Arben Merkoi ICREA & Institut Catal de Nanotecnologia - - PowerPoint PPT Presentation

www.nanobiosensors.org

Arben Merkoçi ICREA & Institut Català de Nanotecnologia Bellaterra, Catalonia, Spain arben.merkoci@icn.cat

2nd JAPANESE- SPANISH BILATERAL SYMPOSIUM “NANOTECHNOLOGIES AND NEW MATERIALS FOR ENVIRONMENTAL CHALLENGES” (SJ-NANO 2013) TSUKUBA (JAPAN), 2013, MARCH 5th

INDEX

• Introduction – Our motivation and detection systems
• Lab-on-a-chip systems: Detection of pesticides and phenols
• Lateral flow / nanomotors based biosensing platforms
• Future perspectives & conclusions

INDEX

• Introduction – Our motivation and detection systems
• Lab-on-a-chip systems: Detection of pesticides and phenols
• Lateral flow / nanomotors based biosensing platforms
• Future perspectives & conclusions

N A N O M A T E R I A L S D E V I C E S

Safety / security Diagnostics Food quality

Environmental monitoring

Other industrial applications

NANOBIOELECTRONICS & BIOSENSORS‘s research aims to integrate nanotechnology methods, tools and materials into low cost, user friendly and efficient (bio)sensors with interest for diagnostics , safety /security and other fields

www.nanobiosensors.org

Chemical Reviews, 111 (5), 3433-3458, 2011. ACS Nano, 2012, DOI: 10.1021/nn301368z E.Morales, A.Merkoçi, Graphene oxide as an optical biosensing platform”, Advanced Materials,. 2012, 24, 3298– 3308

State of the art nanobiosensing technologies

Chemical Reviews, 2012, 112, 5317–5338

• I. Detection after previous dissolving
• II. Direct /onto-electrode detection
• IV. Indirect detection through

nanochannels blocking

• III. Electrocatalytic detections

(silver enhancement, hydrogen evolution)

Au Au Ag 2H+ H2 Ag+ Ag +

Increase the sensitivity and simplicity

Nanomaterials based electrochemical detection tools

2003 2013

TRAC 24 341-349 (2005) ACA 482 149–155 (2003)

NANOPARTICLES & ELECTROCHEMICAL STRIPPING

JACS, 125 3214-3215 (2003) Chemical dissolving followed by stripping analysis Multicoding technology Breast cancer DNA related

Protein detection- direct detection of AuNP

Streptavidin-MB

Human IgG (g/ml)

1e-7 1e-6 1e-5 1e-4 1e-3 1e-2 1e-1 1e+0 1e+1

Abs (492 nm)

1 2 3 4 5 6 7

Peak current (A)

1 2 3 4 5 6 7 DPV of AuNPs Optical anti-Human-HRP-Au Optical anti-Human-HRP

α-human IgG-biotin Au-α-human-HRP Human IgG L.O.D: 52 and 260 pg of human IgG/mL for HRP and electrochemical AuNP-based detections

Analytical Chemistry, 2007, 79, 5232-5240 Analytical Chemistry, 2010, 82, 1151–1156

• Biosens. Bioelectron., 2010, 26, 1710(4pp)
• Electrochem. Commun. , 2010, 12, 1501(3pp)

Catalytic effect of AuNPs towards Ag deposition

Au Au Au Au Ag+ Ag+ Ag+ Au Au

Detection of a-HepB in human serum

L.O.D. : 3 mUI mL -1

Detection of human IgG

L.O.D. : 23 fg mL -1

Indirect gold nanoparticles detection

• Biosens. Bioelectron, 2009, 24, 2475 (8pp)

Catalytic effect of AuNPs towards H2 evolution

Salmonella detection based on diferential voltammetry of AuNP

Biosensors and Bioelectronics 40 (2013) 121–126.

