BIO-MEDICAL ENGINEERING

By-
Dr. Pinku Ranjan
Assistant Professor
ABV-IIITM GWALIOR,
INDIA

1
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan
Biosensors
Sensing means becoming aware of…
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 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Biosensors
• Definition
 An analytical device, which is a combination of biological
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component with a physicochemical detector used for the sensing

an analyze (biological samples)
 Converts biological response into electrical signals
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

What if I can see?

Add instrumentation…
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 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Sensors applications
The primary goal is detect danger so an action can be
taken… Gas concentration Pressure sensing
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sensing

We need a property that correlates to what I want to measure

 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

What I want to measure in medicine?

The concentration of a biomarkers can tell me the

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nature of a disease and what stage it is in

WHITE CELLS INFECTION

KIDNEY
CREATININE
MALFUNCTION
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Sensibility range
PSA (prostate specific antigen) is a biomarker
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related to the existence of prostate cancer…

Antigen Concentration Organ Confinement

PSA 4-10 ng/ml 75%

PSA >10ng/ml >50%

 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Nanosensors
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If I want to measure
something small, I need
something small…
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

What are biosensors?

• A device that uses specific biochemical reactions
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mediated by isolated enzymes, immuno systems, tissues,

organelles or whole cells to detect chemical compounds
usually by electrical, thermal or optical signals.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Principle of Biosensors
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 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Required Characteristics
• Sensitivity • Utility
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• Low detection • Field portability

limits • Ruggedness
• Cost • Reproducibility
• Simplicity • Ease of calibration
• Reliability • Stability
• Speed • Room for
• Accuracy improvement
• Precision
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Types of biosensors

• Based on use of different biological material and sensor

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devices following are the main types –

-Electro-chemical Biosensor
-Amperometric Biosensor
-Thermistor containing Biosensor
-Bioaffinity sensor
-Whole cell Biosensors
-Opto-electronic Biosensor
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Electro-chemical Biosensor
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• This type of biosensor has been developed by using

electronic device like field effect transmitors or light
emitting diode.
• Field effect transmitors measures charge accumulation
on their surface while light emitting diode measures
photo response generated in a silica based chip as an
alternating current.
• The field effect transmitor measures a biochemical
reaction at the surface and induce in to current.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Amperometric Biosensor

• Amperometric biosensors are those which measures the

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reaction of anylate with enzyme and generate electrons

directly or through a mediator.
• Enzyme electrodes are a new type of biosensor which have
been designed for the assay of substrates such as urea,
amino acid, glucose, alcohol and lactic acid. The enzyme is
embedded in thin permeable membrane and produce
oxygen, hydrogen, ammonia, carbon dioxide and other
small molecules. This is detected by specific sensor.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Thermistor Containing Biosensors

• Thermistor is used to record even a small change in

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temperature (between 0.1- 0.001C) during biochemical

reactions.
• By immobilizing enzymes like cholesterol oxidase,
invertase, tyrosinase etc. thermistors have been
developed.
• Thermistors are also developed for Ag-Ab with high
sensitivity in case of thermometric Enzyme Linked
Immunosorbant Assay (ELISA).
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Bioaffinity Sensors

• This measures the concentration of the determinants i.e.

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substrates based on equilibrium binding.

• Such sensors involves the use of radiolabelled,
enzymelabelled or fluorescence labelled substance.
• This type of biosensors are the use of lectin receptors for
saccharides estimation, hormone receptors for hormone,
drug receptors for drug, antibodies receptors for
antigens and nucleic acid for inherited disease and finger
printing.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Whole cell Biosensors

• In this device, either immobilized whole cell of

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microorganisms or their organelles are used.

• Azotobactor is used to measure concentration of nitrate
within 5-10 min.
• The other organism used are Methylomonas for
methane, Trichosporon for acetate and ethanol.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Opto-electronic Biosensors

• In this type of biosensor either enzymes or antibodies

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are immobilized on the surface of a membrane. For

measuring color, biosensor with enzyme and dye is
immobilized to a membrane.
• When substrate is catalysed to yield product, change in
pH of the medium occur. This result in changes in dye
membrane complex. The change in color is measured by
using a light emitting diode and a photodiode.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Evolution of the Medical Biosensor

• Mid 1950’s - Leland C Clark invents an electrode

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that measures dissolved oxygen in the blood of

patients undergoing surgery.
• He surrounded a platinum electrode and
reference electrode with a plastic membrane
permeable to gases. The voltage differential
measured between the two gave the rate at
which the oxygen was being diffused.
ABV-IIITM GWALIOR BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

• Clark extended this electrode idea to include glucose sensors.

• However these never made their way into routine patient care.
This was due to flaws in accuracy when clots formed around
the electrode.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Uses of Biosensors

•
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Environmental Monitoring
• Military
• Law Enforcement
• Medical
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Present Applications of Biosensors

• Medical Care (both clinical and laboratory

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use)
• The determination of food quality
• The detection of environmental pollutants
• Industrial Process Control
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Applications (cont’d.)

