A WIDE REVIEW ON HUMAN BIOSENSORS AND ITS APPLICATIONS

www.wjpps.com │ Vol 10, Issue 11, 2021. │ ISO 9001:2015 Certified Journal │

1998

Anjana et al. World Journal of Pharmacy and Pharmaceutical Sciences

A WIDE REVIEW ON HUMAN BIOSENSORS AND ITS

APPLICATIONS

M. N. Anjana1*

, Sajid S. Sajudeen2, Gifty Jose

2, Besty Antony

2 and Anjali Krishnan R.

2

1Assistant Professor Department of Pharmaceutics, Pushpagiri College of Pharmacy,

Medicity Campus, Thiruvalla, Kerala, 689101, India.

2Final Year B.Pharm Students, Pushpagiri College of Pharmacy, Medicity Campus,

Thiruvalla, Kerala, 689101, India.

ABSTRACT

As the potential threat of bioterrorism increases, there is great need for a

tool that can quick, reliably and accurately detect contaminating bio-

agents in the atmosphere. Biosensing elements are a set of biological

entity, those that are capable of carrying out specific group reactions or

can bind with particular group of compounds, to yield a detectable

signal that is read and transformed by the transducers. There are

mainly three so called generations of biosensors; First generation

biosensors where the normal product of the reaction diffuses to the

transducer and causes the electrical response, second generation

biosensors which involve specific mediators between the reaction and

the transducer in order to generate improved response, and third

generation biosensors where the reaction itself causes the response and

no product or mediator diffusion is directly involved. Biosensors have

many uses in clinical analysis, general health care monitoring. Medically, biosensors can be

used for accurate and precise detection of tumors, pathogens, elevated blood glucose levels in

diabetic patients and other toxins etc. Cells have the ability to modify as per the surrounding

environment for which they are subjected to use as biosensing component. Adhesiveness to

surface is another characteristic advantage that make it is suitable candidate for

immobilization on the matrix surface and attachment of receptors on cell membrane. They are

often used in monitoring treatment effect of drugs, toxic level, drugs of different stress factors

and organic derivatives. This review mainly focused on the types of biosensors and

application of various biosensors in different fields. Biosensors based on ELISA have also

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.632

Volume 10, Issue 11, 1998-2012 Review Article ISSN 2278 – 4357

*Corresponding Author

M. N. Anjana

Assistant Professor

Department of

Pharmaceutics, Pushpagiri

College of Pharmacy,

Medicity Campus,

Thiruvalla, Kerala, 689101,

India.

Article Received on

20 Sept. 2021,

Revised on 10 Oct. 2021,

Accepted on 31 Oct. 2021

DOI: 10.20959/wjpps202111-20584


1999


been developed using labelled antibody or labelled antigen coupled with a suitable transducer.

In this case mostly the biosensors can be employed to detect the bioterrorist attacks like the

intentional use of the biological entities like Bacillus anthracic, Ebola, hepatitis C virus etc.

KEYWORDS: Biosensors, Types of biosensors, Applications.

INTRODUCTION

Biosensors can essentially serve as low-cost and highly efficient devices for this purpose in

addition to being used in other day to day applications.[1]

A chemical sensor is a device that

transforms chemical information, ranging from the concentration of a specific sample

component to total composition analysis, into an analytically useful signal. Biosensors, a

hybrid of physical and chemical sensing technique is among one of the recently described

class of the sensors. Biosensors have been widely used in different scientific disciplines due

to their results. Fluorescence producing biosensors that are encoded by genes have great

importance for researches to study and analyse the complex chemical process going on in the

cells and these kinds of biosensors could be used to target some specific locations in the cell

and it can also be expressed in specific locations in the cell of an organism. In food industry

biosensors can be used for the detection of gasses released from spoiled food, detection of

food contamination or for checking and minimizing the growth of bacteria or fungus in fresh

food. From environmental point of view, these biosensors could be enhanced to detect

pollution in air and presence of any pathogens, heavy metals etc. In military defence system,

they can be used to detect the presence of any harmful biological materials that would remain

undetectable and cause death.[2]

A biosensors is defined as ―self- contained analytical device

that combines a biological element (Biosensing components) with a physicochemical

component (bio transducer component), to generate a measurable signal for detection. A

biosensor is defined as "a self- contained analytical device that combines of an analyte of

biological importance.‖ It consists of three basic components,

a detector to detect the biomolecule and generate stimulus,

a transducer to convert the stimulus to output signal,

a signal processing system to process the output and present it in an appropriate form.


