Sensors International 1 (2020) 100040

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Sensors International
journal homepage: www.keaipublishing.com/en/journals/sensors-international

Biosensor nanoengineering: Design, operation, and implementation for

biomolecular analysis
Buddhadev Purohit b, Pramod R. Vernekar c, Nagaraj P. Shetti c, Pranjal Chandra a, b, *
a
Laboratory of Bio-Physio Sensors and Nanobioengineering, School of Biochemical Engineering, Indian Institute of Technology (BHU) Varanasi, Varanasi, 221005, Uttar
Pradesh, India
b
Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, Assam, India
c
Center for Electrochemical Science and Materials, Department of Chemistry, K.L.E. Institute of Technology, Hubballi, 580030, Karnataka, India

A R T I C L E I N F O A B S T R A C T

Keywords: Robust, reliable, and quantitative detection of biomarkers at ultra-low concentration is of great importance in
Biosensors clinical settings. Biosensor, an analytical device used for sensitive and selective detection of biomarkers offers
Nanoengineering various advantages over the conventional clinical diagnosis, which is both time consuming and not suitable for
Nanomaterials
point of care/onsite diagnosis. A revolution in the understanding and synthesis of nanomaterials in the last couple
Surface functionalization
Smartphone
of decades contributed significantly to the development of the biosensors in terms of sensitivity, catalytic activity,
Miniaturization biocompatibility, and robustness. Additionally, nanomaterials help in miniaturization of the sensing platform and
helping in the commercial success of portable biosensor kits. Surface engineering equally contributed to the
biosensor development by ensuring a reproducible and stable sensing surface, efficient analyte-biorecognition
element interaction, and reduced fouling effect in biological solution. Due to nanomaterial integration and sur-
face engineering, biosensors are now equally sensitive to the lab-based sophisticated instruments to detect a wide
range of molecules of clinical significance. In this review, various types of biosensors, their designs, and their
working principles have been discussed. A detailed account of various types of nanomaterials, their functional-
ization and characterization have also been discussed. The analytical performances of biosensors for both clinical
validation and analyte detection have been incorporated here. The recent trends in advanced biosensors, such as
smartphone interface for biosensing, nanozymes, lab-on-a-chip based detection methods have been discussed.

1. Introduction Microscopy, cell culture, proteomics, or molecular biology-based detec-

tion techniques are though very sensitive, sometimes require days for the
The sensitive detection of biomolecules or chemicals is an important detection of an analyte. All these techniques and instruments are too big
aspect of clinical diagnosis. Particularly, in the case of diseases like to be transported to remote locations and unreliable for detection under
cancer, where early detection can increase the chances of recovery and emergency conditions. There is a requirement of instruments and
survival [1,2]. However, at the earliest stages of a disease, the concen- detection methods, which can be used for quick and sensitive detection
tration of the clinical biomolecules/chemicals is very low. These clinical and can be transported easily. Biosensors, a bioanalytical device capable
molecules, termed as biomarkers confirm the onset of a disease, and their of detecting an ultralow amount of biomarkers with nanomaterials and
concentration reflects the stage of the disease progression. Conventional surface engineering assistance can be used for this purpose [3,4].
methods for the detection of biomarkers include high-end sophisticated The International Union of Pure and Applied Chemistry (IUPAC)
and laboratory-based instruments that require dedicated laboratory defines a biosensor as: “A device that uses specific biochemical reactions
space, skilled human resources, and a significant amount of time. mediated by isolated enzymes, immune systems, tissues, organelles or

* Corresponding author. School of Biochemical Engineering, IIT (BHU), Varanasi, 221005, Uttar Pradesh, India. ,
E-mail addresses: [email protected], [email protected] (P. Chandra).

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https://doi.org/10.1016/j.sintl.2020.100040
Received 14 July 2020; Received in revised form 3 September 2020; Accepted 3 September 2020
Available online 9 September 2020
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B. Purohit et al. Sensors International 1 (2020) 100040

whole cells to detect chemical compounds usually by electrical, thermal In this review, a detailed account of the designing of a biosensor has
or optical signals.” [5] The biosensors are essentially comprised of three been discussed with insight into the uses of nanomaterials, and surface
units i.e.a biorecognition element (BRE), a transducer, and an amplifier engineering for enhancing the efficiency of biosensors. Different types of
and processor. The BRE recognizes the analyte of interest, the transducer biosensors, their analytical performances, and signal amplification stra-
converts the biorecognition event into a measurable signal, which is tegies are also discussed. In recent years, several new technologies have
further processed by a processor and amplifier to get a signal output.A also been incorporated into the biosensors to improve their perfor-
schematic diagram of the fundamental constituents and classification of mances. Some of these methods have been incorporated into the text.
biosensors is shown in Fig. 1. Biosensors are analytical devices, which are
used not only to detect the presence and quantity of an analyte but also to 2. Biosensor
study several other phenomena like cell physiology, cell mechanics, the
effect of drug molecules on cell, etc. [6–8]. So, biosensors are designed From clinical detection [15–17], environmental studies [18], cell
according to the target and transduction mechanism to get a measurable physiology [19]to study the effect of space on astronauts [20,21], bio-
signal with high resolution to distinguish the smallest alternation in sensors have been used successfully in a lot of area of modern research.
concentration of target analyte [9,10]. Also, the biosensors should be The biosensors are designed according to the target analyte as well as the
able to distinguish the target from the non-specific molecules in the transducing mechanism. Based on the uses of labels, biosensors can be
sensing solution. The analytical performance of the biosensors is classified into labeled and label-free biosensors [22]. The labeled bio-
measured in terms of the sensitivity, selectivity, limit of detection, sensors use a reporter or label to detect the analyte, such as enzymes
detection range, reusable capacity, etc. However, the biosensors are (horseradish peroxidase (HRP), catalase, alkaline phosphatase (ALP)),
prone to hindrance in the signal generation due to the effect of sensing electroactive compounds or fluorescent molecules. Though the label
matrix and co-existing molecules [11]. Nanomaterials of different size, helps in signal amplification and increased selectivity for sensing, it in-
shape, and properties, such as metallic nanomaterials, polymers, carbon creases the total cost and sensing time. On the other hand, label-free
nanomaterials, quantum dots, etc. have been used to enhance the effi- techniques are based on the recognition of the target by BREs, and its
ciency of the biosensors and reduce the interference from the sensing simple design favors the development of portable devices. A detailed
matrix. The nanomaterial integration leads to the concept of account of various parts of a biosensor is discussed in the following
point-of-care diagnosis (POC), a portable diagnostic kit for the robust and section.
sensitive onsite detection of the clinically important molecules. Nano-
materials have become an integral part of biosensor designing to achieve 2.1. Sensor matrix/platform
enhanced sensitivity and signal amplification by exploiting its higher
surface area, size-dependent optoelectronic property, electrical conduc- Biosensors are developed on a supporting matrix (sensor matrix),
tivity, high catalytic activity (or sometimes the inertness of material, like which is functionalized to attach BREs for sensitive detection of an an-
AuNP), and biocompatibility.A nanomaterials modified surface can alyte. The nature of the material, fabrication type, and design of a sensor
adsorb more BREs in its surface and also can detect more analytes than a matrix greatly influence the final sensing capacity of a sensor, and
non-modified one [12]. Nanomaterials helped in miniaturization of the carefully chosen based on the analyte and the transducing mechanism
detection device by developing the nanoscale platform, which is portable [23]. Paper, graphite, carbon paste, glassy carbon electrode (GCE),
and can achieve equal sensitivity to the sophisticated instruments [13]. screen printed electrodes (SPE), and indium tin oxide (ITO) are some of
To achieve selectivity, BREs are conjugated onto the nanomaterials the most widely used sensor matrices.
modified sensor surface by introducing suitable immobilization tech- Paper is a low-cost matrix used for the development of optical, elec-
niques. The functionalization strategies also help in increasing the trochemical as well as microfluidic sensing platforms owing to its
detection range of the biosensors by increasing the loading of the BREs on porosity and presence of intrinsic capillary action facilitating the self-
the nanomaterials. These strategies can be covalent interactions or pumping of sample fluids [24]. Choosing a good grade of paper, based
non-covalent interactions. The non-covalent interactions do not destruct on the composition and porosity is important for achieving a desired
the conjugated skeletons, and reduce the loss of electronic properties of sensitivity and stability [25]. A large portion of the commercialized
the nanoparticles (NPs). However, the covalent interactions are preferred biosensors is developed on paper-based dipsticks, lateral flow assay
for its stability and reproducibility of the conjugation surface. Both the (LFA), or μ-PAD [25]. Noble metals (platinum, gold, silver) electrodes are
nanomaterials and the surface chemistry are required for the detection of known for their excellent electron transfer kinetics. Among them, Au is
ultra-low concentration of the analyte and its stability in biological the most commonly used electrode for its stability and inertness with a
samples [14]. working range of
0.1 and 1.3 V.Among various ceramic electrodes, ITO
is the most commonly used due to its excellent electrical conductivity,
wide working window (
0.4–1.9 V), excellent substrate adhesion ca-
pacity, low capacitive current, and stable electrochemical and physical
features, and optical transparency [26–28]. ITO has been progressively
used in biosensors by using salinization, electrophoretic and electro-
chemical deposition, polymerization, and adsorption based modification
strategies to anchor biomolecules. However, ITO is unstable in acidic
environments, and its electron transfer kinetics is less than GCE and
noble metal electrodes. GCEexhibitsa low oxidation rate, chemical
inertness, and wide potential window required for effective biosensing
[29–32]. Bare GCE can effectively catalyze some sensing reactions
without any nanofabrication in the relevant concentration of analytes
[33,34], though GCE is largely known as chemically inert. The
edge-plane sites and defects present on the GCE electrode surface are the
site of such catalysis [35,36]. Screen-printed electrodes are disposable
Fig. 1. Shows the basic design of biosensors with the three most important electrodes developed by printing various types of inks (e.g. carbon, silver,
components, i.e. BREs, transducer, and processor. Based on the transducing and gold inks) on different ceramic or plastic substrates, with tunable
mechanism, biosensors are categorized into optical, mechanical, and electro- shape, pattern, composition and BREs [37,38]. The major advantage of
chemical biosensors. working with SPE is portability, low cost, ease of functionalization, and

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B. Purohit et al. Sensors International 1 (2020) 100040

feasibility of integrating all electrodes (reference, working, and auxiliary achieve sensitivity due to base-pairing rule, and even a single mismatch
electrodes) into a single electrochemical setup. Apart from these, can be detected efficiently to study mutations. Recently, locked nucleic
disposable sensors, such as SIMS are developed due to their low cost, but acids (LNA) and peptide nucleic acids (PNA) are used in genosensors
efficient biosensing capacity [39]. Sensor platforms/matrix should be [59].
chosen carefully, as every matrix has its characteristic pattern and behave 2.2.5. Cell and tissue are used in biosensors for the detection of
differently than others [40–42]. several analytes as well as for monitoring cell physiology and mechanics
[60,61]. Recently, bacteriophage and protein-based phage display is
2.2. Biorecognition elements (BREs) being used to identify analytes based on protein-protein interactions [62,
63]. Non-catalytic transmembrane or soluble protein receptors selec-
The BREs are immobilized on the transducer surface primarily to tively bind to specific molecules with very high affinity and specificity
interact selectively with the analyte molecules, where the biorecognition are also used as BREs.
event is quantified for further sensing applications. BREs can be classified 2.2.6. Molecularly Imprinted Polymers (MIPs) are synthetic
into biocatalytic recognition elements such as enzymes; and bio- polymer with a selective binding site for the target [64,65]. The MIPs are
complexity/bioaffinity recognition element such as an antibody, nucleic synthesized using a molecular template that creates a cavity to capture
acid sequence, aptamer [2,43]. Alternatively, a nanomaterial modified the analyte. However, currently, MIPs are being developed for small
surface is used due to its catalytic behavior to sense analytes, but they molecules only.
lack the selectivity of BREs. The most selective or sensitive BREs are not
always used in the biosensor, but a careful balance between the sensi-
2.3. Transducer
tivity, selectivity, reproducibility, reusability, and physical parameters,
ease of fabrication, etc.is maintained.A systematic BREs decision map
The transducer converts the biorecognition event into a measurable
developed by Morales and Halpern can be used for BREs selection during
signal. Based onthe transducer, biosensors can be classified into optical,
designing of a biosensor (as shown in Fig. 2) [44,45].
mechanical, or electrochemical biosensors.
The most commonly used BREs in biosensors design are as follows:
2.2.1. Enzymes, also known as biocatalyst, are known for their
2.3.1. Optical biosensors
specificity towards the substrate, which can be explained by a lock-and-
Optical biosensors monitor the changes in phase, polarization, or
key or induced fit model. Enzymatic biosensors are biocatalyticin nature
frequency in the optical field of a biorecognition element upon the
i.e. they convert the analyte to a product, which can be measured by the
interaction with the analyte [66]. Based on the transduction mechanism
change in color [46], charge, or capacitance. Enzymes are placed close to
used, an optical biosensor can be categorized into absorption, fluores-
the transducer surface or embedded in conducting nanomaterials for a
cence, luminescence-based biosensors. Adsorption based biosensors
faster and short diffusion path for enhanced signal generation. Glucose
detect the change of amplitude of light when falls on the bare sensing
oxidase is the most commonly used enzyme in biosensor design for
surface to when falls in the presence of analytes. This change in ampli-
glucose detection [47]. HRP, another enzyme, commonly conjugated to
tude can be correlated to quantify the analyte concentration in a solution.
secondary antibodies for signal enhancement [48].
When operated in the visible spectrum of light, this type of biosensors is
2.2.2. Antibody, another protein-based BREs has a unique 3D
termed as colorimetric biosensors. The naked eye based optical detection
recognition pattern on its Fab arm to recognize a particular analyte by
colorimetric biosensors (usually labeled) are generally preferred in low
forming an antigen-antibody immunocomplex. The antibody-antigen
resource settings, where no instrument is required for analyte sensing
immunocomplex can be monitored by a piezoelectric sensor, optical, or
[67]. Analyte concentration can be quantified by using Beer–Lambert's
electrochemical sensor [49–51]. Monoclonal antibodies, polyclonal an-
law as shown in Equation (1).
tibodies, and recombinant antibodies are used in biosensors, where the
proper orientation can increase the sensitivity of the biosensor by many I0
fold [52,53]. log ¼ εCΔx (1)
I
2.2.3. Aptamer, a synthetic oligonucleotide designed by Systemic
Evolution of Ligands by Exponential Enrichment (SELEX) can be devel- where I is the intensity of reflected electromagnetic wave; I0 is the in-
oped to selectively recognize severaltar get analytes such as ions, pro- tensity of the incident electromagnetic wave; ε is the molar extinction
teins, drug molecules, or cells [54–56]. Aptamers are very selective and coefficient of the medium; C is the concentration of the analyte, and Δx is
insensitive to changes in pH, temperature, and ionic strength, for which the change in path length.
they are preferred in biosensors than antibodies. For instance, Mahato et al. developed a naked eye colorimetric optical
2.2.4. Nucleic Acid biosensors or genosensors contains a comple- biosensor where the concentration of the analyte, ALP in milk samples is
mentary sequence stem-loop probe (SLP) or linear probe (LP), in the detected by using the enzymatic behavior of ALP to catalyze a color re-
sensing surface to the target analyte sequence [57,58]. The genosensors agent to change the color of the sample solution [46]. They used a
smartphone to study the color change with references to change in ALP
concentration in the solution as shown in Fig. 3A(b). They found that the
RED color code in RGB profile of the image can be used for quantification
of the analyte (Fig. 3 A(a)), which has a steeper slope with changes in the
color of the sensor surface. Based on the developed plot, they developed a
paper-based kit for ALP detection in milk. Fluorescence-based biosensors
are currently developed to realize the detection of single-molecule or
analyte in a sample. Apart from fluorescence, fluorescence resonance
energy transfer (FRET) and fluorescence recovery after photobleaching
(FRAP) used in biosensors are also based on the Jablonski energy dia-
gram, where the incident light fluoresces at the low energy state and
independent of absorption maxima. The fluorescence-based biosensors
are used for rapid, sensitive, and highly selective detection. Furukawa
Fig. 2. The biorecognition elements decision map that can be used for selecting et al. reported a fluorophore (FAM) tagged aptamer-based biosensor for
the right BREs in the initial design of biosensors (Reprinted with permission the detection of thrombin, where the aptamer-thrombin interaction leads
from ref [44], copyright 2018 American chemical society). to fluorescence as shown in Fig. 3 B (a) [68]. Before thrombin

