These slides use concepts from my (Jeff Funk) course entitled analyzing hi-tech opportunities to analyze the increasing economic feasibility of bio-sensors for measuring cholesterol in humans. Bio-sensors detect the level of cholesterol (and other biological materials) using enzymes, matrices, and transducers. The enzymes, which are held in a matrix, react with the cholesterol and an electric signal is produced from an amperometric transducer. Improvements in sensitivity, response time, shelf life, detection limit, and reusability have been achieved through creating more appropriate biological materials for the enzymes, matrices, and transducers.

1. CHOLESTEROL BIO-SENSOR
• NIHA Agarwalla
• NAUSHAD Rahman
• KARTHIKA Gogulakrishnan
• Sun Chenxi
MT-5009: ANALYZING HI-TECHNOLOGY OPPORTUNITIES
For information on other technologies, please see Jeff Funk’s slide share account (http://www.slideshare.net/Funk98/presentations) or his
book with Chris Magee: Exponential Change: What drives it? What does it tell us about the future?
http://www.amazon.com/Exponential-Change-drives-about-future-
ebook/dp/B00HPSAYEM/ref=sr_1_1?ie=UTF8&qid=1398325920&sr=8-1&keywords=exponential+change

2. CONTENT
• What is Biosensor?
• Why Cholesterol Biosensor?
• Conventional Techniques V/s Biosensor
• Current Trend of CB
• Basic Parameters of CB
• Different Materials of Biosensor
• Commercial Biosensor
• Future CB Biosensor
• Market Analysis

3. BIOSENSOR
• Biosensor is a compact analytical devices or 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.
• Biosensor = Analyte + biorecognation element + transducer .
• A biosensor is an analytical device that detects the level of
glucose, cholesterol, urea or any other chemicals in our body
by using the blood ,urine ,saliva or skin as a sample.
Niha

4. Niha

5. WORKFLOW OF BIOSENSOR
Transducer
Electrochemical Optical Piezoelectric pH Change
Matrix
Electrode Array chip Metal Surface
Immobilization of enzyme
Entrapment Covalent Crosslinking Adsorption
Bio receptor /Biomolecules
Enzymes Antibodies Reagent Other molecules
Analyte
Blood Urea Saliva Skin
Niha

6. CONTD..
Detector
Niha

7. WORKING PRINCIPLE OF BIOSENSOR
Niha

8. CONTD…
Niha

9. EXPLANATION
• Analyte:- It is a sample of Blood ,Urine, Saliva ,Skin
• Analyte is reacted with bio receptor.
• Bio receptors are enzymes, antigens.
• E.g.:- For Glucose detection in blood an enzyme called glucose
oxidase is used.
• For Cholesterol detection in blood an enzyme called
cholesterol oxidase or cholesterol esterase is used.
• Matrix is a solid support which is used to holds the bio
receptor.
• Examples of matrixes are electrodes, array chip or any metal
surface.
Niha

10. EXPLANATION CONTD…
• The only necessary property for matrix that it should not erode or
get affected by pH, temperature or outside environment.
• It is difficult to attach bio receptor to the matrix
• So different immobilization techniques are used for the attachment
• Immobilization techniques are entrapment, adsorption ,cross
linking, covalent bonding etc.
• The bio receptor comes in contact with the analyte and generates
different kinds of signal.
• Signal can be either movement of electron, change in color ,mass
change etc. which is detected by transducer and covert this bio
signals to electrical signals.
• The electrical signal are amplified and is displayed in a monitor.
Niha

11.  Food Analysis
 Study of biomolecules and their interaction
 Drug Development
 Crime detection
 Medical diagnosis (both clinical and laboratory use)
 Environmental field monitoring
 Quality control
 Industrial Process Control
 Detection systems for biological warfare agents
 Manufacturing of pharmaceuticals and replacement organs
APPLICATION OF BIOSENSOR
Niha

12. BIOSENSOR FOR MEDICAL
• Biosensor are used for applications ranging from screening for
disease to allow early intervention, through to the
management of chronic disease and the monitoring of health
and wellbeing.
• Biosensors are an essential tool in the detection and
monitoring of a wide range of medical conditions from cancer
to Parkinson’s disease.
Niha

