Biosensor

Classification
BIOSENSORS, MEMS AND BIO -ROBOTICS
 DEFINITION
 A biosensor is a device that uses a biological component to detect a
chemical substance and converts this detection into a measurable
signal.

 A biosensor is a device that measures biological or chemical reactions

by generating signals proportional to the concentration of an analyte
in the reaction.

Biosensors are employed in applications such as

1. Disease monitoring,
2. Drug discovery,
3. The detection of pollutants,
4. Disease causing microorganisms and
5. Markers that are indicators of a disease in bodily fluids (blood, saliva,
sweat).
Nevertheless, two main characteristics of a biosensor definition have
remained, i.e., the use of a biochemical recognition element, and the
rather simple and compact design that provides real time measurement.
 SCHEMATIC REPRESENTATION OF A BIOSENSOR.

 Analyte- It is a substance of interest that needs

detection. For instance, glucose is an ‘analyte’ in a
biosensor designed to detect glucose.

 Bioreceptor- A molecule that specifically recognises

the analyte is known as bioreceptor. Enzymes, cells,
DNA and antibodies are some examples of
bioreceptors.

 Bio-recognition- It is the process of generation of a

signal (in the form of light, heat, pH, charge or mass
change, etc.) upon interaction of the bioreceptor with
the analyte.
 Biological sensing (recognition) element (Bioreceptors)
 This component is also known as a sensor or detector element and is
responsible for sensing or detecting the presence and/or the
concentration of the target analyte or substance.

 This is a biological component, which serves as a biochemical receptor

that specifically recognises the target analyte.

 Analyte (sample)- It is a substance of interest that needs detection. For

instance glucose is an ‘analyte’ in a biosensor designed to detect glucose.

 When the biological receptor interacts with a target analyte, it generates

a signal in the form of light, heat, pH, charge or mass change.

 This material should be highly specific, stable under storage conditions and
must be immobilized.

 Furthermore, the biological receptor should be capable of selectively

detecting the target compound or analyte in the test sample.

 Biological receptor determines the sensitivity of the entire device through

the generation of the physicochemical signal that is monitored by the
transducer.
 General components of biosensor
Generally, biosensors are composed of three main
components as depicted in Figure. These include

1. A biological recognition-sensing element,

2. Transducer,
3. Signal processing unit.
 Biosensors vs Medical Sensors
Aspect Biosensors Medical Sensors

Definition Analytical devices combining biological and physicochemical Devices designed to monitor physiological
components to detect biological molecules. parameters of the human body.

Components - Biological elements (e.g., enzymes, antibodies, DNA). - Sensor unit measures physical parameters.
- Transducer converts biological signals. - Circuitry processes and outputs data.

Based on biological interactions (e.g., enzyme-substrate, antibody- Measures physical or chemical parameters, often
Detection Mechanism antigen). non-biological.

- Biochemical detection (e.g., glucose monitoring). - Physiological monitoring (e.g., heart rate,
Applications - Pathogen identification. temperature).
- Drug discovery. - Medical imaging (e.g., ultrasound, MRI).

Converts biological responses into electrical, optical, or measurable Measures and outputs physical or physiological
Output signals. data.
- Glucose biosensors. - Pulse oximeters.
Examples - DNA biosensors. - ECG sensors.
- Immunosensors for disease markers. - Blood pressure monitors.
Does not require biological elements; focuses on
Biological Interaction Requires biological recognition elements for detection. physical or chemical sensing.
 Major areas of applications for biosensors.
Biosensors have a very wide range of applications that aim to
improve the quality of life.

This range covers their use for environmental monitoring,

disease detection, food safety, defence, drug discovery and
many more.

 One of the main applications of biosensors is the detection

of biomolecules that are either indicators of a disease or
targets of a drug. For example, electrochemical biosensing
techniques can be used as clinical tools to detect protein
cancer biomarkers [14-16].