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B. Purohit et al. Sensors International 1 (2020) 100040

Fig. 3. (A) Colorimetric detection of ALP by using a color reagent and smartphone interface. (a) Red color code in RGB was found to be more sensitive towards color
changes and used for developing a standard dose-dependent plot for increased ALP concentration(b). (Reused with permission from Refs. [46] copyright 2018 Elsevier
Inc.); (B) Fluorescence-based detection of thrombin, (a) where the addition of thrombin leads to detachment and fluorescence of FAM dye from GO. (b) and (c) shows
the sensing surface before and after the addition of thrombin, respectively, and (c) the change in fluorescence intensity after the addition of thrombin. (Reused with
permission from Ref. [68] copyright 2013 Royal Society of Chemistry); (C) Principle of SPR based sensor, where the sensors detect a change in refractive index (reused
with permission from Ref. [72] copyright 2014 MDPI, Basel, Switzerland under creative commons attribution license). (For interpretation of the references to color in
this figure legend, the reader is referred to the Web version of this article.)

interaction, the fluorescence of the molecules was quenched by GO (as degradation. Another such method, chemiluminescence is used for sen-
shown in Fig. 3 B (b)), and an increase in the fluorescence was correlated sitive detection of various analytes, which doesn't require an incident
to the interaction of increased thrombin-aptamer (as shown in Fig. 3 B light source and suitable for in-vivo sensing [69,70]. Another optical
(c)). The maximum fluorescent intensity at 80 s (as shown in Fig. 3 B (c)), mode of sensing, surface plasmon resonance (SPR) based biosensors
shows the time taken by the sensor to absorb and recognize the target, measure the changes in the refractive index of the sensor surface, which
and the gradual decrease in intensity shows the fluorescence is proportional to the analyte concentration [71]. When a polarized light

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B. Purohit et al. Sensors International 1 (2020) 100040

illuminated on conducting surface at the interface of two media (such as biomolecular interaction [81]. Mechanical biosensors offer some ad-
glass and liquid) at a particular angle, a plasmon is generated resulting in vantages over other biosensors. First, these devices are capable of
a reduction in the intensity of reflected light at a particular angle, called exquisite mass resolution, which can be used to develop bioaffinity based
as resonance angle as shown inFig. 3C [72]. This phenomenon depends biosensors to detect the smallest amount of analyte (e.g. nano-
on the mass on the sensing surface, which can be quantified by a sen- electromechanical systems (NEMS) have achieved zeptogram scale mass
sorgram measuring the shift of reflectivity, angle, or wavelengths against resolution) [82]. Secondly, these devices undergo a measurable
time and use to quantify the amount of analyte on the sensing surface. An displacement against an applied force (convertible to an electric signal),
SPR based sensor consists of an optical detector (to measure intensity which can be used for the quantification of forces at the cellular level [83,
shift), a chip with a gold surface, and a functionalization layer capable of 84]. Third, they have a very fast sensing time, required to study biolog-
ligand immobilization for real time detection. At least one of the inter- ical events. Fourth, the feasibility of detecting an analyte in both liquids,
acting species should be immobilized on the sensing surface for SPR and gaseous phases [85]. Fifth, no need for labeling in these devices,
based detection. Localized SPR (LSPR) of certain metal NPs, such as which is an essential requirement in optical sensing.
AuNP, AgNPs is used for imaging, sensing, as well as photo thermal The mechanical biosensors can be categorized into three major clas-
therapy [73,74].Another set of biosensors use other optical techniques, ses i.e. quartz microbalance, surface acoustic wave, and nano-mechanical
such as optical waveguide interferometric biosensors (optical phase dif- systems. Mechanical biosensors consist of a cantilever probe, a me-
ference measurement) [75], ellipsometric biosensors (measures changes chanical transducer, and the processor. The cantilever probe is modified
in the polarization of light) [76], surface-enhanced Raman scattering to detect the analyte, where its performance, such as protein adsorption
(SERS) biosensors [77] to analyze several cell processor to detect ana- capacity or limit of detection is a function of its shape/size, and pro-
lytes. Fiber optic-based optical biosensors are developed recently for fast portional to the mass of the cantilever [86]. A nano or microscale
detection without using much lab equipment [78–80]. cantilever is preferred to achieve a better LOD. Upon the interaction with
the analyte molecule, the mechanical properties of the cantilever changes
2.3.2. Mechanical biosensors in proportional to mass change, which can be quantified by deflection
In mechanical biosensors, the changes in mechano-physical parame- (static) and resonance (dynamic) modeas shown in Fig. 4 (A) [87]. In
ters such as forces, motion, masses, etc. are quantified after the static mode, the change in cantilever curvature is measured as a result of

Fig. 4. (A) Schematic representation of the mechanical biosensors based on micro-cantilever and its working principles, (a) static mode and (b) dynamic mode
(Reused with permission from Ref. [87] Copyright 2010 Royal Society of Chemistry); (B) a representative image of a cantilever and different types of dynamic mode
based sensing, (b) Transverse mode, (c) torsional mode, (di) lateral mode, and (e) longitudinal mode. (Reproduced with permission from Ref. [88] Copyright 2012
Elsevier Inc.); (C) A schematic representation of (a) the optical read-out and (b) piezo-resistive read-out of mechanical biosensors. (Reprinted with permission from
Ref. [96] Copyright 2006 Elsevier Inc.).

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B. Purohit et al. Sensors International 1 (2020) 100040

mass change due to ligand-analyte interaction. The static mode (Fig. 4 A catalytic BREs based biosensors, such as enzymes, the biocatalysts
(a)), needs a reference electrode, and the bending of the cantilever as a initiate an event increasing the electron transfer to the electrode surfa-
result of biomolecular adsorption can be mathematically explained by ce.Based onthe transducer used, electrochemical biosensors are catego-
Stoney's model as shown in equation (2): rized into amperometry/voltammetry, potentiometry, and
impedometry/conductometry.
Et2
Δσ 1
Δσ 2 ¼ Δz (2)
3ð1
UÞL2 2.3.3.1. Amperometry/voltammetry. In voltammetric/amperometric bio-
Where, (Δσ1- Δσ2) is the surface stress change between the top and sensors, a varying/static potential is applied to a working electrode vs.
the bottom sides; Δz is the cantilever displacement; E is Young's modulus; the reference electrode and the resulting current is measured. The
ν is the Poisson coefficient; and Land t is the length and thickness of applied potential initiates an electrochemical oxidation or reduction
cantilever, respectively. (redox) reaction of the electrolytes present in the solution [99,100]. This
In dynamic mode (also called resonant-mode), the cantilever acts like redox reaction occurs at the working electrode surface and generates
a microbalance (Fig. 4 A(b)), where it measures the changes in resonant electrons at a specific potential based on the nature of the electrolyte.
frequency caused by mass-change. Further, different resonant modes This current response results in a peak proportional to the analyte con-
such as transverse mode, torsional mode, lateral mode, and longitudinal centration in the solution, and the mass transport rate of the reactant
mode are used in dynamic sensing as shown in Fig. 4 (B) [88]. An added molecules from the bulk solution to the electrode acts as the rate-limiting
mass on the cantilever decreases its oscillating frequency, which can be step. The different combinations of parameters, including potential (E),
measured by equation (3): current (I), charge (Q), and time (t), is used to develop different tech-
niques for electrochemistry studies, such as cyclic Voltammetry (CV),
 
1 1 1 chronoamperometry (CA), chronocoulometry (CC),chronopotentiometry
Δm ¼
(3)
0:96π 2 f02 f12 (CP), differential pulse voltammetry (DPV), square wave voltammetry
(SWV), anodic stripping techniques, etc. Among these techniques, CV
where f0and f1 are the fundamental resonance frequency before and after and LSV are used for primary analysis to determine the redox potentia-
the mass adding (m), respectively. l/electrochemical behavior of the analyte, whereas DPV/SWV is used for
The mechanical transducers are grouped into two categories, i.e. sensitive sensing of an analyte.
displacement change based or piezoelectricity based. The displacement-
based transducer uses an optical detector that monitors a reflected laser 2.3.3.2. Potentiometry. Potentiometric biosensors measure the potential
from the cantilever surface, which changes its position as a function of of an electrochemical cell at the working electrode when no current flows
altered mass on it (Fig. 4C (a)). The transducer with a piezoelectric in the EC cell [101]. Potentiometry provides information about the redox
crystal generates piezoelectricity against a stimulus (Fig. 4C (b)), which is activities inside the EC cells. BREs such as enzymes are usually integrated
recorded for biosensing [89]. The (i) mass, (ii) surface stress, (ii) effective into potentiometric sensors, where the catalytically generated ions are
Young's modulus, and (iii) viscoelasticity of the sensor system change as a detected by the electrodes [102]. The relation between the potential and
result of the analyte adsorption, which is monitored by the mechanical the concentration of the analyte is governed by the Nernst Equation as
biosensor. follows:
Wu et al. developed a prostate-specific antigen (PSA) detection
method using a silicon nitride mechanical sensor capable of detecting RT
Ecell ¼ E0cell
lnQ (4)
PSA even in a presence of a very high concentration of non-specific nF
human serum proteins [90]. The sensor can detect the analyte concen-
where Ecell is the potential of the electrochemical cell in zero current; E0
tration sensitively based on the deflection of the cantilever in static mode
cell is the constant potential of the electrochemical cell; R is the real gas
with high specificity. Huber et al. reported a nucleic acid-based me-
constant; T is absolute temperature; n is the number of electron transfer;
chanical biosensor for the detection of mutation in the BRAF gene
F is Faraday constant, and Q is the ratio of ion concentrations at the
without using any label or PCR amplification [91]. Cantilever based
reduced state and oxidized state.
mechanical biosensors have been used in the detection of a wide range of
clinical analytes, such as protein biomarkers [92,93], and cancer cells
2.3.3.3. Impedometry/conductometry. Conductometry biosensors mea-
[94,95] efficiently in various sensing solutions.
sure changes in the electrical conductivity of a sample solution as the
composition of the solution changes during a chemical reaction. In
2.3.3. Electrochemical biosensors
impedimetric biosensors, the impedance is measured across the working
According to IUPAC, an electrochemical (EC) biosensoris defined as
and counter electrode concerning the reference electrode [103,104].
“a self-contained integrated device, which is capable of providing specific
Impedance sensors are most commonly used for label-free detection, as
quantitative or semi-quantitative analytical information” [97]. ECbio-
this method doesn't require any detection tag for sensing of analytes. The
sensors are based on the measurement of changes in current, potential,
attachment of the analytes on the nanomaterial modified surface de-
conductance, or field effect due to the interaction of the target molecule
creases the electron transfer from the solution to the electrode surface
with the BREs on the sensing surface.ECbiosensors are preferred due to
increasing the impedance of the system, which can be calibrated taking
their robustness, selectivity, sensitivity, analytical performances than
known concentrations of the analyte [10]. Mathematically impedance
other biosensors. EC biosensors are based on the three-electrode system
can be expressed as:
i.e. working electrode (e.g. GCE, acts as the transducing element), aux-
iliary/counter electrode (e.g. Pt electrode, acts to complete the circuit), U ðjωÞ
and a reference electrode (e.g. Ag/AgCl electrode, used to establish a Z ðjωÞ ¼ …; ω ¼ 2π f (5)
I ðjωÞ
stable potential) [98].Choosing the right working electrode is one of the
most important tasks in designing an EC biosensor, as nanomaterial where Z is the impedance; U is the potential; I is the current flow through
modification and surface engineering is carried out on the working the circuit, and ω is the angular frequency of sinusoidal potential and f is
electrode. The fundamental principle behind the working of the EC the frequency.
biosensor lies in the transfer kinetics of ions/electrons from the reaction Overall, the electrochemical biosensors offer robust and sensitive
center to the electrode surface. Both in label-free and labeled EC bio- detection of the analytes with the option of device miniaturization and/
sensors, the increase or decrease in electron transfer is measured. In or integrated into a smartphone. Fig. 5. Shows two different classes of