13. CHOLESTEROL BIOSENSOR
• Cardiovascular diseases and cardiac arrest are number one cause of
death globally. One of the most important reasons is
hypercholesterolemia i.e. increased concentration of cholesterol in
blood.
• An estimated 17.3 million people died from CVDs in 2008,
representing 30% of all global deaths .
• The number of people who die from CVDs, mainly from heart
disease and stroke, will increase to reach 23.3. million by 2030
.CVDs are projected to remain the single leading cause of death .
• Estimation of cholesterol level in blood hence is the most important
and challenging task for medical industry.
• Development and Improvement of existing Cholesterol biosensor
has got a worldwide attention.
Niha

14. CONVENTIONAL TECHNIQUES TO
DETERMINE CHOLESTEROL
1.Lieberman-Burchard Test
Lieberman–Burchard is a reagent used in a colorimetric test to
detect cholesterol, which gives a deep green color.
This test uses acetic anhydride and sulfuric acid as reagents.
Niha

15. ADVANTAGE OF BIOSENSOR OVER L-B
TEST
L-B Test Biosensor
Requires lot of reagents Do not require any regent
Time for detection is more than 20 min. Its response time is as less as 5 sec.
It uses acetic acid and sulfuric acid as reagent
which can cause severe burns so require specific
care.
It do not use any such reagent so handling is easy.
For L-B test sample should be extracted from
plasma and this extraction step constitutes a
cumbersome extra step in the assay
Sample can be taken from any part of body. It
simple blood.
Requires pretreatment of sample to avoid optical
interference because of hemoglobin in blood
No pretreatment of sample is required.
Expensive and can only be used by trained people Less expensive and can be used by any people.
Niha

16. CONTD..
L-B Test device :Huge apparatus Bio-sensor : Small and portable .
Niha

17. NON BIOSENSOR TECHNIQUES TO
DETERMINE CHOLESTEROL
2 .High-Performance liquid Chromatography(HPLC)
HPLC used to separate the components in a mixture, to
identify each component, and to quantify each component
HPLC uses mass transfer process involving adsorption
technique to separate different components.
Niha

18. ADVANTAGE OF BIOSENSOR OVER L-B
TEST
HPLC Biosensor
Irreversibly adsorbed compounds not
detected
Do not require any regent
Requires pretreatment of sample to avoid optical
interference because of hemoglobin in blood.
No pretreatment of sample is required.
Complex setup and can only be used by trained
people
Easy to handler it
Bulky and Costly Smaller and Cheaper
NIha

19. CONTD….
HPLC : Bulk Setup Bio-sensor : Small setup
NIha

20. HISTORY OF CHOLESTEROL BIOSENSOR
• First cholesterol biosensor was developed in 1993.
Components Used
Matrix Ppy/Pt covered with
polycarbonate membrane
Bio receptor Cholesterol Oxidase
Immobilization technique Entrapment
Transducer Amperometric
Niha

21. PUBLICATIONS TREND IN CB
0
5
10
15
20
25
30
1993 1995 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013
No of Publication
No of Publication
Niha

22. VARIOUS TYPES OF TRANSDUCER USED IN
CHOLESTEROL BIOSENSOR
Electrochemical
Potentiometric
Amperometric
Cyclic Voltammetry
Optical
Fluorescence
Absorption
Reflection
Niha

23. ELECTROCHEMICAL BASED CB.
• Electrochemical biosensors are normally based on enzymatic
catalysis of a reaction that produces or consumes electrons
and generates signal, which are captured by transducer.
• The signal is proportional to concentration of the analysed
substance.
• Electrochemical are considered to be the most important
cholesterol biosensor.
• Electrochemical sensor may be divided into conductometric,
potentiometric, and amperometric biosensors depending
upon the electrochemical property to be measured by
detector system.
Naushad

24. OPTICAL BASED CB
• An optical fiber-based biosensor is a biosensor that employs
an optical fiber, as a platform for the biological recognition
element, and as a conduit for excitation light and/or the
resultant signal.
• The optical measurement method is critical to the sensitivity
and detection limit of the sensor. Optical transducers use
different types of measurement such as fluorescence,
absorbance ,and chemiluminescence.
Naushad

25. ADVANTAGES OF ELECTROCHEMICAL
OVER OPTICAL
Electrochemical Optical
The color of the sample will not interfere with the
redox reaction in electrochemical method and hence
do not cause any
change in the electrical signal.
Color of the sample interfere with the wavelengths
when using optical method which result in inaccurate
data.
The life time of the reagents used during reaction do
not decrease.
The life time of the reagents can be short under
incident light.
Its response time and sensitivity is very high. Because of the diffusion of analytes, it may cause slow
response time.
No specific reagent is required for electrochemical
biosensor
Fiber Optic Biosensor only works for specific reagent.
Low cost and low power requirement High cost and high power requirement
Naushad

