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. BIO – ELEMENT
2. SENSOR - ELEMENT
Components of biosensor
Fig. 1 Configuration of a biosensor showing biorecognition,
interface, and transduction elements.
History of biosensor development

• Prof Leyland C Clark in 1956 - published his definitive paper on
the oxygen electrode.

• In 1962 Clark and Lyons - enzyme electrode.

• In 1969 –development 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 .
Principle of detection
• A specific “bio” element recognizes a specific
analyte.
• The “sensor” element transduces the change in the
biomolecule into an electrical signal that can be
amplified, 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.
Kinds of biosensor
• El ectrochemi cal bi osensor

• Opti cal bi osensor

• Pi ezoel ectri c bi osensor

• Cal ori metri c bi osensor

APPLICATION OF BIOSENSOR

• Poi nt of car e di agnost i c s .

• Bact er i ol ogi cal det ect i on

• I n Medi cal Car e

• For det er mi nat i on of f ood qual i t y

• Envi r onment al moni t or i ng.

• For I ndust r i al Pr oces s Cont r ol .


BIOSENSOR AND NANOTECHNOLOGY

• Nanotechnology will enable us to design
sensors that are :
much smaller
less power hungry
more sensitive







What Is Nanotechnology?
Richard Feynman’s
(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 atoms
placed 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.

Tools In Nanotechnology

– The main tools used in nanotechnology are four
main microscopes
1– Transmission Electron Microscope (TEM)
2– Atomic Force Microscope (AFM)
3– Scanning Tunneling Microscope (STM)
3– Scanning Electron Microscope (SEM)
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
Nanostructures Map
Graphene oxide
• Graphite when treated with strong oxidizers gives rise to the Graphite oxide,
which is a compound of carbon, oxygen, and hydrogen in variable ratios.

Manufacture of Graphene Oxide
• Graphene Oxide is formed by oxidizing crystalline graphite with a mixture of
sodium nitrate (NaNO
3 ,
sulfuric acid (H
2
SO
4
),and potassium permanganate
(KMnO
4
).The oxidation method is also known as the Hummers method.
• Structurally, the Graphene Oxide is similar to a graphene sheet with its base
having oxygen-containing groups. Since these groups have an high affinity to
water molecules, Graphene Oxide is hydrophilic and can be easily dissolved in
water.



Cont……

• Graphene Oxide is 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 Oxide films 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.





cont…..

• Applications of Graphene Oxide
• Graphene oxide finds application in the
following fields:
• Transparent conductive films
• Paper-like and composite materials
• Energy-related materials
• Biological and medical applications.


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 honeycomb
crystal lattice.

• Graphite itself consists of many graphene sheets stacked together.

• The carbon-carbon bond length in graphene is approximately 0.142 nm.


Graphene production
• Researchers obtained relatively large graphene sheets by
mechanical exfoliation (repeated peeling) of 3D graphite
crystals.

• Another method is to heat silicon carbide to high temperatures
(1100°C) to reduce it to graphene.

• Graphene has excellent properties like:
 Its entire volume is exposed to its surrounding.
 High electrical conductivity.

The functionalized graphene biosystems
Nucleic
acids
Avidin- biotin
Peptide
Cells
Aptamers
Bacteria
Proteins
Nanofabrication methods

BIOSENSOR AND CANCER

• Cancer is an abnormal and an uncontrolled cell growth
due to an accumulation of specific genetic and
epigenetic 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.


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 and
MRI for various cancer detection.
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.


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).
Fig. Application of the biosensor. The use of biosensors to
detect tumor markers in serum has spread widely (Jin, 2011).
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



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.

CHARACTERIZATION OF GRAPHENE COATED
MEMBRANE 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.

Functionalization of graphene
modified membrane
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 4
0
C, followed by rinsing with DI water and
phosphate buffered saline solution (PBS).
• Raman spectroscopy in particular has been found to be a
valuable tool to elucidate the structural properties of
graphene.

Cont…
Fig. 1 (A) Raman map and spectrum of graphene film. The map is
constructed by plotting the peak width at half height of the 2D-band as
the pixel intensity. Scale bar ¼ 0.8 mm. (B) AFM image of the graphene
film. Scale bar ¼ 500 nm.
This journal is ª The Royal Society of Chemistry 2011 J. Mater.
CHARACTERIZATION OF IMMOILIZED GRAPHENE
Electron microscopy (TEM and SEM) and
atomic force microscopy have been used to
characterize the graphene.


FABRICATION OF
NANOIMMUNOSENSOR
(A) Fitting of immobilized membrane into glass cells.
(B) Development of appropriate electronic circuit.
(C)Recording of amperometric signals with proper Ab-
Ag actions.




STANDARDIZATION OF PROTOCOL OF
NANOIMMUNOSENSOR
(A)Recording of amperometric data.



(B)Digitalization of signals and recording.

CONCLUSION

conclusion
• In present work a graphene modified PCTE (polycarbonate Track
Etch)membrane is used in nanoimmunosensors which can be employed to
detect 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.