TECHNICAL SEMINAR
ON
BIOSENSORS
PRESENTED BY GUIDE
VINEETHA M Mr. ANANDA H V
1SG15BT035 Asst Professor.
CONTENTS
 Introduction.
 History
 Principle of biosensor.
 Working of biosensor.
 Types of biosensor.
 Applications of biosensor.
 Advantages and Disadvantages.
Introduction
 Biosensor is an analytical device which uses
enzymes, antibody, tissues that converts biological
response into electrical, thermal or optical signals[1].
 Biosensor consists of two components: first the
“sensing element” and second is the “transducers”.
Sensing element may be either enzymes, antibodies,
DNA, tissues or whole cells which then transduces the
biochemical reaction into electrical signals[1].
History
 Leland C Clark is the father of biosensor[2].
 He invented the glucose biosensor to determine
the glucose level in the sample.
 Enzyme based biosensor was reported by Updike
and Hicks in 1967[2].
Principle of Biosensor
Figure 1: Components of biosensor (3).
Working of Biosensor
Figure 2: Working Of Biosensor [4].
Types of Biosensor:
1. Optical Biosensor
Principle:
 Optical biosensors are the devices that utilize the
principle of optical measurements (absorbance,
fluorescence, chemiluminescence etc.)[6].
Aborbance α Analyte concentration
Components :
 Biorecognition element.
 Light source.
 Optical transducer.
 Display.
Working of Optical Biosensor
Figure 3: Working of Optical Biosensor [5].
Contd
 Applications
Estimation of blood glucose level
 Glucose biosensor comprises of immobilized
enzymes such as glucose oxidase, Horse reddish
peroxidase and chromogen (toulidine)[6].
 The intensity of the colour of the dye can be
measured by using a portable reflectance meter.
Glucose strip production is a very big industry
worldwide [6].
2. Thermal Biosensor:
 Principle:
Heat generated α Analyte concentration[1].
 Components of Thermal Biosensor:
1. Thermal sensor.
2. Enzyme immobilized / free on the thermal sensors.
3. Thermocouple.
4. Display.
Contd
2. Thermal Biosensor Working:
Figure 4: Working Of Thermal Biosensor [7].
Contd
 Applications:
Thermometric biosensors are in use for the estimation of
serum cholesterol. When cholesterol gets oxidized by
the enzyme cholesterol oxidase, heat is generated which
can be measured [6].
Cholesterol catalysis by Cholesterol oxidase [8].
3. Electrochemical Biosensor
Principle:
Electricity or voltage generated α Analyte Concentration
[1].
Components:
 Biorecognition element.
 Transducer.
 Display.
Contd
Figure 5:Working of EC biosensor[9].
5.DNA Biosensor
Principle:
Fluroscence α Concentration of Target DNA [6].
Components:
 DNA immobilized onto chip.
 Transducer.
 Display.
Contd
Figure 6: working of DNA Biosensor [10].
Contd
 Applications:
DNA biosensors are used for detecting DNA and
known as bio detectors. DNA sensors are used to
isolate and detect the strength of single DNA-DNA
interaction.
 Detection of SNP.
 DNA sequencing is very important for drug discovery
and diagnosis of genetic disorders [11].
Applications of Biosensor
1. Glucose Biosensor for Diagnosis of Diabetis
 Diabetis Mellitus is a Metabolic disorder
characterized by hyperglycemia.
 Causes for Diabetis Mellitus :
 Pancreas ineffeciency to produce insulin for
controlling the blood glucose level.
 Mutation in the receptors.
 Two types of Diabetis Mellitus :- IDDM & NIDDM
[12].
Contd
 Principle:
NADPH production α Glucose Concentration.
 Fabrication:
Carbon paste was prepared by mixing carbon powder
and mineral oil in the ratio of 4:1 [13].
 To the Carbon paste RBC [biological component]
extracted from 2 ml of blood sample was mixed
thoroughly for 30 minutes to form uniform
mixture[13].