A B

1m 1m 0.0 0.2 0.4 0.6 0.8 1.0

0.0

• 0.6
• 1.2
• 1.8

6 7 5 4 3 2

I/A E/V vs AgCl

1

A B

10 10

2

10 10

3

10 10

4

10 10

5

10 10

6

10 10

7

0.0

• 0.6
• 1.2
• 1.8

IA Log Log [Salmonella] / CFU mL-1

• 1

Schematic (not in scale)

• f Salmonella detection

DPV using AuNPs electrochemical detection

Protein detection - AuNP & nanochannels

Nanochannels immunoblocking using nanoparticles

200 nm pores RSD of 8% ([CA15-3]: 120 U/mL; n=3) LOD: 52 IU/mL of CA15-3

20nm AuNP Responses to human IgG using 20-nm AuNPs; 80-nm AuNPs; 80-nm AuNP tags anmd Ag enhancement Responses to blood samples spiked with CA15-3

Small, 2011 Effect of AuNP size and Ag enhancement on blocking

Protein detection - AuNP & nanochannels

200 nm nanochannel casted with 50 µL blood Spiked with CA15-3

Ag enhancement

• f 80 nm AuNPs

Small, 2011 RSD of 8% ([CA15-3]: 120 U/mL; n=3) LOD: 52 IU/mL of CA15-3

Cell studies based on CdS QDs

Supernatant Cells Lisis cells Supernatant Cells Lisis cells

A B

1 μA 1 μA a b c a b c

• 1.00
• 0.75
• 0.50

E/V

1 

• 1.00
• 0.75
• 0.50

E/V

1 

CdS QD-SAP interaction with HeLa cells: SWV and CLSM images of cells incubated with QDs incluiding blanks Electrochemical interrogation of cellular uptake of quantum dots decorated with peptide Collaboration with E.Giralt. (UB)

Cells detection based on AuNP & H2 catalysis

Collaboration with Dr. A. González (UV)

• 150
• 100
• 50
• 1.5
• 1.0
• 0.5

E / V i / A

a b c d e f g

• 55
• 45
• 35
• 25
• 15
• 5

t / s i / A

a’ b’ c’ d’ e’ f’ g’

• 150
• 100
• 50
• 1.5
• 1.0
• 0.5

E / V i / A

• 40
• 30
• 20
• 10

t / s i / A

a b a’ b’

A

2H+ H2

B

2H+ H2

• 150
• 100
• 50
• 1.5
• 1.0
• 0.5

E / V i / A

a b c d e f g

• 55
• 45
• 35
• 25
• 15
• 5

t / s i / A

a’ b’ c’ d’ e’ f’ g’

• 150
• 100
• 50
• 1.5
• 1.0
• 0.5

E / V i / A

• 40
• 30
• 20
• 10

t / s i / A

a b a’ b’

A

2H+ H2

B

2H+ H2

A B A B

H+ 2H+ H2 H+ 2H+ H2

B A

a a’ b c b’ c’ d d’

H+ 2H+ H2 H+ 2H+ H2

B A

a a’ b c b’ c’ d d’

AuNP & AuNP-Ab detection

Immunofluorescence analysis by flow cytometry and electrochemical analysis

• f both HMy2 and PC-3 cell lines

agreed.

4000 cells per 700 µL suspension

HMY: Tumoral human B cell line with expressed HLA-DR molecules PC3: Tumoral human prostate cell line

Cancer cell detection (ICN&UV patent) Analytical Chemistry, 2009, 81, 10268–10274

Immunofluorescence Immunoelectrochemistry

Catalytic Nanoparticles for Detection of circulating Cancer Cells (CTC) (Collaboration with Prof. C.Nogues, UAB)

HER based biosensing device developed by Nanobioelectronics & Biosensors Group LEITAT

Simple nanoparticle based technology Small 2012, 8, No. 23, 3605–3612 Nano Lett., 2012, 12 (8), pp 4164–4171