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Fig. 5. Shows two different class of electrochemical biosensor, (A) Schematic representation of the surface modification where an increase in protein concentration (b)
leads to increased Rct value, and calibrated (c) for detection in an unknown sample. (Reused with permission from Ref. [49] Copyright 2020 Elsevier Inc.; (B) shows
the sensing principle and surface modification on GCE for serotonin detection, where an increase in serotonin concentration leads to increased DPV peak value (b)
(Reused with permission from Ref. [105] Copyright 2019 Elsevier Inc.).

electrochemical biosensors. Fig. 5 (A) shows the label-free detection of concentration [105]. Such sensing approaches can be adapted based on
ALP in serum samples, where an increase in the analyte concentration the nature of the analyte, nanomaterial, and electrochemical transducer.
leads to an increase in the impedance value (measured in Resistance to
charge transfer- Rct, unit -Ohm) [49]. The increased immobilization of 3. Nanomaterials assisted surface engineering
ALP on the Au-dendroid/rGO may hinder the charge transfer from the
solution to the SPCE electrode surface, which was then used to develop a According to US Food and Drug Administration (USFDA) and EU
calibration plot against an increasing concentration of ALP. This sensor Commission, NMs are the material with feature size or at least one
was successfully used to detect the ALP concentration with comparable dimension in range of 1–100 nm; whereas according to British Standards
results to clinically followed methods. Fig. 5 (B) shows the principle of Institution, NMs are the material with feature size of 1–1000 nm [106].
serotonin sensing by an Au nanorattle modified GCE electrode. The Nanomaterials are used in many areas of research, such as drug delivery,
electrode catalyzed oxidation of serotonin released electrons into the therapeutics, photo-thermal therapy, as well as biosensing [107,108].
system and results in a current response as a function of serotonin The nanomaterials possess a higher surface to volume ratio, which is used

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B. Purohit et al. Sensors International 1 (2020) 100040

in designing biosensors to make the sensor surface more conducting, catalytic activity, and biocompatibility. Carbon-based nanomaterials
capable of more BREs attachment, and reduce the fouling effects. such as one dimensional (CNT, MWCNT, SWCNT), two dimensional
(GO), three dimensional (fullerene) have been used for the development
of optical as well as electrochemical biosensors [49,122,123]. Among
3.1. Nanomaterials in biosensors them, graphene is regarded as the wonder material [124]and its de-
rivatives such as graphene oxide [125] and reduced graphene oxide
The nanomaterials are used in biosensors fabrication to achieve fast [126]used in biosensors due to its excellent electrical property and ease
movement of charged ions between electrode and target, better electro- of functionalization. Another class of nanomaterials, the luminescent
catalytic activities, fast diffusion of the analyte, pre-concentration of semiconducting nanocrystals, quantum Dots (QD), are used in optical as
analyte molecules near the electrode surface, and anti-fouling effect well as electrochemical biosensors for photo-electrochemical (PEC)
[109–111]. Nanomaterials of different sizes, properties, shapes, and behavior [127–129]. Polymers are another low cost, flexible, and
structures are used to achieve different goals in biosensors, where BREs biocompatible nanomaterial used in biosensing applications due to its
are conjugated by functionalizing the nanomaterials to develop excellent functionalization potential and low toxicity [64]. Conducting
bio-affinity or biocatalytic based biosensor. polymers, such as poly (acetylene), poly (3,4-ethylenedioxythiophene)
Metallic nanoparticles are used for their size-dependent optical, (PEDOT), poly (thiophene) (PTh), poly (p-phenylene vinylene) (PPV),
electromagnetic, and chemical properties. Metal nanoparticles, espe- poly-(pyrrole) (PPy) and poly (aniline) (PANI) though organic in nature,
cially gold nanoparticles (AuNP) is the most widely used NP for bio- are electrically conductive, and extensively used in electrochemical
sensing applications due to its chemical inertness, biocompatibility, biosensors [130,131].
unique size-dependent optical and electronic properties, and ease of
synthesis and functionalization [110,112–114]. AuNP has been used in
combination with other nanomaterials [115,116] or conjugated with 3.2. Nanomaterials functionalization
antibody/aptamer to develop bioaffinity based biosensors [58,117,118].
Metal oxide nanoparticles (MONP), especially of d- and f-block metals Nano-surface engineering of the sensing probe is of great importance,
like Zn, Ce, Mn, and Fe are used for their catalytic activity, adsorption as it is directly responsible for the integrity of the sensing layers. Sensing
ability, biocompatibility, etc. [119,120] However, nanomaterial with two probes involve a nano-bio-interface layer where the BREs are immobi-
or more elements are now developed due to their superior catalytic lized to interact with the analyte molecules to generate a signal. The
behavior than their monometallic counterparts [7,121]. Apart from attachment of the BREs to the nanomaterials should be stable and
metallic nanoparticles, carbon-based nanomaterials are the most consistent enough to generate a uniform signal upon interaction with
commonly used in biosensing applications due to their conductivity, analyte under various physiological environments. A suitable

Fig. 6. (A) The effective biomolecular conjugation over NPs are done to achieve these major objectives, here shown with examples of proteins/peptides; and (B) Five
general bio-conjugation methods, (i) electrostatic interaction, (ii) direct interaction, (iii) secondary interactions, such as biotin-streptavidin (SA) interactions, (iv)
covalent chemical attachment, such as EDC-NHS interaction, (v) encapsulation places the peptide inside the NP material (Reproduced with permission from Refs.
[133] Copyright 2013 American Chemical Society); (C) Chitosan-supported DNA immobilization on electrode surfaces by glutaraldehyde, avidin/biotin, and EDC-NHS
conjugation (Reproduced with permission from Ref. [137] Copyright 2013 American Chemical Society).

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B. Purohit et al. Sensors International 1 (2020) 100040

functionalization strategy of the NPs is followed to ensure a particular biosensing activity. Tables 1 and 2 summarizes the most common
number of biomolecules per NP, proper orientation of the BREs on the nanomaterial characterization techniques and the parameters which can
NPs, relative separation distance of BREs from the NPs, tunable attach- be determined by these instruments.
ment affinity, conserve the optimal function and activity of biomolecules, Among all these techniques, UV–Vis is usually the first characteriza-
and reproducibility of the method in a facile manner with a variety of tion technique performed to monitor the formation of a nanoparticle by
biological molecules [132,133]. These are the major objectives, for observing the characteristic absorption bands [147]. A well-dispersed
which surface engineering is required in biosensors (shown in Fig. 6 (A)). nanomaterial in a solution gives a characteristic absorption band ac-
The conjugation strategies of the BREs over the nanomaterials can be cording to its grain size, concentration, agglomeration state, and refrac-
classified in to covalent (amide coupling reactions, cross-linking, click tive index, and also can be used to determine the stability of the NP in
chemistry, etc.) or non-covalent interactions (electrostatic interaction, colloidal solutions [142].However, electron microscopes are used to
π–π stacking, entrapment in polymers, etc.) [134].A graphical represen- visualize the nanomaterial's size and shape at more details, where elec-
tation of major immobilization strategies is shown in Fig. 6 (B). Among tron beam are used to get resolution at the atomic level. In SEM, the
them, physisorption is a simple immobilization technique, where electron beam is scanned over the surface of NPs, and the scattered
biomolecule such as proteins adsorb on to a nanomaterial surface by electrons are used to get a 3D appearance of the surface features. Though
electrostatic, hydrophobic, van der Waals, or hydrogen bonding inter- atomic-scale resolution may not be achieved by SEM, it is useful to study
action. However, these interactions are sensitive to change in pH, ionic a larger surface area and bulk materials like thin films [148]. TEM can
strength, temperature, and surface condition. Thus, stronger covalent achieve a higher resolution (less than 10
1 nm) than SEM, where trans-
interactions are preferred for biosensor development [135,136]. One mitted electron beams passed through a thin sample and the contrast of
such covalent interaction, AuNPs interact with thiol-containing (–SH, the sample to the surrounding is studied to get the information of the
mercaptans, or sulfhydryl groups) compounds like 11-mercaptoundeca- material [149,150]. SAED is also performed with TEM to know the
noic acid to form a SAM layer, can be used for further BREs functional- crystal structure, interplanar spacing, and other lattice parameters of the
ization. Among all the covalent interactions, glutaraldehyde, EDC-NHS NPs. This diffraction-based technique is often used in combination with
carbodiimide, and biotin-avidin interactions are most commonly used in X-ray based techniques. XRD is an inseparable part of NPs characteriza-
biosensing applications. A schematic representation of all these is shown tion, used to get information on the crystalline structure, nature of the
in Fig. 6 (C)taking chitosan modified Au electrode for DNA immobili- phase, lattice parameters, and crystalline grain size (by Scherrer formula)
zation as an example [137]. Glutaraldehyde acts as a linker between the often required to explain the optical and electronic behavior of the NPs.
hydroxyl group of nanomaterial and the primary amino group of protein The composition of the NPs can be known by comparing the position and
molecule [138–141]. In EDC-NHS carbodiimideconjugation, pro-
teins/antibody are attached to the nanoparticles by the formation of an
amide linkage between the –COOH group on nanoparticle complex and Table 1
–NH2 of the protein. The major advantage of this method is the reaction Various physical techniques are used for nanostructures and probe character-
occurred at physiological pH, and in water forming a strong covalent ization (adapted with permission from Refs. [146] Copyright 2018 Royal Society
bond. Also, nucleic acid synthesized with a free carboxylic acid or pri- of Chemistry).
mary amine group can be functionalized on to nanomaterial surface by Characterization Parameters studied
EDC-NHS chemistry [142]. Biotin-Avidin is one of the strongest in- Techniques
teractions in nature, which is used in affinity-based biosensing. The AFM NP size and shape in 3D mode, dispersion of NPs different
nanoparticle surface is modified with streptavidin molecules, which is matrices.
then incubated with biotin-conjugated affinity probe for the detection of BET Surface area
Cryo-TEM Study complex growth mechanisms, aggregation
the analyte [143,144]. Salinization is another technique where silane
pathways, good for molecular biology and colloid
molecules such as (3-isocyanatopropyl) triethoxysilane (IPTES), 3-ami- chemistry
nopropyltriethoxy silane (APTES) are used to form a SAM layer on a DCS NP size and size distribution
hydroxyl surface to anchor biomolecules [26]. Recently, advanced and Electron tomography Realistic 3D particle visualization, snapshots, video,
quantitative information down to the atomic scale
powerful methods are being developed for reproducible, high-sensitivity,
EDX Elemental composition
multiplexed, high-throughput microfluidic immunoassays, or enzyme DCS NP size and size distribution
assays such as photo activated protein immobilization, electrochemically FTIR Surface composition, ligand binding
activated immobilization, thermally activated immobilization, HRTEM Distinguish monocrystalline, polycrystalline, and
photo-activated protein elution, spatially addressable multiplexed pro- amorphous NPs.
MFM Standard AFM imaging together with the information of
tein immobilization, reversible protein immobilization for repetitive
magnetic moments of single NPs, study magnetic NPs in
assay, and multifunctional immobilization surface for simplified immo- the interior of cells, discriminate from nonmagnetic NPs
bilization procedure [145]. NMR Ligand density and arrangement, electronic core structure,
atomic composition, the influence of ligands on NP shape,
NP size
3.3. Characterization of nanomaterials and sensing probe
SAED crystal structure of NPs
SAXS Particle size, size distribution, growth kinetics
The progress of biosensor as an analytical device was greatly influ- TEM NP size, size monodispersity, shape, aggregation state,
enced by the revolution in the nanotechnology arena in the last few detect and localize/quantify NPs in matrices, study growth
years. The rise of sophisticated techniques, especially electron micro- kinetics
SEM Morphology, dispersion of NPs in cells and other matrices/
scopy, X-ray, and spectroscopy-based methods lead to extensive charac- supports, precision in lateral dimensions of NPs
terization of the NPs. However, researchers face problems during TGA Mass and composition of stabilizers
characterizing the nanomaterials due to the variation in sample prepa- ToF-SIMS, MALDI Chemical information (surface-sensitive) on the functional
ration, lack of universal code for the analysis of the data, lack of suitable group, molecular orientation and conformation, surface
topography
reference material for the calibration of the used analytical tools, and
UV–Vis Optical properties, size, concentration, agglomeration
analysis of the NPs in complex matrices, etc. [146] So, it is important to state, hints on NP shape
characterize the NPs in maximum possible ways to avail reliable infor- XPS Electronic structure, elemental composition, oxidation
mation regarding its shape, size, surface ligand, growth kinetics as well as states, ligand binding (surface-sensitive)
its interaction with the surrounding materials. All these parameters XRD Crystal structure, composition, crystalline grain size
Zeta potential Agglomeration state, dispersion stability,
greatly influence its behavior on the electrode and its subsequent

