Chapter-3

Applications and Uses of Biosensors

 The various types of biosensors such as enzyme-based, tissue-based,
immunosensors, DNA biosensors, thermal and piezoelectric biosensors have been
deliberated here to highlight their indispensable applications in multitudinous fields.
Some of the popular fields implementing the use of biosensors are:
 Food industry to keep a check on its quality and safety, to help distinguish between
the natural and artificial; in the fermentation industry and in the saccharification
process to detect precise glucose concentrations; in metabolic engineering to enable
in vivo monitoring of cellular metabolism.
 Biosensors and their role in medical science including early-stage detection of human
interleukin-10 causing heart diseases, rapid detection of human papilloma virus, etc.
are important aspects.
 Fluorescent biosensors play a vital role in drug discovery and in cancer.
 Biosensor applications are prevalent in the plant biology sector to find out the
missing links required in metabolic processes.
 Other applications are involved in defense, clinical sector, and for marine
applications.
 Applications of Biosensors

Biosensors have been applied in many fields

namely food industry, medical field, marine
sector etc., and they provide better stability and
sensitivity as compared with the traditional
methods.
 In Food Processing, Monitoring, Food Authenticity, Quality and Safety:
An arduous quandary in food processing industry is of quality and safety, maintenance of food products
and processing. Traditional techniques performing chemical experiments and spectroscopy have
shortcomings due to human fatigue, are expensive and time consuming. Alternatives for food
authentication and monitoring with objective and consistent measurement of food products, in a cost-
effective manner, are desirable for the food industry. Thus, development of biosensors in response to the
demand for simple, real-time, selective and inexpensive techniques is seemingly propitious.
Biosensors are used for the detection of pathogens in food. Presence of Escherichia coli in vegetables, is a
bioindicator of faecal contamination in food. E. coli has been measured by detecting variation in pH caused by
ammonia (produced by urease–E. coli antibody conjugate) using potentiometric alternating biosensing systems.
Washing the vegetables such as sliced carrots and lettuce with peptone water provides us with the liquid phase. It
is then separated by amalgamating it in a sonicator, to disaffiliate bacterial cells from food items.

Enzymatic biosensors are also employed in the dairy industry. A biosensor, based on a screen-printed carbon
electrode, was integrated into a flow cell.26 Enzymes were immobilized on electrodes by engulfment in a
photocrosslinkable polymer. The automated flow-based biosensor could quantify the three organophosphate
pesticides in milk.
Fermentation Processes:

In fermentation industries, process safety and product quality are crucial. Thus, effective
monitoring of the fermentation process is imperative to develop, optimize and maintain
biological reactors at maximum efficacy. Biosensors can be utilized to monitor the presence
of products, biomass, enzyme, antibody or by-products of the process to indirectly measure
the process conditions. Biosensors precisely control the fermentation industry and produce
reproducible results due to their simple instrumentation, formidable selectivity, low prices
and easy automation. Nowadays, several kinds of commercial biosensors are accessible;
capable of detecting biochemical parameters (glucose, lactate, lysine, ethanol etc.) and are
widely used in China, occupying about 90% of its market.
Biosensing Technology for Sustainable Food Safety:

The term food quality refers to the appearance, taste, smell, nutritional value, freshness, flavor, texture and
chemicals. Smart monitoring of nutrients and fast screening of biological and chemical contaminants are of
paramount importance, when it comes to food quality and safety. Material science, nanotechnology,
electromechanical and microfluidic systems are striding in to make sensing technology imminent for use in
market. Efforts are being made for developing control systems ensuring food quality and safety and, as a
consequence, human health.

Glucose monitoring becomes indispensable as during storage the food content and composition may get
altered. German studied the electrochemistry of glucose oxidase immobilized on a graphite rod, altered by
gold nanoparticles (AuNPs), which improved its sensitivity.
 Healthcare:
In the discipline of medical science, the applications of biosensors are growing rapidly. Glucose biosensors are widely
used in clinical applications for diagnosis of diabetes mellitus, which requires precise control over blood-glucose levels.
Blood-glucose biosensors usage at home accounts for 85% of the gigantic world market.

Biosensors are being used pervasively in the medical field to diagnose infectious diseases. A promising biosensor
technology for urinary tract infection (UTI) diagnosis along with pathogen identification and anti-microbial
susceptibility is under study.

Identifying end-stage heart failure patients, prone to adverse outcomes during the early phase of left ventricular assisted
device implantation, is important. A novel biosensor, based on hafnium oxide (HfO2), has been used for early-stage
detection of human interleukin (IL)-10. Interaction between recombinant human IL-10 with corresponding monoclonal
antibody is studied for early cytokine detection after device implantation. Fluorescence patterns and electromechanical
impedance spectroscopy characterize the interaction between the antibody–antigen and bio-recognition of the protein is
achieved by fluorescence pattern. Chen et al. applied HfO2 as a greatly sensitive bio-field-effect transistor. HfO2 biosensor
has been functionalized for antibody deposition with detection of a human antigen by electrochemical impedance
spectroscopy.
The biggest dilemma faced today is of heart failure with about one million people suffering from it.
Techniques for detection of cardiovascular diseases include immunoaffinity column assay, fluorometric,
and enzyme-linked immunosorbent assay.41, 42, 43, 44, 45 These are laborious, require qualified personnel
and are time consuming. Biosensors established on electric measurement employ biochemical molecular
recognition for desired selectivity with a particular biomarker of interest.