 Biosensors can also be used as platforms for monitoring

food traceability, quality, safety, and nutritional value
[17,18]. These applications fall in the category of "single
shot" analysis tools, i.e. where cost effective and disposable
sensing platforms are required for the application.
 Major areas of applications for biosensors.
On the other hand an application such as pollution monitoring
[18,19] requires a biosensor to function from a few hours to
several days. Such biosensors can be termed as "long term
monitoring" analysis tools.

Whether it is long term monitoring or single shot analysis,

biosensors find their use as technological advanced devices both in
resource limited settings and sophisticated medical setups:

1. E.G. With applications in drug discovery [20-22];

2. For the detection of a number of chemical and biological

agents that are considered as toxic materials of defence
interest [23];

3. For use in artificial implantable devices such as pacemakers

[24] and other prosthetic devices [25];

4. Sewage epidemiology [26].

 Biological sensing (recognition)
element (Bioreceptors)
 This component can be a tissue, microorganism, organelle, cell
receptor, enzyme, antibody or nucleic acid etc. These can be grouped
into two categories, namely catalytic and non-catalytic receptors.

 The catalytic biological receptors are are special proteins found on the
surface of cells. They act like switches that turn on important processes
inside the cell when they detect specific signals from outside the cell.
These include enzymes, tissues and microorganisms.

 The non-catalytic biosensor devices are used to detect and measure

specific biological molecules without using enzymes or other catalytic
components. Instead of relying on chemical reactions, these sensors
detect changes through physical or chemical interactions between the
target molecule and the sensor surface. They measure analytes such as
steroids, drugs, and toxins etc. These are non-reusable devices which
can only be used once and discarded thereafter. Such receptors include
antibodies, antigens, nucleic acids etc.
 Transducer
 The second segment of the biosensor is the transducer and it is a physical
component.

 Transducer converts the biochemical signal into proportional electrical

signals. The transducer is an element that converts one form of energy into
another. In a biosensor the role of the transducer is to convert the bio-
recognition event into a measurable signal. This process of conversion of the
energy is known as signalisation. Most transducers produce either optical or
electrical signals that are usually proportional to the amount of
analyte/bioreceptor interactions.

 Generally, a transducer is a material that is capable of converting one form of

energy to another. In a biosensor, a transducer is responsible for converting
the biochemical signal received from the biological receptor, which is a result
of the interaction between the target analyte and the biological receptor,
into a measurable and quantifiable signal which can be piezo-electrical,
optical, electrochemical etc.

 The transducer detects and measures the change that occurs during
biological receptor – analyte interaction. An example of a transducer is a pH
sensor in a glucose biosensor.
 Transducer
 An enzyme, known as glucose oxidase, is used as a
biological receptor which binds glucose and converts it
to gluconic acid in the presence of oxygen.

 The pH sensor (transducer) then detects the change in

pH (due to production of gluconic acid) and converts it
into a voltage change.

 The following features are recommended when a

transducer is designed; specificity to the target analyte,
analyte concentration range, response time and
suitability for practical applications.

 Ideally, a transducer should be highly specific to the

analyte, give measurement at the lowest analyte
concentration within the shortest time possible.
 Signal processing Unit
 The output of the transducer will be either
current or voltage relying on the type of
enzyme.

 If the output is voltage, then it is fine.

 But if the output is current, then this current

needs to be converted into equivalent voltage
(using an Op-Amp based current to voltage
converter) before proceeding further.

 The output voltage signal is generally very low

in amplitude and is superimposed on a high
frequency noise signal.

 Thus, the signal is amplified (using an Op-Amp

based Amplifier) and then it is passed through
a Low Pass RC Filter.
 Signal processing Unit
 Signal Processing Unit or a Signal Conditioning
Unit is accountable for performing the process of
amplifying and filtering the signal. The output of
the signal processing unit is termed as an analog
signal.

 This output is equivalent to the biological

quantity being measured.

 The analog signal can be exhibited directly on an

LCD display but usually, this analog signal is
passed to a Microcontroller, where the analog
signal is converted into digital signal. This is done
since it is easy to analyse, process or store a
digital signal.

