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B Y
T E J A S W I N I L . A S A W E
A S S I S T A N T P R O F E S S O R
S I D D H I S I N S T I T U T E O F P H A R M A C Y , T H A N E
BIOSENSORS
BIOSENSORS
 CONTENT:
1.Introduction to biosensor
2. Parts of biosensors
3. Working mechanisms of biosensors
4. Types of Biosensors
5. Application areas of biosensors
What is Biosensor?
 Biosensors are analytical devices that convert a
biological response into electrical signal .
 A biosensor is an analytical device containing an
immobilized biological material (enzyme,
antibody, nucleic acid, hormone, organelle or whole
cell) which can interact with an analyte and
produce physical, chemical or electrical signals that
can be measured.
 An analyte is a compound (e.g. glucose, urea, drug,
pesticide) whose concentration has to be measured.
Father of Biosensor :- Leland C. Clark
The first and the most wide spreadly used
commercial biosensor: the blood
glucose biosensor – developed by Leland C.
Clark in 1962
DIAGRAM BIOSENSOR
Biosensor depends on 2 components
1. Biological component—enzyme, cell etc.
2. Physical component—transducer, amplifier etc.
 The biological component interacts with the analyte
to produce a physical change (a signal) that can be
detected, by the transducer.
 Amplifier enhance the signal while processor convert
physicochemical signal from the receptor.
 The display shows the output.
Working of a Biosensor
• The biological material ( i.e. enzyme) is
immobilized by conventional methods
(adsorption, encapsulation, entrapment,
cross linking or covalent binding).
• Analyte is compound whose
concentration has to be measured.
Ex. Glucose , urea etc
•Bioelement- also called bioreceptor in
which analyte is bound.
Ex. Tissue ,microorganism , enzymes.
•Transducer convert energy to
measurable form.
•Amplifier enhance the signal.
•Display shows the output.
PROPERTIES OF GOOD BIOSENSOR
1. Fast response time.
2. Rapid and continuous measurement.
3. Highly specific for analyte.
4. Mostly devices are tiny and biocompatible.
5. Cheap , small and easy to use.
6. Capable of repeated use.
BIOLOGICAL
BIOSENSOR
PHYSICAL
BIOSENSOR
ENZYME
BASED
Immunosensors
OPTICAL
BIOSENSOR
WHOLE CELL
BASED
PIEZOELECTRI
C
ELECTRO
CHEMICAL
Potentiometric
Amperometric
Calorimetric
THERMA
L
Depending upon transducer biosensors are classified as
DNA Biosensors
ENZYME BASED BIOSENSOR
 ENZYME BASED BIOSENSORS are prepared by
immobilization methods such as ionic bonding,
covalent bonding or van der Waals forces.
 The commonly used enzymes are peroxides,
aminoxidases, polyphenoloxidases, oxidoreductases
etc.
IMMUNOSENSORS
 IMMUNOSENSORS are prepared by antigen antibody reaction .
 antibodies are highly attracted towards their respective antigen
and forms antigen antibody reaction .
 Labelled antigens or antibodies may be used in biosensors based
on enzyme linked immunosorbent assay(ELISA) principles.
 Labelled antigen means attachment of specific tag to antibody for
detection of protein.
 ELISA can be used to detect an antibody to HIV.
DNA BIOSENSORS
 In this single stand nucleic acid molecule is bind
to its complementary strand in a sample.
 DNA biosensors consist of an immobilized DNA
strand to detect the complimentary sequence by
DNA–DNA hybridization.
 Hybridization means two single stranded DNA
bound to form double stranded DNA.
Electrochemical Biosensors:
 Electrochemical biosensors are simple devices
measures the electric current, ionic or conductance
changes carried out by bio electrodes
Amperometric Biosensors:
 These biosensors are based on the movement of electrons
(i.e. determination of electric current) as a result of
enzyme-catalysed redox reactions.
 This contain enzyme electrode or chemically modified
electrodes.
 A redox reaction catalyzed by an enzyme is directly bind
to an electrode .