26. ELECTROCHEMICAL V/S OPTICAL
0.00E+00
5.00E-01
1.00E+00
1.50E+00
2.00E+00
2.50E+00
3.00E+00
3.50E+00
4.00E+00
4.50E+00
5.00E+00
2001 2002 2003 2004 2005 2006 2007 2008
DetectionLevel
Year
Optical Biosensor
Electrochemical
Acrylamine glass
beads
+ ChOx, ChEt, HRP+
Alkylamine glass
beads+ Chox,
Chet, HRP+ Cross-
linking via
Glutaraldehyde
PANI/ITO+
ChOx+
Covalent via
EDC/NHS
TEOS (sol–gel)ITO+
ChOx, ChEt+
Entrapment
Ferrocene
monocarboxylica
cid-PPy/Pt/Pt+
ChOx+
PPy/Pt+ ChOx+
Entrapment FeMC
(physisorbed)-
P(NMPY)-PTS/ITO+
ChOx+ Physical
adso
Naushad
𝜇𝑔/𝑚𝑙

27. 1. Linearity
2. Sensitivity
3. Response time
4. Reusability
5. Shelf life
6. Detection Limit
Basic Parameters to Measure
Efficiency of CB
Naushad

28. SENSITIVITY
0
0.5
1
1.5
2
2.5
1994 1996 1998 2000 2002 2004 2006 2008
Sensitivity(nAM-1)
Year
Improvment In Sensitivity In CB
Sensitivity
1
2
3 4
5
6
7
8
9
Naushad

29. CONTD….
Material No
Tetramethoxy silane sol–gel/poly(1,2-diaminobenzene)/Pt + ChOx+ Entrapment followed by treatment
with Glutaraldehyde vapors+ Ampero. at 0.6 V vs. Ag/AgCl
1
PPy/Pt/Pt+ ChOx+ Entrapment+ Ampero. at 0.5 V vs. Ag/AgCl
2
PPy-p(HEMA)-TEGDA/Pt+ ChOx+ Entrapment+ Ampero. at 0.7 V vs. Ag/AgCl 3
Ferrocene monocarboxylicacid-PPy/Pt/Pt+ ChOx+ Entrapment+ Ampero. at 0.375 V vs. Ag/AgCl 4
PPy/ITO+ ChOx, ChEt+ Entrapment+ Ampero. at 0.5 V vs. Pt wire 5
(a) BSA/dilauroylphosphatidylcholine (DLPC)/rhodium–graphite; (b) Agarose gel-Riboflavin-
DLPC/rhodium–graphite + (a) RfP450scc; (b) Cytochrome P450scc+ (a) Cross-linking via Glutaraldehyde;
(b)+ Electro. (CV) vs. Ag/AgCl
6
3-aminopropyl-modified controlled-pore glass (APCEG)/rotating disk + Chox, ChEt, HRP+ Cross-linking via
Glutaraldehyde+ Ampero. at −0.15 V vs. Ag/AgCl) withTBC 7
Streptavidin/biotin/thioctic acid (SAM)/Au nanowires + ChOx, ChEt+ Covalent via Biotin-avidin+
Voltammetric (SWV) vs. Ag/AgCl 8
FeMC (physisorbed)-P(NMPY)-PTS/ITO+ ChOx+ Physical adsorption+ Electro. (CV) vs. Ag/AgCl 9
Naushad

30. EXPLANATION OF GRAPH
• Sensitivity of an sensor indicates the capacity of the sensor to
respond truly to the change in the output, corresponding to
the change in the input.
• Its depends on various factor like electrode , Enzymes and
Immobilization technique.
Naushad Rahman

31. RESPONSE TIME
0
0.5
1
1.5
2
2.5
3
3.5
1996 1998 2000 2002 2004 2006 2008
ResponseTime(sec)
Year
Response time in sec
Response time in sec
1
2
3
4
5 6
7
8
9
12
11
10
13
14
15
Naushad

