Biosensor = bioreceptor + transducer

A biosensor consists of two components: a bio receptor and a transducer. The bio receptor is a
bio molecule that recognizes the target analyte whereas the transducer converts the recognition
event into a measurable signal. The uniqueness of a biosensor is that the two components are
integrated into one single sensor

What Is a Biosensor

Recent development topics on biosensors

In biosensor development studies, suitable bioreceptor molecule, suitable immobilization method
and transducer should be selected firstly. Biology, biochemistry, chemistry, electrochemistry,
physics, kinetics and mass transfer knowledge is required for this study. Thus we can say that
developing a biosensor is related with a interdisciplinary study. Proportional to the technological
development and increase of interdisciplinary studies biosensors are being more useful and

having more usage areas day by day. Recent development topics which are listed below will be
discussed in this chapter:
Electrochemical biosensor
Fiber-optic biosensor
Carbon Nanotube
Protein Engineering for biosensors
Wireless Biosensors Networks
1. A bioreceptor that is an immobilized sensitive biological element (e.g. enzyme, DNA probe,
antibody) recognizing the analyte (e.g. enzyme substrate, complementary DNA, antigen).
Although antibodies and oligonucleotides are widely employed, enzymes are by far the most
commonly used biosensing elements in biosensors.
2. A transducer is used to convert (bio)chemical signal resulting from the interaction of the
analyte with the bioreceptor into an electronic one. The intensity of generated signal is directly or
inversely proportional to the analyte concentration. Electrochemical transducers are often used to
develop biosensors. These systems offer some advantages such as low cost, simple design o
small dimensions. Biosensors can also be based on gravimetric, calorimetric or optical detection
For example, the glucose concentration in a blood sample can be measured directly by a
biosensor (which is made specifically for glucose measurement) by simply dipping the sensor in
the sample. The simplicity and the speed of measurement is the main advantages of a biosensor.

Fig. 1.1. Biosensor configuration.

To measure the glucose concentration, three different transducers can be used:

An oxygen sensor that measures oxygen concentration

A pH sensor that measures the acid (gluconic acid) production

A peroxide sensor that measures H2O2 concentration.

Note that an oxygen sensor is a transducer that converts oxygen concentration into electrical
current. A pH sensor is a transducer that converts pH change into voltage change. Similarly, a
peroxidase sensor is a transducer that converts peroxidase concentration into an electrical
current.
Considerations in Biosensor Development. Once a target analyte has been identified, the major
tasks in developing a biosensor involves:
1. Selection of a suitable bioreceptor molecule
2. Selection of a suitable immobilization method
3. Selection of a suitable transducer
4. Designing of biosensor considering measurement range, linearity, and minimization of
interference
5. Packaging of biosensor
Requirements for Sensors To be commercially successful, a biosensor has to meet the general
requirements of commercial sensors (Table 1.2). These are:
1. Relevance of output signal to measurement environment
2. Accuracy and repeatability
3. Sensitivity and resolution
4. Dynamic range
5. Speed of response
6. Insensitivity to temperature (or temperature compensation)
7. Insensitive to electrical and other environmental interference
8. Amenable to testing and calibration
9. Reliability and Self-Checking Capability
10. Physical robustness
11. Service requirements
12. Capital cost
13. Running costs and life
14. Acceptability by user
TYPES OF BIOSENSORS
Biosensors can be grouped according to their biological element or their transduction element.
Biological elements include enzymes, antibodies, micro-organisms, biological tissue, and

organelles. Antibody-based biosensors are also called immunosensors. When the binding of the
sensing element and the analyte is the detected event, the instrument is described as an affinity
sensor. When the interactionbetween the biological element and the analyte is accompanied or
followed by a chemical change in which the concentration of one of the substrates or products is
measured the instrument is described as a metabolismsensor. Finally, when the signal is produced
after binding the analyte without chemically changing it but by converting an auxiliary substrate,
the biosensor is called a catalytic sensor.
The method of transduction depends on the type of physicochemical change resulting from the
sensing event. Often, an important ancillary part of a biosensor is a membrane that covers the
biological sensing element and has the main functions of selective permeation and diffusion
control of analyte, protection against mechanical stresses, and support for the biological element.
The most commonly used sensing elements and transducers are described below.