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Table 2 and interactions of NPs [156,157]. Table 3 summarizes the FTIR fre-
Various physical techniques used for nanomaterial characterization and corre- quencies of some of the most common functional groups on the NPs and
sponding parameters tested. (adapted with permission from Ref. [146] Copyright electrode probe [146,158]. Another such technique, Raman spectroscopy
2018 Royal Society of Chemistry. is based on the inelastic scattering of light, used to study the strength of
Parameter Characterization techniques inter/intramolecular bonds, the effect of environmental parameters on
Agglomeration state Zeta potential, DCS, UV–Vis, SEM, TEM NPs, degree of crystallinity, etc. [159,160]. Resonance Raman scattering
3D visualization 3D-tomography, AFM, SEM (RRS) is mostly used to study carbon nanostructures, whereas SERS is
Chemical state/oxidation state XAS, EELS, XPS used predominantly to study the biomolecules adsorption on the surface
Concentration UV–Vis, DCS of metal NPs. For quantitative sensing, Raman is usually performed with
Crystal structure XRD, EXAFS, HRTEM
Dispersion of NP in matrices SEM, AFM, TEM
other techniques [161], such as ToF-SIMS (to study proteins in NPs
Growth kinetics SAXS, NMR, TEM [162]) or plasmonic (to study single molecules electrochemistry [163]).
Elemental-chemical composition XRD, XPS, ICP-MS, SEM-EDX, NMR XPS, another technique, is extensively used to study ligand interactions
Ligand binding/composition/density/ XPS, FTIR, NMR, TGA and surface functionalization of NPs and core-shell structures [164,165].
arrangement/mass, surface
EDX is used for elemental mapping of elements in NPs at the atomic level.
composition
Magnetic properties SQUID, VSM, M€ ossbauer, MFM, FMR, All the above-mentioned characterization techniques are used and vali-
XMCD dated in combination with one another to evaluate the overall property
Optical properties UV–Vis–NIR, PL, EELS-STEM of the NPs.
Shape TEM, HRTEM, AFM, EPLS, FMR Each step of probe modification with NPs or biomolecules leads to an
Size (structural properties) TEM, XRD, DLS, SAXS, HRTEM, SEM,
alternation in its electronic behavior, which can be evaluated by various
AFM, DCS, ICP-MS, UV–Vis, MALDI, NMR
Size distribution DCS, DLS, SAXS, ICP-MS electrochemical techniques. Table 4 summarizes the commonly used
Surface area BET, liquid NMR electrochemical techniques for NPs and probe characterization in an
Surface charge Zeta potential, EPM electrochemical [166]. Based on the applied voltage/excitation signal/-
electrodes, voltammetric techniques are named differently, among which
CV and DPV are the most commonly used techniques in sensing appli-
intensity of the peaks at XRD with the reference available from the In-
cations. CV is used to study the electrochemical behavior of molecules, to
ternational Center for Diffraction Data (ICDD, previously known as Joint
determine the redox potential of an analyte, and for the qualitative
Committee on Powder Diffraction Standards, JCPDS database) [151].
determination [167,168]. The diffusion of electrolyte from the sample to
AFM, another scanning technique is used to get a real topographical
the electrode surface is a rate-limiting step in CV, which can be quantified
image of the NPs surface with high magnification [152,153]. With a
using the Randles-Sevcik equation [104,169]:
horizontal resolution of 0.2 nm and a vertical resolution of 0.1 nm, AFM
is used to measure surface properties such as the frictional force on the Ip ¼ (2.69 x 105) n3/2 ACD1/2 V1/2 (6)
nanoscale, the hardness of surfaces, surface charge distribution, the
surface magnetization of NPs [154,155]. AFM doesn't require any coating (where Ipis the peak current, n is the number of electrons in the redox
before imaging and also doesn't damage the samples. However, a densely reaction, A is the area of the working electrode (in cm2), C is the con-
packed NPs film, as well as a damaged cantilever, can create problems in centration of the electroactive species at the electrode (in mol cm
3), V is
AFM analysis. the scan rate (in V s
1), and D is the diffusion coefficient of the elec-
The nanomaterial surface functionalization as well as probe fabrica- troactive species (in cm2 S
1).
tion is monitored by various characterization techniques, such as FTIR, Also, the ratio of ipc (cathodic) and ipa (anodic peaks), and the dif-
Raman spectroscopy, EDX, elemental mapping, and XPS. A bio- or ference between their potential in CV reveal the reversibility of a reac-
chemically modified probe is studied in FTIR, where a spectrum in the tion. A peak current ratio closer to (
1) and the potential difference to
mid-infrared region (4000–400 cm
1) gives information regarding the 2.3RT/nF, indicates reversibility of the reaction [170]. Where R is the gas
nature and strength of various bonds and functional groups. FTIR is one constant, T is temperature, n is the number of electrons involved in the
of the most important characterization techniques to study the structure reactions, and F is the Faraday's constant. Again, the number of electrons
involved in a reaction can be calculated based on the peak shift of the
scan rate in a voltammogram by using the Laviron equation [171].DPV is
Table 3
a more sensitive technique than CV due to the dual current sampling
FTIR frequencies for some common groups present on the surface of NPs
effect and is used for the quantitative determination of analyte at low
(adapted with permission from Ref. [146] Copyright 2018 Royal Society of
Chemistry, and ref. [158] Copyright 2006 John Wiley & Sons Ltd.).
concentrations [172,173]. In another electrochemical technique, EIS,
alternating current (AC) is used to induce electrochemical reactions on
Vibrational modes Frequency (cm
1)
the working electrode to generally study the electrochemical kinetics of
Methyl C–H asymmetric/symmetricstretch 2970–2950/2880–2860 sensing surface [174,175].The result of this is plotted as the Nyquist plot
Methyl C–H asymmetric/symmetricbend 1470–1430/1380–1370 and Bode plot. From the EIS study, the heterogeneous electron transfer
C¼C alkenyl stretch 1680–1620
Aromatic C–H stretch 3130–3070
rate constant (Ks) can be measured for different electrode by using
O–H hydroxyl group, H-bonded OH stretch 3570–3200 (broad) equation (7) [174,176].
C–O stretch, primary alcohol ~1050
N–H aliphatic primary amine, NH stretch 3400–3380, 3345–3325 RT
Rct ¼ (7)
N–H primary amine, NH bend 1650–1590 n2 F 2 ACKs
C–N, primary amine, CN stretch 1090–1020
Carboxylate 1610–1550/1420–1300 (Where R is the gas constant, T is the temperature, n is the number of
Organic phosphates (P– –O stretch) 1350–1250 electrons involved in the reaction, F is the Faraday constant, A is the area
Aliphatic phosphates (P–O–C stretch) 1050–990 of the electrode, C is the concentration of electrolyte.)
Sulfonates 1365–1340/1200–1100 Apart from these, stripping voltammetry techniques, such as anodic
Organic siloxane or silicone (Si–O–Si) 1095–1075/1055–1020
Organic siloxane or silicone (Si–O–C) 1100–1080
stripping, cathodic stripping, and adsorptive stripping voltammetry is
Thiols (S–H stretch) 2600–2550 used for sensitive detection of the analyte. Current generated as a peak
Thiol or thioether, CH2–S– (C–S stretch) 710–685 during stripping of analyte from the electrode in anodic stripping vol-
Aliphatic chloro-compounds, C–Cl stretch 800–700 tammetry (most commonly used technique) is more sensitive than any
Ammonium ion 3300–3030/1430–1390
other electrochemical detection technique.

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Table 4
Different electrochemical characterization techniques used in biosensing, and their corresponding input and output electrode parameters (Adopted and reused with
permission from Ref. [172] Copyright 2017 Elsevier Inc., and ref. [177] Copyright 2018 Elsevier Ltd.).
Electrochemical Techniques Input Output

Cyclic Voltammetry

Differential Pulse Voltammetry

Square-Wave Voltammetry

Staircase Voltammetry

Electrochemical Impedance Spectroscopy (EIS)

Chronopotentiometry (Constant current)

Cyclic Chronoamperometry

Chronoamperometry (linearly rising current)

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4. Analytical studies using aptamers, antibody, or biotin-avidin like conjugation pairs.

Selectivity is the property to detect a closely related group of molecules
After the probe fabrication, the biosensors are evaluated based on a [182].
set of standard analytical performances [97,178]. Preliminary studies
involving the sensing surface are conducted to assess its sensitivity and f) Response time
selectivity towards the target analyte. A regression plot is developed
based on the signal output with varying concentrations of analyte mol- The time required for a biosensor to generate a signal against an
ecules, which is then used as a standard for further sensing applications analyte concentration or the time taken by sensor to capture a detectable
[179]. Based on the regression plot/Calibration plot, these analytical number of analyte molecules on its surface. Microfluidics, analyte cap-
performances of biosensors are evaluated, such as: ture by beads distributed in solution, nano/micromotors are some of the
strategies used to reduce the response time of a biosensor [183].
a) Sensitivity
g) Recovery time
The sensitivity of a biosensor is defined as the slope of the regression
line, indicating a change in the output signal to fluctuation in analyte The time required by a biosensor to be ready for the second sensing
concentration. Biosensor with a steeper slope is more sensitive and cycle after completing the first one is called recovery time.
capable of distinguishing a very small change in analyte concentration in An ideal biosensor has a wide LDR, low LOD, high sensitivity, high
a solution. Sensitivity can be expressed as follows: selectivity, fast response time, low recovery time, and long shelf life.
Biosensors are designed to detect the analyte sensitively in the entire
YQ
YP clinically significant range.
Sensitivity ¼ (8)
XQ
XP
h) Real sample analysis
where, XP and XQ are two analyte concentrations in the calibration plot, and
YP and YQ are their corresponding signals.
Biosensors are ideally developed for the detection of analytes of
clinical importance in various biological fluids or samples. So, after the
b) Limit of detection (LOD)
validation of the analytical performances of the biosensor, the clinical
samples are analyzed by the biosensors. The concentration of the analyte
The minimum amount of analyte that can give a distinguishable
is quantified by using the previously developed calibration plot. How-
signal in the biosensing mechanism is known as LOD. LOD is generally
ever, sometimes, when the clinical sample is not available, spike-
calculated by using Equation (9):
recovery and standard addition methods are followed to mimic the real
3SDBlank sample. The mimic solution is prepared by adding co-existing or struc-
LOD ¼ (9) turally similar molecules. In the spike-recovery method, a known con-
Slope
centration of the analyte is injected into the sensing solution and its
where LOD is the limit of detection; SDBlank is the standard deviation of blank concentration is measured by the senor taking the calibration curve as
and the slope signifies the sensitivity of the probe for the particular analyte. the standard [115,123]. Mathematically this can be explained as equa-
tion (11):
c) Limit of quantification (LOQ)
ðObservedÞ
%Recovery ¼  100 (11)
The minimum amount of target analyte that can be detected by a Expected
biosensor is known as LOQ. LOQ can be expressed mathematically using (where Observed is the value of analyte concentration observed after
Equation (10): the spiking of analyte; Neat is the value of analyte concentration present
in the sample before the spiking; and Expected is the exact concentration
10SDBlank
LOQ ¼ (10) of the analyte spiked into the sample.)
Slope
If the recovery response is close to the spiked value, then there is no
It should be noted that the LOQ could be equal to or higher than the effect of coexisting molecules on the signal generation. If the recovery
LOD, but it could not be lower than the LOD [180]. response is significantly higher than the spiked value, it is due to false-
positive results. Similarly, a very low recovery response is a false posi-
d) Linear dynamic range (LDR) tive result which may be due to the interaction of the sensing probe with
a non-specific analyte. In the standard addition method, a calibration plot
LDR is the range of analyte concentration in the regression plot, is developed by adding a known amount of analyte in the sensing solu-
where the altered signal is directly proportional to the fluctuation in tion. Then the unknown sample is treated with the sensor and the analyte
analyte concentrations. Biosensors are designed accordingly to have a concentration in it can be calculated by tracing the points to zero con-
wider LDR to cover the clinically significant concentration of the analyte centrations in the standard plot.
[181].
Apart from the sensing parameters based on the regression line, other 5. Probe development for biosensing
analytical parameters used to evaluate biosensor performances are:
A biosensor probe is designed to achieve optimum analytical perfor-
e) Selectivity mances in a wide range of physical parameters like pH, ionic strength,
temperature,etc. The successive steps involved in sensing probe designing
Selectivity is the property of a biosensor to distinguish the target are as follows:
analyte from other molecules, and produce a signal output against the
target analyte only. Specificity and selectivity are two important pa- a) Sensing matrix selection: Based on the transducers type GCE, ITO, or
rameters often used interchangeably in biosensor literature. Specificity paper-like matrix is chosen.
is the ideal condition, where the biosensors recognize only a single b) Surface engineering: It includes the selection of proper nanomaterial
target analyte even in a complex solution. Specificity can be achieved by and chemical functionalization strategies.

