Biosensor

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 Sensor
 A sensor is an instrument that responds to a physical stimulus
(such as heat, light, sound, pressure, magnetism, or motion)

 It collects and measures data regarding some property of a

phenomenon, object, or material

 Sensors are an important part to any measurement and automation

application

 The sensor is responsible for converting some type of physical

phenomenon into a quantity measurable by a data acquisition (DAQ)
system

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“Biosensor” once refered to any device which responds to chemical
species in biological samples or using biological components.

A sensor which incorporates a biological

element

Analyte SIGNAL
Biomolecule Transducer

3
 A sensor that integrates a biological element with a physiochemical
transducer to produce an electronic signal proportional to a single analyte
which is then conveyed to a detector.
 Biosensors Timeline

 Concept -- Leland Clark (1956)

 Urea Sensor -- Guibault & Montalvo (1969)

 Glucose Analyser -- Yellow Springs Instr. Co. (1973)

 Enzyme Thermistor – Mosbach (1974)

 Microbial Electrodes – Divis (1975)  Fibre-Optic Oxygen Sensor – Lubbers & Opitz (1975)

 Glucose-Controlled Insulin Infusion System Biostator

– Clemens et al. (1976)

 Immnosensor – Liedberg et al. (1982)

 Enzyme Electrode – MediSense Inc. (1987)

 BIAcore – Pharmacia, Sweden (1990)

 Glucose Biosensor

Glucose
Glucose +O2 Gluconic acid + H2O2
oxidase
Conventional Transducers
OPTICAL METHOD

ELECTROCHEMICAL METHODS

Typical pH Glass Electrode Radiometer Clark-Type Electrode

 Detector

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 The component used to bind the target molecule.

Must be highly specific, stable under storage conditions, and immobilized.

Microorganism
Tissue
Cell
Organelle
Nucleic Acid
Enzyme
Enzyme Component
Receptor
Antibody

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Acts as an interface, measuring the physical change that occurs
with the reaction at the bioreceptor then transforming that
energy into measurable electrical output.
 Signals from the transducer are
passed to a microprocessor where
they are amplified and analyzed.

The data is then converted to

concentration units and transferred to
a display or/and data storage device.

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measures change in mass

measures change in electric distribution

measures change in light intensity

measures change in heat

 Piezo-Electric Biosensors

Some piezo-electric devices utilize crystals, such

as quartz, which vibrate under the influence of an
electric field. The frequency of this oscillation
depends on their thickness and cut.

Others use gold to detect the specific angle

at which electron waves (surface plasmons) electronics.howstuffworks.com

are emitted when the substance is exposed

to laser light.

The change in frequency is proportional to the mass of

absorbed material.
 Electrochemical Biosensors

•Amperometric for applied current: Movement of e- in redox reactions

detected when a potential is applied between two electrodes.

•Potentiometric for voltage: Change in distribution of charge is detected

using ion-selective electrodes, such as pH-meters.

•Conductimetric for impedance

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 Optical Biosensors

•Colorimetric for color: Measure change in light

adsorption as reactants are converted to
products.

•Photometric for light intensity: Photon output

for a luminescent or fluorescent process can be
detected with photomultiplier tubes or
photodiode systems.

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 Calorimetric Biosensors

If the enzyme catalyzed reaction is exothermic, two thermistors may be used to

measure the difference in resistance between reactant and product and, hence,
the analyte concentration.

www4.tsl.uu.se/~Atlas/DCS/DCSIL/therm.html
• Nanotechnology opened doors to new ways of identifying and
quantifying biomolecules through use of nanosensors and nanoprobes.

• Tools are capable of monitoring biomolecular processes within single

cells.

• Use in biological and medicinal research.

• Two major categories, (i) biosensors (consists of biological

recognition element called bioreceptor (Ab, enzyme, cell) and a signal
transducer (ii) chemical sensors (chemical recognition element eg pH
sensitive) and a signal transducer).