Enzymes are proteins with high catalytic activity and selectivity towards substrates They have
been used for decades to assay the concentration of diverse analytes. Their commercial
availability at high purity levels makes them very attractive for mass production of enzyme
sensors. Their main limitations are that pH, ionic strength, chemical inhibitors, and temperature
affect their activity. Most enzymes lose their activity when exposed to temperatures above 60deg.
Most of the enzymes used in biosensor fabrication are oxidases that consume dissolved oxygen
and produce hydrogen peroxide Enzymes have been immobilized at the surface of the transducer
by adsorption, covalent attachment, entrapment in a gel or an electrochemically generated
polymer, in bilipid memb-ranes or in solution behind a selective membrane. Enzymes are
commonly coupled to electro-chemical and fiber optic transducers.
Antibodies are proteins that show outstanding selectivity. They are produced by blymphocytes in
response to antigenic structures, that is, substances foreign to theorganism. Molecules larger than
about 10 kDa can stimulate an immune response. Smaller molecules like vitamins or steroids can
be antigenic (also called haptens) but they do not cause an immune response unless they are
conjugated to larger ones like bovine serum albumin. Many antibodies are commercially
available and commonly used in immunoassays. Antibodies are usually immobilized on the
surface of the transducer by covalent attachment by conjugation of amino, carboxyl, aldehyde, or

sulfhydryl groups. The surface of the transducer must be previously functionalized with an
amino, carboxyl, hydroxyl, or other group. A review of conjugation techniques can be found
elsewhere. Antibodies sharesimilar limitations with enzymes. Furthermore, binding may not be
reversible and regeneration of the surface may require drastic changes in conditions like low pH,
high ionic strength, detergents, etc. Therefore, efforts are being made to produce low cost, single
use sensors. Probably the main potential advantage of immunosensors over traditional
immunoassays is that they could allow faster and in-field measurements. Immunosensors usually
employ optical or acoustic transducers.
Microbes The use micro-organisms as biological elements in biosensors is based on the
measurement of their metabolism, in many cases accompanied by the consump-tion of oxygen or
carbon dioxide, and is, in most cases, measured electrochemically. Microbial cells have the
advantage of being cheaper than enzymes or antibodies, can be more stable, and can carry out
several complex reactions involving enzymes and cofactors. Conversely, they are less selective
than enzymes, they have longer response and recovery times, and may require more
frequent calibration. Micro-organisms have been immobi-lized, for example, in nylon nets,
cellulose nitrate membranes, or acetyl cellulose. Other biological elements such as animal of
vegetable tissue and membranes as well as organelles and nucleic acids have been researched but
are out of the scope of this article. A summary of some biological elements and transducers used
in the fabrication of biosensors is presented
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
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.
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.
Essential properties of a biosensor:
(i) Specificity: a biosensor should be specific to the analyte which it interact.
(ii) Durability: it should withstand repeated usage.
(iii) Independent nature: It should not be affected by variations in the environment like
temperature, pH etc.
(iv) Stability in results: the results produced by interaction should be corresponding to the
concentration of analyte.
(v) Ease of use and transport: it should be small in size so that it can be easily carried and used.
1.They can measure nonpolar molecules thatdo not respond to most measurement devices
2.Biosensors are specific due to the immobilized system used in them
3.Rapid and continuous control is possible with biosensors
4.Response time is short (typically less than a minute) and
5.Practical
There are also some disadvantages of biosensors:
1.Heat sterilization is not possible because of denaturaziation of biological material,
2.Stability of biological material (such as enzyme, cell, antibody, tissue, etc.), depends on
the natural properties of the molecule that can be denaturalized under environmental
conditions (pH, temperature or ions)
3.The cells in the biosensor can become intoxicated by other molecules that are capable of
diffusing through the membrane.

Transducer elements
Electrochemical.
Amperometric and potentiometric transducers are the most commonly used electrochemical
transducers. In amperometric transducers, the potential between the two electrodes is set and the
current produced by the oxidation or reduction of electroactive species is measured and
correlated to the concentration of the analyte of interest. Most electrodes are made of metals like
platinum, gold, sliver, and stainless steel, or carbon-based
materials that are inert at the potentials at which the electrochemical reaction takes place.
However, because some species react at potentials where other species are present, either a
selective membrane is used or an electron mediator that reacts at lower potential is incorporated
into the immobilization matrix or to the sample containing the analyte. Potentiometric
transducers measure the potential of electrochemical cells with very low current. Field effect

transistors (FET) are potentiometric devices based on the measurement of potential at an

insulator electrolyte interface. The metal gate of a FET can besubstituted by an ion selective
membrane to make a pH transducer (pH ISFET). Enzymes have been immobilized on the surface
of such pH ISFET to produce enzyme- sensitized field effect transistors (ENFET). A complete
description of such sensors can be found elsewhere.
Fiber optic biosensor

Optic Fiber
The core and cladding parts play a very important role particularly on the light transmission.
Their refractive indices are n1 and n2, respectively (Fig 5). In the Fig 6a the core and cladding
interference act as mirror because of their different refractive indices. The series of internal
reflections transmit the light from one end of the fiber to the other one
The basic system of a fiber optic biosensor consists of a light source, an optical fiber, sensing
material and a detector. An optical fiber transmits the light and also acts as the substrate for the
sensing material. Detector measures the output signal. (Fig8) Some light source of opticalbiosensors are tungsten lamp, deuterium lamp, xenon lamp, LEDs, Laser, Laser diodes and some
light detectors for optic biosensors are avalanche photodiodes, photodiodes, photomultipliers,
charge- coupled devices

