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GRAPHENE MODIFIED NANOPOROUS
MEMBRANE BASED
NANOIMMUNOSENSOR FOR THE
SELECTIVE DETECTION OF CANCER
BIOMARKERS
SPEAKER-
ID.NO. -
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CONTENTS
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CONTENTS
INTRODUCTION TO BIOSENSOR
BIOSENSOR AND NANOTECHNOLOGY
BIOSENSOR AND CANCER
OBJECTIVES
CONCLUSION
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WHAT IS BIOSENSOR?
BIOSENSOR IS A SENSING DEVICE WHICH CAN
CONVERT A BIOLOGICAL RESPONSE INTO AN
ELECTRICAL SIGNAL.
THE NAME BIOSENSOR SIGNIFIES THAT THE
DIVECE IS A COMBINATION OF TWO PARTS :
1. BIOELEMENT
2. SENSOR - ELEMENT
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Components of biosensor
Fig. 1 Configuration of a biosensor showing biorecognition,
interface, and transduction elements.
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Principle of detection
A specific bio element recognizes a specificanalyte.
The sensor element transduces the change in the
biomolecule into an electrical signal that can beamplified, displayed, and analyzed.
The bioelement may be an enzyme, antibody, living
cells, tissue, etc. The sensing element may be electric current,
electric potential, intensity, mass, conductance,
impedance, temperature and so on.
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Kinds of biosensor
Electrochemical biosensor
Optical biosensor
Piezoelectr ic biosensor
Calor imetr ic biosensor
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History of biosensor development
Prof Leyland C Clark in 1956 - published his definitive paper onthe oxygen electrode.
In 1962 Clark and Lyons - enzyme electrode.
In 1969development of first potentiometric biosensor.
In 1974 - the use of thermal transducers for biosensors.
In 1975 - Divis suggested that bacteria could be used as the
biological element in microbial electrodes .
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Cont..
Since the early 1970 - building of immunosensor.
Peterson in 1980 - First fibre optic pH sensor.
In 1982 - First fibre optic-based biosensor for glucose detection.
In 1983 - First surface plasmon resonance (SPR) immunosensor.
In 1984 - First mediated amperometric biosensor.
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APPLICATION OF BIOSENSOR
P o i n t o f c a r e d i a g n o s t i c s .
B a c t e r i o lo g i c a l d e t e c t i o n
I n M e d i c a l C a r e
F o r d e t e r m i n at i o n o f f o o d q u a l i t y
E n v i r o n m e n t a l m o n i t o r i n g .
F o r I n d u s t r i a l P r o c e s s C o n t r o l .
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If I want to measure
something small, I
need something
small
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BIOSENSOR AND NANOTECHNOLOGY
Nanotechnology will enable us to design
sensors that are :
much smaller
less power hungry
more sensitive
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What Is Nanotechnology?
Richard Feynmans
(1918-1988)
A nanometre is 1/1,000,000,000 (1 billionth) of a metre, which is around
1/50,000 of the diameter of a human hair or the space occupied by 3-4 atomsplaced end-to-end.
A few carbon atoms on the
surface of highly oriented
pyrolytic graphite (HOPG). Image
obtained by Scanning Tunneling
Microscope (STM).
nanotechnology is a field to understand, create, and
use structures, devices and systems that have
fundamentally new properties and functions because
of their nanoscale structure.
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Tools In Nanotechnology
The main tools used in nanotechnology are four
main microscopes
1Transmission Electron Microscope (TEM)
2Atomic Force Microscope (AFM)
3Scanning Tunneling Microscope (STM)
3Scanning Electron Microscope (SEM)
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NANOMATERIALS Nanostructured materials have been successfully used in the last
years for the construction of fast, accurate and sensitive sensors as
they have excellent properties.
Carbon nanotubes, nanowires, and nanochannels, Quantum dots,
nanoparticles are all examples of nanomaterials.
(The small size of allows for a greater surface to volume ratio)
Carbon
nanotubes
Fullerene
Dendrimers
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Nanostructures Map
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Graphene oxide
Graphite when treated with strong oxidizers gives rise to the Graphite oxide,
which is a compound of carbon, oxygen, and hydrogenin variable ratios.
Manufacture of Graphene Oxide
Graphene Oxideis formed by oxidizing crystalline graphite with a mixture of
sodium nitrate (NaNO3 , sulfuric acid (H2SO4),and potassium permanganate(KMnO4).The oxidation method is also known as the Hummers method.
Structurally, the Graphene Oxideis similar to a graphene sheet with its base
having oxygen-containing groups. Since these groups have an high affinity to
water molecules, Graphene Oxideis hydrophilic and can be easily dissolved in
water.
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Cont
Graphene Oxideis a poor conductor but when it undergoes treatment using
heat, light, or chemical reduction, most of graphene's properties are restored.
Chemical reduction is normally done using hydrazine.