T Vo-Dinh et al; Sensors and Actuators; B 74 (2001) 2-11

 Why Nanosensors ???
 Particles that are smaller than the characteristic lengths associated with the
specific phenomena often display new chemistry and new physics that lead to
new properties that depend on size

 When the size of the structure is decreased, surface to volume ratio increases
considerably and the surface phenomena predominate over the chemistry and
physics in the bulk

 The reduction in the size of the sensing part and/or the transducer in a sensor is
important in order to better miniaturise the devices

 Science of nano materials deals with new phenomena, and new sensor devices
are being built that take advantage of these phenomena

 Sensitivity can increase due to better conduction properties, the limits of

detection can be lower, very small quantities of samples can be analysed, direct
detection is possible without using labels, and some reagents can be eliminated.
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 Size and Compatibility

Nano sensors deliver real-time information about the antibodies to antigens, cell receptors to their
glands, and DNA and RNA to nucleic acid with a complimentary sequence

Sensitivity of the conventional biosensors is in the range between 10 3 and 104 colony forming units
(CFU)/ml. The dimensional compatibility of nanostructured materials renders nanotechnology as an
obvious choice derived from its ability to detect ~ 1 CFU/ml sensitivity

Reduced detection time than conventional methods

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 Nanosensors
Recognition principle Transducing mechanism
Luminescence
DNA Absorption
Acoustic wave Optical Polarization
Antibody/antigen Microbalance
Mass Fluorescence
Resonant
Molecular recognition Electrochemical
Enzymatic Piezoelectric
Biomimetic Thermal
Applications Conductometric
Antibodies
Volatile gases & Amperometric
Enzyme
Environmental vapors ions Potentiometric
Organelle
Vitamins Food Ion-sensitive
pH Medical diagnosis
Immunological
analytes
Cell or tissue

Scheme 1. Representation of recognition process and application of Nanosensor

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 Functional similarities and technical differences of
natural and artificial (Manmade) Noses
Odour Analysis with components of human nose

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 Odorant molecules produce ionic or electronic

signals through the action of receptors in a
membrane (or of sensor/transducer units)
 The signals determine the components of a
“feature vector” and are subsequently evaluated by
means of pattern recognition in the brain

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Dr. Michael Strano at the University of] Illinois, "We have developed molecular sheaths
around the nanotube that respond to a particular chemical and modulate the
nanotube's optical properties."

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SPOT-NOSED Project: A layer of olfactory proteins on a nanoelectrode could react
with low-concentration odorants. This technology could be used by doctors to
diagnose diseases at earlier stages.

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 Current Research
Nanosphere lithography (NSL) derived triangular Ag nanoparticles were used to
detect streptavidin down to one picomolar concentrations.

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Nanowerk

Nanosensors in environmental analysis

Nanosensors in environmental analysis

 Current Research

The School of Biomedical

Engineering has developed an
anti-body based piezoelectric
nanobiosensor to be used for
anthrax,HIV hepatitis detection.

http://www.biomed.drexel.edu/TechPort/Contents/Lec/Overview/Lec_BioSensors.pdf
 Electronic Nose
General structure of a chemical sensor

Three figures of merit related to gas sensing technology:

• Reversibility
• Sensitivity
• Selectivity

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 H2S nanosensor
• In each sensing cell, two electrodes, source and
drain, are made to have a nanoscale gap between
them

• A typical gap-width of ca. 40–60 nm has been

achieved

• Au nanoparticles are placed randomly over the

gap area

• Adsorption of H2S molecules onto the

nanoparticles may significantly change the
hopping behaviour of electrons through the
particles

Measurement of the I–V curves before and after the H2S exposure (3
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 Optical Nanosensors
 Luminescence (specifically, fluorescence) is commonly used in optical chemical-sensing
techniques

 The inherent sensitivity of fluorescence analysis is well known and, because of this, it is
often used for trace analysis

 Owing to the small size of the sensors, the sampling volume was reduced by more than six
orders of magnitude over conventional sensors, making it ideal for subcellular measurements

Example
Nanobiosensors for the detection of nitric oxide via the fluorescence detection of cytochrome c’, or
fluorescently labeled cytochrome c’ (a variant of cytochrome c in which the heme group is bound by
two cysteines)

Salient features:
 Fast response time (<1 s), reversible, and linear up to 1 mM nitric oxide

 Detection limit is 20 µM, making the sensor useful for some biological samples

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 Probes Encapsulated By Biologically Localized Embedding (PEBBLEs)
Optical sensors that contain dyes whose fluorescence is quenched in the presence of the
analyte to be determined