Tailoring graphene production toward biosensors applications

Merkoçi et al. Carbon 2012, 50:2987 Graphene Oxide as an Optical Biosensing Platform Merkoçi et al. Adv Mater 2012 DOI: 10.1002/adma.201200373

• roll to roll
• ink-jet printing
• screen-printing
• graphene

composites / inks

WATER POLLUTION

Pesticides Phenols Heavy metals Bacteria Toxins etc. Water, air and soil pollution causes 40 % of deaths worldwide

http://www.news.cornell.edu

Water-related diseases are one of the leading causes of death

• worldwide. Over 3 million people

die each year.

http://worldsavvy.org/monitor

Monitoring water quality should be done periodically to check for aquatic problems in-field sensing systems are necessary

Micro-Klean™ Industrial Wastewater Treatment Systems QuickShip Water Treatment Equipment www.ge-energy.com

Solutions are needed for smart systems that can detect pollutants and evaluate the efficiency of their removal

In-situ smart sensors and evaluators of the pollutants removal efficiency

• High sensitivity for various potential pollutants
• Versatility in evaluating pollutants removal

efficiency

• Easy to be integrated
• Cost / efficiency

Pesticides Phenols Heavy metals Bacteria Others Detect pollutants and gives qualitative & quantitative information for their destruction/removing strategies.

Nanomaterials based biosensing devices Nanomaterials with high and selective adsorbing / photacatalytic properties

Sensitivity Stability Versatility Cost / efficiency

Pesticides Phenols Heavy metals Bacteria Others

INDEX

• Introduction – Our motivation and detection systems
• Lab-on-a-chip systems: Detection of pesticides and phenols
• Lateral flow / nanomotors based biosensing platforms
• Future perspectives & conclusions

Lab-on-a-chip for ultrasensitive detection of carbofuran by enzymatic inhibition with replacement of enzyme using magnetic beads.

a b c I III f e d II g

A B

a b c I III f e d II g

A B Lab Chip, 9, 213–218, 2009

Pesticides detection

6 5 4 3 2 1

Magnet Enzyme-labelled magnetic beads Pesticide ATCh TCh

III II I II II II II II I I I I I III III III III III d d d d d d

6 5 4 3 2 1

Magnet Enzyme-labelled magnetic beads Pesticide ATCh TCh

III II I II II II II II I I I I I III III III III III d d d d d d

6 5 4 3 2 1

Magnet Magnet Enzyme-labelled magnetic beads Enzyme-labelled magnetic beads Pesticide Pesticide ATCh ATCh TCh TCh

III II I II II II II II I I I I I III III III III III d d d d d d

60 s injection 90 s injection % Inhibition [Carbofuran] (ppb)

L.O.D: 0,34 ppb (90” inhibit. time; 5% inhibit)

Lab Chip, 9, 213–218, 2009.

100 %   

blank inhibition blank

S S S I

Pesticides detection

Phenolic compounds detection

REACTION OF CATECHOL &TYROSINASE USING A BIO-CONJUGATE ON SCREEN PRINTING ELECTRODE (SPE) Catechol O-Quinone Tyrosinase MWCNT Magnetic nanoparticle (size: 100 nm)

+2H+ +2e-

O2 SPE Magnet Advanced Functional Materials , 2010

Tyrosinase MWCNT Magnetic nanoparticle (size: 100 nm)

SPE Magnet

y = 0.0019x + 0.0078 R2 = 0.9987

0.0000 0.0200 0.0400 0.0600 0.0800 0.1000 0.1200 0.1400 10 20 30 40 50 60 70 80

[Catechol], M Current, A

Test with dummy cell (WE(a)) 250.000 500.000 750.000 1000.000 1250.000

• 6
• 0.150x10
• 6
• 0.125x10
• 6
• 0.100x10
• 6
• 0.075x10
• 6
• 0.050x10
• 6
• 0.025x10

t / s i / A

[catechol]=10 M LOD= 5.4 nM LOQ= 17.9 nM

Advanced Functional Materials , 2010

REACTION OF CATECHOL &TYROSINASE USING A BIO-CONJUGATE ON SCREEN PRINTING ELECTRODE (SPE)