The carbon paste is packed at the tip of 5 cm glass
tube and electrical circuit consisting of copper wire is
then inserted at the other end of glass tube [13].
Contd
 Instrumentation:
Glucose biosensor is a three electrode system. It consists
of reference electrode, working electrode and auxiliary
electrode [13].
 Ag/AgCl is used as reference electrode and platinum
wire is used as auxiliary electrode. These electrodes are
placed in a electrochemical cell of 40ml capacity [13].
Contd
 Working Principle:
Glucose biosensor consists of Hexo kinase, G-6-PD and RBCs.
When the biosensor comes in contact with glucose in the sample.
RBC is pentrates into RBC by enzyme Glucose permease.
Then glucose is catalysed by hexokinase to produce Glucose 6 Phosphate.
G-6-P is catalysed by G-6-PD into 6-Phospogluconate with the oxidation of NADP
into NADPH2 [13].
Contd
Figure 7: NADPH Production in the biosensor [13].
2.PAPER BASED BIOSENSOR FOR WATER
ANALYSIS
 Presence of contaminants in water is a major concern,
contaminants such as pesticides, micro organisms, heavy
metals and chemicals released by various industries [14].
 Detection of contaminants in water is important before
releasing the water into the environment. Development of
biosensors helps to detect water contaminants in fast, accurate
and rapid detection[14].
 Paper based biosensor is the developing technique used for
analysing the water quality[14].
 Paper analytical technique in combination with biosensor
serves as a new technique for qualitative and quantitative
analysis of water [14].
 The key advantage of this technique is portable, low cost and
efficient[14].
Contd
 According to World Health Organization [WHO] and
Environment Protection Act [EPA], the Maximum
contaminant level [MCL] are been prescribed for water
released by various industries.
SL.
NO
COMPONENT MCL RANGE [µg L-1]
1 Arsenic 10
2 Mercury 2-6
3 Lead 10
4 Uranium 30
5 Cadmium 3-5
TABLE 1: List of contaminats and their allowable ranges in water [14].
Contd
 Instrumentation:
SL
NO
PAPER PROPERTIES
1 Cellulose filter paper Hydrophilic, permeable to aqueous
solution and available with various
pore size and thickness
2 Chromatography paper Separation of nanoparticles and
applicable for agglutination based
assay.
3 Nitrocellulose membrane Hydrophobic and retention of
components due to electrostatic
interaction.
4 Printing paper Microfluidic channel creation for
sample analysis.
TABLE 2: Paper types used in biosensors fabrication [14].
Contd
 Fabrication:
FIG 8: A Represents Wax Printing [14].
Contd
 Applications:
Ethinylestradiol is a synthetic harmone which is used in
oral contraceptives.
This is the most commonly present in pharmaceutical
effluent. Increasing concentration of ethinylestradiol
in water to has a greater potential to effect the aquatic
biota.
Paper based biosensor is developed to determine the
amount of ethinyl estradiol in water.
Contd
Paper biosensor immobilized with Anti-EE2.
Water containing EE2 conjugated with HRP is used as sample.
Anti –EE2 specifically interacts with EE2 conjugated with HRP.
Substrate 10-Acetyl-3,7-dihydroxyphenoxazine and H2O2.
Production of chromogen Resorufin(blue dye).
Detected by LED Fluorescence detector.
Transduced by Optical transducer.
Amount of EE2 present in water is displayed
Advantages
 High sensitivity.
 Eco- friendly.
 Simple fabrication.
 Simple monitoring device in situ.
 Mass production is easier.
 Highly specific to analyte.
 Low cost.
 Fast response.
 Accurate measurement.
Diasdavantages
 Poor reproducibility.
 Temperature dependent system.
 pH influences the performance of the biosensor.
 Requires sample preparation.
References
1. Saraju P Mohanty et.al. “Biosensors : A Tutorial
Review”,IEEE Potentials, 25[2], 35-40 [2006].