2000


Fig 1: schematic representation of biosensor.[3]

CLASSIFICATION OF BIOSENSORS

Biosensors may be classified according to the biological specificity conferring mechanism, or

to the mode of the signal transduction or, alternatively, a combination of the two, these might

also be described as amperometric, potentiometric, field effect, or conductivity sensors.

Alternatively they could be termed, for example as amperometric enzyme sensors Inczedy et

al., 1998.[4]

As an example, the former biosensors may be considered as enzyme or immune

sensors. For classification, several approaches can be utilized,

a. Depending upon the used transduction principle, biosensors should be distributed in

groups of electrochemical, mass dependent, optical, radiation sensitive and so on.

b. Enzymes, nuclic acid, proteins, saccharides, oligonucleotides, ligands etc. are various sets

of biosensors which could be acquired if bio element is considered as the basis

categorization.

c. Following the type of detected analyte, classes of DNA , glucose, toxins, mycotoxins,

drugs or enzymes based biosensors could achieved.[5]

SOME COMMON TYPES OF BIOSENSORS

1. Resonant Biosensors;

In this type of biosensor, an acoustic wave transducer is coupled with an antibody (bio-

element). When the analyte molecule (or antigen) gets attached to the membrane, the mass of

the membrane changes. The resulting change in the mass subsequently changes the resonant

frequency of the transducer. This frequency change is then measured Optical-detection

Biosensor.

2. Optical biosensors

The output transduced signal that is measured is light for this type of biosensor. The biosensor

can be made based on optical diffraction or electrochemiluminescence. In optical diffraction

based devices, a Silicon wafer is coated with a protein via covalent bonds. The wafer is


2001


exposed to UV light through a photo-mask and the antibodies become inactive in the exposed

regions. When the diced wafer chips are incubated in an analyte, antigen-antibody bindings

are formed in the active regions, thus creating a diffraction grating. This grating produces a

diffraction signal when illuminated with a light source such as laser. The resulting signal can

be measured or can be further amplified before measuring for improved sensitivity.[6]

Fig 3: optical biosensor.[6]

3. Thermal-detection Biosensors

This type of biosensor is exploiting one of the fundamental properties of biological reactions,

namely absorption or production of heat, which in turn changes the temperature of the

medium in which the reaction takes place. They are constructed by combining immobilized

enzyme molecules with temperature sensors. When the analyte comes in contact with the

enzyme, the heat reaction of the enzyme is measured and is calibrated against the analyte

concentration. The total heat produced or absorbed is proportional to the molar enthalpy and

the total number of molecules in the reaction. The measurement of the temperature is

typically accomplished via a thermistor, and such devices are known as enzyme thermistors.

Their high sensitivity to thermal changes makes thermistors ideal for such applications. Unlike

other transducers, thermal biosensors do not need frequent recalibration and are insensitive to

the optical and electrochemical properties of the sample. Common applications of type of

biosensor include the detection of pesticides and pathogenic bacteria.[7]

4. Ion-Sensitive Biosensors

These are semiconductor FETs having an ion-sensitive surface. The surface electrical potential

changes when the ions and the semiconductor interact. This change in the potential can be

subsequently measured. The Ion Sensitive Field Effect Transistor (ISFET) can be constructed

by covering the sensor electrode with a polymer layer. This polymer layer is selectively


2002


permeable to analyte ions. The ions diffuse through the polymer layer and in turn cause a

change in the FET surface potential. This type of biosensor is also called an ENFET (Enzyme

Field Effect Transistor) and is primarily used for pH detection.[8]

5. Electrochemical Biosensors

Electrochemical biosensors are mainly used for the detection of hybridized DNA, DNA-

binding drugs, glucose concentration, etc. The underlying principle for this class of

biosensors is that many chemical reactions produce or consume ions or electrons which in

turn cause some change in the electrical properties of the solution which can be sensed out

and used as measuring parameter. Electrochemical biosensors can be classified based on the

measuring electrical parameters as.[9]

a. Conductimetric,

b. Amperometric

c. Potentiometric.