 Signal processing unit is the associated part

which consists of Signal Conditioning Unit, a
Processor or Micro-controller and a Display Unit.
 Main components of the biosensors
Enzymes- Enzymes are biological molecules, typically proteins, that act as catalysts to accelerate chemical reactions. In biosensors, they are
used for their high specificity to target analytes, making them ideal for detecting specific substances.

Antibodies are specialized proteins produced by the immune system that recognize and bind specifically to antigens (foreign molecules,
pathogens, or biomarkers). In biosensors, antibodies are used for their exceptional specificity in detecting target analytes.

Nucleic acids, such as DNA (Deoxyribonucleic Acid) and RNA (Ribonucleic Acid), are biological molecules responsible for storing and
transmitting genetic information. In biosensors, nucleic acids are used for their ability to hybridize (bind) specifically with complementary
sequences, enabling highly precise detection of target analytes.

Cells, as biological recognition elements in biosensors, are living units capable of detecting and responding to specific analytes through
cellular reactions. They can be bacterial, yeast, plant, or mammalian cells, depending on the application.

Tissues refer to a collection of biological cells organized to perform specific functions. In biosensors, tissues serve as biological recognition
elements, offering a natural environment for biochemical reactions and enabling the detection of specific analytes.
 ENZYMES-
 Enzymes are proteins that help speed up chemical
reactions in our bodies.

 Enzymes are essential for digestion, liver function and

much more. Too much or too little of a certain enzyme can
cause health problems. Enzymes in our blood can also help
healthcare providers check for injuries and diseases.

 All enzymes are proteins, but not all proteins are enzymes.

 One of the most important roles of enzymes is to aid in

digestion. Digestion is the process of turning the food we
eat into energy.

 For example, there are enzymes in our saliva, pancreas,

intestines and stomach. They break down fats, proteins and
carbohydrates. Enzymes use these nutrients for growth and
cell repair.

Enzymes also help with:

 Breathing.
 Building muscle.
 Nerve function.
 Ridding our bodies of toxins.
 WHAT ARE THE DIFFERENT TYPES OF
 There are ENZYMES?
thousands of individual enzymes in
the body. Each type of enzyme only has one job.

 All enzymes are proteins, but not all proteins are enzymes.

 Enzymes are a specialized subset of proteins with catalytic

functions.

For example,

 The enzyme sucrase breaks down a sugar

called sucrose.
 Lactase breaks down lactose, a kind of sugar
found in milk products.

Some of the most common digestive enzymes are:

 Carbohydrase breaks down carbohydrates into

sugars.
 Lipase breaks down fats into fatty acids.
 Protease breaks down protein into amino acids.
 Parts of Enzymes
What are the parts of an enzyme?
Each enzyme has an “active site.” This area has a unique shape. The substance an enzyme works
on is a substrate. The substrate also has a unique shape. The enzyme and the substrate must fit
together to work.

How do temperature and pH affect enzymes?

Enzymes need the right conditions to work. If conditions aren’t right, enzymes can change shape.
Then, they no longer fit with substrates, so they don’t work correctly.
Each enzyme has an ideal temperature and pH:

 pH: Enzymes are sensitive to acidity and alkalinity. They don’t work properly if an environment
is too acidic or basic. For example, an enzyme in the stomach called pepsin breaks down
proteins. If your stomach doesn’t have enough acid, pepsin can’t function optimally.

 Temperature: Enzymes work best when your body temperature is normal, about 98.6°F
(36.6°C). As temperature increases, enzyme reactions increase. But if the temperature gets too
high, the enzyme stops working. That’s why a high fever can disrupt bodily functions.
 Enzyme based biosensors
 Enzyme based biosensors are most popular biosensors
due to its specialist features such as specificity, rate
enhancement, regulation and mild working condition.
 Enzyme biosensors are useful tools for monitoring rapid
changes in metabolite levels in realtime, include pure
enzyme preparations or biological processes.
 They have been derived on immobilization processes such
as van der Waals forces, ionic or covalent bonding.
 The well-known enzymatic biosensors today are glucose
biosensors.
 Enzymatic biosensors are known for their prolonged use
and reusability due to the fact that enzymes used as
biological receptors cannot be consumed.
 Thus, the detection limit and the lifetime of enzyme
based biosensors is greatly enhanced by the stability of
the enzyme.
 SCHEMATIC ILLUSTRATION OF ENZYMATIC BIOSENSORS

