Applied Biotechnology Innovation Centre Electrochemical Nanopatterning and Microsystems Ioanis Katakis Department of Chemical Engineering, ATIC Technology Innovation Centre, Universitat Rovira i Virgili, Tarragona, Spain November 29, 2010 NANOJASP 2010, Barcelona
Electrochemical Nanopatterning and Microsystems. Ioanis Katakis Department of Chemical Engineering, ATIC Technology Innovation Centre, Universitat Rovira i Virgili, Tarragona, Spain November 29, 2010 NANOJASP 2010, Barcelona. CONTROLLING RATES OF REACTION AND MASS TRANSPORT. - PowerPoint PPT Presentation
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Applied Biotechnology Innovation Centre
Electrochemical Nanopatterning and
Microsystems
Ioanis Katakis
Department of Chemical Engineering, ATIC Technology Innovation Centre, Universitat Rovira i Virgili, Tarragona, Spain
November 29, 2010
NANOJASP 2010, Barcelona
Applied Biotechnology Innovation Centre
DEFINING TERMS
HOLDING EVERYTHING TOGETHER
THE TRANSDUCER
THE BIOMOLECULE(S)THE TRANSDUCTION
CHEMISTRY
MODULATING ACTIVITIES
CONTROLLING RATES OF REACTION AND MASS TRANSPORT
• The generic bioelectronic element
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MODULE 1
--
MODULE 2
++ + MODULE 3
-- -
MODULE 4
Os
Os Os+ +
MODULE 5
MODULE 6...
-
-+
+
+
Os
Os
Os
+
+-
-
-
• Each module can be any of the components. You aim at flexibility and control of outcome.
MODULAR IMMOBILISATION...
• Narváez et al. Biosens Bioelectr 15:43-52 (2000)
• Narváez et al. J Electroanal Chem 430:227-33 (1997)
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• Rational manipulation of bioregeneration kinetics
Au / MPS / RP / PSS / FDH
+
+Os
Os
Os
S SO3-
S SO3-
SO3-
SO3-
SO3-
ORDERED IMMOBILISATION: Multilayer self-assembled redox polyelectrolyte-FDH architecture in gold electrodes
average catalytic current = 1.04 µA RSD = 16 % n = 6
0 0.1 0.2 0.3 0.4 0.5
E / V vs Ag/AgCl
0
0.4
0.8
1.2
I / µ
AAu / MPS / RP / PSS / (B / PSS)n / FDH
S SO3-
S SO3-
+
+Os
Os
Os
SO3-
SO3-
SO3-
SO3-
SO3-
SO3-+
+
+
+
Fructose modular electrodes
…TO CONTROL PROPERTIES
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SUPRAMOLECULAR ARCHITECTURES FOR COMPLEX BIOSENSING, AMPLIFICATION, AND
CATALYSIS TASKS
• Popescu et al. J Electroanal Chem 464:208-14 (1999)
Lipid
Os-Phendion-surfactant
GLDH
NAD+
Cholesterol
Substrate
Product
Dehydrogenase
NAD(P)+
NAD(P)H
Mediatorred
Mediatorox
Electrode e-
APPLY JUDICIOUSLY MOLECULAR ENGINEERING
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• CONTACT AND CONTACT-LESS SPOTTING WELL DEVELOPED FOR 100+ mm RESOLUTION
• BIOMOLECULE PHOTOLITHOGRAPHY FOR HIGH DENSITY APPLICATIONS
BUT WHAT ABOUT PATTERNING?
WHAT IF WE COULD USE ELECTROCHEMICALLY-DRIVEN METHODS FOR BOTH PATTERNING AND DETECTION?
IDE: Higher signal was obtained from the electrode with wild probe (1.7A), 0.4A was obtained from the mutated probe and 0.3 A from the control without target.
Also in CE higher signal was obtained
from the electrode with wild probe
(50nA), while 0nA was obtained from
electrode where mutated probe was
immobilised
BIOPHOTOLITHGRAPHY: HYBRIDISATION
Applied Biotechnology Innovation Centre
• HCG amperometric detection through sandwich assay
1st electrode
(Sample) 2nd electrode
(Control)
AuAu
HRP
H2O2 e-
e1
deprotection
Incubation with redox
polymer and anti-HCG in
e1
BSA blocking in
e1
Incubation of HCG target
and biotinylated anti-HCG
in e1
e2 deprotectionIncubation with redox
polymer and anti-HCG in
e2
BSA blocking in
e2
Incubation of biotinylated
anti-HCG in e2 (control)
Incubation of both
electrodes with
streptavidine-HRP
Amperometric detection of HRP
IDE: lower signal was obtained comparing with DNA, however as in DNA wafers the control was lower (2nA) than the sample (40nA)
Also in CE there is a lower
response from the control,
nevertherless the signal is lower.
BIOPHOTOLITHGRAPHY: MOLECULAR RECOGNITION(1)
Applied Biotechnology Innovation Centre
• T4 amperometric detection through competition assay
1st electrode
(Control)
2nd electrode
(Sample)
AuAu
H2O2 e-
e1
deprotection
Incubation with redox
polymer and BSA-T4 in e1BSA blocking in
e1
Incubation with anti-T4 in e1
(Control)e2 deprotection
Incubation with redox
polymer and BSA-T4 in e2BSA blocking in
e2
Incubation with T4 and anti-T4
in e2
Incubation of both
electrodes with anti-
Rabbit Igg-HRP
Amperometric detection of HRP
HRP
A competition assay was carried
out to detect T4. 94.2nA was
obtained from the control and
47.3nA from the sample
BIOPHOTOLITHGRAPHY: MOLECULAR RECOGNITION(2)
Applied Biotechnology Innovation Centre
SHSH SH
SH
HRP
e-
H2O2
SH
SHSH
SH
A difference of 1,5µA between sampleand blank and a limit of detection of 6.31fmoles was obtained
MODULATING ACTIVITY: RECOGNITION TO SENSING
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H2O2
e-
HRP
H2O2
e-
- 57% and 23% of signal displaced in colourimetric and electrochemical displacement- Fast response: 2 minutes in electrochemical displacement
AND FROM SENSING TO FACILE SENSING
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Os+Os+
Os+Os+
Os+Os+
Os+Os+
Os+Os+
Os+Os+
- - --- -
+ + +++ +- - --- - }N1
+ + +++ +
+ + +++ + }N2
- - --- -
Glucose oxidase produces H2O2
Peroxidase uses H2O2 and consumes electrons at E2
Glucose oxidase produces electrons at E1
e-
e-
TOWARDS THE ULTIMATE NANOMACHINE(?): SELF POWERED, SELF PROPELLED, SELF PROPAGATING
INTEGRATING TECHNOLOGIES FOR MORE FUNCTIONS
• Pescador et al Langmuir (2008)
Applied Biotechnology Innovation Centre
Using same principles for versatile microsystem operation