2. Payoshnee Bhalinge et.al. “Biosensors: nanotools of
detection- A review”, Int J of Healthcare and biomedical
research, 04, 26-39 [2016].
3. Nicola stefano fracchilla, Silvia Artuso Agostino
cortelezzi. “Biosensors in Clinical Practice : Focus on
Oncohematology”, Journal of sensors, 13(5) , 6423-6447
[2013].
4. https://www.eduhk.hk/biotech/eng/classrm/explain/ge
ne16.jpg.
5. https://pubs.rsc.org/services/images/RSCpubs.ePlatfor
m.Service.FreeContent.ImageService.svc/ImageService/
Articleimage/2016/RA/c5ra15269k/c5ra15269k-f5_hi-
res.gif
Contd
6.http://www.biologydiscussion.com/enzymes/biosens
ors/biosensors-features-principle-and-types-with-
diagram/10240.
7.https://www.cell.com/cms/attachment/538394/37414
51/gr1.Journalpg
8. http://file.scirp.org/Html/1-2270015/203ec242-b8aa-
4353-a3b6-fc9f95baa105.jpg
9. Jitendra kumar et.al. biosensor for environmental
and clinical monitoring, BARC NEWS LETTER, 324,
34-38 [2012].
10.https://ars.els-cdn.com/content/image/1-s2.0-
S016599361200146X-gr1
Contd
11. C Bartic et.al. “Organic thin film transistor as transducer for
bioanalytical applications”, Journal of Analytical and
Bioanalytical Chemistry, 384 354-365 [2006].
12.AL. Galant, R. C. Kaufman, and J. D. Wilson, “Glucose:
detection and analysis,” Journal of Food Chemistry, 88, 149–160
[2015].
13.Amandeep Kaur and Neelam Verma. “Electrochemical
biosensor for monitoring insulin in normal individual and
diabetic mellitus patients, European journal of experimental
biology, 2[2], 389-395 [2012].
14.Celine I L Justino et.al. “Recent progress in environmental
monitoring: A Review”, J of sensor basel, 17, [2017].
15.
Acknowledgment
I would like to thank My Guide Mr. Ananda
H V, HOD Dr. Veena S More and all the
Teaching Staff for the constant support.
THANK
YOU ……

Biosensors: Their types

  • 1.
    TECHNICAL SEMINAR ON BIOSENSORS PRESENTED BYGUIDE VINEETHA M Mr. ANANDA H V 1SG15BT035 Asst Professor.
  • 2.
    CONTENTS  Introduction.  History Principle of biosensor.  Working of biosensor.  Types of biosensor.  Applications of biosensor.  Advantages and Disadvantages.
  • 3.
    Introduction  Biosensor isan analytical device which uses enzymes, antibody, tissues that converts biological response into electrical, thermal or optical signals[1].  Biosensor consists of two components: first the “sensing element” and second is the “transducers”. Sensing element may be either enzymes, antibodies, DNA, tissues or whole cells which then transduces the biochemical reaction into electrical signals[1].
  • 4.
    History  Leland CClark is the father of biosensor[2].  He invented the glucose biosensor to determine the glucose level in the sample.  Enzyme based biosensor was reported by Updike and Hicks in 1967[2].
  • 5.
    Principle of Biosensor Figure1: Components of biosensor (3).
  • 6.
    Working of Biosensor Figure2: Working Of Biosensor [4].
  • 7.
    Types of Biosensor: 1.Optical Biosensor Principle:  Optical biosensors are the devices that utilize the principle of optical measurements (absorbance, fluorescence, chemiluminescence etc.)[6]. Aborbance α Analyte concentration Components :  Biorecognition element.  Light source.  Optical transducer.  Display.
  • 8.
    Working of OpticalBiosensor Figure 3: Working of Optical Biosensor [5].
  • 9.
    Contd  Applications Estimation ofblood glucose level  Glucose biosensor comprises of immobilized enzymes such as glucose oxidase, Horse reddish peroxidase and chromogen (toulidine)[6].  The intensity of the colour of the dye can be measured by using a portable reflectance meter. Glucose strip production is a very big industry worldwide [6].
