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Biosensors
Biosensors are receiver-transducer systems that can sense the biological activities and
provide data related to it.
Analytes are the cells, antibodies, enzymes or any other biological materials that are of
interest for specific biosensors. These analytes are sensed by the receptors. The latter are
very sensitive parts connected to the analytes that make the transducers become
activated. Each transducer converts the input of the receptors into the signal of interest.
The transducer output can be electrical, heat, magnetic, optical etc, as per the choice of
biosensor and its application.
The systematic workflow of biosensors is presented in figure below…..
The general structure of biosensors is schematically shown in Figure below. They consist of
two main parts, the receptor and the transducer (this does not mean that they are physically
separated). The receptor interacts with the analyte selectively, while the transducer
produces an electrical or optical signal as a result of the former interaction. This signal carries
information about the concentration of the analyte.
Receptor parts contain the biologically active components that are capable for specific
chemical reactions with the analyte. Nature has created an almost endless variety of
biological compounds that may act as receptors. According to the type of receptor,
biosensors can be distinguished into the following groups which are discussed next
section ( in types of Biosensors)
Enzymatic Biosensors
Biosensors that were developed first belong to the group of enzymatic sensors. A great
number of different types have been developed since their appearance. The greatest
efforts have been made in the field of glucose sensors, where devices with a possible
operation period of several months could be fabricated. Meanwhile, a great
development has been achieved in enzyme immobilization techniques. Without giving
a detailed theoretical description of biocatalytic processes, the most important
definitions and reaction models are summarized.
Theory / Principle of Enzymatic Biosensors

Enzymes are high-molecular-weight proteins synthesized inside living cells. They influence
the reaction speed of chemical processes. With hormones and vitamins, they form the
biocatalytic system of living things. The reagent, the chemical transition of which the
enzyme catalyzes specifically, is called its substrate. Enzymatic analysis is a special branch of
analytical chemistry, the goal of which is to detect enzymes or substrates by means of their
specific reaction. Enzymatic sensors employ enzymes to measure the concentration of their
substrate in the analyte of interest.
Enzyme Immobilization Techniques
Immobilization means the conversion of an enzyme from a water-soluble, mobile state

into a water-insoluble state without affecting its catalytic activity. Coimmobilization of
several enzymes or enzyme cosubstrate/mediator systems is often necessary. Since the
immobilization method strongly influences how the enzyme molecules are exposed to
environmental effects and how they react to them, it determines the stability and lifetime
of the sensor.
One key issue of biosensor fabrication is the appropriate stable immobilization of enzyme
molecules onto a transducer surface or into a functional membrane. A great number of
techniques have been developed such as; encapsulation of a free solubleenzyme, physical
adsorption and entrapment, direct cross-linking, covalent binding, and embedment of
enzymatic molecules into conducting polymers.
Operation Characteristics of enzymatic biosensors
Types of Enzymatic Biosensors
Enzymes constitute a group of more than 2000 proteins having so-called biocatalytic
properties. These properties give the enzymes the unique and powerful ability to
accelerate chemical reactions inside biological cells. Most enzymes react only with specific
substrates even though they may be contained in a complicated mixture with other
substances. It is important to keep in mind, however, that soluble enzymes are very
sensitive both to temperature and pH variations and they can be inactivated by many
chemical inhibitors. For practical biosensor applications, these enzymes are normally
immobilized by insolubilizing the free enzymes via entrapment into an inert and stable
matrix such as starch gel, silicon rubber, or polyacrylamide. This process is important to
ensure that the enzyme retains its catalytic properties and can be reusable.
There are many types of enzymatic biosensors depending upon their sensing principle.
Some of them are listed below:
i) Glucose Sensors
ii) Urea Sensors
iii)Sucrose Sensor
iv) Alcohol Sensor
v) Free fatty acids Sensors
vi) ADP Sensor
vii) ATP Sensor
Glucose Sensors
A typical example of an enzyme-based sensor is a glucose sensor that uses the enzyme
glucose oxidase. Glucose plays an important role in metabolic processes. In patients
suffering from diabetes mellitus, the pancreas does not produce sufficient amounts of
insulin to control adequately the level of glucose in their blood. Therefore, to manage
the disease, these patients must monitor and regulate their blood glucose level on a
regular basis by medication and insulin injections. Currently available glucose sensors are
based on an immobilized enzyme, such as glucose oxidase, which acts as a catalyst.
Glucose is detected by measuring electrochemically either the amount of gluconic acid or
hydrogen peroxide (H2O2) produced or by measuring the amount of oxygen consumed,
according to the following chemical reaction:
Biocatalytic enzyme-based sensors generally consist of an electrochemical gassensitive

transducer or an ion-selective electrode with an enzyme immobilized in or on a
membrane that serves as the biological mediator. The analyte diffuses from the bulk
sample solution into the bio-catalytic layer where an enzymatic reaction takes place.
The electroactive product that is formed (or consumed) is usually detected by an ion-
selective electrode. A membrane separates the basic sensor from the enzyme if a gas is
consumed (such as O2) or is produced (such as CO2 or NH3). Although the concentration of
the bulk substrate drops continuously, the rate of consumption is usually negligible. The
decrease is detected only when the test volume is very small or
when the area of the enzyme membrane is large

enough. Thus, this electrochemical analysis is
nondestructive, and the sample can be reused.
Measurements are usually performed at a
constant pH and temperature either in a stirred
medium solution or in a flowthrough solution.

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Biosensors

Theory / Principle of Enzymatic Biosensors

Immobilization means the conversion of an enzyme from a water-soluble, mobile state

Biocatalytic enzyme-based sensors generally consist of an electrochemical gassensitive

when the area of the enzyme membrane is large

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