The fundamental principle of operation of an

enzyme electrode is shown in Figure 2.3, in which
the substrate to be determined diffuses into the
enzyme layer, where the enzymatic reaction
occurs (Equation 2.1):

resulting in a product ( P or ) or consuming co-

reactant which can be measured by transducers.
The most commonly used enzyme biosensors in
the clinical sector have been designed for urea,
lactate, glucose, glutamate, and cholesterol
 Antibody based sensors
 Antibody-based biosensors or immunosensors have
revolutionized diagnostics for the detection of a plethora
of analytes such as disease markers, food and
environmental contaminants, biological warfare agents
and illicit drugs.

 Antibodies are ideal biorecognition elements that provide

sensors with high specificity and sensitivity.

 Antibodies are large Y-shaped proteins produced by

plasma cells that are utilized by the immune system to
identify and target pathogens such as bacteria and
viruses.

 Their small size, high stability and easy genetic

manipulation make recombinant antibody fragments
valuable and robust tools for the fabrication of
immunosensors.
 Antibody based sensors
 Capture Antibody (Capture Ab)--The biosensor platform
contains immobilized capture antibodies (Ab), which are highly
specific to the target analyte (antigen/biomarker). These
antibodies are fixed on the biosensor support surface and
function to selectively bind with the analyte.

 Analyte/Antigen/Biomarker Binding-- The target analyte

(antigen or biomarker) from the sample binds specifically to
the capture antibodies, forming an immune complex. This
selective binding ensures only the target molecule is detected,
reducing false readings.

 Detection Antibody with Reporter Molecule- A secondary

antibody (detection Ab), labeled with a reporter
molecule/particle, binds to the captured analyte. The reporter
molecule could be an enzyme, fluorophore, nanoparticle, or
electrochemical tag that facilitates signal generation.

 Detector- The interaction between the analyte and antibodies

generates an output signal.
 Applications of Antibody-Based Biosensors

Medical diagnostics- Detection of disease biomarkers (e.g., COVID-19, cancer, HIV)

Food safety- Identification of pathogens or toxins

Environmental monitoring- Detection of contaminants in water or air

Biotechnology research- Protein and antigen analysis

 The Cell
The cell is the basic structure of the body. The human body is built of
billions and trillions of cells. Cells of various organs vary according to their
function. Each cell contains the hereditary material and can make copies of
itself by reproducing and multiplying. After a specific life span, the old cells
die off. Parts of the cell are called organelles. Human cells contain the
following major parts:

1. Nucleus – This is the central part of the cell that carries the blueprint
for the cell functioning and tells the cell when to grow, reproduce and
die. It also houses DNA (deoxyribonucleic acid).
2. Mitochondria – These are the cell's powerhouses and produce energy
for the various activities of the cell.
3. Cytoplasm – This is a jelly-like fluid within the cell in which the other
organelles float.
4. Endoplasmic reticulum (ER) – This helps process the molecules
(e.g., proteins) created by the cell.
5. Ribosomes – These lie over the ER and process the genetic
instructions or the blueprints within the DNA and create new proteins.
These can also float freely in the cytoplasm.
6. Lysosomes and peroxisomes – These help in digesting foreign
bacteria that invade the cell, rid the cell of toxic substances
7. Cell membrane – This is the outer lining of the cell.
8. The Golgi complex- is an essential organelle in eukaryotic cells,
responsible for the modification, sorting, and packaging of proteins and
lipids for transport to their appropriate destinations
 The Chromosomes
 Within the nucleus, the DNA strands are tightly
packed to form chromosomes. During the cell
division, the chromosomes are visible.

 Each chromosome has a constriction point called

the centromere from where two arms are
formed.

 The short arm of the chromosome is labeled the

“p arm.” The long arm of the chromosome is
labeled the “q arm.”Each pair of chromosomes is
shaped differently by the location of the
centromere and the size of the p and q arms.