It is possible to deposit Graphene Oxidefilms on any substrate, and then
convert it into a conductor. These coatings may be used in solar cells, flexible
electronics, chemical sensors, liquid crystal devices
Graphene, which is a conductor, graphene oxide is a semiconductors and can
replace silicon in electronics applications.
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cont..
Applications of Graphene Oxide
Graphene oxidefinds application in the
following fields:
Transparent conductive films
Paper-like and composite materials
Energy-related materials Biological and medical applications.
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Graphene
Graphene, is one of the allotropic forms of carbon.
It is a one-atom-thick planar sheet of carbon atoms that are densely packed in a honeycombcrystal lattice.
Graphite itself consists of many graphene sheets stacked together.
The carbon-carbon bond length in graphene is approximately 0.142 nm.
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Graphene production
Researchers obtained relatively large graphene sheets bymechanical exfoliation (repeated peeling) of 3D graphitecrystals.
Another method is to heat silicon carbide to high temperatures(1100C) to reduce it to graphene.
Graphene has excellent properties like:
Its entire volume is exposed to its surrounding.
High electrical conductivity.
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Nanofabrication methods
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BIOSENSOR AND CANCER
Cancer is an abnormal and an uncontrolled cell growth
due to an accumulation of specific genetic andepigenetic defects.
Biosensor technology has the potential to provide: fast and accurate detection.
reliable imaging of cancer cells.
monitoring of angiogenesis and cancer metastasis.
ability to determine the effectiveness of anticancer
chemotherapy agents.
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Preexisting technology
Existing cancer screening methods include:
(1) the CA 15.3 test and mammography to detect breast
cancer in women.
(2) prostate-specific antigen (PSA) level detection in blood
sample for men to detect prostate cancer.
(3) blood detection for colon cancer.
(4) endoscopy, CT scans, X-ray, ultrasound imaging andMRI for various cancer detection.
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Cont
These traditional diagnostic methods however
are not very powerful methods :-
as they can not detect cancer at very early
stages.
some of the screening methods are quite costly
and not available for many people.
so use of biosensors to detect cancer biomarkers
in serum has spread widely.
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Biomarkers
In terms of cancer, the analyte being detected by
the biosensor is a tumor biomarker.
A biomarker is an indicator of a biological state of
disease.
Biomarkers can be DNA, RNA, or protein (i.e.,
hormone, antibody, oncogene, or tumor
suppressor).
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OBJECTIVES
Surface modification of nanoporous membrane by
graphene oxide
Characterization of graphene coated nanoporous
membrane by SEM
Immobilization of antibody on nanoporous membrane
Characterization of immobilized nanoporous
membrane.
Fabrication of nanoimmunosensor.
Detection of sample.
Standardization of protocol of nanoimmunosensors
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Surface modification of nanoporous
membrane by graphene oxide
(1)Coating of the graphene oxide onto the
nanoporous membrane by drop coating method.
Drop wise graphene is poured onto the membrane.
A thin film is obtained onto the membrane surface.
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CH R CTERIZ TION OF GR PHENE CO TED
MEMBR NE BY SEM
The membranes categorized in two different classes-
(1) PCTE membrane was used as such
(2) PCTE membrane coated with graphene nanolayer over
the one surface of membrane with the help of drop coating
method.
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Functionalization of graphene
modified membrane
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cont...
Graphene is incubated with linker molecule in
dimethylformamide (DMF).
The linker-modified graphene then incubated with
antibody in Na2CO3-NaHCO3 buffer solution (pH 9.0)
overnight at 40
C, followed by rinsing with DI water andphosphate buffered saline solution (PBS).
Raman spectroscopy in particular has been found to be a
valuable tool to elucidate the structural properties of
graphene.
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Cont
Fig. 1 (A) Raman map and spectrum of graphene film. The map isconstructed by plotting the peak width at half height of the 2D-band as
the pixel intensity. Scale bar 0.8 mm. (B) AFM image of the graphene
film. Scale bar 500 nm.
This journal is The Royal Society of Chemistry 2011 J. Mater.
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CHARACTERIZATION OF IMMOILIZED GRAPHENE
Electron microscopy (TEM and SEM) and
atomic force microscopy have been used to
characterize the graphene.
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ST ND RDIZ TION OF PROTOCOL OF
N NOIMMUNOSENSOR
(A)Recording of amperometric data.
(B)Digitalization of signals and recording.
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CONCLUSION
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conclusion
In present work a graphene modified PCTE (polycarbonate Track
Etch)membrane is used in nanoimmunosensors which can be employed todetect specific cell membrane-associated target antigens.
The morphology of graphene modified nanoporous membrane was
characterized by Scanning Electron Microscopy and chemical analysis was
completed by FTIR (Fourier Transform Infrared Spectroscopy) prior to use in
nanobiosensor.
This antibody immobilized membrane structure was then tested with a no.
of few antigens and cross checked by structurally related antigens for
specificity.
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