TOWARDS SUSTAINABLE, WIRELESS, AUTONOMOUSNANOCELLULOSE-BASED QUANTITATIVE DRUG OFABUSE BIOSENSING PLATFORM

Inkjet printing of GO-MB composite

The inkjet parameters of drop spacing, temperature, waveform and number of layers

have been tailored, and the final settings are shown in the table.

Graphene Oxide / Methylene Blue Ink (GO-MB)

For the ink formulation, we explored in detail the GO-MB interaction

mechanism based on π-π-stacking and ionic interactions as well as the

specific antibody attachment.

For preparation of antibody (AB) GO-MB inks, the native MB is partly

exchanged with antibodies conjugated with MB.

GO-MB ink formulation and characterization (no antibody)

Eva Melnik1// Eva.melnik@ait.ac.at // Christoph Steininger1, Steffen Kurzhals1, Giorgio Mutinati1

1 AIT Austrian Institute of Technology GmbH, Center for Health and Bioresources, Molecular Diagnostics, Giefinggasse 4, 1210 Vienna, Austria.

Parameters selected

Drop spacing 5

Plate temperature

40°C

Number of layers

2

Drug of Abuse Electrochemical Sensor

In the project GREENSENSE we merge molecular

detection and printed electronics in the form of a fully-

integrated biosensing platform using nanocellulose (NC).

The biosensing platform with the newly developed printed

(DoA) biosensors will integrate different NC-based printed

electronic components (supercapacitor and/or a primary

battery, display and NFC antenna) and a single microchip

to enable wireless communication and electrochemical

readout.

Assay: We target label-free electrochemical assays [1-3]

employing a redox-active layer system modified with

capture molecules. In this type of assay, specific binding

events reduce the electron transport through the layer

system to the electrode.

Our approach is the

development of an inkjet

printable composite

based on graphene oxide

(GO) modified with

antibody-methylene blue

conjugates (MB-AB).

[1] Li, Yan; et al. (2011) Anal. Sci. 27(7), 727.

[2] Du, Yan; et al. (2010) Biosens. & Bioelect. 25(8), 1902.

[3] Kong, Fen-Ying; et al. (2013) Chem. Comm. 49(11),1052.

Drug of Abuse Detection

Commercially available detection devices:

The drug of drug of abuse (DoA) is highly interesting for many fields of

applications e.g. employment setting (safety-sensitive positions in transport,

construction or sports) or the criminal justice system. The existing devices use

either urine or saliva as sample liquid and employ lateral flow based tests

systems. The appearance of a colored line indicates if a certain drug (e.g.

cocaine, morphine, tetrahydrocannabinol, amphetamine, secobarbital) was

consumed.

New challenges:

For the improvement of the existing systems, the project Greensense aims at:

• quantitative electrochemical detection of drugs of abuse in urine and saliva

• measurement time <10 min

• detection in the ng/mL range

This project has received funding from the European Unions Horizon 2020 research and innoviation

programme under grant agreement No 761000. This publication only reflects the authors views and the

European Union is not liable for any use that may be made of the information contained therein.

HTTP://GREENSENSE-PROJECT.EU/

0,0 0,5 1,0 1,5 2,0 2,5 3,0 3,5

0,0

0,2

0,4

0,6

0,8

1,0

1,2

Peak c

urr

ent

diffe

rence (D

I p) [µ

A/m

m2]

anti-IgG HRP (L) used for immobilization [µM]

ΔIP = ΔIP(𝑚𝑎𝑥)[𝐿]

𝐾𝑑+[𝐿]

2.) activity

and sensing

properties

1.) GO-MB

ink formulation and

characterization

Effect of NaCl concentration Effect of pH values (in 50mM

NaCL)

Effect of glucose (in 50mM

NaCL)

-0.5 -0.4 -0.3 -0.2 -0.1 0.0

0

200

400

600

800

1000

1200

I no

rma

lize

d [nA

/mm

2]

Potential [V]

0mM 1min

0mM 10min

5mM 1min

5mM 10min

9mM 1min

9mM 10min

13mM 1min

13mM 10min

17mM 1min

17mM 10min

20mM 1min

20mM 10min

Differential Pulse Voltammetry (DPV) of MB-GO ink (no antibody)

Inkjet printed MB-GO on

working electrode of screen

printed three-electrode sensor

Differential pulse voltammetry

(scan rate:0.5V/s, step: 0.005V,

Epuls: 0.05V, tpuls=0.005s)

AgCl reference electrode

Carbon paste counter electrode

Carbon paste working electrode

-0.5 -0.4 -0.3 -0.2 -0.1 0.0

0

200

400

600

800

1000

1200

1400

1600

1800

2000 pH5

pH6

pH7

pH8

pH9

I no

rma

lize

d [nA

/mm

2]

Potential [V]

0 2 4 6 8 10

800

850

900

950

1000

1050

1100

1150

1200

pH 5

pH 6

pH 7

pH 8

I norm

aliz

ed (

nA

/mm

2)

Time [min]

-0.5 -0.4 -0.3 -0.2 -0.1 0.0

0

200

400

600

800

1000

1200

1400

1600

I no

rmaliz

ed [

nA

/mm

2]

Potential [V]

10mM NaCl

31mM NaCl

51mM NaCl

69mM NaCl

85mM NaCl

100mM NaCl

114mM NaCl

127mM NaCl

0 20 40 60 80 100 120 140 160 180

-0.42

-0.40

-0.38

-0.36

-0.34

-0.32

-0.30

-0.28

measurement sensor 1

measurement sensor 2

measurement sensor 3

measurement sensor 4

Pote

ntial [V

]

Conc. NaCl [mM]

0 20 40 60 80 100 120 140 160 180

400

600

800

1000

1200

1400

measurement sensor 1

measurement sensor 2

measurement sensor 3

measurement sensor 4

I norm

aliz

ed [n

A/m

m2]

Conc. NaCl [mM]

5 6 7 8 9

1100

1200

1300

1400

1500

1600

1700

I norm

aliz

ed (

nA

/mm

2)

first measurement sensor 1

first measurement sensor 2

first measurement sensor 3

first measurement sensor 4

pH

0 10 20 30 40 50 60

650

700

750

800

850

900

950

1000

1050

I no

rma

lize

d [

nA

/mm

2]

Time [min]

Glucose addition

0

2

4

6

8

10

12

14

16

18

20

Glu

cose

con

c [m

M]