 Humans typically have 23 pairs of

chromosomes, for a total of 46. Twenty-two of
these pairs, called autosomes, look the same in
both males and females.

 The 23rd pair is called the gender

chromosomes and differs between males and
females. Females have two copies of the X
 The Genes
 Genes are hereditary material that lies within
the cell nucleus.

 Genes, which are made up of DNA, act as

instructions to make molecules called proteins.

 The Human Genome Project has estimated that

humans have between 20,000 and 25,000
genes. Every person has two copies of each
gene, one inherited from each parent.

 These are mostly similar in all people, but a

small number of genes (less than 1 percent of
the total) are slightly different between people,
and this forms the basis of paternity tests and
DNA analysis.
 DNA

 DNA, or deoxyribonucleic acid, is the hereditary material in

humans and almost all other organisms.

 Nearly every cell in a person’s body has the same DNA.

 Most DNA is located in the cell nucleus (where it is called nuclear

DNA), but a small amount of DNA can also be found in the
mitochondria (where it is called mitochondrial DNA or mtDNA).

DNA contains four chemical bases:

1. Adenine (A)
2. Guanine (G)
3. Cytosine (C)
4. Thymine (T).
 DNA VS RNA

Feature DNA (Deoxyribonucleic Acid) RNA (Ribonucleic Acid)

 DNA is the "master blueprint" that Full Form Deoxyribonucleic Acid Ribonucleic Acid
stores genetic instructions. Structure Double-stranded (double helix) Single-stranded
Sugar Deoxyribose (lacks one oxygen) Ribose (has an extra oxygen)
 RNA is the "messenger and worker"
Adenine (A), Thymine (T), Cytosine Adenine (A), Uracil (U), Cytosine
that helps make proteins from DNA Nitrogen Bases
(C), Guanine (G) (C), Guanine (G) (No Thymine)
instructions. Stores genetic information and Helps in protein synthesis and
Function
instructions for cell activities carries messages from DNA
 DNA has Thymine (T), while RNA has Mostly in the nucleus (some in Found in the nucleus and
Location
Uracil (U) instead. mitochondria) cytoplasm
Stability Very stable (long-term storage) Less stable (short-lived)
 DNA is double-stranded and stable, Three main types: mRNA
while RNA is single-stranded and Types Only one type (messenger RNA), tRNA (transfer
RNA), rRNA (ribosomal RNA)
temporary.
Replication Can self-replicate Cannot self-replicate
 DNA
DNA base pairs
DNA bases pair up with each other, A with T and C
with G, to form units called base pairs. Each base
is also attached to a sugar molecule and a
phosphate molecule.

DNA in humans contains around 3 billion bases,

and these are similar in two persons for about 99%
of the total bases. These bases are sequenced
differently for various information that needs to
be transmitted. This is similar to how different
sequences of letters form words and sequences of
words form sentences.

Nucleotides and the double helix

A base, sugar, and phosphate in combination are
called a nucleotide.

Nucleotides are arranged in two long strands

that form a spiral called a double helix. This looks
like a twisted ladder, and the base pairs form
the ladder's rungs, and the sugar and
 How DNA-Based Biosensors Work

1. Probe Immobilization on the Sensor Surface- A single-

stranded DNA (ssDNA) probe is attached to the biosensor
surface. This probe is specifically designed to recognize and
hybridize with a complementary target DNA sequence.

2. Target DNA Recognition via Hybridization (The probe binds

with complementary target DNA)- When a sample
containing the target DNA is introduced, complementary
base pairing occurs between the probe and target DNA
strands. This hybridization is highly specific due to the
Watson-Crick base pairing (A-T and G-C).

3. Signal Transduction (Conversion of Hybridization into a

Detectable Signal)-Once hybridization occurs, the DNA
biosensor converts this event into a measurable signal using
different transduction methods:
 DNA based biosensors
 To develop DNA biosensors, immobilization of the
probe becomes the most crucial step.

 The strong pairing of lined up nucleotide strands

between bases in their complementary parts
influences biosensors based on DNA, RNA, and
peptide nucleotide acids to be the most sensitive
tool.