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B. Purohit et al. Sensors International 1 (2020) 100040

c) Selection and immobilization of BREs: Antibody, enzyme, or aptamer matrix [199]. The sensor can sense IL-6 sensitively even after exposing
is selected based on the nature of the analyte, and immobilized the electrode to human serum samples for one month. The pores in the
properly to retain its optimum function. BSA-GA excluded the bigger protein molecules, and the conjugated
d) Concepts of signal enhancement: Various strategies are adopted to antibody interacted with the specific antigen only.
amplify the sensing signal, which is essential for the detection of very
low quantity of analyte [118,184]. In voltammetry based biosensors, 6.3. Nanozymes
the NPs are used to increase the conductivity and catalytic activity of
the sensor for signal enhancement [115]. Another method uses Nanozymes are the nanomaterials that exhibit enzyme-like properties
enzyme tagged secondary antibodies to catalyze an additional reac- of catalyzing a particular set of reactions [200]. Though the natural en-
tion to enhance the signal and helps in getting a quantitative result. It zymes are far more diverse and superior in specificity towards the sub-
should be noted that this enzyme enhances the signal, but it has no strate, nanozymes are developed for their tunable catalytic activities, low
direct role in recognizing the analyte. A multi-enzyme cascade system cost, high stability, robustness to harsh environments, ease of function-
is also used to enhance the signal [185]. Different groups have also alization [201,202]. Since the first reported enzymatic activity of nano-
reported the use of stimulator/irritant to commonly extract a sizeable particles [203], more and more nanozymes have been used by exploiting
amount of analyte directly from the body tissue to be detected by the their unique physicochemical properties such as fluorescence, electricity,
sensor [186,187]. paramagnetic properties for biosensor development. Various nano-
e) Then a concentration (or dose) dependent plot is developed, which is materials such as carbon-based nanomaterials (e.g. CNT, SWNTs, GO,
used as a standard for further studies. carbon nanodots), metal NPs (e.g. AuNPs), metal oxide-based nano-
f) The next step involved the assessment of the fouling effect of co- materials (e.g. CeO2) have been used as nanozymes and based on their
existing molecules on signal generation. If the sensing matrix has a catalytic mechanism categorized into two classes i.e. oxidoreductase
negative effect on signal generation, antifouling surfaces or method- family and hydrolase family [200,204,205].
ologies are adopted to minimize the reduction in signal.
g) Real-time sensing application: The sensor probe is calibrated for the 6.4. Smartphone interface
quantitative detection of an analyte in a complex biological solution
in the presence of many coexisting molecules. The number of smartphone users worldwide is around 1.75 billion,
and using the advanced features of these devices for biosensing data
6. Recent trends in biosensor studies collection and processing at a very low cost can be proven unparalleled
[206]. A smartphone can act as a detector or/and as an interface for an
6.1. Calibration-free sensors instrument for biosensing applications [207]. The high-resolution cam-
era can be used directly for optical detection (e.g. microscopic
The biosensors are desired to function in complex systems without bio-imaging, fluorescence, colorimetric biosensing) or coupling with
any calibration on-site or sample preparation step, which may help optical refractometric fabrication for SPR based detection. Miniaturized
semiskilled persons to operate the biosensors easily. The error in signal and portable electrochemical biosensors have been developed by using
generation during the calibration-free sensors should be minimum, and inbuilt circuits or Bluetooth connected circuits to detect multiple mole-
various strategies are adopted to obtain a low level of error percentage cules [207–210].A smartphone-based application (commonly called as
[188]. Visentin et al. developed a calibration-free SPR based immuno- ‘App’) can increase the user acceptability for onsite detection.
sensor to detect Human Leucocyte Antigens (HLA) in serum [189]. Li
et al. developed a calibration-free aptamer-based biosensor, where the 6.5. Wearable biosensors
binding induced signal change is insensitive to variations in sensor sur-
face fabrication [190]. A ratio of responsive and nonresponsive square Wearable biosensors offer continuous, real-time sensing information,
wave voltammetry signal was used for the determination of cocaine, and most often use non-invasive samples for sensing a wide range of
doxorubicin, and kanamycin. The group developed another analytes [211]. Recently, electrochemical and optical-based wearable
calibration-free phenylalanine biosensor, where the specific binding of sensors are focused mostly on integrating flexible electronics, wireless
aptamer with a complex of phenylalanine and a rhodium-based receptor data transmission, self-powered sensors, and skin implantable sensors,
was monitored using a redox indicator to determine the analyte con- etc. [212]. Gao et al. developed a fully integrated wearable sensor for
centration using square wave voltammetry [191]. Adsorption and fouling human perspiration monitoring, which can detect glucose, lactose, Naþ,
on the sensor surface by biological and chemical molecules are the major Kþ simultaneously [213]. The sensor is connected via Bluetooth and
limitations for calibration-free sensor development, though the sensor real-time sensing data can be accessed directly by smartphone (Fig. 7 B).
technologies need to improve much more to be truly calibration-free. Elsherif et al. developed a hydrogel-based wearable contact lens-based
sensor to monitor tear glucose levels in real-time [214]. Recently,
6.2. Anti-fouling surface-based sensors several tattoo-based biosensing methods have been reported using
minimally invasive procedure using various dyes which change color in
The adsorption/precipitation/polymerization of non-specific bio- response to change in pH or any other stimuli [215,216]. Yetisen et al.
molecules, macromolecules, or chemicals (such as proteins, lipids, or developed a minimally invasive dermal tattoo based colorimetric
cells) to the sensor surface cause large noise, hinder in analyte – BREs biosensor sensitive enough to detect changes in glucose, pH changes on
interactions, and lead to non-specific signal output. This phenomenon, the outer skin as shown in Fig. 7 A [217].They used enzymes, dyes, and
termed as fouling is a universal problem in in-vivo sensing in all types of other chemicals to monitor the analyte concentrations onsite covering
biosensors. Hydrophilic surface coating, the release of nitric oxide, and the physiologically important range.
innovation in reference electrode are now used to avoid such drawbacks
[192,193]. An anti-fouling layer on the sensor surface not only increases 6.6. Lab on a chip
the reliability of the sensor result but also prolong the sensor shelf life.
Various polymers (poly ethylene glycol (PEG)) [194], conductive Another miniaturized detection platform, Lab-on-a-chip (LOC) is a
hydrogels [195], peptides [196], SAM layer [197], ternary DNA mono- compact millimeter to centimeter size single chip with several integrated
layers [198] have been used to reduce the fouling effects. Sabate del Río analytical techniques to detect an analyte in a very small amount of
et al. used BSA-glutaraldehyde (GA) cross-linked porous material sample and with a fast analysis time [218,219]. Microfluidics is an in-
embedded with Au nanorods to reduce the fouling effect in the sensing tegral part of LOC, where it helps to detect an analyte in a very small

13
B. Purohit et al. Sensors International 1 (2020) 100040

Fig. 7. (A) An injectable dermal tattoo based biosensors for sensing of pH, glucose, and albumin concentrations (Reused with permission from Ref. [217] Copyright
2019 Wiley-VCH Verlag GmbH & Co.); (B) wearable smart wristband to monitor several analytes in the skin (Reused with permission from Refs. [213] Copyright 2016
Macmillan Publishers Limited); (C) the skin-interfaced microfluidic lab-on-a-chip device for sensing of chloride, sodium, and zinc, and its detection by a smartphone
(Reused with permission from Ref. [222] Copyright 2018 National Academy of Sciences).

volume of the sample [220]. Paper-based microfluidics, centrifugal chips, Acknowledgments

digital nucleic acid detection chips are developed for ions, pathogens,
disease detections [221–223]. Sekine et al. developed a smartphone Dr. Pranjal Chandra thanks Prof. Pramod Kumar Jain, Director
assisted microfluidics device to detect chloride, zinc, and sodium in IIT(BHU) for encouragement and providing the necessary facility for
sweat [222]. They used fluorescence molecules that specifically develop completion of this work.
color upon interaction with sweat analyte, which can be quantified with
the smartphone as shown in Fig. 4C. Lab-on-a chip offers low cost, simple, References
and real-time detection with very low sample volume.
[1] R. Etzioni, N. Urban, S. Ramsey, M. McIntosh, S. Schwartz, B. Reid, J. Radich,
G. Anderson, L. Hartwell, Early detection: the case for early detection, Nat. Rev.
7. Future prospects and conclusion Canc. 3 (2003) 243.
[2] B. Purohit, A. Kumar, K. Mahato, S. Roy, P. Chandra, Cancer cytosensing
Biosensors are bioanalytical devices developed for sensitive detection approaches in miniaturized settings based on advanced nanomaterials and
biosensors, in: Nanotechnol. Mod. Anim. Biotechnol., Elsevier, 2019,
of analytes quantitatively. From its inception from Clark electrode [224] pp. 133–147, https://doi.org/10.1016/B978-0-12-818823-1.00009-0.
to modern-day biosensors, the research field has advanced greatly [225]. [3] R. Prakash, P. Chandra, Nanobiomaterial Engineering: Concepts and Their
Optical, mechanical, and electrochemical are the major classes of bio- Applications in Biomedicine and Diagnostics, Springer, 2020.
[4] P. Chandra, Nanobiosensors for Personalized and Onsite Biomedical Diagnosis,
sensors, where novel innovations are being widely implemented to make Institution of Engineering and Technology, 2016, https://doi.org/10.1049/
it more efficient and prepared for applications in everyday life. All these PBHE001E.
biosensors are developed to sensitively detect a very low amount of ana- [5] N. Fracchiolla, S. Artuso, A. Cortelezzi, Biosensors in clinical practice: focus on
oncohematology, Sensors 13 (2013) 6423–6447, https://doi.org/10.3390/
lyte in a complex biological matrix with low sample volume and without s130506423.
fouling effect. Several strategies are followed to make the biosensor stable [6] P. Chandra, N.X. Son, H.-B. Noh, R.N. Goyal, Y.-B. Shim, Investigation on the
and effective for a longer period. Nanomaterials and surface chemistry are downregulation of dopamine by acetaminophen administration based on their
simultaneous determination in urine, Biosens. Bioelectron. 39 (2013) 139–144.
widely utilized for the effective attachment of BREs molecules and
[7] A. Kumar, B. Purohit, K. Mahato, R. Mandal, A. Srivastava, P. Chandra, Gold-iron
creating an anti-fouling surface to retain the sensitivity for a longer time. bimetallic nanoparticles impregnated reduced graphene oxide based nanosensor
However, the biosensors need to address many challenges before being for label-free detection of biomarker related to non-alcoholic fatty liver disease,
widely applied in day to day life. The nanomaterials used in biosensors Electroanalysis 31 (2019) 2417–2428, https://doi.org/10.1002/
elan.201900337.
need to be functional and working at physiological pH, room or body [8] P. Chandra, H.-B. Noh, Y.-B. Shim, Cancer cell detection based on the interaction
temperature, under ambient conditions, and remain stable for a longer between an anticancer drug and cell membrane components, Chem. Commun. 49
period.Nanomaterials fabrication increases the effective surface area and (2013) 1900, https://doi.org/10.1039/c2cc38235k.
[9] A. Carpenter, I. Paulsen, T. Williams, Blueprints for biosensors: design, limitations,
signal intensity, but not it's signal to noise ratio [226], which should be and applications, Genes 9 (2018) 375, https://doi.org/10.3390/genes9080375.
addressed to develop better biosensor. The enzymatic biosensors should [10] K. Mahato, P.K. Maurya, P. Chandra, Fundamentals and commercial aspects of
come up with innovations to use enzymes, especially where the catalysis nanobiosensors in point-of-care clinical diagnostics 3 (2018) 1–14, https://
doi.org/10.1007/s13205-018-1148-8. Biotech. 8.
center is embedded in its 3D structure or require co-factor for catalysis. [11] G. Rocchitta, A. Spanu, S. Babudieri, G. Latte, G. Madeddu, G. Galleri, S. Nuvoli,
Low-cost, anti-fouling surfaces, and mass production of biosensors without P. Bagella, M. Demartis, V. Fiore, R. Manetti, P. Serra, Enzyme biosensors for
compromising on the sensitivity should be prioritized. biomedical applications: strategies for safeguarding analytical performances in
biological fluids, Sensors 16 (2016) 780, https://doi.org/10.3390/s16060780.
[12] P. De Luna, S.S. Mahshid, J. Das, B. Luan, E.H. Sargent, S.O. Kelley, R. Zhou, High-
Declaration of competing interest curvature nanostructuring enhances probe display for biomolecular detection,
Nano Lett. 17 (2017) 1289–1295, https://doi.org/10.1021/acs.nanolett.6b05153.
[13] K.M. Koo, E.J.H. Wee, Y. Wang, M. Trau, Enabling miniaturised personalised
The authors declare that they have no known competing interests that diagnostics: from lab-on-a-chip to lab-in-a-drop, Lab Chip 17 (2017) 3200–3220,
could influence this work. https://doi.org/10.1039/C7LC00587C.