d. Glucose Biosensors

The most commercially successful biosensors are amperometric glucose biosensors. These

biosensors have been made available in the market in various shapes and forms such as

glucose pens, glucose displays, etc. The first historic experiment that served as the origin of

glucose biosensors was carried out by Leland C. Clark. He used platinum (Pt) electrodes to

detect oxygen. The enzyme glucose oxidase (GOD) was placed very close to the surface of

platinum by physically trapping it against the electrodes with a piece of dialysis membrane.

The enzyme activity changes depending on the surrounding oxygen concentration. Glucose

reacts with glucose oxidase (GOD) to form gluconic acid while producing two electrons and

two protons, thus reducing GOD. The reduced GOD, surrounding oxygen, electrons and

protons (produced above) react to form hydrogen peroxide and oxidized GOD (the original

form). This GOD can again react with more glucose. The higher the glucose content, more

oxygen is consumed. On the other hand, lower glucose content results in more hydrogen

peroxide. Hence, either the consumption of oxygen or the production of hydrogen peroxide

can be detected by the help of platinum electrodes and this can serve as a measure for glucose

concentration. Disposable amperometric biosensors for the detection of glucose are also

available. The typical configuration is a button- shaped biosensor consisting of the following

layers: metallic substrate, graphite layer, isolating layer, mediator modified membrane,

immobilized enzyme membrane (GOD), and a cellulose acetate membrane. This biosensor


2003


uses graphite electrodes instead of platinum electrodes (as originally used by Clark). The

isolating layer is placed on the graphite electrodes which can filter out certain interfering

substances (ascorbic acid, uric acid) while allowing the passage of hydrogen peroxide and

oxygen.[10]

The amperometric reading of the biosensor (current versus glucose concentration)

shows that the relationship is linear up to a specific glucose concentration. In other words

current increases linearly with glucose concentration, hence it can be used for detection. The

current and future applications of glucose biosensors are very broad due to their immediate use

in diabetic self-monitoring of capillary blood glucose.

Fig 7: Glucose biosensor.[10]

6. A Biosensor to Monitor Cell Morphology

Another type of biosensor can be used to monitor cell morphology in tissue culture

environments. The sensing principle used is known as Electric Cell- substrate Impedance

Sensing (ECIS). In this process, a small gold electrode is immersed in a tissue culture medium.

When cell get attached and spread on the electrodes, the impedance measured across the

electrodes changes. This changing impedance can be used forn understanding the cell behavior

in the culture medium. The attachment and spreading behavior of the cells are important

factors for this type of biosensor. Cancerous cells can usually grow and reproduce (mitosis)

freely in a medium without being attached to any substrate/surface. Normal cells, on the other

hand, need to be attached to a surface before they grow. After attachment the shape of the cells

becomes flat and no longer remains spherical. The principle of measurement is as follows: The


2004


cells are grown on gold electrodes. The electrodes are immersed in a tissue culture medium

which works as electrolyte. A voltage is applied through a resistance and the magnitude and

phase of the voltage are measured with a lock-in- amplifier.[11]

Since the current is constant,

the measured magnitude and phase can be assumed to be proportional to impedance

(resistance and capacitance). After some time, it is found that the resistance and capacitance

values fluctuate very often. This happens when cells are alive and moving. This type of

biosensor has several advantages: It is less time consuming compared to conventional

methods, it is possible to automate and quantify cell morphology measurements, and the

fluctuating pattern can be used as signature for a cell.