 Senses ions (H+, Ca2+, K+, Na+, Mg2+, Zn2+, Cu2+, Cl−) in cellular environments due to
their small size (20 to 600 nm in diameter) and protect the sensing elements (i.e. fluorescent
dyes) by encapsulating them within an inert matrix

 The selectivity and sensitivity of these nanosensors are comparable to those of

macroscopic ion selective optodes, and electrodes, while the response time and absolute
detection limit are significantly better

X In most practical applications, these PEBBLE sensors have been problematic because of
signal fluctuations that were not directly caused by the concentration of the analyte. These
fluctuations can be due to light scattering or to fluctuations in the excitation source (i.e. the
higher the excitation power, the greater the intensity of the fluorescence)

 Ratiometric PEBBLE sensors overcome this problem. In this kind of sensor, a fluorescent
indicator dye and a fluorescent reference dye are encapsulated inside the inert matrix. The
sensor response is based on intensity ratios between the indicator and reference dyes

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 Exceptional properties of
carbon nanotubes
 CNT have a high length-to-radius ratio, which allows for greater control
over the unidirectional properties of the materials produced

 they can behave as metallic, semiconducting or insulating material

depending on their diameter, their chirality, and any functionalization or
doping

 they have a high degree of mechanical strength. In fact they have a

greater mechanical strength and flexibility than carbon fibres

 their properties can be altered by encapsulating metals inside them to

make electrical or magnetic nanocables or even gases, thus making them
suitable for storing hydrogen or separating gases

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 Schematic structure of a Carbon Nanotube-Field Effect Transistor

 Electrical properties of CNT are sensitive to the effects of charge transfer and chemical
doping by various molecules

 The electronic structures of target molecules near the semiconducting nanotubes cause
measurable changes to the nanotubes’ electrical conductivity

 Nanosensors based on changes in electrical conductance are highly sensitive but they
are also limited by factors such as their inability to identify analytes with low adsorption
energies, poor diffusion kinetics and poor charge transfer with CNTs

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 CNT based Nano Sensors
System Target Salient feature Reference
species
Single wall nanotubes (SWNT) NH3 and Sensitive to 200 ppm of NO2, and 1% of NH3. Science 287 (2000) 1801.
NO2
Single wall nanotubes (SWNT) N2, He, Gas concentrations as low as 100 ppm can be detected Appl. Phys. Lett. 83
O2, and Ar (2003) 2280.

Multi-wall carbon nanotubes NH3 Gas concentrations as low as 10 ppm can be detected. J. Phys. D: Appl. Phys. 36
(MWCNTs) The sensor showed a reversible response of few (2003) L109.
minutes.

Poly(o-anisidine) (POAS) HCl Nine times increase in sensitivity compared to uncoated Chemical Physics Letters,
coated CNT CNT Volume 383( 5-6) ( 2004)
617.

MWNT-SiO2 CO2, O2 Sensor response time is approximately 45 s, 4 min, and IEEE Sens. J. 2 (2002) 82.
and NH3 2 min for CO2,O2, and NH3, respectively.
The sensor response is reversible for O2 and CO2 , but
irreversible for NH3

Single-walled carbon nanotube β-D- Senses β-D-glucose in solution phase by two distinct Nat. Mater. 4 (2005)
(SWNT) glucose mechanisms of signal transduction: fluorescence and 86.
charge transfer.

Poly(vinylferrocene) (PVF) Glucose Glucose concentration in real blood sample can be The Analyst 131 (2006)
derivatised MWCNTs determined. 670.

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 Real-time detection of proteins and DNA
Detection of Proteins

(A) Schematic of a biotin-modified Si nanowire and subsequent binding of streptavidin to the modified
surface. (B) Plot of conductance versus time for a biotin-modified Si nanowire, where region 1 corresponds
to the buffer solution, region 2 corresponds to the addition of 250 nM streptavidin, and region 3
corresponds to pure buffer solution. (C) Conductance versus time for an unmodified Si nanowire, where
regions 1 and 2 are the same as in (B).