 The electrons are transferred from the substrate to the
electrode via enzyme and redox mediator.
 It is a good example of “Amperometric Biosensors” ,
widely used throughout the world by diabetic patients.
Blood glucose monitoring is the use of a glucose meter for
testing the concentration of glucose in the blood . Important
in diabetes management, a blood glucose test is typically
performed by piercing the skin.
Potentiometric Biosensors
 In these biosensors, changes in ionic concentrations
are determined by use of ion- selective electrodes.
 pH electrode is the most commonly used ion-
selective electrode .
 These biosensors consist of membrane containing
immobilized enzyme and surrounding by pH
meter.
 The other important electrodes are ammonia-
selective and CO2 selective electrodes.
•The potential difference obtained between the
potentiometric electrode and the reference electrode can
be measured.
•Potential difference is proportional to the concentration
of the substrate.
•The major limitation of potentiometric biosensors is the
sensitivity of enzymes to PH , ammonia and carbon
dioxide .
Conductimetric Biosensors
 The changes in ionic species is transferred into electrical conductivity (
electrical current pass through the solution )in Conductimetric Biosensors.
 Conductivity measurement is based on the biocatalytic reaction of sample on
an electrode.
 The conductimetric transducer consist of reference electrode and working
electrode.
 Both electrodes are coated with nata de coco membrane and enzyme is
immobilized only on working electrode
 Nata de coco, a bacterial cellulose as a result of coconut water fermentation,
is a conductive polymer with a electrical conductivity of 553 μS/cm( Micro
Siemens per cm) and has high mechanical stability.
 Example is Urea biosensor utilizing immobilized urease.
 urea biosensors are very successfully used during dialysis and renal surgery
Thermometric or Calorimetric Biosensors
 Biological reactions are associated with the
production of heat in thermometric biosensors.
 They are more commonly referred to as thermal
biosensors or calorimetric biosensors.
 Thermometric biosensors are in use for the
estimation of serum cholesterol.
 Thermometric biosensors can be used as a part of
Enzyme-Linked Immuno-Sorbent Assay (ELISA)
and the new technique is referred to as
Thermometric ELISA (TELISA).
ELISA
 ELISA stands for enzyme-linked immuno sorbent
assay.
 It is a commonly used laboratory test to detect
antibodies in the blood.
 This test is often used to see if any one have
been exposed to viruses or other substances
that cause infection.
A diagrammatic representation of a thermal biosensor is in
Fig.
It consists of a heat insulated box
fitted with heat exchanger
(aluminum cylinder).
The reaction takes place in a small
enzyme packed bed reactor.
As the substrate enters the bed, it
gets converted to a product and
heat is generated.
The difference in the temperature
between the substrate and product
is measured by Thermistors.
Even a small change in the
temperature can be detected by
thermal biosensors.
Optical Biosensors
 Optical biosensors are the devices that utilize the
principle of optical measurements (absorbance,
fluorescence, internal reflection spectroscopy etc.
 It determines changes in light absorption between the
reactants and products.
 Optical biosensors primarily involve enzyme and
antibodies as the transducting elements.
 these biosensors usually do not require reference
sensors, as the comparative signal can be generated using
the same source of light as the sampling sensor.
 Fiber optic biosensor is most common example of
Biosensor.
Fig.Represents the Fibre Optic Lactate Biosensor.
In this changes in molecular O2 concentration
is measured by determining the quenching
(reducing ) effect of O2 on a fluorescent dye.
O2 has a quenching (reducing) effect on the
fluorescence. Hence the amount of fluorescent
generated by dyed film is depend on oxygen.
Lactate present in reaction mixture utilize
oxygen and decrease in the reducing effect of
oxygen resulting to increase in fluorescent
output measured.
The following reaction is catalyzed by the
enzyme lactate monooxygenase.
lactate
O2 +LACTATE CO2+Acetate+H2O
monooxygenase
Piezoelectric Biosensors
 Piezoelectric biosensors are based on the principle of acoustics
(sound vibrations), hence they are also called as acoustic biosensors.