  • 10.
    2. Thermal Biosensor: Principle: Heat generated α Analyte concentration[1].  Components of Thermal Biosensor: 1. Thermal sensor. 2. Enzyme immobilized / free on the thermal sensors. 3. Thermocouple. 4. Display.
  • 11.
    Contd 2. Thermal BiosensorWorking: Figure 4: Working Of Thermal Biosensor [7].
  • 12.
    Contd  Applications: Thermometric biosensorsare in use for the estimation of serum cholesterol. When cholesterol gets oxidized by the enzyme cholesterol oxidase, heat is generated which can be measured [6]. Cholesterol catalysis by Cholesterol oxidase [8].
  • 13.
    3. Electrochemical Biosensor Principle: Electricityor voltage generated α Analyte Concentration [1]. Components:  Biorecognition element.  Transducer.  Display.
  • 14.
    Contd Figure 5:Working ofEC biosensor[9].
  • 15.
    5.DNA Biosensor Principle: Fluroscence αConcentration of Target DNA [6]. Components:  DNA immobilized onto chip.  Transducer.  Display.
  • 16.
    Contd Figure 6: workingof DNA Biosensor [10].
  • 17.
    Contd  Applications: DNA biosensorsare used for detecting DNA and known as bio detectors. DNA sensors are used to isolate and detect the strength of single DNA-DNA interaction.  Detection of SNP.  DNA sequencing is very important for drug discovery and diagnosis of genetic disorders [11].
  • 18.
    Applications of Biosensor 1.Glucose Biosensor for Diagnosis of Diabetis  Diabetis Mellitus is a Metabolic disorder characterized by hyperglycemia.  Causes for Diabetis Mellitus :  Pancreas ineffeciency to produce insulin for controlling the blood glucose level.  Mutation in the receptors.  Two types of Diabetis Mellitus :- IDDM & NIDDM [12].
  • 19.
    Contd  Principle: NADPH productionα Glucose Concentration.  Fabrication: Carbon paste was prepared by mixing carbon powder and mineral oil in the ratio of 4:1 [13].  To the Carbon paste RBC [biological component] extracted from 2 ml of blood sample was mixed thoroughly for 30 minutes to form uniform mixture[13]. The carbon paste is packed at the tip of 5 cm glass tube and electrical circuit consisting of copper wire is then inserted at the other end of glass tube [13].
  • 20.
    Contd  Instrumentation: Glucose biosensoris a three electrode system. It consists of reference electrode, working electrode and auxiliary electrode [13].  Ag/AgCl is used as reference electrode and platinum wire is used as auxiliary electrode. These electrodes are placed in a electrochemical cell of 40ml capacity [13].
  • 21.
    Contd  Working Principle: Glucosebiosensor consists of Hexo kinase, G-6-PD and RBCs. When the biosensor comes in contact with glucose in the sample. RBC is pentrates into RBC by enzyme Glucose permease. Then glucose is catalysed by hexokinase to produce Glucose 6 Phosphate. G-6-P is catalysed by G-6-PD into 6-Phospogluconate with the oxidation of NADP into NADPH2 [13].
  • 22.
    Contd Figure 7: NADPHProduction in the biosensor [13].
  • 23.
    2.PAPER BASED BIOSENSORFOR WATER ANALYSIS  Presence of contaminants in water is a major concern, contaminants such as pesticides, micro organisms, heavy metals and chemicals released by various industries [14].  Detection of contaminants in water is important before releasing the water into the environment. Development of biosensors helps to detect water contaminants in fast, accurate and rapid detection[14].  Paper based biosensor is the developing technique used for analysing the water quality[14].  Paper analytical technique in combination with biosensor serves as a new technique for qualitative and quantitative analysis of water [14].  The key advantage of this technique is portable, low cost and efficient[14].
  • 24.
