion
Triggerthe
biological
cascade resulting
in
foreign
body
response
Centraltodiagnosticarray/sensordevicedesign&
performance
Initiateotherbioadhesion:e.g.,marinefouling,bacterial
adhesion
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FundamentalsonProteins
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ProteinFunctions
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ProteinFunctions(cont.)
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ProteinFunctions(cont.)
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ProteinStructure
Proteinshavemultiplestructurallevels.
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ProteinStructure
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ProteinStructure
AAsidegroups
havevariable
chemical
character
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ProteinStructure
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SyntheticPolymersvs.Proteins
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ProteinAdsorptiononBiomaterialSurfaces
In
as
short
a
time
as
can
be
measured
after
implantation
in
a
living
system
(
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Background
A) Proteinactivity
variesinadsorbed
vs.solvatedstate
Why???
ProteinAdsorptiononBiomaterialSurfaces
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Background
B. Entropicforces
ProteinAdsorptiononBiomaterialSurfaces
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Background
B. Entropicforces
ProteinAdsorptiononBiomaterialSurfaces
Gaininentropyprobably
isthemaindrivingforce
duringspontaneous
proteinadsorption!!
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Background
C. Adsorbedproteins
initiatephysiological
responsesto
biomaterials
ProteinAdsorptiononBiomaterialSurfaces
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ModelsforProteinAdsorption:LangmuirModel
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ModelsforProteinAdsorption:LangmuirModel
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ModelsforProteinAdsorption:LangmuirModel
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ModelsforProteinAdsorption:LangmuirModel
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ModelsforProteinAdsorption:Scatchard Plot
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ModelsforProteinAdsorption:Scatchard Plot
SurfaceSensitive
MeasurementsNecessary
1. Ellipsometry
2. SurfacePlasmon
Spectroscopy
3. QuartzCrystal
Microbalance
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Summary:ProteinAdsorption
Proadsorptionfactors
Dehydrationoftheinterface
Increaseoftheproteininteriormobility
Columbicinteractions
VanderWallinteractions
Antiadsorption
factors
Strongwaterbindingtointerface
Proteinrigidity
Surfacemobility
Lownetcharge
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BiomaterialsLecture
18
Surfaces:InteractionsofProteins
with
Surfaces:
Part
2
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ProteinSurfaceInteractions:Part2
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Vroman Effect
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Observations
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IrreversibleAdsorption
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Restructuring
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MultilayerFormation
M t f Ad b d P t i
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MeasurementofAdsorbedProtein1) Techniquesforquantifyingadsorbedamounts
M t f Ad b d P t i
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MeasurementofAdsorbedProtein
Measurement of Adsorbed Protein
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MeasurementofAdsorbedProtein
Measurement of Adsorbed Protein
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InSituEllipsometry
2) Techniquesforquantifyingadsorbedamounts
MeasurementofAdsorbedProtein
Ellipsometry
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Ellipsometry
Elli
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Ellipsometry
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SurfacePlasmonResonanceSpectroscopy(SPR)
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SurfacePlasmonResonanceSpectroscopy(SPR)
f l ( )
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SurfacePlasmonResonanceSpectroscopy(SPR)
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20 40
0.0
0.2
0.4
0.6
0.8
1.0
Reflectivity
IncidentAngle/Deg
Simulatedcurveswithincreasingthickness
Gold Film
Glass Slide
He-Ne Laser
632.8nm
0
Detector
Polarizer
Prism
Adsorbed Molecules
Evanescent wave optical technique
sensitive
to
changes
in
thickness
and
optical
propertiesofthinandultrathinfilmsAngstromsensitivity
Nondestructivetosamples
Attenuated totalreflection (ATR) setup ina Kretschmann configuration,opticsare
awayfrom
the
sample
and
subphase
Surfaceplasmon excitationobserved inreflectivityangularscan
Relativelyinsensitivetoenvironmentalchanges temperature,viscosity,etc.
Multiskop
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SurfacePlasmonResonanceSpectroscopy(SPR)
Surface Plasmon Resonance Spectroscopy (SPR)
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SurfacePlasmonResonanceSpectroscopy(SPR)
Quartz Crystal Microbalance (QCM)
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QuartzCrystalMicrobalance(QCM)
Time
Frequency
F
BeginningofAdsorption Equilibration
Injection
Sauerbrey Equation
PersonalComputer
FrequencyCounter
OscillatingCircuit
Piezoelectric
Quartzcrystal
Scatchard Equation
Howtomeasurebindingconstants?
qqq
AFmF 22
ooFFKNKcFF bb
Sauerbrey,G.,Z.Phys. 1959, 155,206.Sauerbrey,
G.,
Ann.N.Y.Acad.Sci. 1949, 51,660672.
QCM D (Dissipation)
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QCMD(Dissipation)
Simultaneousmonitoringoffrequency(F)and dissipation(D)atmultipleharmonicsofthequartzresonator
Dissipationrevealsqualitativeinformationabouttheviscoelasticpropertiesoftheadsorbedlayer
Dissipationofaviscoelasticpolymer
layeron
aquartz
resonator
is
heavily
influencebyitsstructure. RigidfilmsshowsmallDvalueswhilesoftorflexiblefilmsshowlargerDvalues
Modelingtheviscoelasticpropertiesusing
aVoight model
can
yield
quantitativeinformation
stored
dissipated
E
ED
2
LargerEdissipated yields
moreviscoelasticresponse
Dissipation
http://www.qsense.com/$2/qcmd.swf
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Mustuseatechniquesensitivetothestructureoftheprotein
CircularDichroism
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Circular Dichroism
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CircularDichroism
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