Innovation and Development of Study Field

Nanomaterials at the Technical University of Liberec

nano.tul.cz
These materials have been developed within the ESF
project: Innovation and development of study field
Nanomaterials at the Technical University of Liberec
 Technical University of Liberec

Nanomaterials for
biosensors

2
 Outlook

What is biosensor
Biosensor components
Nanomaterials in biosensors
Thin films
Nanoparticles
Carbon based biosensors

3
 What is biosensor

A biosensor is a self-contained integrated device, which is capable of

providing specific quantitative or semi-quantitative analytical information
using a biological recognition element (biochemical receptor) which is
retained in direct spatial contact with a transduction element.

Schematic of biosensor
4 Ibtisam E. Tothill, World Mycotoxin Journal, 2011, 4 (4) 361-374
http://www.rpi.edu/dept/chem-eng/Biotech-Environ/BIOSEN/biosen.htm
 What is biosensor

Biosensors usually yield a digital electronic signal which is

proportional to the concentration of a specific analyte or group of
analytes.

While the signal may in principle be continuous, devices can be

configured to yield single measurements to meet specific market
requirements.

5 Biosensors & Bioelectronics (2005)

 What is biosensor

Biosensors and affinity sensor devices have been shown to have the
ability to provide rapid, cost effective, specific and reliable quantitative
and qualitative analysis.

6 Ibtisam E. Tothill, World Mycotoxin Journal, 2011, 4 (4) 361-374

Biosensor construction
 Applications of biosensors
Glucose monitoring in
diabetes patients
Other medical health related
targets
Environmental applications
Remote sensing of airborne
bacteria
Detection of pathogens
Determining levels of toxic
substances
Detection of toxic metabolites
7
Detection and determining
of organophosphate
Routine analytical
measurements
Determination of drug
residues in food
Drug discovery and
evaluation of biological
activity of new compounds
Protein engineering in
biosensors
 Biosensor components

8 http://en.wikipedia.org/wiki/File:Biosensor_System.jpg
 Nanomaterials in biosensors

The use of nanomaterials and structures such as semiconductors and

conducting polymer nanowires, and nanoparticles (carbon nanotubes,
silica nanoparticles, dendrimers, noble metals nanoparticles, gold
nanoshells, superparamagnetic nanoparticles quantum dots, polymeric
nanoparticles) for biosensor applications is expanding rapidly.

9 Ibtisam E. Tothill, World Mycotoxin Journal, 2011, 4 (4) 361-374

 Nanomaterials in biosensors

Unique and novel physical and/or chemical characteristics

of nanomaterials can aid the design of bio-sensors with
improved analytical characteristics.

High surface / volume ratio

Novel elctro-optical properties
Increased catalytical activity
Enhanced electron transfer

Silicon nanowire sensors

10 M. Ming-Cheng Cheng, Current Opinion in Chemical Biology 2006, 10:1119
.
 Nanomaterials in biosensors
Organic based:
Fullerenes
Carbon nanotubes
Dendrimers
Liposomes
Inorganic:
Quantum dots
Metal nanorods
Metal nanoparticles
Examples of nanoparticles used in
sensors developments
11 Ibtisam E. Tothill, World Mycotoxin Journal, 2011, 4 (4) 361-374
.
 Nanomaterials in biosensors
- the controlling parameters

Cost Detection limit

Selectivity Sensitivity

Reproducibility Stability
Nikos. A. Chaniotakis, Nanomaterials in the design of chemical sensors and biosensors, A bottom up approach, E-MRS
12
Spring 2006
 Nanomaterials in biosensors
- disciplines involved in design
Chemistry
Inorganic
Physical
Inorganic
Materials Macromolecular Biology

Polymers DNA

DEVICES
Nanoparticles Enzymes
Semiconductors Cells

Biosensors
13 Nikos. A. Chaniotakis, Nanomaterials in the design of chemical sensors and biosensors, A bottom up approach, E-MRS
Spring 2006
 Thin films for biosensing

Nanostructured thin films have opened the possibility to fabricate

electrochemical sensors and biosensors with high power of detection
due to intrinsic properties associated with their dimensions at
nanoscale level.

The aim objective behind the utilization of these materials is to combine

the high power of detection with preservation of the structural integrity
of the biomolecules and, also, maintaining their biocatalytic activity.