7. DNA Detection

The category of biosensors used for DNA detection is also known as biodetectors. The

objective is to isolate and measure the strength of single DNA–DNA or antibody–antigen

bonds, which in turn helps in detecting and characterizing single molecules of DNA or

antigen. In one method, multiple copies of the sample DNA are created using polymerase

chain reaction (PCR). On the other hand, FABS (Force Amplified Biological Sensor), BARC

(Bead Array Counter), and FDA (Force Differentiation Assay) biosensors can perform many

such measurements in a single easy operation. In these cases magnetic microbeads are used to

pull on DNA– DNA or antibody–antigen bonds with a known force, and the strengths of the

presumed bonds are tested by observing with a micromechanical sensor (FABS), or with a

magneto resistive sensor (BARC) whether the beads -detach from the surface. This kind of

biosensor is extremely useful in the detection of Antrax, Ricin, Botulinum and other

pathogens. The FABS is needed for monitoring the concentration of various biological agents

that may possibly be present in the environment. FABS is designed in such a way that it is

fully automated, compact and rugged, and can be implemented remotely. FABS can detect

various biologically active materials like toxins, proteins, viruses, and bacteria, in low

concentrations. To accomplish this it uses a sandwich assay technique, in which antibodies

against a particular protein, virus, or bacterium are covalently bound to a solid surface. The

sample solution flows over the surface, and the antibodies capture the virus present in the

sample. Next, super paramagnetic beads, also coated with an antibody against the virus, flow

through the liquid and bind to the analyte. After washing away excess beads, a number of

beads remain bound to the surface through the virus. By determining the number of beads,

the concentration of virus in the original sample is calculated. The biodetectors are used to

identify a small concentration of DNA (of microorganisms like viruses or bacteria) in a large


2005


sample. This relies on comparing sample DNA with DNA of known microorganisms (probe

DNA). Since the sample solution may contain only a small number of bio organism molecules,

multiple copies of the sample DNA need to be created for proper analysis. This is achieved by

the help of polymerase chain reaction (PCR). PCR starts by splitting samples of double-helix

DNA into two parts by heating it. If the reagents contain proper growth enzymes, then each

of these strands will grow the complementery missing part and from the double helix

structure again. This happens when the temperature is lowered. Thus, in one heating/cooling

cycle the amount of sample DNA is doubled (one cycle time is one about minute). Typically,

25-40 cycles are needed to produce approximately a billion copies. This amount is sufficient

for optical detection. The advantages of this type of biosensor are: (i) many times faster than

conventional PCR. (ii) more efficient in the number of DNA copies produced, (iii) easily

designed to use small volumes, and (iv) economical.[12]

Fig 9: Biosensors in DNA detection.[12]

PRINCIPLE OF BIOSENSOR

The desired biological material (usually a specific enzyme) is immobilized by conventional

methods (physical or membrane entrapment, noncovalent or covalent binding). This

immobilized biological material is in intimate contact with the transducer. The analyte binds

to the biological material to form a bound analyte which in turn produces the electronic

response that can be measured. In some instances, the analyte is converted to a product which

may be associated with the release of heat, gas (oxygen), electrons or hydrogen ions. The

transducer can convert the product linked changes into electrical signals which can be


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amplified and measured.[13,14]

WORKING OF BIOSENSORS

The electrical signal from the transducer is often low and superimposed upon relatively high

and noisy (containing a high frequency signal component of an apparently random nature,

due to electrical interference pre generated with in the electronic component of the

transducer) baseline.[15]

The signal processing normally involves substracting a reference

baseline signal, derived from a similar transducer without any biocatalyst membrane from the

sample signal, amplifying the resultant signal difference and electronically filtering

(smoothing) out the unwanted signal noise. The relatively slow nature of the biosensor

response considerably eases the problem of electrical noise filtration. The analogue signal

produced at this stage maybe output directly but is usually converted to a digital signal and

passed to a microprocessor stage where the data is processed, manipulated to desired units

and output to a display devices or data store.

APPLICATIONS OF BIOSENSORS IN VARIOUS FIELDS

The advantages of biosensors include low cost, small size, quick and easy use, as well as a

sensitivity and selectivity greater than the current instruments.[16]

The most popular example

is glucose oxidase-based sensor used by individuals suffering from diabetes to monitor

glucose levels in blood. Biosensors have found potential applications in the industrial

processing and monitoring, environmental pollution control, also in agricultural and food

industries. The introduction of suitable biosensors would have considerable impact in the

following areas.[17]

