BIOSENSORS

UNIT-1
Dr K. Chandrasekhar
NIT Warangal
Telangana
India
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What is Biosensor?
A sensor is a converter that measures a physical quantity and
converts it into a signal which can be read by an observer or
by an instrument.
A biosensor is an analytical device which converts a
biological response into an electrical signal.
 A biosensor is an analytical device, used for the detection of
an analyte, that combines a biological component with a
physicochemical detector.
A device incorporating a biological sensing element either
intimately connected to or integrated within a transducer.
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FATHER OF THE BIOSENSOR

Professor Leland C Clark Jnr 1918–2005

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A historical perspective
1916 ► First report on immobilization of proteins : adsorption of invertase on
activated charcoal
1922 ► First glass pH electrode.
1956 ► Clark published his definitive paper on the oxygen electrode.
1962 ► First description of a biosensor: an amperometric enzyme electrodre for
glucose (Clark).
1969 ► Guilbault and Montalvo – First potentiometric biosensor: urease
immobilized on an ammonia electrode to detect urea.
1970 ► Bergveld – Ion selective Field Effect Transistor (ISFET).
1975 ► Lubbers and Opitz described a fibre-optic sensor with immobilised
indicator to measure carbon dioxide or oxygen.
1975 ► First commercial biosensor ( Yellow springs Instruments glucose biosensor).
1975 ► First microbe based biosensor, First immunosensor.
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A historical perspective
1976 ► First bedside artificial pancreas (Miles).
1980 ► First fibre optic pH sensor for in vivo blood gases (Peterson).
1982 ► First fibre optic-based biosensor for glucose.
1983 ► First surface plasmon resonance (SPR) immunosensor.
1984 ► First mediated amperometric biosensor: ferrocene used with glucose
oxidase for glucose detection.
1987 ► Blood-glucose biosensor launched by MediSense ExacTech.
1990 ► SPR based biosensor by Pharmacia BIACore.
1992 ► Hand held blood biosensor by i-STAT
1996 ► Launching of Glucocard
1998 ► Blood glucose biosensor launch by LifeScan FastTake.
1998 ► Roche Diagnostics by Merger of Roche and Boehringer Mannheim.
Current ► Nanoparicles, Quantom dots, nanowire, nanotube, etc.
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• A nanoparticle is a microscopic particle with at least one dimension

less than 100 nm.

• Quantum dots (QD) are very small semiconductor particles, only

several nanometres in size, so small that their optical and electronic
properties differ from those of larger particles.

• A nanowire is a nanostructure, with the diameter of the order of a

nanometer (10−9 meters). It can also be defined as the ratio of the
length to width being greater than 1000.

• A nanotube is a nanoscale material that has a tube-like structure.

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Basic Characteristics of a Biosensor

1. LINEARITY: Linearity of the sensor should be high forthe detection of

high substrate concentration.

2. SENSITIVITY: Value of the electrode response per substrate

concentration.

3. SELECTIVITY: Chemicals Interference must be minimised for

obtaining the correct result.

4. RESPONSE TIME: Time necessary for having 95% of the response.

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A successful biosensor must possess at least some of the
following beneficial features

1. The biocatalyst must be highly specific for the purpose of the analyses,
be stable under normal storage conditions and, except in the case of
colorimetric enzyme strips and dipsticks (see later), show good
stability over a large number of assays (i.e. much greater than 100).

2. The reaction should be as independent of such physical parameters as

stirring, pH and temperature as is manageable. This would allow the
analysis of samples with minimal pre-treatment. If the reaction
involves cofactors or coenzymes these should, preferably, also be co-
immobilised with the enzyme.
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3. The response should be accurate, precise, reproducible and linear over

the useful analytical range, without dilution or concentration. It should
also be free from electrical noise.

4. If the biosensor is to be used for invasive monitoring in clinical

situations, the probe must be tiny and biocompatible, having no toxic or
antigenic effects. If it is to be used in fermenters it should be sterilisable.
This is preferably performed by autoclaving but no biosensor enzymes
can presently withstand such drastic wet-heat treatment. In either case, the
biosensor should not be prone to fouling or proteolysis.
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5. The complete biosensor should be cheap, small, portable and

capable of being used by semi-skilled operators.

6. There should be a market for the biosensor. There is clearly little

purpose developing a biosensor if other factors (e.g. government
subsidies, the continued employment of skilled analysts, or poor
customer perception) encourage the use of traditional methods and
discourage the decentralisation of laboratory testing.
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Basic priciple of Biosensors
Basic principle of biosensor involved in three element: -

1. First: The biological recognization element which is highly

specific towards the biological material analytes produces.
2. Second: Transducesrs detect and transduces signal from
biological target - receptor molecule to electrical signal.
3. Third: After transduction signals from biological to
electrical signal where its amplification is necessary and
takes place and read out in detector, after processing the
values are displayed for monitor and controlling the system .
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Bioreceptor Molecular
Analyte recognition