 Piezoelectric crystals form the basis of these biosensors. The
crystals with positive and negative charges vibrate with
characteristic frequencies.
 Adsorption of certain molecules on the crystal surface alters the
resonance frequencies which can be measured by electronic devices.
 A biosensor for formaldehyde has been developed by incorporating
formaldehyde dehydrogenase.
 A biosensor for cocaine in gas phase has been created by attaching
cocaine antibodies to the surface of piezoelectric crystal.
Whole Cell Biosensors
 In the whole cell biosensors or microbial biosensors
,immobilized whole cell of microorganism or their
organelles are used.
 These may employed live or dead microbial cell.
 The microbial cells are easily available with less cost .
 They are less sensitive to variation in pH and
temperature compared to enzymes.
Application of Biosensors
 The four applications are:
1. Applications in Medicine and Health
2. Applications in food Industry
3. Applications in Pollution Control
4. Applications in Military.
1.Applications in Medicine and Health
 Biosensors are successfully used for the quantitative estimation
of several biologically important substances in body fluids e.g.
glucose, cholesterol, urea.
 Glucose biosensor is used for diabetic patients for regular
monitoring of blood glucose.
 Blood gas monitoring for pH, pCO2 and pO2 is carried out
during critical care and surgical monitoring of patients.
 Several toxic compounds produced in the body can also be
detected.
2. Applications in food Industry
 Biosensors can be used for monitoring of fermentation
products and estimation of various ions.
 Thus, biosensors help for improving the fermentation
conditions for a better yield.
 Now a days, biosensors are employed to measure the
odour and freshness of foods.
 Freshness of stored fish can be detected by ATPase.
 ATP is not found in spoiled fish and this can be detected
by using ATPase.
 Pharmaceutical company has developed immobilized
cholesterol oxidase system for measurement of
cholesterol concentration in foods (e.g. butter).
3. Applications in Pollution Control
(Environmental Monitoring)
 Biosensors are very helpful to monitor
environmental (air, water) pollution.
 The concentrations of pesticides can be measured
by biosensors.
 Biosensor have wide application for measurement
estimation and control of water, soil, air
contamination .
 Ex. Determination of pesticides by potentiometric
biosensor.
4. Applications in Military
( Defense)
 Biosensors have been developed to detect the toxic
gases and other chemical agents used during war.
 DNA Biosensor is used to detect criminals.
Thank You

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BIOSENSOR.pptx

  • 1. B Y T E J A S W I N I L . A S A W E A S S I S T A N T P R O F E S S O R S I D D H I S I N S T I T U T E O F P H A R M A C Y , T H A N E BIOSENSORS
  • 2. BIOSENSORS  CONTENT: 1.Introduction to biosensor 2. Parts of biosensors 3. Working mechanisms of biosensors 4. Types of Biosensors 5. Application areas of biosensors
  • 3. What is Biosensor?  Biosensors are analytical devices that convert a biological response into electrical signal .  A biosensor is an analytical device containing an immobilized biological material (enzyme, antibody, nucleic acid, hormone, organelle or whole cell) which can interact with an analyte and produce physical, chemical or electrical signals that can be measured.  An analyte is a compound (e.g. glucose, urea, drug, pesticide) whose concentration has to be measured.
  • 4. Father of Biosensor :- Leland C. Clark The first and the most wide spreadly used commercial biosensor: the blood glucose biosensor – developed by Leland C. Clark in 1962
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  • 7. Biosensor depends on 2 components 1. Biological component—enzyme, cell etc. 2. Physical component—transducer, amplifier etc.  The biological component interacts with the analyte to produce a physical change (a signal) that can be detected, by the transducer.  Amplifier enhance the signal while processor convert physicochemical signal from the receptor.  The display shows the output.
  • 8. Working of a Biosensor • The biological material ( i.e. enzyme) is immobilized by conventional methods (adsorption, encapsulation, entrapment, cross linking or covalent binding). • Analyte is compound whose concentration has to be measured. Ex. Glucose , urea etc •Bioelement- also called bioreceptor in which analyte is bound. Ex. Tissue ,microorganism , enzymes. •Transducer convert energy to measurable form. •Amplifier enhance the signal. •Display shows the output.