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 Detection of DNA

(D) Schematic of a Si nanowire sensor surface modified with PNA receptor before and after duplex
formation with target DNA. (E) Si nanowire DNA sensing where the arrow corresponds to the addition
of a 60 fM complementary DNA sample and the inset shows the device conductance following
addition of 100 fM mutant DNA. (F) Conductance versus DNA concentration, where data points
shown in red and blue are obtained from two independent devices.

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 Nanosensors for drug discovery
(A)Illustration of tyrosine kinase function,
where ATP binds to the kinase active site
and then phosphate is transferred to a
tyrosine (Tyr) residue of the substrate
protein.
(B)Detection of ATP binding and small-
molecule inhibition using a Si nanowire
sensor device functionalized with the
Abelson tyrosine kinase Abl.
(C)Structures of small molecules investigated
for the inhibition of ATP binding to Abl.
(D)Normalized conductance versus time data
recorded from Abl modified Si nanowire
devices using solutions containing 100 nM
ATP and 50 nM small molecule Gleevec
(red), A1 (blue), A2 (green), A3 (pink), and
biotin (black).

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 Magnetic Nanoparticles as Nanosensors
 Upon target binding, these nanosensors cause changes in the spin - spin relaxation
times of neighboring water molecules, which can be detected by magnetic resonance
techniques

 These magnetic nanosensors have been designed to detect specific mRNA,

proteins, enzymatic activity, and pathogens (e.g., virus) with sensitivity in the low
femtomolar range (0.5 ± 30 fmol)

3 - 5 nm monocrystalline iron oxide

nanoparticles (MION)

Diagram of the magnetic nanosensors acting as magnetic relaxation switches.

Superparamagnetic nanoparticles self-assemble in the presence of a target with a corresponding
decrease in the solution T2 relaxation time. Self-assembled nanoparticles can be dispersed by the
action of an enzyme, temperature or pH change depending on the nature of the bond holding the
nanoassembly together.
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 Magnetic nanosensors for detecting DNA
 Cluster formation of magnetic nanoparticles, upon addition of a complementary
oligonucleotide, resulted in a quick and significant decrease in the spin- spin relaxation times (T2)
of neighboring water molecules, at 40 ºC (Figure A)
 When a non complementary oligonucleotide was used, no change in T2 was observed
 At 1.5 T, significant differences were readily apparent by MRI between the samples in the low
femtomole range (0.5 - 2.7 fmol)

A. Temporal change of water T2 relaxation times with B. T2 changes (T2) of an aqueous solution
(□) and without (♦) complementary oligonucleotide. of nanosensors as a function of temperature
Insert shows the effect of increasing concentrations of cycling
oligonucleotides on water T2

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Magnetic nanosensors for measuring various enzymatic activities
including restriction endonucleases, methylases, and proteases

NMR imaging of enzymatic activity

 BamHI (from Bacillus amyloli) is a type II restriction endonuclease, having the capacity for
recognizing short sequences (6 b.p.) of DNA and specifically cleaving them at a target site.

 Restriction endonuclease detection using a pair of nanosensors with complementary

oligonucleotides (1 and 2)

 When mixed in solution the oligonuleotides hybridize causing the nanosensors to cluster (3).

 BamHI sensitive nanoassembly, with a corresponding decrease in T2, is formed upon treatment
with BamHI, the nanoassemblies disassemble, and an increase in T2 is observed (4)

 Treatment with other restriction endonucleases (EcoRI, HindIII, DpnI) do not cause a change in
T2 of the solution (5, 6, 7)

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 Metal oxide nano-crystals for sensing
 Metal oxides possess a broad range of electronic,
chemical, and physical properties that are often highly
sensitive to changes in their chemical environment.

 The sensing properties of semiconductor metal oxide

(nano-belts, nano-wires or nano-ribbons) assures improved
selectivity and stability due to there crystallinity

 Their peculiar characteristics and size effects make

them interesting both for fundamental studies and for
potential nano-device applications, leading to a third
generation of metal oxide gas sensors

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 A schematic summary of the kinds of quasi-one-
dimensional metaloxide nanostructures

(A) nanowires and nanorods; (B) core-shell structures with metallic inner
core, semiconductor, or metal-oxide; (C) nanotubules/nanopipes and
hollow nanorods; (D) heterostructures; (E) nanobelts/nanoribbons; (F)
nanotapes, (G) dendrites, (H) hierarchical nanostructures; (I) nanosphere
assembly; (J) nanosprings.

