AYEBALE CHARITY

20/U/0412
What is a biosensor?
• A biosensor is an analytical device that combines the specificity of a
biological sensing element for the analyte of interest with a transducer
to produce a signal proportional to target analyte concentration.

• Biosensors are analytical devices that convert a biological response

into an electrical signal.
• This signal can result from a change in proton concentration, release or
uptake of gases such as ammonia or oxygen, light emission, absorption or
reflectance, heat emission, mass change, etc.

• The transducer converts this signal into a measurable response such as

current, potential, temperature change, absorption of light, or mass
increase through electrochemical, thermal, optical, or piezoelectric means.

• This signal can be further amplified, processed, or stored for later analysis.

• Components that may be used to construct a biosensor include organisms,

tissues, cells, organelles, membranes, enzymes, receptors, nucleic acids,
antibodies, organic molecules.
Types of biosensors
Enzyme-based biosensors
• These are based on biological recognition. In order to operate, the enzymes
must be available to catalyze a specific biochemical reaction and be stable
under the normal operating conditions of the biosensor.

• They offer specificity and catalytic properties for signal amplification. Their
use spans oxido-reductases, hydrolases, and lyases in diverse applications.
Recent advancements enhance sensitivity, stability, and shelf life through
genetic modifications. Improved electrochemical interfaces and sampling
Silica, quartz/crystal and glass biosensors

• Silicon nanomaterials have greater potential for technological

advances in biosensor applications due to their biocompatibility,
abundance, electronic, optical, and mechanical properties.

• Silicon nanomaterials have no toxicity which is an important

prerequisite of biomedical and biological applications.
Nanomaterials-based biosensors

• Nanomaterials such as gold, silver, silicon, carbon-based materials, such

as graphite, grapheme, are used for developing biosensor
immobilization, which allows the nano molecules to interact with the
target analyte, enabling the biosensor to detect and quantify them
accurately.

• Nanoparticle-based materials provide great sensitivity and specificity for

developing electrochemical and other types of biosensors.
Antibody-based biosensors

• Antibody-based biosensors utilize the high specificity and affinity of

antibodies to detect target molecules. These biosensors typically
involve immobilizing antibodies onto a surface, such as a chip or
membrane, where they capture and bind to specific antigens present in
the sample. The interaction between the antibody and antigen
generates a measurable signal, which can be quantified to determine
the concentration or presence of the target molecule.
Cell-based biosensors

• Cell-based biosensors for environmental applications can be organized according

to cell type. For example, bacteria, yeast, algae and tissue culture cells.

• Cell-based biosensors employ living cells as sensing elements to detect various

analytes. These biosensors utilize the natural response of cells to specific stimuli,
such as changes in pH, ion concentration, or the presence of specific molecules.

• The cells are typically engineered to express reporter genes or proteins whose
expression levels correlate with the presence or concentration of the target
analyte. The detection mechanism often involves monitoring changes in cell
morphology, viability, fluorescence, or electrical activity.
DNA biosensors

• DNA biosensors utilize the unique properties of DNA molecules for

the detection of various analytes. These biosensors typically involve
immobilizing DNA strands onto a surface, where they can specifically
bind to complementary sequences present in the sample.

• The binding event induces a detectable signal, which can be measured

to quantify the presence or concentration of the target analyte. DNA
biosensors can detect a wide range of targets, including nucleic acids,
proteins, small molecules, and even ions.
Receptor-based biosensor
• Receptor-based biosensors utilize biological receptors, such as proteins or
enzymes, to specifically bind to target molecules of interest.

• These biosensors typically involve immobilizing the receptor onto a surface,

where it captures and interacts with the target analyte. The binding event
generates a measurable signal, which can be transduced and quantified to
determine the presence or concentration of the target molecule.

• Receptor-based biosensors offer high specificity and sensitivity, making them

valuable tools in various fields including medical diagnostics, environmental
monitoring, and drug development.
Transducers - Principles of Operation

Biosensors based on electrochemical transducers

• There are three types of electrochemical biosensors: potentiometric,

amperometric, and conductometric.

• Conductometric sensors are usually nonspecific and have a poor

signal-to-noise ratio; as yet they have not been applied to bioprocess
monitoring
• Conventional potentiometric biosensors consist of an ion-selective electrode
(ISE; pH, ammonium, and fluoride) or a gas-sensing electrode (CO, and
NH,) coated with an immobilized enzyme layer. The enzymatic reaction with
the analyte generates a change in potential due to ion accumulation or
depletion.

• Potentiometric transducers measure the difference in potential that is

generated across an ion-selective membrane separating two solutions at
virtually zero current flow.

• Examples of applications of potentiometric probes include the detection of

glucose, urea, and creatinine.
Amperometric transducers
• Operate at a fixed potential with respect to a reference electrode and the current
generated by the oxidation or reduction of species at the surface of the working
electrode is measured.

• They are based on redox enzymes; thus, their appeal is due to the availability of
a large number of oxido-reductase enzymes that can act on fatty acids, sugars,
amino acids, aldehydes, and phenols

• Merits include high sensitivity, wide dynamic range, rapid response time,
selectivity and long-term stability.

• Demerits include calibration requirements, electrode fouling, enzyme

Amperometric biosensors based Hydrogen peroxide-based biosensors.

