ENZYME PROBES –

BIOSENSORS
Parmar Amita
Roll No. 30
M.Sc. Microbiology Sem 3
Department of Biosciences
 CONTENTS OF THIS PRESENTATION
• Introduction of biosensors

• Basic components of enzyme biosensors

• Types of biosensors

• Enzyme biosensors for COVID -19 detection in the air

• Standard M nCoV real-time detection kit

• Applications of biosensors

• Conclusion

• References & bibliography

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 INTRODUCTION
• What is a Biosensor ?
A biosensor is an analytical device, used for the
detection of a chemical substance, that
combines a biological component with a
physicochemical detector.
The term ‘biosensor’ was introduced by
Cammann in 1977.

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FATHER OF BIOSENSORS
The first 'true' biosensor was
developed by Leland C. Clark, Jr in
1956 for oxygen detection.

He is known as the 'father of

biosensors' and his invention of the
oxygen electrode bears his name:
'Clark electrode’

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 A biosensor is a device
composed of two elements :
• A bioreceptor that is an immobilized
sensitive biological element
recognizing the analyte.

• A transducer is used to convert

(bio)chemical signal resulting from the
interaction of the analyte with the
bioreceptor into an electronic one.
(General structure of a biosensor)
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 TYPES OF BIOSENSORS
• Based on Bioreceptors :

Enzyme biosensor

Microbial Biosensor

Affinity biosensor

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 Enzyme Biosensors :
An enzyme biosensor is an
analytical device that
combines an enzyme with a
transducer to produce a
signal proportional to target
analyte concentration.

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 Enzyme based biosensors are divided into several categories based on the
transducer types :

(1) (2) (3) (4)

Electrochemical Enzymes-based Enzyme based Enzyme-based

Biosensors optical biosensors Thermistors Piezoelectric
biosensors

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 (1) Electrochemical sensors :

They are one of the most extensively used

biosensors whose working mechanisms are
based upon the electrochemical properties of
transducers and analytes .

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The majority of the current glucose biosensors are of the
electrochemical type. The use of electrochemical biosensors provides
advantages such as simplicity, rapidity, low cost, and high sensitivity.

(A typical design of an enzyme modified

(Glucose oxidase Biosensor)
electrochemical biosensor)

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 (2) Enzymes-based optical biosensors :

The optical transducers of enzyme-based biosensors measure changes in optical

properties such as fluorescence intensity, light absorption, reflectance,
chemiluminescence, evanescent wave, reflective index, and Raman scattering, resulting
from the interaction of a biocatalyst with a target analyte.

One of the earliest examples of an optical biosensor for clinical applications is a test strip
for glucose in urine, commercialized in 1957 .

The working principle of the sensor utilized a cellulose pad coimmobilized with GOx and
peroxidase in a cascade manner.
Firstly, GOx catalyzes the oxidation of glucose to gluconic acid and hydrogen peroxide.

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The second immobilized peroxidase enzyme then catalyzes the reaction
between the formed hydrogen peroxide and orthotolidine, to yield a deep blue-
colored product. The change in blue color could be visually determined by the
eye, and was used as a semiquantitative measurement of glucose
concentration in urine.

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 (3)Enzyme-Based Thermistors

Almost all enzymatically catalyzed reactions are exothermic, which may be used as a
basis for measuring the rate of reaction and the analyte concentration.

A thermistor is a type of resistor resistance that is dependent on temperature, and it

measures changes in temperature by molar enthalpy change (∆H) in the form of
electrical signals, such as resistance

Thermistor-based calorimeters, popularly known as enzyme thermistors (ET), use

thermistors to measure electrical changes due to changes in temperature following a
biocatalytic reaction, and this system is especially exploited for quantification
purposes.

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The main drawbacks of enzyme
thermistor is non-specificity,
because thermal signal is
dependent only on the underlying
reaction.

The thermometric enzyme-linked

immunosorbent assay (TELISA),
for the assay of endogenous and
exogenous compounds in
biological fluids has been
developed by Mattiasson et al.

The figure shows the schematic

illustration of the TELISA method
for both direct competitive (A)
and sandwich (B) formats.
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(4) Enzyme-Based Piezoelectric Biosensors

● The most common type of

piezoelectric biosensor is
quartz crystal microbalance
(QCM), which is able to
determine nanograms of
material.
● The sensor consists of a thin
wafer of quartz-sensing
crystal plated with metallic
electrodes on either sides of
the crystal by means of vapor
deposition.