  • 9. PROPERTIES OF GOOD BIOSENSOR 1. Fast response time. 2. Rapid and continuous measurement. 3. Highly specific for analyte. 4. Mostly devices are tiny and biocompatible. 5. Cheap , small and easy to use. 6. Capable of repeated use.
  • 11. ENZYME BASED BIOSENSOR  ENZYME BASED BIOSENSORS are prepared by immobilization methods such as ionic bonding, covalent bonding or van der Waals forces.  The commonly used enzymes are peroxides, aminoxidases, polyphenoloxidases, oxidoreductases etc.
  • 12. IMMUNOSENSORS  IMMUNOSENSORS are prepared by antigen antibody reaction .  antibodies are highly attracted towards their respective antigen and forms antigen antibody reaction .  Labelled antigens or antibodies may be used in biosensors based on enzyme linked immunosorbent assay(ELISA) principles.  Labelled antigen means attachment of specific tag to antibody for detection of protein.  ELISA can be used to detect an antibody to HIV.
  • 13. DNA BIOSENSORS  In this single stand nucleic acid molecule is bind to its complementary strand in a sample.  DNA biosensors consist of an immobilized DNA strand to detect the complimentary sequence by DNA–DNA hybridization.  Hybridization means two single stranded DNA bound to form double stranded DNA.
  • 14. Electrochemical Biosensors:  Electrochemical biosensors are simple devices measures the electric current, ionic or conductance changes carried out by bio electrodes
  • 15. Amperometric Biosensors:  These biosensors are based on the movement of electrons (i.e. determination of electric current) as a result of enzyme-catalysed redox reactions.  This contain enzyme electrode or chemically modified electrodes.  A redox reaction catalyzed by an enzyme is directly bind to an electrode .  The electrons are transferred from the substrate to the electrode via enzyme and redox mediator.  It is a good example of “Amperometric Biosensors” , widely used throughout the world by diabetic patients.
  • 16. Blood glucose monitoring is the use of a glucose meter for testing the concentration of glucose in the blood . Important in diabetes management, a blood glucose test is typically performed by piercing the skin.
  • 17. Potentiometric Biosensors  In these biosensors, changes in ionic concentrations are determined by use of ion- selective electrodes.  pH electrode is the most commonly used ion- selective electrode .  These biosensors consist of membrane containing immobilized enzyme and surrounding by pH meter.  The other important electrodes are ammonia- selective and CO2 selective electrodes.
  • 18. •The potential difference obtained between the potentiometric electrode and the reference electrode can be measured. •Potential difference is proportional to the concentration of the substrate. •The major limitation of potentiometric biosensors is the sensitivity of enzymes to PH , ammonia and carbon dioxide .
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  • 20. Conductimetric Biosensors  The changes in ionic species is transferred into electrical conductivity ( electrical current pass through the solution )in Conductimetric Biosensors.  Conductivity measurement is based on the biocatalytic reaction of sample on an electrode.  The conductimetric transducer consist of reference electrode and working electrode.  Both electrodes are coated with nata de coco membrane and enzyme is immobilized only on working electrode  Nata de coco, a bacterial cellulose as a result of coconut water fermentation, is a conductive polymer with a electrical conductivity of 553 μS/cm( Micro Siemens per cm) and has high mechanical stability.  Example is Urea biosensor utilizing immobilized urease.  urea biosensors are very successfully used during dialysis and renal surgery
  • 21. Thermometric or Calorimetric Biosensors  Biological reactions are associated with the production of heat in thermometric biosensors.  They are more commonly referred to as thermal biosensors or calorimetric biosensors.  Thermometric biosensors are in use for the estimation of serum cholesterol.  Thermometric biosensors can be used as a part of Enzyme-Linked Immuno-Sorbent Assay (ELISA) and the new technique is referred to as Thermometric ELISA (TELISA).