• They typically consist of enzymes that

on measurement of oxygen
produce or consume hydrogen peroxide in
depletion. the presence of the target analyte, along with

• These utilize oxygen-sensitive an electrode for amperometric detection.

Chemically modified amperometric

electrodes to detect changes in
biosensors.
oxygen concentration resulting
• These involve the modification of electrode
from enzymatic reactions.
surfaces with specific chemicals to enhance

their performance sensitivity, selectivity and

stability.
Biosensors based on thermal effects
Biosensors based on optical effects
• Thermal enzyme sensors are based •
In these, the change in optical properties
on the principle that the heat such as UV/vis absorption, bio- and
evolved in an enzymatic reaction chemiluminescence, reflectance, and
can be utilized to calorimetrically fluorescence caused by the interaction of
determine the amount of substrate the biocatalyst with the target analyte is

reacted. monitored optically.

• Thus, thermometric indicators or • Fiber optic-based biosensors offer

transducers require only a single advantages of compactness, flexibility,

resistance to electrical noise, and a small

reaction step producing sufficient or
probe size.
measurable heat.
Biosensors based on surface acoustic
wave/piezoelectric transducers
• Piezoelectric materials can be used as sensor transducers due to their ability to
transmit acoustic waves. If a recognition element (biological or non-
biological) is immobilized on the surface of a piezocrystal, its binding with the
analyte of interest causes a change in the resonant frequency of the crystal,
which is then correlated to the mass change.

• Can be used in gas phase analysis to measure ammonia, nitrous oxides,

hydrocarbons, methane, etc. by coating the surface with selective adsorbents

• The advantages are that they are inexpensive but they may suffer from poor
sensitivity and problems of nonspecific binding
Integration of biosensors in on-line process
monitoring schemes.
• Any scheme for on-line monitoring and control of a bioprocess must include
three essential components: an analytical device (biosensor), a suitable
configuration employing an in situ or ex situ arrangement for contacting the
fermentation broth with the biosensor, a control system with the necessary
hardware and software in order to employ a suitable control strategy.

• Biosensors may be integrated into an on-line scheme for the monitoring of

substrates and products in the bioreactor using either in situ monitoring or
flow injection analysis.
In situ monitoring
• Currently, pH and dissolved oxygen and carbon dioxide measurement during
cell cultivation in a bioreactor are made using sterilizable, in situ probes.
Fiber optic-based sensors have been used in situ for NADH measurements in
bioreactors

• In order to maximize the response, conditions of operation for these detectors

need to be optimized with respect to temperature, pH, and ionic strength.

• The requirements for sterilization in bioreactors and problems with microbial

growth on the detector surface make in situ applications for such biosensors
difficult.
Flow Injection Analysis (FIA)
• In FIA, the liquid sample to be analyzed is injected into a moving, non-
segmented carrier stream of buffer solution flowing continuously past a
detector. The sample forms a zone that is transported across the detector. The
latter continuously records the change in either absorbance, current,
electrode potential, or any other physical parameter due to the passage of
this zone.

• The advantages of FIA include reduced risk of contamination, automatic

recalibration of the sensor to counter the drift, short response time,
requirement of small sample quantity, multicomponent monitoring by array
The essential components of a FIA system are:
• selector valve to switch the injection flow between a sample to be
analyzed and calibration standards
• multichannel peristaltic pump module or multiple peristaltic pumps to
transport carrier buffer and appropriate solution to the injection valve
• injection valve with a fixed-volume loop to inject samples or standards
into a carrier stream at programmed intervals
• Detector transducer system interfaced to a recorder or computer.
Applications of biosensors Biodefense biosensing
applications

• Biological warfare agent

detection

• Environmental surveillance

• Monitoring bio-threat agents

• Detection of pathogens

• Identification of biological
toxins
In food processing In fermentation industries

• Assure food safety • Monitoring biomass and cell

viability
• Detect heavy metals
• Monitoring metabolites
• Detection of pathogens in
• Controlling PH and oxygen levels
food.
• Detecting contaminants
• Detection and quantification of
• Quality control and process
sweeteners
optimization
In medical field Fluorescent biosensors
• Glucose biosensors are used for diagnosis of • Fluorescent biosensors are imaging agents,
diabetes mellitus, which requires precise for use in cancer and drug discovery.
control over blood-glucose levels. • They have enabled insights into the role
• To diagnose infectious diseases
and regulation of enzymes at cellular level
• A novel biosensor, based on hafnium oxide has
• They are employed in probing gene
been used for early stage detection of human
expression, protein localization, and
interleukin.
conformation in fields such as signal
• Quantitative measurement of cardiac markers in
transduction, transcription, cell cycle and
undiluted serum.
apoptosis
• Microfluidic impedance assay for controlling
endothelin induced cardiac hypertrophy
In metabolic engineering Biosensors in plant biology

• Metabolite monitoring • Nutrient monitoring in soil, water,

• Pathway optimization or plant tissues

• Pesticide detection on crops

• Feedback control
• Disease diagnosis
• Bioprocess monitoring
• Environmental monitoring such
• High-throughput screening
as air quality, pollution levels

• GMO detection

• Plant phenotyping