When the reaction occurs between sensing element and the analyte, there is a change both its
physico-chemical and optical properties. This transduction mechanism, generates optical signals,
is related with analyte concentration. To measure the optical signals, the difference between
incident and output light is determined at the location where the sensing element is fixed. Output
light is send to detector by fiber. Collected light (reflected, emitted, absorbed light) is measured
on the detector.
The Fiber Optic Biosensor have some advantages and disadvantages which are shown
below.
The Advantages of Fiber Optic Biosensor [18, 19]
1. There is no need reference electrode in the system
2. It can be easily moved, because there is no reagent in contact of any optical fiber
3. There are no electrical safety hazards and electrical interference
4. It is less dependent than temperature compared with electrode
5. It can be found in-vivo measurement applications because of easy miniaturization
6. Multiple analytes can be determined thanks to guide the light in different wavelengths
at the same time.
7. It can be used for the most of chemical analytes because of its spectroscopic properties.
The Disadvantages of Fiber Optic Biosensor
1. The life time of the reagents can be short under incident light
2. Because of the diffusion of analytes, it may cause slow response time
3. Fiber Optic Biosensor only works for spesific reagent.
4. Optimized commercial accessories have limited availability when using them with
optical fibers.

Acoustic.
Electro acoustic devices used in biosensors are based on the detection of a change of mass
density, elastic, viscoelastic, electric, or dielectric properties of a membrane made of chemically
interactive materials in contact with a piezoelectric material. Bulk acoustic wave (BAW) and
surface acoustic wave (SAW) propagation transducers are commonly used. In the first, a crystal
resonator, usually quartz is connected to an amplifier to form an oscillator whose resonant
frequency is a function of the properties of two membranes attached to it. The latter is based on
the propagation of SAWs along a layer of a substrate covered by the membrane whose properties

affect the propagation loss and phase velocity of the wave. SAWs are produced and measured by
metal inter digital transducers deposited on the piezoelectric substrate as shown.

Applications:
A biosensor has a wide range of applications in different fields.
Medicinal Application: biosensors have been used in various diagnostic procedures to determine
various tests.
Industrial application: various manufacturing processes can be monitored by biosensors to
provide assistance with regard to increase the quality and quantity of product obtained.
Environmental application: it helps in measuring the toxicity of water bodies, microbial
contamination of natural resources helping in developing steps towards a cleaner environment.
Military application: it helps to detect explosives, drugs etc., aiding in defence of the people.
Another breakthrough in the field of biosensors was the production of a product called smart
skin. It is a kind of biosensor which detects any chemical or biological attack nearby and warns
the person using the same.
Drug development: a biosensor called nano sensors has been developed which detects and
analyse the binding of proteins to its targets which has proved very useful in drug designing.
The application of biosensor areas [4] are clinic, diagnostic, medical applications, process
control, bioreactors, quality control, agriculture and veterinary medicine, bacterial and viral
diagnostic, drag production, control of industrial waste water, mining, military defense
industry [5], etc.
A few advantages of biosensors are listed below:

1. They can measure nonpolar molecules that do not respond to most measurement devices
2. Biosensors are specific due to the immobilized system used in them
3. Rapid and continuous control is possible with biosensors
4. Response time is short (typically less than a minute) and
5. Practical
There are also some disadvantages of biosensors:
1. Heat sterilization is not possible because of denaturaziation of biological material,
2. Stability of biological material (such as enzyme, cell, antibody, tissue, etc.), depends on
the natural properties of the molecule that can be denaturalized under environmental
conditions (pH, temperature or ions)
3. The cells in the biosensor can become intoxicated by other molecules that are capable of
diffusing through the membrane.
Applications
There are many potential applications of biosensors of various types. The main requirements for
a biosensor approach to be valuable in terms of research and commercial applications are the
identification of a target molecule, availability of a suitable biological recognition element, and
the potential for disposable portable detection systems to be preferred to sensitive laboratorybased techniques in some situations. Some examples are given below:

Glucose monitoring in diabetes patients historical market driver

Other medical health related targets

Environmental applications e.g. the detection of pesticides and river water contaminants
such as heavy metal ions[39]

Remote sensing of airborne bacteria e.g. in counter-bioterrorist activities

Remote sensing of water quality in coastal waters by describing online different aspects
of clam ethology (biological rhythms, growth rates, spawning or death records) in groups
of abandoned bivalves around the world[6]

Detection of pathogens[40]

Determining levels of toxic substances before and after bioremediation

Detection and determining of organophosphate

Routine analytical measurement of folic acid, biotin, vitamin B12 and pantothenic acid as
an alternative to microbiological assay

Determination of drug residues in food, such as antibiotics and growth promoters,

particularly meat and honey.

Drug discovery and evaluation of biological activity of new compounds.

Protein engineering in biosensors[41]