32. TABLE
1 PPy/Pt covered with polycarbonate membrane +ChOx +
Entrapment+Ampero. at 0.7 V vs. SCE
8
PPy/Pt+ ChOx+ Entrapment+ Ampero. at 0.7 V vs. Ag/AgCl
2 2-aminoethanethiolate/Au +ChOx, ChEt for TC, ChOx for FC
+Cross-linking via Glutaraldehyde +Amperometric at 0 V vs.
SCE with thionin
as electron mediator
9
ITO glass+ Cytochrome P450SCC (5 g in 40 LB)+ LB films+
Electro. (CV) vs. Ag/AgCl
3
Hexadecyl mercaptan (SAM)/Au+ Molecular imprints+
Template+ Electro. (CV) vs. Ag/AgCl
10 BSA/dilauroylphosphatidylcholine (DLPC)/rhodium–
graphite; (b) Agarose gel-Riboflavin-DLPC/rhodium–
graphite + (a) RfP450scc; (b) Cytochrome P450scc+ (a)
Cross-linking via Glutaraldehyde; (b)+ Electro. (CV) vs.
Ag/AgCl
4 TEOS derived solgel+ ChOx, HRP+ Entrapment+ Ampero. at
0.75 V vs. Ag/AgCl
11 Sol–gel/CNT-Pt/graphite electrode+ ChOx+ Entrapment+
Electro. (CV) and Ampero. −0.2 V vs.
5 Poly(ethylene imine)(PEI)/poly(styrene sulfonate)/ITO +
ChOx+ LBL deposition of PEI and ChOx + Ampero. at 0.6 V vs.
Ag/AgCl
12
1-hexadecanethiol (SAM)/Au+ Molecular imprinted layer+
Templete + Voltammetry (CV) vs. Ag/AgCl
6
PPy- doecylbenzene sulfonate/ITO+ ChOx+ Physical
adsorption+ Electro. (CV) vs. Ag/AgCl
13 MWCN/Screen Printed Carbon Electrode+ Chox, Chet,
HRP, K4 Fe(CN)6+ Physical adsorption+ Ampero. at 0.3 V
vs. Ag/AgCl
7 (a) Octadecylsilica (ODS)/TEOS sol–gel for cholesterol in
aqueous micelle solution; (b) ODS/HECMC/PVA gel for
cholesterol in hydrophobic organic solvent + ChOx+
Entrapment+ Optical oxygen transducer
14
Multilayer of Pt-MWCNT-CHIT and poly(sodium-p -
styrenesulfonate) onto Au + ChOx+ Cross-linking via
Glutaraldehyde+ Ampero. at 0.1 V vs. SCE
15 ZnO/Au+ ChOx+ Physical adsorption+ Electro. (CV) vs.
Ag/AgCl
Naushad Rahman

33. EXPLANATION OF GRAPH
• Response Time:- Time taken by a sensor to detect the
cholesterol in a sample.
• Lower the response time higher is the efficiency of the
biosensor.
NIha

34. REUSABILITY
0
20
40
60
80
100
120
140
160
180
200
1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008
Reusability(Times)
Year
Reusability
RE
6
1
2
3
4
5

35. TABLE
Component Component
1
PANI/Pt+ ChOx+ Electro. doping+ Ampero. at
0.6 V vs. SCE and 1% tritonX-100 5
Poly(dialyldimethylammonium chloride)
(PDDA)/MWCNTs/Au + ChOx covered by o-PDD+
LBL deposition of PDDA and ChOx+ Ampero. at
0.7 V vs. SCE
2 Acrylamine glass beads+ ChOx, ChEt, HRP+
Covalent via diazotization+ Spectro. at 520 nm
using APZ+ phenol 6
PANI/ITO+ ChOx+ Covalent via EDC/NHS+ Spectro.
at 500 nm using o-dianisidine
3 Silisic sol–gel/PB/GC+ ChOx+ Entrapment+
Electro. (CV) at −0.05 V vs. Ag/AgCl
4 Polyacrylonitrile fiber+ ChOx+ Cross-linking via
Glutaraldehyde+ Spectro. at 520 nm using APZ+
phenol
Naushad Rahman

36. EXPLANATION OF GRAPH
• Reusability means number times sensor can be use before
losing its precision level .
• It depends upon the electrode quality .
• As sensor electrode will erode by interfacing element and
reduce its surface area to volume ration will decrease.
Naushad Rahman

37. SHELF LIFE
Naushad
0
20
40
60
80
100
120
2000 2001 2002 2003 2004 2005 2006 2007 2008
ShelfLife(Days)
Year
Improvment in Shelf Life
1
2
3
4 5
6
7