Roberto A. S. Luz et al. Nanobioelectrochemistry From implantable biosensors to green power generation,
14 http://www.springer.com/978-3-642-29249-1
 Thin films for biosensing

The role of the control of depositing monolayers of organic films and

their final properties was first studied by Irving Langmuir and Katherine
Blodgett in the beginning of XX Century.

LangmuirBlodgett films has

been considered an important
path for biosensors fabrication
and many kinds of architecture
has been reported in the last
decades as very promising
approaches for biosensors
development. Schematic of LangmuirBlodgett monolayer film

Roberto A. S. Luz et al. Nanobioelectrochemistry From implantable biosensors to green power generation,
15 http://www.springer.com/978-3-642-29249-1
http://en.wikipedia.org/wiki/Langmuir%E2%80%93Blodgett_film
 Thin films for biosensing

Deposition of a LangmuirBlodgett Formation of an ordered monolayer

multilayer at the airwater interface

16 F. Davis, S.P.J. Higson / Biosensors and Bioelectronics 21 (2005) 120

 Thin films for biosensing
Polysilicon nanowire biosensors fabricated using a top-down process
may be used to determine the binding constant of two inflammatory
biomarkers. Nanofabrication process is based on simple and mature
photolithography, thin film technology, and plasma etching, enabling an
easy route to mass manufacture.

Antibody-functionalized nanowire
sensors were used to detect the
proteins interleukin-8 (IL-8) and
tumour necrosis factor-alpha (TNF-
) over a wide range of
concentrations, demonstrating
excellent sensitivity and selectivity.

17 Reprinted with permission from Mohammad M. A. Hakim et al. Nano Lett., 2012, 12 (4), pp 18681872
Copyright (2012) American Chemical Society
 Thin films for biosensing

DNA hybridization and receptorligand binding to microfabricated

cantilevers produce surface stress changes that may be measured
directly for detection of analytes.

A biosensor is made by
functionalizing one side of the
cantilevers with receptor
molecules and then detecting
the mechanical bending
induced by the binding of a
ligand.

A section of a microfabricated silicon

cantilever array
18 C. Jianrong et al. / Biotechnology Advances 22 (2004) 505518 .
 Thin films for biosensing

A force amplified biological sensor (FABS) capable of detecting biological

species such as cells, proteins, toxins, and DNA at low concentrations.

A cantilever-beam force
transducer senses the
presence of magnetic
beads, the number of
which is proportional to
the concentration of
analyte in the sample.

Concept of a force amplified biological sensor (FABS)

19 C. Jianrong et al. / Biotechnology Advances 22 (2004) 505518

 Nanoparticle based biosensors

Schematic of some of the optical and electrical detection alternatives that are being
used for DNA, proteins and cells analysis thanks to the use of nanoparticles
20 A. Merkoc / Biosensors and Bioelectronics 26 (2010) 11641177
 Nanoparticle based biosensors
Example of nanostructured electrocatalytic membrane
synthesized using a combination of three methods.
PAMAM dendrimers with cobalt hexacyanoferrates-modified gold
nanoparticles were alternated with poly(vinylsulfonic acid) layers on
ITO (indium tin oxide) electrodes.

This film was used as substrate for

immobilization of glucose oxidase in
the presence of bovine serum
albumin and glutaraldehyde as cross
- linker. The modified electrode was
successfully applied as a biosensor
for the amperometric measurement
Schematic representation of reaction
of glucose. of glucose at ITO(PVS/PAMAM
Au)3@CoHCFGOx electrode
21 F.N. Crespilho et al. Electrochemistry Communications 8 (2006) 16651670
.
 Nanoparticle based biosensors
A magnetic probe captures a
target using either monoclonal
antibody or complementary
oligonucleotide. Target-specific
gold nanoparticles sandwich
the target and account for
target identification and
amplification. The bar-code
oligonucleotides are released,
and detected using the
scanometric method. The
target can be DNA, RNA or
protein.

22 M. Ming-Cheng Cheng, Current Opinion in Chemical Biology 2006, 10:1119

 Carbon based biosensors

The utilization of carbon nanomaterials allows to increase the charge

transfer in bioelectrochemical devices. These includes the
modification of electrodes with several kinds of carbon at nanometre
range like carbon powders, carbon nanotubes, graphene sheets and
carbon capsules.