14
B. Purohit et al. Sensors International 1 (2020) 100040

[14] J.I. Reyes-De-Corcuera, H.E. Olstad, R. García-Torres, Stability and stabilization of glutamate, Biosens. Bioelectron. 19 (2004) 1521–1528, https://doi.org/10.1016/
enzyme biosensors: the key to successful application and commercialization, j.bios.2003.12.004.
Annu. Rev. Food Sci. Technol. 9 (2018) 293–322, https://doi.org/10.1146/ [42] M.B. Rooney, D.C. Coomber, A.M. Bond, Achievement of near-reversible behavior
annurev-food-030216-025713. for the [Fe (CN) 6] 3-/4-redox couple using cyclic voltammetry at glassy carbon,
[15] S. Mahapatra, P. Chandra, Clinically practiced and commercially viable nanobio gold, and platinum macrodisk electrodes in the absence of added supporting
engineered analytical methods for COVID-19 diagnosis, Biosens. Bioelectron. 165 electrolyte, Anal. Chem. 72 (2000) 3486–3491.
(2020) 112361, https://doi.org/10.1016/j.bios.2020.112361. [43] A. Kumar, B. Purohit, K. Mahato, P. Chandra, Advance engineered nanomaterials
[16] K. Mahato, S. Kumar, A. Srivastava, P.K. Maurya, R. Singh, P. Chandra, in point-of-care immunosensing for biomedical diagnostics, Immunosensors,
Electrochemical immunosensors: fundamentals and applications in clinical 2019, pp. 238–266.
diagnostics, in: Handb. Immunoass. Technol., Elsevier, 2018, pp. 359–414. [44] M.A. Morales, J.M. Halpern, Guide to selecting a biorecognition element for
[17] N.P. Shetti, S.J. Malode, S. Roy, P. Chandra, K.R. Reddy, S. Chatterjee, biosensors, Bioconjugate Chem. 29 (2018) 3231–3239, https://doi.org/10.1021/
Electroanalytical techniques for investigating biofilms: applications in biosensing acs.bioconjchem.8b00592.
and biomolecular interfacing, in: Nanomater. Diagnostic Tools Devices, Elsevier, [45] J.A.B. Tech, A. Prasad, K. Mahato, P.K. Maurya, P. Chandra, Analytical &
2020, pp. 293–329, https://doi.org/10.1016/B978-0-12-817923-9.00011-0. Bioanalytical Techniques Biomaterials for Biosensing Applications, 2016, p. 7,
[18] M. Mahmoudpour, J. Ezzati Nazhad Dolatabadi, M. Torbati, A. Homayouni-Rad, https://doi.org/10.4172/2155-9872.1000e124.
Nanomaterials based surface plasmon resonance signal enhancement for detection [46] K. Mahato, P. Chandra, Paper-based miniaturized immunosensor for naked eye
of environmental pollution, Biosens. Bioelectron. 127 (2019) 72–84, https:// ALP detection based on digital image colorimetry integrated with smartphone,
doi.org/10.1016/j.bios.2018.12.023. Biosens. Bioelectron. 128 (2019) 9–16, https://doi.org/10.1016/
[19] J. Shi, E.S. McLamore, D. Marshall Porterfield, Nanomaterial based self- j.bios.2018.12.006.
referencing microbiosensors for cell and tissue physiology research, Biosens. [47] Y. Liu, M. Wang, F. Zhao, Z. Xu, S. Dong, The direct electron transfer of glucose
Bioelectron. 40 (2013) 127–134, https://doi.org/10.1016/j.bios.2012.06.059. oxidase and glucose biosensor based on carbon nanotubes/chitosan matrix,
[20] A. Roda, M. Mirasoli, M. Guardigli, M. Zangheri, C. Caliceti, D. Calabria, Biosens. Bioelectron. 21 (2005) 984–988.
P. Simoni, Advanced biosensors for monitoring astronauts' health during long- [48] M. Kondzior, I. Grabowska, Antibody-electroactive probe conjugates based
duration space missions, Biosens. Bioelectron. 111 (2018) 18–26, https://doi.org/ electrochemical immunosensors, Sensors 20 (2020), 2014.
10.1016/j.bios.2018.03.062. [49] K. Mahato, B. Purohit, A. Kumar, P. Chandra, Clinically comparable impedimetric
[21] A. Roda, M. Zangheri, M. Guardigli, F. Di Nardo, L. Anfossi, C. Baggiani, immunosensor for serum alkaline phosphatase detection based on
M. Benassai, E. Carrubba, G. Neri, V. Zolesi, P. Simoni, M. Mirasoli, electrochemically engineered Au-nano-Dendroids and graphene oxide
Chemiluminescence biosensor for non-invasive crew health monitoring at the nanocomposite, Biosens. Bioelectron. 148 (2020) 111815, https://doi.org/
international space station, Aerotec. Missili Spaz. 99 (2020) 103–109, https:// 10.1016/j.bios.2019.111815.
doi.org/10.1007/s42496-020-00052-4. [50] P. Chandra, H. Noh, R. Pallela, Y. Shim, Ultrasensitive detection of drug resistant
[22] O.A. Sadik, A.O. Aluoch, A. Zhou, Status of biomolecular recognition using cancer cells in biological matrixes using an amperometric nanobiosensor, Biosens.
electrochemical techniques, Biosens. Bioelectron. 24 (2009) 2749–2765, https:// Bioelectron. 70 (2015) 418–425, https://doi.org/10.1016/j.bios.2015.03.069.
doi.org/10.1016/j.bios.2008.10.003. [51] R. Pallela, P. Chandra, H.B. Noh, Y.B. Shim, An amperometric nanobiosensor using
[23] E. Cesewski, B.N. Johnson, Electrochemical biosensors for pathogen detection, a biocompatible conjugate for early detection of metastatic cancer cells in
Biosens. Bioelectron. 159 (2020) 112214, https://doi.org/10.1016/ biological fluid, Biosens. Bioelectron. 85 (2016) 883–890, https://doi.org/
j.bios.2020.112214. 10.1016/j.bios.2016.05.092.
[24] K. Mahato, A. Srivastava, P. Chandra, Paper based diagnostics for personalized [52] S. Sharma, H. Byrne, R.J. O'Kennedy, Antibodies and antibody-derived analytical
health care: emerging technologies and commercial aspects, Biosens. Bioelectron. biosensors, Essays Biochem. 60 (2016) 9–18, https://doi.org/10.1042/
96 (2017) 246–259, https://doi.org/10.1016/j.bios.2017.05.001. EBC20150002.
[25] K. Mahato, B. Purohit, A. Kumar, P. Chandra, Based Biosensors for Clinical and [53] K. Kramer, B. Hock, Antibodies for biosensors, in: Ultrathin Electrochem. Chemo-
Biomedical Applications: Emerging Engineering Concepts and Challenges, 2020. And Biosens., Springer, 2004, pp. 3–22.
[26] E.B. Aydın, M.K. Sezgintürk, Indium tin oxide (ITO): a promising material in [54] W. Zhou, P.-J. Jimmy Huang, J. Ding, J. Liu, Aptamer-based biosensors for biomedical
biosensing technology, TrAC Trends Anal. Chem. (Reference Ed.) 97 (2017) diagnostics, Analyst 139 (2014) 2627, https://doi.org/10.1039/c4an00132j.
309–315, https://doi.org/10.1016/j.trac.2017.09.021. [55] Y. Zhu, P. Chandra, C. Ban, Y.B. Shim, Electrochemical evaluation of binding
[27] L. Yang, Y. Li, AFM and impedance spectroscopy characterization of the affinity for aptamer selection using the microarray chip, Electroanalysis 24 (2012)
immobilization of antibodies on indium–tin oxide electrode through self- 1057–1064, https://doi.org/10.1002/elan.201100734.
assembled monolayer of epoxysilane and their capture of Escherichia coli O157: [56] U. Farooq, Q. Yang, M.W. Ullah, S. Wang, Principle and development of phage-
H7, Biosens. Bioelectron. 20 (2005) 1407–1416. based biosensors, in: Biosens. Environ. Monit., 2019. IntechOpen.
[28] M. Mizuhashi, Electrical properties of vacuum-deposited indium oxide and indium [57] S. Bansal, A. Jyoti, K. Mahato, P. Chandra, R. Prakash, Highly sensitive in vitro
tin oxide films, Thin Solid Films 70 (1980) 91–100. biosensor for enterotoxigenic Escherichia coli detection based on ssDNA anchored
[29] H.E. Zittel, F.J. Miller, A glassy-carbon electrode for voltammetry, Anal. Chem. 37 on PtNPs-chitosan nanocomposite, Electroanalysis 29 (2017) 2665–2671.
(1965) 200–203. [58] I. Bhatnagar, K. Mahato, K.K.R. Ealla, A. Asthana, P. Chandra, Chitosan stabilized
 Nikolic, V.M. Jovanovic, Glassy
[30] A. Dekanski, J. Stevanovic, R. Stevanovic, B.Z. gold nanoparticle mediated self-assembled glip nanobiosensor for diagnosis of
carbon electrodes, Carbon N. Y. 39 (2001) 1195–1205, https://doi.org/10.1016/ invasive aspergillosis, Int. J. Biol. Macromol. 110 (2018) 449–456.
S0008-6223(00)00228-1. [59] S. Chung, P. Chandra, J.P. Koo, Y.-B. Shim, Development of a bifunctional
[31] W. Zhang, S. Zhu, R. Luque, S. Han, L. Hu, G. Xu, Recent development of carbon nanobiosensor for screening and detection of chemokine ligand in colorectal
electrode materials and their bioanalytical and environmental applications, Chem. cancer cell line, Biosens. Bioelectron. 100 (2018) 396–403.
Soc. Rev. 45 (2016) 715–752. [60] N. Gupta, V. Renugopalakrishnan, D. Liepmann, R. Paulmurugan, B.D. Malhotra,
[32] R.L. McCreery, Advanced carbon electrode materials for molecular Cell-based biosensors: recent trends, challenges and future perspectives, Biosens.
electrochemistry, Chem. Rev. 108 (2008) 2646–2687. Bioelectron. 141 (2019) 111435, https://doi.org/10.1016/j.bios.2019.111435.
[33] M. Lu, R.G. Compton, Voltammetric pH sensing using carbon electrodes: glassy [61] P. Wang, G. Xu, L. Qin, Y. Xu, Y. Li, R. Li, Cell-based biosensors and its application
carbon behaves similarly to EPPG, Analyst 139 (2014) 4599–4605, https:// in biomedicine, Sensor. Actuator. B Chem. 108 (2005) 576–584, https://doi.org/
doi.org/10.1039/c4an00866a. 10.1016/j.snb.2004.11.056.
[34] B.R. Kozub, N.V. Rees, R.G. Compton, Electrochemical determination of nitrite at a [62] J.W. Lee, J. Song, M.P. Hwang, K.H. Lee, Nanoscale bacteriophage biosensors
bare glassy carbon electrode; why chemically modify electrodes? Sensor. Actuator. beyond phage display, Int. J. Nanomed. 8 (2013) 3917–3925, https://doi.org/
B Chem. 143 (2010) 539–546, https://doi.org/10.1016/j.snb.2009.09.065. 10.2147/IJN.S51894.
[35] C.E. Banks, T.J. Davies, G.G. Wildgoose, R.G. Compton, Electrocatalysis at graphite and [63] U. Farooq, Q. Yang, M. Wajid Ullah, S. Wang, Principle and development of phage-
carbon nanotube modified electrodes: edge-plane sites and tube ends are the reactive based biosensors, in: Biosens. Environ. Monit., 2019, p. 13, https://doi.org/
sites, Chem. Commun. (2005) 829–841, https://doi.org/10.1039/b413177k. 10.5772/intechopen.86419. IntechOpen.
[36] A. Gennaro, A.A. Isse, C.L. Bianchi, P.R. Mussini, M. Rossi, Is glassy carbon a really [64] K. Spychalska, D. Zając, S. Baluta, K. Halicka, J. Cabaj, Functional polymers
inert electrode material for the reduction of carbon-halogen bonds? Electrochem. structures for (Bio)Sensing application—a review, Polymers 12 (2020) 1154,
Commun. 11 (2009) 1932–1935, https://doi.org/10.1016/j.elecom.2009.08.021. https://doi.org/10.3390/polym12051154.
[37] H.M. Mohamed, Screen-printed disposable electrodes: pharmaceutical [65] H. Montaseri, P.B.C. Forbes, Molecularly imprinted polymer coated quantum dots
applications and recent developments, TrAC Trends Anal. Chem. (Reference Ed.) for fluorescence sensing of acetaminophen, Mater. Today Commun. 17 (2018)
82 (2016) 1–11, https://doi.org/10.1016/j.trac.2016.02.010. 480–492, https://doi.org/10.1016/j.mtcomm.2018.10.007.
[38] O.D. Renedo, M.A. Alonso-Lomillo, M.J.A. Martínez, Recent developments in the [66] S.M. Borisov, O.S. Wolfbeis, Optical Biosensors, Chem. Rev. 108 (2008) 423–461,
field of screen-printed electrodes and their related applications, Talanta 73 (2007) https://doi.org/10.1021/cr068105t.
202–219, https://doi.org/10.1016/j.talanta.2007.03.050. [67] X. Wang, F. Li, Z. Cai, K. Liu, J. Li, B. Zhang, J. He, Sensitive colorimetric assay for
[39] A.J. Killard, Disposable sensors, Curr. Opin. Electrochem. 3 (2017) 57–62, https:// uric acid and glucose detection based on multilayer-modified paper with
doi.org/10.1016/j.coelec.2017.06.013. smartphone as signal readout, Anal. Bioanal. Chem. 410 (2018) 2647–2655.
[40] B. Serra, S. Jim enez, M.L. Mena, A.J. Reviejo, J.M. Pingarr on, Composite [68] K. Furukawa, Y. Ueno, E. Tamechika, H. Hibino, Protein recognition on a single
electrochemical biosensors: a comparison of three different electrode matrices for graphene oxide surface fixed on a solid support, J. Mater. Chem. B. 1 (2013)
the construction of amperometric tyrosinase biosensors, Biosens. Bioelectron. 17 1119–1124, https://doi.org/10.1039/c2tb00167e.
(2002) 217–226, https://doi.org/10.1016/S0956-5663(01)00269-X. [69] H.-W. Yeh, H.-W. Ai, Development and applications of bioluminescent and
[41] R.D. O'Neill, S.C. Chang, J.P. Lowry, C.J. McNeil, Comparisons of platinum, gold, chemiluminescent reporters and biosensors, Annu. Rev. Anal. Chem. 12 (2019)
palladium and glassy carbon as electrode materials in the design of biosensors for 129–150, https://doi.org/10.1146/annurev-anchem-061318-115027.

15
B. Purohit et al. Sensors International 1 (2020) 100040

[70] R. Nie, X. Xu, X. Cui, Y. Chen, L. Yang, A highly sensitive capillary-based applied chemistry: physical chemistry division, commission I.7 (biophysical
immunosensor by combining with peroxidase nanocomplex-mediated signal chemistry); analytical chemistry division, commission V.5 (electroanalytical,
amplification for detection of procalcitonin in human serum, ACS Omega 4 (2019) biosens, Bioelectron. 16 (2001) 121–131, https://doi.org/10.1016/s0956-
6210–6217, https://doi.org/10.1021/acsomega.9b00249. 5663(01)00115-4.