38. TABLE
1 Tributylmethyl phosphonium
chloride polymer
membrane/pyrolitic graphite
electrode + ChOx, HRP+
Entrapment+ Ampero. at −0.28 V vs.
Ag/AgCl
4
PPy/ITO+ ChOx, ChEt+ Entrapment+
Ampero. at 0.5 V vs. Pt wire
2 Acrylamine glass beads+ ChOx,
ChEt, HRP+ Covalent via
diazotization+ Spectro. at 520 nm
using APZ+ phenol
5
Sol–gel/CNT-Pt/graphite electrode+
ChOx+ Entrapment+ Electro. (CV) and
Ampero. −0.2 V vs.
3 Silisic sol–gel/PB/GC+ ChOx+
Entrapment+ Electro. (CV) at −0.05 V
vs. Ag/AgCl
6 BSA/TEOS/ITO+ ChOx, HRP+
Covalent+ Ampero. at 0.75 V vs.
Ag/AgCl
7 BSA/Polycarbonate/Oxygen
electrode+ ChOx, ChEt+ Cross-linking
via Glutaraldehyde+ Polarographic at
−0.7 V vs. Ag/AgCl
Naushad Rahman

39. EXPLANATION OF GRAPH
• Shelf Life:- Duration till which one can use electrode to detect
cholesterol.
• Higher the shelf life higher is the efficiency of the biosensor.
Naushad

40. DETECTION LIMIT
Naushad
0
10
20
30
40
50
60
70
1992 1994 1996 1998 2000 2002 2004 2006 2008
DetectionLimit(mgdl)
Year
Detection Limit
DL
2
1
3
4
5
6 7
8
9
1310
11
12

41. TABLE
Naushad Rahman
Component Component
1 PPy/Pt covered with polycarbonate membrane
+ChOx + Entrapment+Ampero. at 0.7 V vs. SCE
7 Poly(o-phenylenediamine)/PPy/Pt/Pt+ ChOx+
Entrapment in PPy+ Ampero. at 0.5 V vs. Ag/AgCl
2 HRP-hydroxymethyl ferrocene-carbon paste
electrode +ChOx ChEt+In solution
+Chronoamperometry at 0 V vs. SCE
8 Acrylamine glass beads+ ChOx ChEt HRP+
Covalent via diazotization+ Spectro. at 520 nm
using APZ+ phenol
3 Laponite clay nanoparticls-poly((12-pyrrol-1-
yldodecy)triethylammonium tetrafluoroborate)/Pt
disk electrode + ChOx ChEt for TC ChOx for FC
+Entrapment +Ampero. at 0.53 V vs. Ag/AgCl
9
PPy/Pt+ ChOx+ Entrapment+ Ampero. at 0.7 V vs.
Ag/AgCl
4 Glassy carbon+ ChOx+ In solution + Electro. at 0 V
vs. SCE using thionin as mediator
10 ITO glass+ Cytochrome P450SCC (5 g in 40 LB)+ LB
films+ Electro. (CV) vs. Ag/AgCl
5 Tetramethoxy silane sol–gel/poly(12-
diaminobenzene)/Pt + ChOx+ Entrapment followed
by treatment with Glutaraldehyde vapors+
Ampero. at 0.6 V vs. Ag/AgCl
11
Silica sol–gel- chitosan (CHIT)-MWCNT/PB/GC +
ChOx+ Entrapment+ Ampero. at −0.05 V vs.
Ag/AgCl
6
Hexadecyl mercaptan (SAM)/Au+ Molecular
imprints+ Template+ Electro. (CV) vs. Ag/AgCl
12 CHIT/NiNPs/histidine/MWCNT/GC electrode +
ChOx+ Cross-linking via Glutaraldehyde+ Ampero.
−0.2 V vs. SCE
13 TEOS (sol–gel)ITO+ ChOx ChEt+ Entrapment+
Spectro. at 500 nm using 4-APP + phenol

42. EXPLANATION OF GRAPH
• DL is the lowest quantity of a substance that can be
distinguished from the absence of that substance within a
stated confidence limit.
• Lower the DL greater is the selectivity of the CB.
Naushd

43. DIFFERENT MATERIALS FOR
AMPEROMETRIC BIOSENSORS
Metallic Materials Thermoplastic
Polymeric Material
Thermosetting
Polymeric Material
Applications flow through reusable
sensors
Moderate
temperature with no
strong solvents, used
as working electrode
Used in combination
with polar solvents,
used as working
electrode
Advantages lower electronic noise low detection limits, high sensitivities, lower
applied potential, reduction of background,
efficient electron transfer
Disadvantages Moderate
performance
commercial production of pure and defect-
free polymers is difficult
and costly
Examples Gold, Ion, Silver Carbon, conductive polymer, sol-gel polymer,
functional polymer
Chenxi