 That probes, which are short oligonucleotides capable

of hybridization with individual areas of the target
nucleotide sequence, together with various chemical
composition and conformational arrangements, were
employed in the development of DNA biosensors.

 Extremely high sensibility and selectivity is needed to

maximize the hybridization efficiency and minimize
non-specific binding.
 Signal Transduction (Conversion of Hybridization into a
Detectable Signal)-

A. Electrochemical DNA Biosensors- Detect changes in electrical properties upon

hybridization. Ex. Impedance Spectroscopy, Voltammetry, Conductometric Sensing

B. Optical DNA Biosensors- Detect hybridization using light-based techniques.

C. Piezoelectric DNA Biosensors-- Utilize a quartz crystal microbalance (QCM) to measure

mass changes.DNA hybridization increases the mass on the sensor, altering its
resonance frequency.
D. Nanomaterial-Enhanced DNA Biosensors-Integrate nanomaterials like carbon
nanotubes, gold nanoparticles, and graphene for signal amplification.Example: Gold
nanoparticles aggregate upon DNA binding, leading to a visible color change.
 Aptamer-Based and DNA based Sensors
 Aptamer-based sensors, or aptasensors, are biosensors that use aptamers
instead of antibodies or enzymes for target recognition.

 Aptamers are short, single-stranded DNA or RNA sequences that fold into
unique 3D structures to selectively bind to specific molecules, such as proteins,
small molecules, viruses, or even cells.

 Aptasensors- Can detect anything – proteins, viruses, bacteria, toxins, drugs,

and even metal ions. They use special DNA or RNA pieces (aptamers) that
change shape when they bind to a target, creating a signal. Aptasensors are
more flexible because they can detect a wide range of molecules, not just DNA.
Aptamers are stronger, last longer, and can work in harsh conditions. They are
used in medical tests (e.g., detecting viruses like COVID-19), food safety (e.g.,
toxin detection), and pollution monitoring (e.g., detecting heavy metals).

 DNA Biosensors- Detect only DNA or RNA sequences by matching

complementary strands (like a zipper closing). They Use matching DNA strands;
when they find their perfect pair, a signal is produced. DNA Biosensors are best
for genetic tests, like detecting viruses or mutations. DNA probes are more
delicate and can break down more easily. They are mostly used in genetic tests
(e.g., checking for inherited diseases or detecting bacteria through DNA).
 Cell-Based Biosensors

 Cell-based biosensors are devices that use living

cells as biological recognition elements to detect
chemical, biological, or environmental changes.

 These biosensors leverage the natural response of

cells to external stimuli, converting biological
activity into measurable signals (electrical, optical,
or mechanical).

 Unlike traditional biosensors that use enzymes,

antibodies, or aptamers, cell-based biosensors
provide real-time, dynamic, and long-term
monitoring of a wide range of analytes, including
toxins, pathogens, and pharmaceuticals.
 Types of Cell-Based Biosensors

1. Whole-Cell Biosensors- Use genetically engineered cells to produce a detectable signal in response to specific
chemicals. Example: E. coli engineered to glow in the presence of arsenic.

2. Stem Cell-Based Biosensors- Use human stem cells to test new drugs and detect toxic substances.Example:
Monitoring drug effects on heart cells for cardiotoxicity.

3. Bacterial Biosensors- Bacteria are used to sense environmental pollutants and toxins.Example: Pseudomonas
putida bacteria that change color in response to heavy metals.

4. Neuronal Cell-Based Biosensors- Use nerve cells to detect neurotoxins (e.g., botulinum toxin).Example: Detecting
pesticides that affect nerve function.

5. Microbial Fuel Cells (MFCs)- Use bacteria to generate electricity when exposed to organic matter.Example:
Wastewater treatment sensors that detect pollution levels.
 Functions of Proteins
Proteins are essential molecules made up of amino acids and are involved in nearly every biological process in
living organisms. They act like the building blocks, machines, and messengers of the body.

Proteins are vital for every function of life, acting as builders, messengers, defenders, and engines of the body.