CLINICAL AND DIAGNOSTIC APPLICATIONS

Among wide range of applications of biosensors, the most important application is in the

field of medical diagnostics. The electrochemical variety is used now in clinical biochemistry

laboratories for measuring glucose and lactic acid. One of the key features of this is the

ability for direct measurement on undiluted blood samples. Consumer self-testing, especially

self-monitoring of blood components is another important area of clinical medicine and

healthcare to be impacted by commercial biosensors. Nowadays reusable sensors also permit

calibration and quality control unlike the present disposable sticks where only measurement

can be carried out. Such testing will improve the efficiency of patient care, replacing the often

slow and labour intensive present tests.[17]


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BIOSENSORS: THE NEW WAVE IN CANCER DIAGNOSIS

In medicine biosensors can be used to monitor blood glucose levels in diabetics, detect

pathogens, and diagnosis and moniter cancer. The military has a strong interest in the

development of biosensors as counter bioterrorism devices that can detect elements of

chemical and biological warfare to avoid potential exposure or infection. The vision of the

Future for biosensors even include chip scale devices placed in the human body, for

monitoring vital signs, correcting abnormalities or even signaling a call for help in an

emergency. Thus, by measuring levels of certain proteins expressed and/ or secreted by tumor

cells, biosensors can detect whether a tumor is present, whether it is benign or cancerous cells.

Biosensors that can detect multiple analytes may prove particularly useful in cancer diagnosis

and monitoring, since most types of cancer involved multiple biomarkers. The ability of

biosensors to test for multiple markers at once not only help with diagnosis, but also saves time

and financial resources. The earlier cancer can be detected, the better the chance of a cure.

Currently, many cancers are diagnosed only after they have metastasized throughout the

body. Effective, accurate methods of cancer detection and clinical diagnosis are urgently

needed. Biosensors are devices that are designed to detect a specific biological analyte by

essentially converting a biological entity (ie, protein, DNA, RNA) into an electrical signal

that can be detected and analyzed. The use of biosensors in cancer detection and monitoring

holds vast potential. Biosensors can be designed to detect emerging cancer biomarkers and to

determine drug effectiveness at various target sites. Biosensor technology has the potential to

provide fast and accurate detection, reliable imaging of cancer cells, and monitoring of

angiogenesis and cancer metastasis, and the ability to determine the effectiveness of

anticancer chemotherapy agents.[18]

CANCER BIOMARKERS

The National Cancer Institute (NCI) defines a biomarker as ―a biological molecule found in

blood, other body fluids, or tissues that is a sign of a normal or abnormal process or of a

condition or disease. A biomarker may be used to see how well the body responds to a

treatment for a disease or condition.[19]

Biomarkers can be of various molecular origins,

including DNA (ie, specific mutation, translocation, amplification, and loss of

heterozygosity), RNA, or protein (ie, hormone, antibody, oncogene, or tumor suppressor).

Cancer biomarkers are potentially one of the most valuable tools for early cancer detection,

accurate pretreatment staging, determining the response of cancer to chemotherapy treatment,

and monitoring dis-ease progression.[19]

Some of the major cancer biomarkers is presented


2008


below.

A. Prostate-specific antigen (PSA)

It was one of the first tumor biomarkers to be identified and put into routine clinical use and

for screening and diagnosis of prostate cancer. Studies have shown that above-normal PSA

levels correlate directly with prostate cancer. A normal level of PSA is 4.0 ng/mL. A study

conducted by Smith found that roughly 30% of men with a PSA level between 4.1 and 9.9

ng/mL had prostate cancer.

B. CA 15-3

CA 15-3 is an important biomarker analysed in breast cancer patients. Other biomarkers that

are linked to breast cancer are carcinoembryonic antigen (CEA), BRCA1, BRCA2, and CA

27.29.17,27 CA 15-3 is used clinically most often to moni-tor patient therapy in cases of

advanced breast cancer. In patients with breast cancer, it has been shown that CA 15-3

concentrations increase by 10% in Stage I cancer, 20% in Stage II, 40% in Stage III, and 75%

in Stage IV breast cancer.

C. Cancer antigen 125

Elevated cancer antigen (CA) 125 is most commonly associated with ovarian cancer and is

also linked to cancers of the uterus, cervix, pancreas, liver, colon, breast, lung and digestive

tract. Several nonpathological conditions such as menstruation and pregnancy can also result

in increased levels of CA 125.22 CA 125 is elevated in 90% of women with advanced ovarian

cancer and in 40% of patients with intra-abdominal malignancy. However, it should also be

noted that in Stage 1 ovarian cancer, 50% of patients will have normal CA 125 levels.