44. METAL
Chitosan-gold
 High electron transfer rate (Electron transfer rate constant was
estimated to be 15.6 s−1)
 Enhanced stability
 Moderate bioactivity
 magnetic Fe3O4/chitosan
 Fast response to H2O2 and excellent linear relationships
 Excellent bioactivity
 Long-time stability and good reproducibility
Chenxi

45. CARBON NANOTUBES
• Accuracy, efficiency and detection limit of a biosensor can be
increased with the use of carbon nanotubes immobilized on
bare electrodes
• High electron transfer, high surface area, minimization of the
surface fouling, high stability, excellent adsorptive and
biocompatibility.
Chenxi

46. SOL-GEL POLYMERIC MATERIALS
Sol-gel materials provide a versatile way for immobilization due
to the presence of inorganic M–O or M–OH–M bridges forming
a continuous network containing a liquid phase which can then
be dried out to form a solid, porous polymeric matrix
Excellent sensitivity
Good reproducibility
Remarkable stability
Rapid response
Chenxi

47. CONDUCTIVE POLYMERIC MATERIALS
Superior reusability
Excellent thermal stability
Large surface to volume ratio
High conductivity, sensitivity
Good biocompatibility
Wide linear range
Graphene/chitosan
Chenxi

48. FUNCTIONAL POLYMERIC MATERIAL
These are polymeric materials which possess different
functional groups, for example, thiols, amines, carboxylic
acids and others
Low detection limit, wide detection range
Fast response
Good stability.
Anti-interference ability
Chenxi

49. COMPARISON OF SELECTED
MATERIALS
Chenxi

50. INVASIVE CB
• Invasive biosensors are the one in which blood sample is
required and is obtained from the finger tip by pricking.
• These sample is placed in the test strip and the test strip is
inserted into the biosensor device and amount of cholesterol
is detected.
NIHA

51. NON INVASIVE BIOSENSOR.
• Non Invasive biosensor are the one in which no pricking is
required it uses skin cholesterol to measure the cholesterol
level in human body.
• Skin contains over 11% of the body cholesterol and ages in
parallel with vascular connective tissue. As arterial walls
accumulate cholesterol, so do the skin tissues.
• A high skin cholesterol level is a reliable predictor of higher
cholesterol accumulation in the arteries and, accordingly, can
be used in combination with other risk factors to assess risk of
coronary artery disease.
NIHA

52. CONTD..
• The simple test is conducted by placing a drop of digitonin, which binds selectively to the
cholesterol in the skin, on the palm of the hand. This liquid also contains an enzyme linked to
the digitonin by a copolymer. After a one-minute incubation period, the area is blotted dry to
remove any unbound digitonin solution. A second drop of liquid is then added, containing a
substrate for the horseradish peroxidase enzyme. When combined, a blue color change
occurs in direct proportion to the amount of digitonin that is bound to skin cholesterol. After
two minutes, a hand-held spectrophotometer (color reader) is placed over the drop to
measure the precise blue color, which indicates the skin cholesterol value.
PREVU* Skin Cholesterol Test
NIha

53. INVASIVE V/S NON INVASIVE
Invasive Non Invasive
It requires sample so finger pricking is done
which is very painful
No pricking is required so painless
cholesterol determination
It require patient preparation such as
fasting before cholesterol determination
No prior patient preparation is required.
More sensitive and more accurate Less Sensitive and less accurate
Latest cholesterol biosensor can detect
total cholesterol, LDL,HDL and triglycerides
separately.
It can detect only total cholesterol
It is the present. It can be the future as high research is done
in the non invasive biosensor.
NIha

54. Commercial Cholesterol biosensor
Electrochemical techniques are simple, relatively
cheap, and rapid as compared to other methods; but
also have a potential for further improvement.
Other methods.
colorimetric, high performance liquid
chromatography (HPLC) using gel electrophoretic
chip, capillary electrophoresis, spectrophotometric
and fluorometric
1st generation of Commercial cholesterol
biosensors use cholesterol esterase or
cholesterol oxidase .They depend on the
presence of oxygen for the enzyme –
catalysed reaction and the production and
measurement of hydrogen peroxide.
•2nd Generation use electron mediators
which help to transfer electrons from the
enzyme to electrode surface
.Ferro/ferricyanide, ferrocene, conducting
organic salts and redox dyes were used as
mediators.
Karthika