Basic priciple of
BIOSENSING Biosensors Transducer
PRINCIPLE

Data recognition
Measurements
and display
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Components of a Biosensor

Detector
Introduction to Biosensors: 1st and 2nd Components

Bioreceptor Transducer

Absorption
Antibody Fluorescence
Optical Interference

potentiometric
Enzyme Electrochemical amperometric
conductimetric

Nucleic Acid (DNA) Mass based

Cell Temperature based

Dielectric properties
Electric & Permeability properties
MIP
Magnetic Voltage or Current
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(BIORECEPTORS)
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1st Components: Bioreceptors
 In a biosensor, the bioreceptor is designed to interact with the
specific analyte of interest to produce an effect measurable by the
transducer.
 High selectivity for the analyte among a matrix of other chemical
or biological components is a key requirement of the bioreceptor.
 While the type of biomolecule used can vary widely, biosensors
can be classified according to common types of bioreceptor
interactions involving: antibody/antigen, enzymes/ligands, nucleic
acids/DNA, cellular structures/cells, or biomimetic materials.
Introduction to Biosensors: 1st and 2nd Components 28-8-2018

Bioreceptor Transducer

Absorption
Antibody Fluorescence
Optical Interference

potentiometric
Enzyme Electrochemical amperometric
conductimetric

Nucleic Acid (DNA) Mass based

Cell Temperature based

Dielectric properties
Electric & Permeability properties
MIP
Magnetic Voltage or Current
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Types of Bioreceptors:
 Antibody/antigen interactions
 Artificial binding proteins
 Enzymatic interactions
 Affinity binding receptors
BIORECEPTORS  Nucleic acid interactions
 Epigenetics
 Organelles
 Cells
 Tissue
 Types of Transducers
Transducer

Absorption
Fluorescence Measure changes in light intensity
Optical Interference

potentiometric
Electrochemical amperometric Measure changes in electric distribution
conductimetric

Mass based Measure changes in mass of the analyte

(Piezoelectric detection)

Temperature based Measure changes in temperature of the

(Calorometric detection) analyte
Dielectric properties
Electric & Permeability properties Measure changes in Electric and Magnetic
Magnetic Voltage or Current properties of analyte
Transducer Types of Biosensors
Absorption
Optical
Fluorescence
Interference
1. Optical Biosensor

Electrochemical
potentiometric
amperometric 2. Potentiometric Biosensor
conductimetric

Mass based
3. Amperometric Biosensor
(Piezoelectric detection)

Temperature based
4. Piezo-electric Biosensor
(Calorometric detection)

Electric &
Dielectric properties
Permeability properties
5. Calorimetric Biosensor
Magnetic Voltage or Current
1. Optical Biosensor
These sensors rely on a change in the refractive index, absorbance and
fluorescence properties of analyte molecules or a chemo-optical
transducing medium.
2. Potentiometric Biosensors
In these sensors, the biological recognition element converts the
recognition process into a potential signal to provide an analytical signal.
3. Amperometric Biosensor
Amperometric sensors (AS) measure the current change resulting from
the oxidation and reduction of an electroactive species while a constant
potential is being applied.
The change in the current is related to the concentration of the species in
solution.
4. Piezo-electric Biosensor
• Piezoelectric sensors are sensitive mass-to-frequency transducers that
are of interest for potential applications in analytical chemistry.
• A piezoelectric device is sensitive to changes in the mass, density, or
viscosity of samples in contact with its active surface.

5. Calorimetric Biosensor
• The fundamental principle of a calorimetric biosensor relates to the
measurement of the changes in temperature in the reaction between the
biorecognition element and a suitable analyte.
• The calorimetric biosensors measure heat evolution (enthalpy change)
following biochemical reactions.
Main technical definitions:
1. Calibration
2. Selectivity
3. Sensitivity
4. Reproducibility
5. Detection limits
6. Response time
1. Calibration
• The first consists in measuring the biosensor response to
interfering substance addition: a calibration curve for each
interfering substance is plotted and compared to the analyte
calibration curve, under identical operating conditions.

2. Selectivity
• Selectivity is expressed as the ratio of the signal output with
the analyte alone to that with the interfering substance alone,
at the same concentration as that of the analyte.
3. Response time/Speed
• Transient response time corresponds to the time necessary for
the first derivative of the output signal to reach its maximum
value (dR/dt)max following the analyte addition.

4. Reproducibility
• Reproducibility is a measure of the scatter or the drift in a
series of observations or results performed over a period of
time.
• It is generally determined for the analyte concentrations
within the usable range.
5. Detection limit
• The limits of detection (LOD) and of quantification (LOQ)
take into account the blank and the signal fluctuation (noise).
(The working concentration range, which may considerably extend the linear
concentration range, is determined by the lower and upper limits of quantification.)

6. Sensitivity
• The sensitivity of the sensor is defined as the slope of the
output characteristic curve or, more generally, the minimum
input of physical parameter that will create a detectable
output change.
(In some sensors, the sensitivity is defined as the input parameter change required to
produce a standardized output change.)
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