[71] P. Damborský, J. Svitel, J. Katrlík, Optical biosensors, Essays Biochem. 60 (2016) [98] B.R. Eggins, Chemical Sensors and Biosensors, John Wiley & Sons, 2008.
91–100, https://doi.org/10.1042/EBC20150010. [99] S.G. Meirinho, L.G. Dias, A.M. Peres, L.R. Rodrigues, Voltammetric aptasensors for
[72] Y. Yanase, T. Hiragun, K. Ishii, T. Kawaguchi, T. Yanase, M. Kawai, K. Sakamoto, protein disease biomarkers detection: a review, Biotechnol. Adv. 34 (2016)
M. Hide, Surface plasmon resonance for cell-based clinical diagnosis, Sensors 14 941–953, https://doi.org/10.1016/j.biotechadv.2016.05.006.
(2014) 4948–4959, https://doi.org/10.3390/s140304948. [100] V.K. Gupta, R. Jain, K. Radhapyari, N. Jadon, S. Agarwal, Voltammetric techniques
[73] S. Zhang, Q. Huang, L. Zhang, H. Zhang, Y. Han, Q. Sun, Z. Cheng, H. Qin, S. Dou, for the assay of pharmaceuticals-A review, Anal. Biochem. 408 (2011) 179–196,
Z. Li, Vacancy engineering of Cu 2- x Se nanoparticles with tunable LSPR and https://doi.org/10.1016/j.ab.2010.09.027.
magnetism for dual-modal imaging guided photothermal therapy of cancer, [101] J. Ding, W. Qin, Recent advances in potentiometric biosensors, TrAC Trends Anal.
Nanoscale 10 (2018) 3130–3143. Chem. (Reference Ed.) 124 (2020) 115803, https://doi.org/10.1016/
[74] R. Ahmad Mohamed Ali, D. Mita, W. Espulgar, M. Saito, M. Nishide, j.trac.2019.115803.
H. Takamatsu, H. Yoshikawa, E. Tamiya, Single cell analysis of neutrophils NETs [102] D.S. Juang, C.H. Lin, Y.R. Huo, C.Y. Tang, C.R. Cheng, H.S. Wu, S.F. Huang,
by microscopic LSPR imaging system, Micromachines 11 (2020) 52. A. Kalnitsky, C.C. Lin, Proton-ELISA: electrochemical immunoassay on a dual-
[75] P. Kozma, F. Kehl, E. Ehrentreich-F€ orster, C. Stamm, F.F. Bier, Integrated planar gated ISFET array, Biosens. Bioelectron. 117 (2018) 175–182, https://doi.org/
optical waveguide interferometer biosensors: a comparative review, Biosens. 10.1016/j.bios.2018.06.012.
Bioelectron. 58 (2014) 287–307. [103] P. Chandra, Y.N. Tan, S.P. Singh, Next Generation Point-of-care Biomedical
[76] Y. Fei, Y.-S. Sun, Y. Li, H. Yu, K. Lau, J.P. Landry, Z. Luo, N. Baumgarth, X. Chen, Sensors Technologies for Cancer Diagnosis, Springer Singapore, Singapore, 2017,
X. Zhu, Characterization of receptor binding profiles of influenza a viruses using https://doi.org/10.1007/978-981-10-4726-8.
an ellipsometry-based label-free glycan microarray assay platform, Biomolecules 5 [104] A.J. Bard, L.R. Faulkner, J. Leddy, C.G. Zoski, Electrochemical Methods:
(2015) 1480–1498. Fundamentals and Applications, wiley, New York, 1980.
[77] Y. Pang, C. Wang, L. Lu, C. Wang, Z. Sun, R. Xiao, Dual-SERS biosensor for one- [105] K. Mahato, B. Purohit, K. Bhardwaj, A. Jaiswal, P. Chandra, Novel electrochemical
step detection of microRNAs in exosome and residual plasma of blood samples for biosensor for serotonin detection based on gold nanorattles decorated reduced
diagnosing pancreatic cancer, Biosens, Bioelectron 130 (2019) 204–213. graphene oxide in biological fluids and in vitro model, Biosens. Bioelectron. 142
[78] N. Polley, S. Basak, R. Hass, C. Pacholski, Fiber optic plasmonic sensors: providing (2019) 111502, https://doi.org/10.1016/j.bios.2019.111502.
sensitive biosensor platforms with minimal lab equipment, Biosens. Bioelectron. [106] J. Jeevanandam, A. Barhoum, Y.S. Chan, A. Dufresne, M.K. Danquah, Review on
132 (2019) 368–374, https://doi.org/10.1016/j.bios.2019.03.020. nanoparticles and nanostructured materials: history, sources, toxicity and
[79] Y.J. Zhang, J.C. Hsu, J.H. Tsao, Y.S. Sun, Fabrication of a bare optical fiber-based regulations, Beilstein J. Nanotechnol. 9 (2018) 1050–1074.
biosensor, Micromachines 10 (2019) 1–8, https://doi.org/10.3390/mi10080522. [107] A. Kumar, S. Roy, A. Srivastava, M.M. Naikwade, B. Purohit, K. Mahato,
[80] Y. Zhao, R. jie Tong, F. Xia, Y. Peng, Current status of optical fiber biosensor based V.G.M. Naidu, P. Chandra, Nanotherapeutics: a novel and powerful approach in
on surface plasmon resonance, Biosens. Bioelectron. 142 (2019) 111505, https:// modern healthcare system, in: Nanotechnol. Mod. Anim. Biotechnol., Elsevier,
doi.org/10.1016/j.bios.2019.111505. 2019, pp. 149–161.
[81] J. Tamayo, P.M. Kosaka, J.J. Ruz, A.  San Paulo, M. Calleja, Biosensors based on [108] A. Baranwal, A.K. Chiranjivi, A. Kumar, V.K. Dubey, P. Chandra, Design of
nanomechanical systems, Chem. Soc. Rev. 42 (2013) 1287–1311, https://doi.org/ commercially comparable nanotherapeutic agent against human disease-causing
10.1039/c2cs35293a. parasite, Leishmania, Sci. Rep. 8 (2018) 8814, https://doi.org/10.1038/s41598-
[82] T. Braun, M.K. Ghatkesar, N. Backmann, W. Grange, P. Boulanger, L. Letellier, H.- 018-27170-1.
P. Lang, A. Bietsch, C. Gerber, M. Hegner, Quantitative time-resolved [109] S. Sharma, N. Singh, V. Tomar, R. Chandra, A review on electrochemical detection
measurement of membrane protein–ligand interactions using microcantilever of serotonin based on surface modified electrodes, Biosens. Bioelectron. 107
array sensors, Nat. Nanotechnol. 4 (2009) 179. (2018) 76–93, https://doi.org/10.1016/j.bios.2018.02.013.
[83] W.J. Polacheck, C.S. Chen, Measuring cell-generated forces: a guide to the [110] B. Purohit, A. Kumar, K. Mahato, P. Chandra, Electrodeposition of metallic
available tools, Nat. Methods 13 (2016) 415–423. nanostructures for biosensing applications in health care, J. Sci. Res. 64 (2020)
[84] J. Rajagopalan, M.T.A. Saif, MEMS sensors and microsystems for cell 68–73, https://doi.org/10.37398/JSR.2020.640109.
mechanobiology, J. Micromech. Microeng. 21 (2011) 54002. [111] A. Kumar, B. Purohit, P.K. Maurya, L.M. Pandey, P. Chandra, Engineered
[85] M. Rodahl, F. H€ oo€k, A. Krozer, P. Brzezinski, B. Kasemo, Quartz crystal nanomaterial assisted signal-amplification strategies for enhancing analytical
microbalance setup for frequency and Q-factor measurements in gaseous and performance of electrochemical biosensors, Electroanalysis 31 (2019) 1615–1629,
liquid environments, Rev. Sci. Instrum. 66 (1995) 3924–3930, https://doi.org/ https://doi.org/10.1002/elan.201900216.
10.1063/1.1145396. [112] K. Mahato, S. Nagpal, M.A. Shah, A. Srivastava, P.K. Maurya, S. Roy, A. Jaiswal,
[86] A.K. Gupta, P.R. Nair, D. Akin, M.R. Ladisch, S. Broyles, M.A. Alam, R. Bashir, R. Singh, P. Chandra, Gold nanoparticle surface engineering strategies and their
Anomalous resonance in a nanomechanical biosensor, Proc. Natl. Acad. Sci. U.S.A. applications in biomedicine and diagnostics 3 (2019) 1–19, https://doi.org/
103 (2006) 13362–13367, https://doi.org/10.1073/pnas.0602022103. 10.1007/s13205-019-1577-z. Biotech. 9.
[87] M. Alvarez, L.M. Lechuga, Microcantilever-based platforms as biosensing tools, [113] K. Mahato, A. Kumar, B. Purohit, S. Mahapatra, A. Srivastava, P. Chandra,
Analyst 135 (2010) 827–836, https://doi.org/10.1039/b908503n. Nanomaterial functionalization strategies in bio-interface development for
[88] B.N. Johnson, R. Mutharasan, Biosensing using dynamic-mode cantilever sensors: modern diagnostic devices, in: Biointerface Eng. Prospect. Med. Diagnostics Drug
a review, Biosens. Bioelectron. 32 (2012) 1–18, https://doi.org/10.1016/ Deliv., Springer, 2020, pp. 195–214.
j.bios.2011.10.054. [114] A. Kumar, B. Purohit, K. Mahato, S. Mahapatra, A. Srivastava, P. Chandra, Bio-
[89] M. Pohanka, The piezoelectric biosensors: principles and applications, Int. J. nano-interface engineering strategies of AuNPs passivation for next-generation
Electrochem. Sci. 12 (2017) 496–506, https://doi.org/10.20964/2017.01.44. biomedical applications, in: Biointerface Eng. Prospect. Med. Diagnostics Drug
[90] G. Wu, R.H. Datar, K.M. Hansen, T. Thundat, R.J. Cote, A. Majumdar, Bioassay of Deliv., Springer, 2020, pp. 215–231.
prostate-specific antigen (PSA) using microcantilevers, Nat. Biotechnol. 19 (2001) [115] A. Kumar, B. Purohit, K. Mahato, S. Roy, A. Srivastava, P. Chandra, Design and
856–860, https://doi.org/10.1038/nbt0901-856. development of ultrafast sinapic acid sensor based on electrochemically
[91] F. Huber, H.P. Lang, N. Backmann, D. Rimoldi, C. Gerber, Direct detection of a nanotuned gold nanoparticles and solvothermally reduced graphene oxide,
BRAF mutation in total RNA from melanoma cells using cantilever arrays, Nat. Electroanalysis 32 (2020) 59–69, https://doi.org/10.1002/elan.201900406.
Nanotechnol. 8 (2013) 125–129, https://doi.org/10.1038/nnano.2012.263. [116] A. Baranwal, P. Chandra, Clinical implications and electrochemical biosensing of
[92] D.K. Agarwal, A. Prasad, M. Vinchurkar, S. Gandhi, D. Prabhakar, S. Mukherji, monoamine neurotransmitters in body fluids, in vitro, in vivo, and ex vivo models,
V.R. Rao, Detection of heart-type fatty acid-binding protein (h-FABP) using Biosens. Bioelectron. 121 (2018) 137–152, https://doi.org/10.1016/
piezoresistive polymer microcantilevers functionalized by a dry method, Appl. j.bios.2018.09.002.
Nanosci. 8 (2018) 1031–1042, https://doi.org/10.1007/s13204-018-0723-y. [117] P. Chandra, W. Choon, A. Koh, H. Noh, Y. Shim, Biosensors and Bioelectronics in
[93] D.K. Agarwal, A. Kushagra, M. Ashwin, A.S. Shukla, V. Palaparthy, Sensitive vitro monitoring of i-NOS concentrations with an immunosensor : the inhibitory
detection of cardiac troponin-I protein using fabricated piezoresistive effect of endocrine disruptors on i-NOS release, Biosens. Bioelectron. 32 (2012)
microcantilevers by a novel method of asymmetric biofunctionalization, 278–282, https://doi.org/10.1016/j.bios.2011.11.027.
Nanotechnology 31 (2020) 115503, https://doi.org/10.1088/1361-6528/ [118] M.H. Akhtar, K.K. Hussain, N.G. Gurudatt, P. Chandra, Y.-B. Shim, Ultrasensitive
ab5a18. dual probe immunosensor for the monitoring of nicotine induced-brain derived
[94] C. Li, M. Zhang, Z. Zhang, J. Tang, B. Zhang, Microcantilever aptasensor for neurotrophic factor released from cancer cells, Biosens, Bioelectron 116 (2018)
detecting epithelial tumor marker Mucin 1 and diagnosing human breast 108–115.
carcinoma MCF-7 cells, Sensor. Actuator. B Chem. 297 (2019) 126759, https:// [119] J.M. George, A. Antony, B. Mathew, Metal oxide nanoparticles in electrochemical
doi.org/10.1016/j.snb.2019.126759. sensing and biosensing: a review, Microchim. Acta. 185 (2018) 358, https://
[95] H. Etayash, K. Jiang, S. Azmi, T. Thundat, K. Kaur, Real-time detection of breast doi.org/10.1007/s00604-018-2894-3.
cancer cells using peptide-functionalized microcantilever arrays, Sci. Rep. 5 [120] P.R. Solanki, A. Kaushik, V. V Agrawal, B.D. Malhotra, Nanostructured metal
(2015) 1–13, https://doi.org/10.1038/srep13967. oxide-based biosensors, NPG Asia Mater. 3 (2011) 17–24.

[96] L.G. Carrascosa, M. Moreno, M. Alvarez, L.M. Lechuga, Nanomechanical [121] R. Mandal, A. Baranwal, A. Srivastava, P. Chandra, Evolving trends in bio/
biosensors: a new sensing tool, TrAC Trends Anal. Chem. (Reference Ed.) 25 chemical sensor fabrication incorporating bimetallic nanoparticles, Biosens.
(2006) 196–206, https://doi.org/10.1016/j.trac.2005.09.006. Bioelectron. 117 (2018) 546–561.
[97] D.R. Thevenot, K. Toth, R.A. Durst, G.S. Wilson, Electrochemical biosensors: [122] E. Heydari-Bafrooei, A.A. Ensafi, Typically used carbon-based nanomaterials in the
recommended definitions and classification1International union of pure and fabrication of biosensors, in: Electrochem. Biosens., Elsevier, 2019, pp. 77–98.