55. http://pubs.acs.org/action/showImage?doi=10.1021%2Fcr068123a&iName=master.img-005.jpg&type=master
Commercial Cholesterol biosensor
3rd generation of cholesterol
biosensors: Use direct electron
transfer between enzymes and
electrodes in the absence of mediator.
The distance between immobilised
enzyme and electrode should be as
small as possible. Such biosensors are
more sensitive.
A wide range of cholesterol biosensors are available in market today using a
range of matrix ,immobilisation and transducer technologies.
Karthika

56. COMMERCIAL CB
• Cholesterol Bio sensors used in Clinical analysis are usually
more accurate than home kits.
• Cholesterol biosensors used in Labs devices like Autoanalyzer
which is capable of doing dozen of analyses simultaneously by
a single machine from a small amount of serum.
Karthika

57. COMMERCIAL CHOLESTROL
BIOSENSOR
cholesterol oxidase for assay of total and free
cholesterol in serum by continuous-flow
analysis.
Desktop version
http://en.wikipedia.org/wiki/AutoAnalyzer
Hitachi 7070
http://japancare.trustpass.alibaba.com/product/
124017974-
103336093/Auto_Analyzer_HITACHI_7070_91
1_.html
Evolution of Commercial biosensors
Standalone
Huge devices
Portable
Home
kits
Karthika

58. DESKTOP VERSION OF CB
Karthika

59. CONTD..
Karthika

60. HOME KITS
Detects as little as 200 nM (80 ng/ml) cholesterol
Helps accurately measure cholesterol content of 0.01 µl of
human serum
Detects both free cholesterol and cholesteryl esters.
High sensitivity and excellent linearity of the assay at low
levels of cholesterol (0–0.015 µg⁄ml).
Karthika

61. AMPLEX® RED ASSAYS DETECTION LIMIT
Different product use different reagent (polyethylene glycol cholesteryl ether
, Cholesterol oxidase , horseradish peroxidase ) and electrode combinations.
Karthika
high sensitivity and
excellent linearity of
the assay at low
levels of cholesterol
(0–0.015 µg⁄ml)
𝜇𝑔/𝑚𝐿
Fluorescence

62. COMMON TECHNOLOGY IN
COMMERCIAL CB
Most common technique in Home Kits
Cholesterol Kit uses cholesterol oxidase to produce hydrogen
peroxide, which is then detected by the reagent in the
presence of horseradish peroxidase (HRP).
Enzyme coupled technology
Measure by reflectance of light(photometric).
An electrochemical reaction which generates an electrical
current proportional to the amount of Cholesterol-
Electrochemical biosensors.
Karthika

63. CONTD..
Karthika

64. FEATURES ON WHICH SUCCESS OF CB
DEPENDS
• Be stable under normal storage conditions and show good stability
over a large number of assays (i.e. much greater than 100)
• The reaction should be as independent of physical parameters as
pH and temperature
• The response should be accurate, precise, reproducible and linear
over the useful analytical range. It should also be free from
electrical noise.
• Should be cheap , small, portable
• Capable of being used by unskilled person.
• Screen Printing played an important role in biosensors
commercialization.
• Variation of enzymes and electrode materials had improved
performance in sensitivity storage and shelf life.
Karthika

65. COMMERCIAL CB CONTD..
Karthika

66. CONTD..
Karthika

67. CONTD…
Karthika

68. CONTD..
Karthika

69. FUTURE CHOLESTEROL BIOSENSOR
Non invasive Biosensors and Wearable Biosensors are the
future of CB.
Wearable Biosensor
• These are the devices that will monitor continuously the
physiological signals.
• They rely on wireless sensor ,the data are recorded and is
used to monitor patients health condition.
• They are very helpful to athletes, handicapped, old aged and
to professional people.
NIha

70. WEARABLE BIOSENSOR
• Examples of wearable biosensor:
Shoes Biosensor
Shirt BiosensorRing Biosensor
NIha

71. WEARABLE BIOSENSOR
• Ring sensor Can measure the heart rate and
oxygen saturation rate.
• Shirt Sensor can measure body temperature, heart
rate and respiration rate.
• Their main applications are :-
 Chronic Surveillance of abnormal heart failure
 In cardio-vascular dieses to measure hyper tension.
Wireless Monitoring for people in hazardous
operations.
NIha