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 Oxide based Nanosensors
Metal oxide Target species Salient features Reference

V2O5 nanofibres 1-Butylamine, toluene, Extremely high sensitivity was measured for 1- Sens. Actuators B 106
propanol butylamine (below 30 ppb) and moderate sensitivity (2005) 730.
for ammonia. In contrast, only very little sensitivity
was observed for toluene and 1-propanol vapours.

SnO2 nanobelts CO, NO2, ethanol Sensitivity at the level of a few ppb Appl. Phys. Lett. 81
(2002) 1869.
In2O3 nanowires NH3, NO2 The response times have been determined to be 5 s for Appl. Phys. Lett. 82
100-ppm NO2 and 10 s for 1% NH3, and the lowest (2003) 1613.
detectable concentrations are 0.5 ppm for NO2 and
0.02% for NH3

ZnO nanowires Ethanol Sensitive to ethanol concentration is in the range of 1– App. Phys. Lett. 84
100 ppm. Sensitivity increases sharply as the (2004) 3654.
temperature is raised from 200 to 300 °C

MoO3 nanorods Ethanol and CO The detection limit for ethanol and CO is lower than Chem. Phys. Lett. 407
30 ppm (2005) 368
Cd-doped ZnO Relative humidity Cd-doped ZnO nanowires show a clear positive App. Phys. Lett. 84
nanowire temperature coefficient of resistance effect, which is (2004) 3085.
quite abnormal as compared to pure ZnO nanowires

SnO2 Dimethyl Sensitive to 53 ppb DMMP and can be improved via App. Phys. Lett. 86
methylphosphonate doping nanobelts with catalytic additives. (2005) 063101.
(DMMP)

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 Nano-sensors can warn of tsunamis

New technologies like nano-sensors with the ability to detect even a

one-centimetre rise in waves could forewarn coastal areas of tsunamis,
said M. Palaniswami, professor of Department of Electrical and Electronic
Engineering, University of Melbourne.

Nanotechnological, inexpensive sensors that can detect invisible,

odorless hydrogen leaks and sound the alarm wirelessly could help
safeguard future vehicles and refueling stations based on the gas,
experts told UPI's Nano World.

The Silicon AFM sensor with a tip radius of less than 10 nm contributed
to many scientific breakthroughs

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Nanosensor Probes Single Living Cells

The nano-needle of 50-nm-diameter silver-coated optical fiber that

carries a helium-cadmium laser beam.

Attached to the optical fiber tip are monoclonal antibodies that

recognize and bind to benzo(a)pyrene tetrol (BPT).

The laser light, which has a wavelength of 325 nm, excites the
antibody-BPT complex at the fiber tip, causing the complex to
fluoresce.

The newly generated light travels up the fiber into an optical detector.

The layer of silver is deposited on the fiber wall to prevent the laser
A nanosensor probe
excitation light and the fluorescence emitted by the antibody-BPT carrying a laser beam (blue)
complex from escaping through the fiber. penetrates a living cell to
detect the presence of a
The ben-zo[a]pyrene (carcinogen) metabolite reacts with the cell's product indicating that the
DNA, forming a DNA adduct, which can be hydrolyzed into a product cell has been exposed to a
called benzo(a)pyrene tetrol (BPT). cancer-causing substance

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 CNTs as Nanosensors!

Sensors based on carbon nano-tubes can improve the

detection of vapors from explosives.

Sensors arranged in arrays will be tuned to respond to

the presence of specific explosives and biological and
chemical agents.

Each nano-tube will be anchored to a metalized silicon

substrate at one end of the tube and chemically
functionalized to bind only to a specific molecule at the
other end.

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 The tube experiences a lowering of
the frequency when an extra mass is
attached to the functionalized end of
the tube.
 The presence of a mass of an
adsorbed agent, such as Anthrax, on
the free end of the nano-tubes will
produce a measurable frequency
shift.
 The selective binding of agents to
the chemically functionalized nano-
tubes will allow the array to sense the
presence of different BCX agents.

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The possibilities are endless!

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