Roberto A. S. Luz et al. Nanobioelectrochemistry From implantable biosensors to green power generation,
23 http://www.springer.com/978-3-642-29249-1
 Carbon based biosensors
carbon nanotubes
Chemical sensor consisting of a single-walled carbon nanotube field
effect transistor (swCN-FET) with a nanoscale layer of single stranded
DNA (ssDNA) adsorbed to the tube's outerwall. These sensors have
been able to detect methanol, trimethylamine, propionic acid, dimethyl
methylphosphonate (a simulant of sarin), and dinitrotoluene (a
derivative of TNT) at the ppm level.

DNA-Functionalized Carbon Nanotube Chemical Sensor

24 http://www.lrsm.upenn.edu/~nanophys/biosensors.html
.
 Carbon based biosensors
carbon nanotubes
Penicillin biosensor based on a capacitive field-effect structure
functionalized with a dendrimer / carbon nanotube multilayer.

Nanoarchitecture based on capacitive field effect transistor-modified with LBL

of PAMAM and CNTs as sensing platforms for penicillin G detection

25 Siqueira Jr, J.R., et al. Biosens. Bioelectron. 25 (2009) 497501

 Carbon based biosensors
carbon nanotubes
Glucose biosensor using CNT-
nanoelectrode ensembles. The CNT
array was fabricated by growing CNTs
directly on the patterned catalysts. The
density of the CNT array can be
adjusted by changing the density of
the catalysts. The GOx molecules
were attached to the open-ended tips
of the CNTs by forming amide linkages
between their amine residues and
carboxylic-acid groups on the CNT Glucose sensor based on carbon-
tips via carbodiimide chemistry. nanotube (CNT) nanoelectrode ensembles

26 Jun Wang, Yuehe Lin, Trends in Analytical Chemistry, Vol. 27, No. 7, 2008
 Carbon based biosensors
buckypaper
Mediator-free electrochemical biosensor based on buckypaper with
enhanced stability and sensitivity for glucose detection.

Buckypaper is a thin
sheet made from an
aggregate of carbon
nanotubes.

(a) a titanium plate sputtered with a thin layer of gold; (b)

gold interaction with chitosan; (c) attachment of buckypaper to
the gold surface by chitosan; (d) activation of buckypaper; (e)
immobilization of enzymes; (f) the fabricated biosensor Buckypaper made
27 Ahmadalinezhad et al. / Biosensors and Bioelectronics 30 (2011) 287293 of carbon nanotubes
http://en.wikipedia.org/wiki/Buckypaper
 Carbon based biosensors
fullerenes
Fullerenes is a very promising family of electroactive compounds.
There are some unique characteristics that make this new type of
compound very promising as mediators in amperometric biosensors.
They have multiple redox states in a wide range of potentials, very
low solubility in aqueous solutions, and are stable in many redox
forms.

Fullerene C60
V.G. Gavalas, N.A. Chaniotakis / Analytica Chimica Acta 409 (2000) 131135
28 Nikos. A. Chaniotakis, Nanomaterials in the design of chemical sensors and biosensors, A bottom up approach, E-MRS
Spring 2006
 Carbon based biosensors
fullerenes

Operational principles of light induced fullerene mediated biosensor with

operational potential decreased to + 100 mV

29 Nikos. A. Chaniotakis, Nanomaterials in the design of chemical sensors and biosensors, A bottom up approach, E-MRS
Spring 2006
 Quantum dots based biosensors

In semiconducting nanomaterials such as CdS and CdSe, electrons in

the basal state can be promoted to an excited level by shinning light
with energy equal or higher than the bandgap. The electrons can then
return to their initial state by losing energy in the form of light. As a
consequence, nanocrystals of semiconducting materials are highly
fluorescent, a feature that is particularly interesting for sensing and in
vivo imaging.

Quantum Dots with emission maxima in a 10-nm step

http://en.wikipedia.org/wiki/Quantum_dot
30 R. de la Rica et al. / Advanced Drug Delivery Reviews 64 (2012) 967978
 Quantum dots based biosensors
(A) Schematic illustration of optical coding based on wavelength and intensity
multiplexing. Large spheres represent polymer microbeads, in which small
coloured spheres (multicolour quantum dots) are embedded according to
predetermined intensity ratios.

(B) Ten distinguishable

emission colours of ZnS-
capped CdSe QDs excited
with a near-UV lamp.

(C) Fluorescence of a
mixture of CdSe/ZnS QD-
tagged beads emitting
single-color signals.

31 A. Merkoc / Biosensors and Bioelectronics 26 (2010) 11641177