72. WEARABLE BIOSENSOR
Advantage:-
Continuous Monitoring
Easy to use
Reduce Hospitalization Fee
Disadvantage:-
Initial Cost is high
Limited number of physiological parameter is
measured
Niha

73. EFFECT OF TECHNOLOGY IN
DEVELOPMENT OF CB
Two most important technology that will play a significant role
for the development of CB is
Nanotechnology
Microsystem Technology.
Various kind of nanomaterial are applied to cholesterol
biosensor such as gold nanoparticles, carbon nanotubes,
Nanowires, Graphene and Quantum dots.
Niha

74. WHY NANO TECHNOLOGY??
• These materials are generally used because of their unique
physical, chemical, mechanical, magnetic and optical
properties,
• They markedly enhance the sensitivity and specificity of
detection.
• They have great potential in the detection of DNA, RNA,
proteins, glucose ,pesticides and other small molecules from
clinical samples, food industrial samples, as well as
environmental monitoring.
Niha

75. MULTI ANALYTE DETECTION
They also help in Multi analyte Detection.
• Development of sensors capable of determining several
analyte simultaneously can represent an interesting tool in
clinical industry.
• This will help to reduce the cost and will also lessen the
diagnostic time.
NIha

76. MINIATURIZATION
They also led to Miniaturization of biosensor.
• Miniaturization allows the handling of low-volume samples, a
reduction in reagent consumption and waste generation, and
increases sample throughput .
• Miniaturization can benefit Biosensor by making it
inexpensive and easy-to-handle analytical devices.
• Nanowire are smaller than the red blood cell whose diameter
is 6.8 micro meter.
• Use of Nanowire Biosensor can help to develop implantable
biosensor too.
NIha

77. EFFECT OF NANOPARTICLE ON
SENSITIVITY
0
5
10
15
20
25
30
35
40
45
50
Sensitivity(mV)
Sensitivity(mV)
NIha

78. EFFECT OF NANOPARTICLE ON
DETECTION LIMIT
0
0.2
0.4
0.6
0.8
1
1.2
1.4
Detection Limit(microM)
Detection Limit(microM)
NIha

79. EFFECT OF MWCNT ON DETECTION
LIMIT
0
0.1
0.2
0.3
0.4
0.5
0.6
2000 2002 2004 2006 2008 2010 2012 2014
Detection Limit(micromol)
Detection Limit(micromol)
NIha

80. EFFECT OF GRAPHENE ON DL AND
SENSITIVITY
NIHA
0
0.5
1
1.5
2
2.5
3
0
5
10
15
20
25
30
35
40
2009.5 2010 2010.5 2011 2011.5 2012 2012.5 2013 2013.5 2014 2014.5
LimirOFDetection(μM)
Sensitivity(μAmM1
cm-2)
YEAR
Sensitivity
LOD (μM)

81. TOTAL MARKET OF BIOSENSOR
• The biosensor market is dominated by only a few products
• For medical diagnostics, approximately 90% of biosensors are
glucose monitors, blood gas monitors, and electrolyte or
metabolite analyzers
• Half of all biosensors produced worldwide are glucose
monitors Sales are projected at $1.28 billion in the US in 2012
• The majority of the remaining market includes biosensors
directed at environmental control, fermentation monitoring,
alcohol testing, and food control
NIha

82. GROWTH IN THE USAGE OF
BIOSENSOR
NIha

83. MARKET ANALYSIS OF BIOSENSOR
• According to a new market report published by Transparency
Market Research “Biosensors Market - Global Industry
Analysis, Size, Share, Growth, Trends and Forecast, 2012 -
2018,” in 2011, the global biosensors market was valued at
USD 9.9 billion and it is expected to grow at a CAGR of 9.6%
from 2012 to 2018 to reach a market of USD 18.9 billion by
2018.
• In 2011, the biosensors market was valued to be USD 9,973.5
million and is expected to grow at a CAGR of 9.6% from 2012
to 2018.
NIha

84. FURTHER ANALYSIS
• The development of new biosensors and devices based on
this technology is a highly capital intensive exercise.
• Although miniaturization allows for economies of scale to be
achieved in the actual manufacturing of biosensors, huge
capital investment is required for research and development.
• Though the U.S. remains the largest market in the world,
Asian countries namely India and China are witnessing fast
growth and are predicted to emerge as dominating markets in
the near future.
NIha

85. FURTHER TREND
NIha

86. Thank You