The various other biosensors applications include: quantitative measurement of cardiac markers in
undiluted serum, microfluidic impedance assay for controlling endothelin-induced cardiac hypertrophy,
immunosensor array for clinical immunophenotyping of acute leukemias, effect of oxazaborolidines on
immobilized fructosyltransferase in dental diseases; histone deacylase (HDAC) inhibitor assay from
resonance energy transfer, biochip for a quick and accurate detection of multiple cancer markers and
neurochemical detection by diamond microneedle electrodes.
Fluorescent Biosensors

Fluorescent biosensors are imaging agents, for use in cancer and drug discovery. They have enabled
insights into the role and regulation of enzymes at cellular level. GFP-based and genetically encoded
FRET biosensors play a vital role.

Fluorescent biosensors are small scaffolds onto which one or several fluorescent probes are mounted
(enzymatically, chemically or genetically) through a receptor. The receptor identifies a specific
analyte or target, thereby transducing a fluorescent signal which can be readily detected and
measured.46, 47 Fluorescent biosensors can probe ions, metabolites, and protein biomarkers with great
sensitivity and can also report the presence, activity or status of the target (serum, cell extracts) in
complex solution. They are employed in probing gene expression, protein localization, and
conformation in fields such as signal transduction, transcription, cell cycle and apoptosis. Indication
of arthritis, inflammatory diseases, cardiovascular and neurodegenerative diseases, viral infection,
cancer and metastasis is done using these sensors.
Fluorescent biosensors are used in drug discovery programmed for the identification of drugs by
high throughput, high content screening approaches, for post screening analysis of hits and
optimization of leads. These are considered potent tools for preclinical evaluation and clinical
validation of therapeutic potential, biodistribution and pharmacokinetics of candidate drugs.
Fluorescent biosensors are effectively employed for early detection of biomarkers in molecular and
clinical diagnostics, for monitoring disease progression and response to treatment/therapeutics, for
intravital imaging and image guided surgery.

A genetically-encoded FRET biosensor developed for detection of Bcr-Abl kinase activity was used
on cancer patient cells to assess Bcr-Abl kinase activity and to establish an interrelation with the
disease status in chronic myeloid leukaemia. This probe was further employed to regulate response
to therapy, and to observe the onset of drug-resistant cells, permitting prediction for alternative
therapeutics.
Biodefense Biosensing Applications:
Biosensors can be used for military purposes at times of biological attacks. The main motive of such
biosensors is be to sensitively and selectively identify organisms posing threat in virtually real time called
biowarfare agents (BWAs) namely, bacteria (vegetative and spores), toxins and viruses. Several attempts to
device such biosensors has been done using molecular techniques which are able to recognize the chemical
markers of BWAs.

Nucleic acid-based sensing systems are more sensitive than antibody-based detection methods as they provide
gene-based specificity, without utilizing amplification steps to attain detection sensitivity to the required
levels.

The human papilloma virus HPV (double stranded DNA virus) has been categorized into two types: HPV 16
and 18; and is related to invasive cervical cancer. HPVs can be rapidly detected using a novel leaky surface
acoustic wave peptide nucleic acid biosensor with double two-port resonators. This probe directly detects HPV
genomic DNA without polymerase chain reaction amplification, and can also bind to the target DNA
sequences with a lot of efficacy and precision.
In Metabolic Engineering:
Environmental concerns and lack of sustainability of petroleum-derived products are gradually exhorting need for
development of microbial cell factories for synthesis of chemicals. Researchers view metabolic engineering as the
enabling technology for a sustainable bioeconomy.57 They have also envisioned that a substantial fraction of fuels,
commodity chemicals and pharmaceuticals will be produced from renewable feedstocks by exploiting microorganisms
rather than relying on petroleum refining or extraction from plants. The high capacity for diversity generation also
requires efficient screening methods to select the individuals carrying the desired phenotype. The earlier methods were
spectroscopy-based enzymatic assay analytics however they had limited throughput. To circumvent this obstacle
genetically encoded biosensors that enable in vivo monitoring of cellular metabolism were developed which offered
potential for high-throughput screening and selection using fluorescence-activated cell sorting (FACS) and cell survival,
respectively.

FRET sensors comprised a pair of donor and acceptor fluorophores, and a ligand-binding peptide was sandwiched
between the two. When it was bound by a ligand of interest the peptide underwent a conformational change thereby a
FRET change. Though they had high orthogonality, temporal resolution, and ease of construction, FRET sensors were
merely able to report the copiousness of metabolites concerned and were unable to exert downstream regulation to the
signal.
Biosensors in Plant Biology

Revolutionary new technologies in the areas of DNA sequencing and molecular imaging, have lead to advancements in plant
science. Traditional methods of mass spectroscopy for gauging insights into cellular and subcellular localization, and
measure of ion and metabolite levels had unprecedented precision but lacked the key information regarding location and
dynamics of enzyme substrates, receptors and transporters. However, this information can be easily successfully tapped using
biosensors. To measure a dynamic process under physiological conditions, we need to device tactics to visualize the actual
process, for instance, the conversion of one metabolite into another or triggering of signalling events. This visualization can
be done by sensors which respond dynamically.

Biosensors can be utilized to identify missing components pertinent to metabolism, regulation, or transport of the analyte.
FRET sensor for sucrose, responsible for the identification of proteins, performs a transport step in phloem loading-
sucrose efflux from the mesophyll. Fluorimeter-based assays with FRET sugar sensors successfully recognize sugar
transporters that can function immediately after exposure of starved yeast cells to glucose. Similar assays identify genes
that affect cytosolic or vacuolar pH in yeast and justify that biosensors can be applied in genetic screens provided imaging
technologies of suitable throughput are available.