    Contd  According toWorld Health Organization [WHO] and Environment Protection Act [EPA], the Maximum contaminant level [MCL] are been prescribed for water released by various industries. SL. NO COMPONENT MCL RANGE [µg L-1] 1 Arsenic 10 2 Mercury 2-6 3 Lead 10 4 Uranium 30 5 Cadmium 3-5 TABLE 1: List of contaminats and their allowable ranges in water [14].
  • 25.
    Contd  Instrumentation: SL NO PAPER PROPERTIES 1Cellulose filter paper Hydrophilic, permeable to aqueous solution and available with various pore size and thickness 2 Chromatography paper Separation of nanoparticles and applicable for agglutination based assay. 3 Nitrocellulose membrane Hydrophobic and retention of components due to electrostatic interaction. 4 Printing paper Microfluidic channel creation for sample analysis. TABLE 2: Paper types used in biosensors fabrication [14].
  • 26.
    Contd  Fabrication: FIG 8:A Represents Wax Printing [14].
  • 27.
    Contd  Applications: Ethinylestradiol isa synthetic harmone which is used in oral contraceptives. This is the most commonly present in pharmaceutical effluent. Increasing concentration of ethinylestradiol in water to has a greater potential to effect the aquatic biota. Paper based biosensor is developed to determine the amount of ethinyl estradiol in water.
  • 28.
    Contd Paper biosensor immobilizedwith Anti-EE2. Water containing EE2 conjugated with HRP is used as sample. Anti –EE2 specifically interacts with EE2 conjugated with HRP. Substrate 10-Acetyl-3,7-dihydroxyphenoxazine and H2O2. Production of chromogen Resorufin(blue dye). Detected by LED Fluorescence detector. Transduced by Optical transducer. Amount of EE2 present in water is displayed
  • 29.
    Advantages  High sensitivity. Eco- friendly.  Simple fabrication.  Simple monitoring device in situ.  Mass production is easier.  Highly specific to analyte.  Low cost.  Fast response.  Accurate measurement.
  • 30.
    Diasdavantages  Poor reproducibility. Temperature dependent system.  pH influences the performance of the biosensor.  Requires sample preparation.
  • 31.
    References 1. Saraju PMohanty et.al. “Biosensors : A Tutorial Review”,IEEE Potentials, 25[2], 35-40 [2006]. 2. Payoshnee Bhalinge et.al. “Biosensors: nanotools of detection- A review”, Int J of Healthcare and biomedical research, 04, 26-39 [2016]. 3. Nicola stefano fracchilla, Silvia Artuso Agostino cortelezzi. “Biosensors in Clinical Practice : Focus on Oncohematology”, Journal of sensors, 13(5) , 6423-6447 [2013]. 4. https://www.eduhk.hk/biotech/eng/classrm/explain/ge ne16.jpg. 5. https://pubs.rsc.org/services/images/RSCpubs.ePlatfor m.Service.FreeContent.ImageService.svc/ImageService/ Articleimage/2016/RA/c5ra15269k/c5ra15269k-f5_hi- res.gif
  • 32.
  • 33.
    Contd 11. C Barticet.al. “Organic thin film transistor as transducer for bioanalytical applications”, Journal of Analytical and Bioanalytical Chemistry, 384 354-365 [2006]. 12.AL. Galant, R. C. Kaufman, and J. D. Wilson, “Glucose: detection and analysis,” Journal of Food Chemistry, 88, 149–160 [2015]. 13.Amandeep Kaur and Neelam Verma. “Electrochemical biosensor for monitoring insulin in normal individual and diabetic mellitus patients, European journal of experimental biology, 2[2], 389-395 [2012]. 14.Celine I L Justino et.al. “Recent progress in environmental monitoring: A Review”, J of sensor basel, 17, [2017]. 15.
  • 34.
    Acknowledgment I would liketo thank My Guide Mr. Ananda H V, HOD Dr. Veena S More and all the Teaching Staff for the constant support.
  • 35.