 Proteins provide shape, strength, and structure to cells and tissues. They provide strength to skin, bones, and
tendons.

 Proteins called enzymes speed up chemical reactions in the body. They break down starch into sugar.

 Proteins carry molecules around the body and store essential substances. Hemoglobins transport oxygen in the
blood.

 Some proteins act as hormones, controlling body processes such as Insulin regulates blood sugar levels

 Proteins help protect the body from pathogens like viruses and bacteria. Antibodies identify and neutralize
harmful invaders.

 Proteins are essential for muscle contraction and movement.

 Proteins transmit signals between cells to coordinate actions.

 In extreme conditions (e.g., starvation), proteins can be broken down for energy.
 Protein based sensors
 Protein-based sensors are specialized molecular devices or
biological tools that use proteins to detect and measure
specific molecules, ions, or environmental changes.

 These sensors rely on the unique binding capabilities and

structural flexibility of proteins to interact with target
substances, producing measurable signals.

1. The protein in the sensor is designed to recognize and bind

to a specific molecule (like sugar, calcium, or a virus).

2. When the target molecule attaches to the protein, the

protein often changes its shape.

3. This change produces a signal, like light (fluorescence), an

electrical current, or another measurable effect.
 The first biosensor
 The development of the first biosensor is closely associated
with the name of L. C. Clark, who first suggested the oxygen
probe (1956) for measuring oxygen in the blood and then
described the ‘‘enzyme electrode’’ consisting of this probe
and two dialysis membranes carrying a small portion of the
aqueous solution of glu cose oxidase in between them (Fig.
1.2) (Clark and Lyons 1962).

 The enzyme catalyzed the oxidative conversion of glucose

to gluconic acid. Thus, the oxygen content in the proximity
of the electrode surface decayed proportionally to the
glucose concentration.

Fig. 1.2 First ‘‘enzyme electrode’’ with glucose

oxidase solution entrapped in the sandwich-
type membranes
 Classification of biosensors based on principle

 The most commonly applied classification of

biosensors is based on the type of
transduction element used in the sensor.

 These biosensors are grouped into three main

categories, amperometric, potentiometric,
optical, acoustic, piezoelectric, and
calorimetric biosensors.

 The working principles of each of the three

biosensors are different and can thus be
implemented in a variety of applications.
 Classification of biosensors based on principle

Based on the working principles of Transducer of the biosensor, classification is done as

follows
 Amperometric Biosensor: Based on the movement of electrons produced in the redox reaction.
 Potentiometric Biosensor: Based on the changes in the distribution of charge causing an electrical potential
to be produced.
 Optical Biosensor: Light output during the reaction or light absorption difference between the reactants and
products.
 Acoustic Biosensor: Utilizes acoustic or mechanical waves as a detection mechanism to obtain medical,
biochemical, and biophysical information about the analyte of interest
 Piezoelectric Biosensor: Piezoelectric effect due to the mass of the reactants
 Calorimetric Biosensors: Heat output input by the reaction
 Challenges in biosensing research

 Clear cut advantages over existing methods for analyses of that analyte.

 Identification of the market that is interested in biosensor of certain analyte of interest.

 Testing the performance of the biosensor both in use and after storage. Response of a biosensor
after six months storage is the absolute minimum for any practical commercial application.

 Stability, costs and ease of manufacturing each component of the biosensor.

 Hazards and ethics associated with the use of developed biosensor.

 For the fabrication of a biosensor for non-specialist markets, the
following conditions are required
 The desired analyte should be specific and stable under a normal storage condition.

 The sensor should be accurate, precise and show high sensitivity in a reproducible way, and linearity must
be obtained with different concentrations.

 Physical parameters such as pH, temperature should be optimised, which will lead to sample analysis with
minimal pre-treatment.

 The biosensor should be small and biocompatible so that it can be used for invasive monitoring in clinical
diagnostics.

 The fabricated biosensor should be portable, cost-effective, small,and capable of being used by semi-
skilled operators.

 The biosensor should provide real-time analysis so that it can be employed for rapid measurements of
analytes from human samples.