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 Approaches in Improving Enzyme Usage in Biosensors
● Biological Modification
(1) Site-Directed Mutagenesis

(a) Enzyme Amino Acid Substitution

(b) Enzyme Amino Acid Removal
(c) Non-Natural Amino Acid Incorporation
(d) Enzymatic Addition of a Genetic Tag

(2) Fusion Protein Technology

● Chemical Modification
(1) Site-Specific Chemical Modification
(2) Nonspecific Modification of the Enzyme Surface
(3) Chemical Cross-Linking
(4) Use of Polymers

● Multi-Enzyme System 15
 ENZYME BIOSENSORS FOR COVID -19 DETECTION IN THE AIR :
A team of researchers from Empa (Zürich,
Switzerland) and ETH Zurich (Zürich, Switzerland)
usually work on measuring, analyzing and reducing
airborne pollutants such as aerosols and artificially
produced nanoparticles,

has succeeded in developing a novel sensor for

detecting the new coronavirus that could be used to
measure the concentration of the virus in the
environment.

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Biosensors for COVID-19 are mostly designed
on the surface nucleoproteins, which binds to
the host angiotensin-converting enzyme 2
(ACE-2) receptor and the internal genetic
material .

Nanomaterials such as gold and carbon have

gained vast interest in sensor technology and
have produced promising devices for sensing
the virus and its biomolecules.

These nanomaterials fused with analyte such as

complementary single-stranded nucleic acid (Image: New biosensor detects COVID-19
aptamer could be a new strategy for detecting virus in air)
SARS-CoV-2 in clinical samples.

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The sensor combines two different effects to detect the virus safely and
reliably: an optical and a thermal one.

The sensor is based on tiny structures of gold, so-called gold nanoislands,

on a glass substrate.

Artificially produced DNA receptors that match specific RNA sequences of

the SARS-CoV-2 are grafted onto the nanoislands.

The receptors on the sensor are the complementary sequences to the

virus' unique RNA sequences, which can reliably identify the virus.

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(Schematic structure of SARS-Cov-2 and its possible targets for diagnosing)

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 STANDARD M nCoV Real-Time Detection kit
Information Detail
Test time Within 90 mins
Nasopharyngeal swab,
Specimen
Oropharyngeal swab, Sputum
Storage temperature -25~ -15℃

BENEFITS :
Fast & Easy High sensitivity & specificity

One step reaction for identification &
detection of COVID-19
One step real time RT-PCR
Provide all reagents required for PCR
Designed according to WHO interim
guidance for laboratory testing of
novel coronavirus COVID-19
nCoV primers/probes, ORF 1ab,E
gene
Provide internal control
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This kit is based on TaqMan probe real-time
fluorescent PCR technology.

Cycle conditions :

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 APPLICATIONS OF BIOSENSORS
1) Applications in Medicine and Health as diagnostic
Tools
2) As Analytical Tools for the Assessment of Food
Quality and Food Safety
3) Applications in Pollution Control
4) Applications in defence
5) Applicaction in agriculture and so on.

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 ENZYME BIOSENSORS PRODUCING
COMPANIES WORLDWIDE
• Systea S.P.A (New Delhi ,India )
• Metrohm AG (Banglore, India)
• Enveo (Bhubneshwar , India)
• Aptomar AS (Mumbai , India)
• Abbott Laboratories (Chicago , United States)
• ABTECH Scientific Inc. (Richmond, United States)
• Affymetrix(California, United States )
• Bayer AG( Leverkuen ,Germany )
• Biacore International Ab( Upsala , Sweden)
• Cygnus, Inc( Ponderay, United States)
• Diagnoswiss(zi Les Ilettes –Switzerland)
• Lifescan(California, United States)
• Neogen Corporation( Michigan , United States)

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 CONCLUSION
Enzyme-based biosensing has proven to be a valuable technique for the qualitative and
quantitative analysis of a variety of target analytes in biomedicine, environmental, food
quality control, agricultural, and pharmaceutical industry.

In comparison with conventional analytical methodologies, enzyme-based biosensors offer

significant benefits, such as miniaturization, real-time diagnosis capability, high sensitivity
and specificity, minimum sample preparation, and high-throughput, bedside clinical testing
and portability.

With the development of computer software and hardware technology, sensor technology
will also develop, and the biological system of closed-loop control will be possible. In the
future, biosensors will be widely used in many fields, such as food detection, medical care,
disease diagnosis, environmental monitoring, fermentation industry, and so on. 24
 REFERENCES :
• Hoang Hiep Nguyen 1,2,†, Sun Hyeok Lee 1,2,†, Ui
Jin Lee 1,3, Cesar D. Fermin 4 and Moonil Kim.
Immobilized Enzymes in Biosensor Applications.
Sensors 2020, 20(6), 1721

• www.empa.ch , A new biosensor for the COVID-19

virus

• Anastasios Economou , Steve Karapetis , Georgia-

Paraskevi Nikoleli , Dimitrios P. Nikolelis . Enzyme-
based Sensors . Advances in Food Diagnostics
(pp.231-250)

• Rachel Samson1,2 · Govinda R. Navale1,2 ·

Mahesh S. Dharne . Biosensors: frontiers in rapid
detection of COVID-19. 3 Biotech 10, pp. 385
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 THANK YOU
MAY YOU ALL HAVE A GREAT DAY !