D. Cancer-testis antigens

Cancer-testis (CT) antigens are a unique class of cancer biomarker. They are highly

expressed in many tumors, but not in normal cells, except for germ cells of the testis. Thus,

they have been heavily pursued as potential immunogenic targets for cancer immunotherapies

(ie, cancer vaccines), and autoantibodies to antigens have been pursued as cancer biomarkers.

E. RCAS1 (EBAG-9)

Receptor-binding cancer antigen expressed on SiSo cells (RCAS1) has been shown to be

overexpressed in nearly 100% (98.4%) of gastric carcinomas and correlates closely with

gastric tumor progression. RCAS1 has also been implicated as a marker of poor prognosis in


2009


gallbladder, esophageal, breast, and endometrial cancer and with cancer relapse in laryngeal

and pharyngeal cancer.

F. Proteomics and new cancer biomarkers

The convergence of three major areas of research—advances in proteomic technologies (ie,

LC-MS/MS, MALDI-MS), sequencing of the human genome, and the development of

sophisticated software algorithms for comparing and analyzing large data sets derived from

MS data—has resulted in rapid progress in the identification of new cancer biomarkers. The

power of combining these techniques is that proteomic profiles can be generated using tumor

tissue biopsies or plasma pro-teins as the source material. Current MS techniques can gener-

ate hundreds of thousands of individual mass spectra from a given protein sample.

Industrial Applications

Along with conventional industrial fermentation producing materials, many new products are

being produced by large scale bacterial and eukaryotes cell culture. The monitoring of these

delicate and expensive processes is essential for minimizing the costs of production; specific

biosensors can be designed to measure the generation of a fermentation product.

Environmental Monitoring

Environmental water monitoring is an area in which whole cell biosensor may have

substantial advantages for the increasing number of pollutants finding their way into the

groundwater systems and hence into drinking water important targets for pollution biosensors

now include anionic pollutants such as nitrates and phosphates. The area of biosensor

development is of great importance to military and defense applications such as detection of

chemical and biological species used in weapons.

Agricultural Industry

Enzyme biosensors based on the inhibition of cholinesterases have been used to detect traces

of organophosphates and carbamates from pesticides selective and sensitive microbial sensors

for measurement of ammonia and methane have been studied. However, the only

commercially available biosensors for wastewater quality control are biological oxygen

demand (BOD) analyzers based on microorganisms like the bacteria Rhodococcus

erythropolis immobilized in collagen or polyacrylamide.


2010


Food Industry

Biosensors for the measurement of carbohydrates, alcohols, and acids are commercially

available. These instruments are mostly used in quality assurance laboratories or at best,

online coupled to the processing line through a flow injection analysis system. Their

implementation inline is limited by the need of sterility, frequent calibration, analyte dilution,

etc. Potential applications of enzyme based biosensors to food quality control include

measurement of amino acid, amines, amides, heterocyclic compounds, carbohydrates, gases,

cofactors, alcohol and phenol. Biosensors can be used in industries such as wine beer, yogurt,

and soft drinks producers. A biosensors is made up of three components; a recognition

elements, a signal transducer, a signal processor.

CONCLUSION

Biosensors offers amazing possibilities for the future these include glucose monitoring, food

analysis, clinical and diagnostic applications, environmental monitoring, bacterial

monitoring, and application in tissue engineering, for diagnosis of cancer. Although the

complexity and diversity of cancer has posed many challenges in medical field, biosensors

technology has the potential to provide fast, accurate results, while maintaining cost

effectiveness. Biosensors are the most reliable and cost effective tool available at present.

Accurate and early diagnosis of cancer will help in reducing the death rates. Further

biomarkers can also be used to monitor treatment progression and thus biosensors can

provide information about the effectiveness of the treatment. There is no doubt that this

decade will extensively involve biosensors due to their above mentioned uniqueness.

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A WIDE REVIEW ON HUMAN BIOSENSORS AND ITS APPLICATIONS

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