Presentation on theme: "Applied Biotechnology Innovation Centre Electrochemical Nanopatterning and Microsystems Ioanis Katakis Department of Chemical Engineering, ATIC Technology."— Presentation transcript:
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
Applied Biotechnology Innovation Centre 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)
Applied Biotechnology Innovation Centre Rational manipulation of bioregeneration kinetics Fructose modular electrodes …TO CONTROL PROPERTIES
Applied Biotechnology Innovation Centre SUPRAMOLECULAR ARCHITECTURES FOR COMPLEX BIOSENSING, AMPLIFICATION, AND CATALYSIS TASKS Popescu et al. J Electroanal Chem 464:208-14 (1999) Substrate Product Dehydrogenase NAD(P) + NAD(P)H Mediator red Mediator ox Electrode e - APPLY JUDICIOUSLY MOLECULAR ENGINEERING
Applied Biotechnology Innovation Centre 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?
Applied Biotechnology Innovation Centre Campàs & Katakis Int J Env Anal (2004), PCT/EP2003/000262 (2002) Campàs & Katakis Sens. & Actuators B (2006) gold colloid oligonucleotide colloidal gold- oligonucleotide selective deposition deposition conjugation NANOCOLLOID SYNTHESIS AND MODIFICATION FOR NANOPATTERNING AND ARTIFICIAL INTELLIGENCE PATTERNING BIOLOGICAL PROPERTIES
Applied Biotechnology Innovation Centre Electrochemistry on SPE e-e- ARE THEY FUNCTIONAL? Concept works but high exists non-specific adsorption
Applied Biotechnology Innovation Centre GOx Adding properties (transduction) to nanopatterns MOLECULAR ENGINEERING OF “NANO”...
Applied Biotechnology Innovation Centre Au PEG SH PEG SH PEG SH PEG SH Thioctic acid SAM e1 (-1.2V) desorption of Thioctic acid e1 (+0.8V) selective adsorption of HRP-Os-Au e2 -1.2V) selective desorption The deprotection of the second electrode avoids the non-specific adsorption detected. e2 (+1.2V) deposition of Gox-Os-Au There is only a 3,4% of non-specific response from the second electrode H2O2H2O2 e- Amperometric detection of HRP (e1) Non-specific adsorption was detected on the second electrode Glucosee- Amperometric detection of GOx (e2) 5% of non-specific response was detected from the first electrode Amperometric detection of electrodeposited biomolecules …AND IMPROVING SELECTIVITY
Applied Biotechnology Innovation Centre S S H O O O O O S S H O O O O O S S H O O O O O S S H O O O O O S S O O O O O O Au O O O O O H S S S S H O O O O O S S H O O O O O S S H O O O O O S S H O O O O O +700mV ONE STEP FURTHER: PATTERNS AT MOLECULAR LEVEL
Applied Biotechnology Innovation Centre Peak around +0.6V Electrochemical deprotection was nearly complete within one scan DOES IT WORK?
Applied Biotechnology Innovation Centre EQCM data shows hope (but still 30% non specificity) IS IT SELECTIVE?
Applied Biotechnology Innovation Centre Resist coating First laser exposure First biomolecule coating Second laser exposure Second biomolecule coating Polyelectrolyte blocking Laser ablation or lithography work equally well AND YET ANOTHER METHOD OF PATTERNING
Applied Biotechnology Innovation Centre Layer optimisation: GOX(first layer) SAOX (second layer) GOX 2h. POS+GOX SOX 2h. POS+SOX After third layer of polyelectrolyte the response for enzyme decrease 30%. BIOPHOTOLITHGRAPHY: CATALYSIS(1)
Applied Biotechnology Innovation Centre 1st electrode (Sample) 2nd electrode (Control) Au Deprotection of e1 with UV Incubation of e1 with redox polymer and streptavidine Immobilisaion of biotinylated mutated capture probe in e2 Deprotection of e2 with UV Incubation of e2 with redox polymer and streptavidine Immobilisation of biotinylated wild capture probe in e1 Incubation of both electrode with the biotinylated target Incubation of both electrode with streptavidine-HRP Amperometric detection of both electrodes HRP H2O2e- BSA blocking in e1 BSA blocking in e2 IDE: Higher signal was obtained from the electrode with wild probe (1.7 A), 0.4 A 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) Au HRP H2O2e- 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 deprotection Incubation 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) Au H2O2e- e1 deprotection Incubation with redox polymer and BSA-T4 in e1 BSA blocking in e1 Incubation with anti-T4 in e1 (Control) e2 deprotection Incubation with redox polymer and BSA-T4 in e2 BSA 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 SH HRPHRP e-e- H2O2H2O2 A difference of 1,5µA between sample and blank and a limit of detection of 6.31fmoles was obtained MODULATING ACTIVITY: RECOGNITION TO SENSING
Applied Biotechnology Innovation Centre H 2 O 2 e- HRPHRP H 2 O 2 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
Applied Biotechnology Innovation Centre Os+ - - - - - - + + + + + + - - - - - - }N1}N1 + + + + + + + + + + + + }N2}N2 - - - - - - Glucose oxidase produces H 2 O 2 Peroxidase uses H 2 O 2 and consumes electrons at E 2 Glucose oxidase produces electrons at E 1 e-e- 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 ELECTRODEPOSITION AS PART OF OPERATION Mata at al Electroch. Acta (2009)
Applied Biotechnology Innovation Centre THE TEAM Panagiotis Argitis NCSR DEMOKRITOS Mònica Campàs Mònica Mir Srujan Dondapati Pablo Lozano Universitat Rovira i Vrigili
Applied Biotechnology Innovation Centre Our work is financed by: MICROPROTEIN (Patterning and arraying) HEALTHY AIMS (Fuel cells) CELSITIVE (Pathogen Detection) CIDEM and our Clients URV THE MONEY
Applied Biotechnology Innovation Centre THANK YOU FOR YOUR ATTENTION