Phenolic compounds detection

Tyrosinase-magnetic nanoparticles & CNTs based biosensor

ON-OFF EFFECT INDUCED BY THE MAGNET ON OFF ON OFF

Tyrosinase MWCNT Magnetic nanoparticle (size: 100 nm)

SPE Magnet Advanced Functional Materials , 2010

REACTION OF CATECHOL &TYROSINASE USING A BIO-CONJUGATE ON SCREEN PRINTING ELECTRODE (SPE)

Phenolic compounds detection

Pump 1 Pump 2

PDMS SPE

Fluidic System for Phenol Detection

Optical and SEM Characterization of CaCO3 microparticles

PDMS SPE

50µM Phenol

Electrochemical enzyme- based biosensors constitute promising technology for the in situ monitoring of phenolic compounds

CaCO3/Tyr Biosensor for phenol detection

Electrophoresis 2013

INDEX

• Introduction – Our motivation and detection systems
• Lab-on-a-chip systems: Detection of pesticides and phenols
• Lateral flow / nanomotors based biosensing platforms
• Future perspectives & conclusions

Lateral Flow µPAD microfluidic paper-based analytical device

http://www.chimicabioanalitica.unito.it/immu noassay.htm

• Anal. Chem. 2010, 82, 3(8pp)

PAPER BASED NANOBIOSENSORS

Lateral flow / nanoparticles biosensing platforms

What can nanoparticles bring?

• More stability
• Multidetection capability
• Higher sensitivity
• Novel / versatile detection platforms

A 3-Cent HIV Test by Harvard

Dipstick

Sample without Cd-EDTA sample pad conjugate pad test line control line absorption pad AuNPs BSA EDTA Cd Cd-EDTA-BSA-AuNPS A281G5 mAb Anti-BSA mAb

A B C

samplewith Cd-EDTA

LFIA (Cadmium determination in drinking water)

Cd-EDTA conventional biosensor device Typical image of the cadmium detection by the lateral flow device

a b c

50 100 150 200 250 300 350 400 2000 10 20 30 40 50 60 70

Colour intensity difference Cd concentration (ppb)

a b c

Collaboration with Prof. D.Blake, Univ. Of Tulane, USA

50 10 20 30 40 50 60 70

Colour intensity difference

a b c

Blake D A et al. J. Biol. Chem. 1996;271:27677

Enhancing of biosensing

Template-based catalytic microengines (no need for clean room)

Nano/micromotors (Collaboration with J.Wang, USA)

ACS Nano, 2012, 6, 4445-4451

Superhydrophobic Alkanethiol-Coated Microsubmarines for Effective Removal of Oil

2H2O2(l)  2H2O(l) + O2 Nano Letters, 12, 396−401. 2012

Bacterial Isolation by Lectin-Modified Microengines

Nanoscale, 2013, In Press

Coupling nanomotors effect with biosensing

INDEX

• Introduction – Our motivation and detection systems
• Lab-on-a-chip systems: Detection of pesticides and phenols
• Lateral flow / nanomotors based biosensing platforms
• Future perspectives & conclusions

CONCLUSION / Research outcomes

• Nanotechnology allows us to develop smaller , easy to use and cost-effective

devices such as biosensors or lab-on-a-chip

• Nanoparticle based biosensing systems are shown to be high sensitive and

cost effective devices with interest for environmental applications between

• ther industries
• Simple cost/efficient paper based devices as well as nano/micromotors as

novel material for enhancing of biosensing technology are promising alternatives in environmental monitoring.

• Further improvement and more efficient designs of pollutants detection

systems using nanostructurated detectors including integration of pollutants sensing and removal/destruction at the same platform are still necessary.

Thank you! Any question?

Nanobioelectronics and Biosensors Group Catalan Institute of Nanotechnology (ICN) Read more at: www.nanobiosensors.org

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