  • 22. ELISA  ELISA stands for enzyme-linked immuno sorbent assay.  It is a commonly used laboratory test to detect antibodies in the blood.  This test is often used to see if any one have been exposed to viruses or other substances that cause infection.
  • 23. A diagrammatic representation of a thermal biosensor is in Fig. It consists of a heat insulated box fitted with heat exchanger (aluminum cylinder). The reaction takes place in a small enzyme packed bed reactor. As the substrate enters the bed, it gets converted to a product and heat is generated. The difference in the temperature between the substrate and product is measured by Thermistors. Even a small change in the temperature can be detected by thermal biosensors.
  • 24. Optical Biosensors  Optical biosensors are the devices that utilize the principle of optical measurements (absorbance, fluorescence, internal reflection spectroscopy etc.  It determines changes in light absorption between the reactants and products.  Optical biosensors primarily involve enzyme and antibodies as the transducting elements.  these biosensors usually do not require reference sensors, as the comparative signal can be generated using the same source of light as the sampling sensor.  Fiber optic biosensor is most common example of Biosensor.
  • 25. Fig.Represents the Fibre Optic Lactate Biosensor. In this changes in molecular O2 concentration is measured by determining the quenching (reducing ) effect of O2 on a fluorescent dye. O2 has a quenching (reducing) effect on the fluorescence. Hence the amount of fluorescent generated by dyed film is depend on oxygen. Lactate present in reaction mixture utilize oxygen and decrease in the reducing effect of oxygen resulting to increase in fluorescent output measured. The following reaction is catalyzed by the enzyme lactate monooxygenase. lactate O2 +LACTATE CO2+Acetate+H2O monooxygenase
  • 26. Piezoelectric Biosensors  Piezoelectric biosensors are based on the principle of acoustics (sound vibrations), hence they are also called as acoustic biosensors.  Piezoelectric crystals form the basis of these biosensors. The crystals with positive and negative charges vibrate with characteristic frequencies.  Adsorption of certain molecules on the crystal surface alters the resonance frequencies which can be measured by electronic devices.  A biosensor for formaldehyde has been developed by incorporating formaldehyde dehydrogenase.  A biosensor for cocaine in gas phase has been created by attaching cocaine antibodies to the surface of piezoelectric crystal.
  • 27. Whole Cell Biosensors  In the whole cell biosensors or microbial biosensors ,immobilized whole cell of microorganism or their organelles are used.  These may employed live or dead microbial cell.  The microbial cells are easily available with less cost .  They are less sensitive to variation in pH and temperature compared to enzymes.
  • 28. Application of Biosensors  The four applications are: 1. Applications in Medicine and Health 2. Applications in food Industry 3. Applications in Pollution Control 4. Applications in Military.
  • 29. 1.Applications in Medicine and Health  Biosensors are successfully used for the quantitative estimation of several biologically important substances in body fluids e.g. glucose, cholesterol, urea.  Glucose biosensor is used for diabetic patients for regular monitoring of blood glucose.  Blood gas monitoring for pH, pCO2 and pO2 is carried out during critical care and surgical monitoring of patients.  Several toxic compounds produced in the body can also be detected.
  • 30. 2. Applications in food Industry  Biosensors can be used for monitoring of fermentation products and estimation of various ions.  Thus, biosensors help for improving the fermentation conditions for a better yield.  Now a days, biosensors are employed to measure the odour and freshness of foods.  Freshness of stored fish can be detected by ATPase.  ATP is not found in spoiled fish and this can be detected by using ATPase.  Pharmaceutical company has developed immobilized cholesterol oxidase system for measurement of cholesterol concentration in foods (e.g. butter).
  • 31. 3. Applications in Pollution Control (Environmental Monitoring)  Biosensors are very helpful to monitor environmental (air, water) pollution.  The concentrations of pesticides can be measured by biosensors.  Biosensor have wide application for measurement estimation and control of water, soil, air contamination .  Ex. Determination of pesticides by potentiometric biosensor.
  • 32. 4. Applications in Military ( Defense)  Biosensors have been developed to detect the toxic gases and other chemical agents used during war.  DNA Biosensor is used to detect criminals.