• Biosensors in process control will be able to measure materials in

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the process flow of temperature, pressure and the acidity

readings.
• The development of biosensors in industry can improve
manufacturing techniques, which would allow for a wider range
of sensing molecules to be produced at a cheaper rate.
• In the field of medicine, tumor cells are used as a biosensor to
monitor chemotherapeutic drug susceptibilities.
• Biosensors also play a role in the manufacturing of
pharmaceuticals and replacement organs such as an artificial
pancreas for diabetics.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Applications (cont’d.)
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• It is found that biosensors are widely used in environmental

testing by using a “flow-through” system to monitor wastewater
(a flow-through system includes instruments using surface
plasmon resonance).
• These censors are not only restricted to the use of molecules.
Rather living microorganisms such as bacteria can be used to
detect substances such as vitamin B12.
• Conclusively, biosensors play a part in the field of medicine,
environmental quality, and industry mainly by identifying
material and the degree of concentration present.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Example of biosensors
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Pregnancy test

Detects the hCG protein in urine. Interpretation and

data analysis performed by the user

Glucos monitoring device (for diabetes patients)

Monitors the glucose level in the blood.

Interpretation and data analysis performed by a
microprocessor.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Example of biosensors
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Infectous disease biosensor from RBS

Data analysis and interpretatoin

performed by a microprocessor

Old time coal miners’ biosensor

Data analysis and interpretation performed by the coal miner.
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Research Biosensors
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Biacore Biosensor platform

General and flexible, good tool for
development of specific biosensors

For a comprehensive list of research biosensor suppliers

see: www.realtimebiosensor.com
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Working Principle
 Enzyme substrate specificity

 Bio & Sensor elements coupled by Membrane & Matrix Entrapment (Fig. – 3),
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Physical adsorption, Covalent Bonding

Fig. - 1 Fig. - 2

Fig. - 3
ABV-IIITM GWALIOR BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Resonant/ Piezoelectric Biosensors

 Analyte - Antigen
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 Bioelement – Antibody

 Transducer – Acoustic wave transducer

 Signal – Change in frequency/mass measured
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Optical Detection Biosensor

 Analyte - Antigen

 Bioelement – Antibody
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 Transducer – Silicon wafer exposed to UV light

 Signal – Light measured

UV light
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Thermal Detection Biosensor

 Analyte - Antigen
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 Bioelement – Immobilized enzyme

 Transducer – Thermistor

 Signal – Heat measured

 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Ion Sensitive Biosensor (ISFET)

 Analyte – Antigen (Ion)

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 Bioelement – Antibody
 Transducer – Semiconductor – Field Effect
Transistor (FET)
 Signal – pH change measured
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Electrochemical Biosensors
Conductimetric Amperometric Potentiometric
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Glucose Biosensors

Principle – Change in chemical reaction produce or

consume ions/ electrons, which causes electrical change
and it is measured.

Glucose biosensors first invented by Leland C. Clark

(Used Platinum eletrodes)
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Glucose Biosensors
 Analyte – Glucose (Substrate)
 Bioelement – Immobilized Enzyme Membrane – Glucose Oxidase (GOD)
 Transducer – Graphite electrode
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 Signal – Change in chemical ion measured

 Strip – Glucose Oxidase GOD

 GOD reacts with glucose and produce
gluconic acid
 More glucose, more oxygen is consumed
 Less glucose, more hydrogen peroxide is
consume
 A mediator replaces oxygen, accepting the
electron and passes to the electrode
 This generates current
 This current is reported as glucose
concentration
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Application

DNA Detection
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 Uses – Detection of Anthrax, Botulinum, Harmful pathogens

 Objective – Isolate single DNA-DNA, Antigen-antibody bond

 Methods

 1. Polymerase Chain Reaction (PCR) – multiple copies of DNA

 2. Force Amplified Biological Sensor(FABS)

 3. Force Differentiation Assay (FDA)

 4. Bead Array Counter (BAC)

 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Application

Pollution Detection
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 Uses – Detection of specific xenobiotic pollutants

 Objective – Isolate specific pollutants, ex; lux gene coupled ( rDNA technology)
- reporter gene - fluorescence
 Methods

 1. Spectrometry (UV, IR, Visible)

 2. Chromatography (GC, TLC, HPLC)

 3. Immunoassay (ELISA)
 4. Monoclonal antibodies
 BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan

Application

Food Industry
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 Uses
 Measurement of carbohydrates, alcohols
 Sterility, food safety in meats, beverages (Beer, soft drinks) etc.
 Objective – Used in quality assurance
 Methods
 1. Flow injection system
ABV-IIITM GWALIOR BIO-MEDICAL ENGINEERING Dr. Pinku Ranjan