16
B. Purohit et al. Sensors International 1 (2020) 100040

[123] B. Purohit, A. Kumar, K. Mahato, P. Chandra, Novel sensing assembly comprising [149] M. Naito, T. Yokoyama, K. Hosokawa, K. Nogi, Nanoparticle Technology
engineered gold dendrites and MWCNT-AuNPs nanohybrid for acetaminophen Handbook, Elsevier, 2018.
detection in human urine, Electroanalysis 32 (2020) 561–570, https://doi.org/ [150] J. Park, K. An, Y. Hwang, J.-G. Park, H.-J. Noh, J.-Y. Kim, J.-H. Park, N.-M. Hwang,
10.1002/elan.201900551. T. Hyeon, Ultra-large-scale syntheses of monodisperse nanocrystals, Nat. Mater. 3
[124] K.S. Novoselov, A.K. Geim, S. V Morozov, D. Jiang, Y. Zhang, S. V Dubonos, I. V (2004) 891–895, https://doi.org/10.1038/nmat1251.
Grigorieva, A.A. Firsov, Electric field effect in atomically thin carbon films, [151] C.N.R. Rao, K. Biswas, Characterization of nanomaterials by physical methods,
Science 306 (2004) 666–669, 80-. Annu. Rev. Anal. Chem. 2 (2009) 435–462, https://doi.org/10.1146/annurev-
[125] J. Kailashiya, N. Singh, S.K. Singh, V. Agrawal, D. Dash, Graphene oxide-based anchem-060908-155236.
biosensor for detection of platelet-derived microparticles: a potential tool for [152] P.J. de Pablo, Atomic force microscopy of virus shells, in: Semin. Cell Dev. Biol.,
thrombus risk identification, Biosens. Bioelectron. 65 (2015) 274–280. 2018, pp. 199–208.
[126] J. Lee, J. Kim, S. Kim, D.-H. Min, Biosensors based on graphene oxide and its [153] M.S. Bull, R.M.A. Sullan, H. Li, T.T. Perkins, Improved single molecule force
biomedical application, Adv. Drug Deliv. Rev. 105 (2016) 275–287, https:// spectroscopy using micromachined cantilevers, ACS Nano 8 (2014) 4984–4995.
doi.org/10.1016/j.addr.2016.06.001. [154] J.S. Lee, J. Song, S.O. Kim, S. Kim, W. Lee, J.A. Jackman, D. Kim, N.-J. Cho, J. Lee,
[127] N. Zhang, L. Zhang, Y.-F. Ruan, W.-W. Zhao, J.-J. Xu, H.-Y. Chen, Quantum-dots- Multifunctional hydrogel nano-probes for atomic force microscopy, Nat. Commun.
based photoelectrochemical bioanalysis highlighted with recent examples, 7 (2016) 1–14.
Biosens. Bioelectron. 94 (2017) 207–218, https://doi.org/10.1016/ [155] D.J. Müller, Y.F. Dufrene, Atomic force microscopy as a multifunctional molecular
j.bios.2017.03.011. toolbox in nanobiotechnology, in: Nanosci. Technol. A Collect. Rev. From Nat.
[128] F.A.A. Manan, W.W. Hong, J. Abdullah, N.A. Yusof, I. Ahmad, Nanocrystalline Journals, World Scientific, 2010, pp. 269–277.
cellulose decorated quantum dots based tyrosinase biosensor for phenol [156] D.-H. Tsai, M. Davila-Morris, F.W. DelRio, S. Guha, M.R. Zachariah, V.A. Hackley,
determination, Mater. Sci. Eng. C 99 (2019) 37–46, https://doi.org/10.1016/ Quantitative determination of competitive molecular adsorption on gold
j.msec.2019.01.082. nanoparticles using attenuated total reflectance–fourier transform infrared
[129] H. Bhardwaj, C. Singh, R.K. Kotnala, G. Sumana, Graphene quantum dots-based spectroscopy, Langmuir 27 (2011) 9302–9313, https://doi.org/10.1021/
nano-biointerface platform for food toxin detection, Anal. Bioanal. Chem. 410 la2005425.
(2018) 7313–7323, https://doi.org/10.1007/s00216-018-1341-y. [157] B.C. Smith, Fundamentals of Fourier Transform Infrared Spectroscopy, CRC press,
[130] N. Aydemir, J. Malmstr€ om, J. Travas-Sejdic, Conducting polymer based 2011.
electrochemical biosensors, Phys. Chem. Chem. Phys. 18 (2016) 8264–8277, [158] J. Coates, Interpretation of infrared spectra, A practical approach, in: Encycl. Anal.
https://doi.org/10.1039/c5cp06830d. Chem., John Wiley & Sons, Ltd, Chichester, UK, 2006, pp. 10815–10837, https://
[131] F. Ghorbani Zamani, H. Moulahoum, M. Ak, D. Odaci Demirkol, S. Timur, Current doi.org/10.1002/9780470027318.a5606.
trends in the development of conducting polymers-based biosensors, TrAC Trends [159] M.S. Dresselhaus, P.C. Eklund, Phonons in carbon nanotubes, Adv. Phys. 49
Anal. Chem. (Reference Ed.) 118 (2019) 264–276, https://doi.org/10.1016/ (2000) 705–814, https://doi.org/10.1080/000187300413184.
j.trac.2019.05.031. [160] J.R. Ferraro, Introductory Raman Spectroscopy, Elsevier, 2003, ISBN 978-0-12-
[132] I. Medintz, Universal tools for biomolecular attachment to surfaces, Nat. Mater. 5 254105-6.
(2006), https://doi.org/10.1038/nmat1776, 842–842. [161] R. Goodacre, D. Graham, K. Faulds, Recent developments in quantitative SERS:
[133] K.E. Sapsford, W.R. Algar, L. Berti, K.B. Gemmill, B.J. Casey, E. Oh, M.H. Stewart, moving towards absolute quantification, TrAC Trends Anal. Chem. (Reference Ed.)
I.L. Medintz, Functionalizing nanoparticles with biological molecules: developing 102 (2018) 359–368, https://doi.org/10.1016/j.trac.2018.03.005.
chemistries that facilitate nanotechnology, Chem. Rev. 113 (2013) 1904–2074, [162] A. Rafati, A. Boussahel, K.M. Shakesheff, A.G. Shard, C.J. Roberts, X. Chen,
https://doi.org/10.1021/cr300143v. D.J. Scurr, S. Rigby-Singleton, P. Whiteside, M.R. Alexander, M.C. Davies,
[134] A. Sassolas, L.J. Blum, B.D. Leca-Bouvier, Immobilization strategies to develop Chemical and spatial analysis of protein loaded PLGA microspheres for drug
enzymatic biosensors, Biotechnol. Adv. 30 (2012) 489–511. delivery applications, J. Contr. Release 162 (2012) 321–329, https://doi.org/
[135] K. Nakanishi, T. Sakiyama, Y. Kumada, K. Imamura, H. Imanaka, Recent advances 10.1016/j.jconrel.2012.05.008.
in controlled immobilization of proteins onto the surface of the solid substrate and [163] E. Cortes, P.G. Etchegoin, E.C. Le Ru, A. Fainstein, M.E. Vela, R.C. Salvarezza,
its possible application to proteomics, Curr. Proteonomics 5 (2008) 161–175, Monitoring the electrochemistry of single molecules by surface-enhanced Raman
https://doi.org/10.2174/157016408785909622. spectroscopy, J. Am. Chem. Soc. 132 (2010) 18034–18037.
[136] F. Rusmini, Z. Zhong, J. Feijen, Protein immobilization strategies for protein [164] A.G. Shard, A straightforward method for interpreting XPS data from core–shell
biochips, Biomacromolecules 8 (2007) 1775–1789, https://doi.org/10.1021/ nanoparticles, J. Phys. Chem. C 116 (2012) 16806–16813.
bm061197b. [165] D.D. Sarma, P.K. Santra, S. Mukherjee, A. Nag, X-ray photoelectron spectroscopy: a
[137] W. Suginta, P. Khunkaewla, A. Schulte, Electrochemical biosensor applications of unique tool to determine the internal heterostructure of nanoparticles, Chem.
polysaccharides chitin and chitosan, Chem. Rev. 113 (2013) 5458–5479, https:// Mater. 25 (2013) 1222–1232.
doi.org/10.1021/cr300325r. [166] Y. Zhu, T. Gao, X. Fan, F. Han, C. Wang, Electrochemical techniques for
[138] I. Migneault, C. Dartiguenave, M.J. Bertrand, K.C. Waldron, Glutaraldehyde: intercalation electrode materials in rechargeable batteries, Acc. Chem. Res. 50
behavior in aqueous solution, reaction with proteins, and application to enzyme (2017) 1022–1031.
crosslinking, Biotechniques 37 (2004) 790–802. [167] P.T. Kissinger, W.R. Heineman, Cyclic voltammetry, J. Chem. Educ. 60 (1983)
[139] E. Povedano, F.H. Cincotto, C. Parrado, P. Díez, A. Sanchez, T.C. Canevari, 702–706, https://doi.org/10.1021/ed060p702.
S.A.S. Machado, J.M. Pingarr on, R. Villalonga, Decoration of reduced graphene [168] N. Elgrishi, K.J. Rountree, B.D. McCarthy, E.S. Rountree, T.T. Eisenhart,
oxide with rhodium nanoparticles for the design of a sensitive electrochemical J.L. Dempsey, A practical beginner's guide to cyclic voltammetry, J. Chem. Educ.
enzyme biosensor for 17β-estradiol, Biosens. Bioelectron. 89 (2017) 343–351, 95 (2018) 197–206, https://doi.org/10.1021/acs.jchemed.7b00361.
https://doi.org/10.1016/j.bios.2016.07.018. [169] B. Purohit, K. Mahato, A. Kumar, P. Chandra, Sputtering enhanced peroxidase like
[140] S.K. Chan, T.S. Lim, A straw-housed paper-based colorimetric antibody-antigen activity of a dendritic nanochip for amperometric determination of hydrogen
sensor, Anal. Methods. 8 (2016) 1431–1436, https://doi.org/10.1039/ peroxide in blood samples, Microchim. Acta. 186 (2019) 658, https://doi.org/
c5ay01828e. 10.1007/s00604-019-3773-2.
[141] T.H.V. Kumar, A.K. Sundramoorthy, Electrochemical biosensor for methyl [170] S.J. Hendel, E.R. Young, Introduction to electrochemistry and the use of
parathion based on single-walled carbon nanotube/glutaraldehyde crosslinked electrochemistry to synthesize and evaluate catalysts for water oxidation and
acetylcholinesterase-wrapped bovine serum albumin nanocomposites, Anal. reduction, J. Chem. Educ. 93 (2016) 1951–1956, https://doi.org/10.1021/
Chim. Acta 1074 (2019) 131–141, https://doi.org/10.1016/ acs.jchemed.6b00230.
j.aca.2019.05.011. [171] E. Laviron, General expression of the linear potential sweep voltammogram in the
[142] M. Yüce, H. Kurt, How to make nanobiosensors: surface modification and case of diffusionless electrochemical systems, J. Electroanal. Chem. Interfacial
characterisation of nanomaterials for biosensing applications, RSC Adv. 7 (2017) Electrochem. 101 (1979) 19–28, https://doi.org/10.1016/S0022-0728(79)80075-
49386–49403, https://doi.org/10.1039/c7ra10479k. 3.
[143] A. Jain, K. Cheng, The principles and applications of avidin-based nanoparticles in [172] Y.S. Choudhary, L. Jothi, G. Nageswaran, Electrochemical Characterization,
drug delivery and diagnosis, J. Contr. Release 245 (2017) 27–40, https://doi.org/ Elsevier Inc., 2017, https://doi.org/10.1016/B978-0-323-46140-5.00002-9.
10.1016/j.jconrel.2016.11.016. [173] M.N. Catrinck, L.L. Okumura, A.A. Silva, A.A. Saczk, M.F. Oliveira, New and
[144] C.M. Dundas, D. Demonte, S. Park, Streptavidin-biotin technology: improvements sensitive electroquantification of sulfentrazone in soil by differential-pulse
and innovations in chemical and biological applications, Appl. Microbiol. voltammetry, J. Braz. Chem. Soc. 26 (2015) 1751–1759.
Biotechnol. 97 (2013) 9343–9353, https://doi.org/10.1007/s00253-013-5232-z. [174] J.R. Macdonald, Impedance spectroscopy, Ann. Biomed. Eng. 20 (1992) 289–305.
[145] D. Kim, A.E. Herr, Protein immobilization techniques for microfluidic assays, [175] N. Jaffrezic-Renault, Impedimetric Measurements and the Biomatrix, Elsevier,
Biomicrofluidics 7 (2013), https://doi.org/10.1063/1.4816934. 2018, https://doi.org/10.1016/B978-0-12-409547-2.13489-4.
[146] S. Mourdikoudis, R.M. Pallares, N.T.K. Thanh, Characterization techniques for [176] S. Vandarkuzhali, N. Gogoi, S. Ghosh, B. Prabhakara Reddy, K. Nagarajan,
nanoparticles: comparison and complementarity upon studying nanoparticle Electrochemical behaviour of LaCl3 at tungsten and aluminium cathodes in
properties, Nanoscale 10 (2018) 12871–12934, https://doi.org/10.1039/ LiCl–KCl eutectic melt, Electrochim. Acta 59 (2012) 245–255, https://doi.org/
C8NR02278J. 10.1016/j.electacta.2011.10.062.
[147] E.N. Kaufmann, in: Elt.N. Kaufmann (Ed.), Characterization of Materials, 2 Volume [177] P.S. Nnamchi, C.S. Obayi, Electrochemical Characterization of Nanomaterials,
Set, Charact. Mater. 2 Vol. Set, 2003, p. 1392, 1392. ISBN 0-471-26882-8. Wiley- Elsevier Ltd., 2018, https://doi.org/10.1016/B978-0-08-101973-3.00004-3.
VCH, January 2003. [178] J. Inczedy, T. Lengyel, A.M. Ure, A. Gelencser, A. Hulanicki, Others, Compendium
[148] C. Shi, D.K. Luu, Q. Yang, J. Liu, J. Chen, C. Ru, S. Xie, J. Luo, J. Ge, Y. Sun, Recent of Analytical Nomenclature, Hoboken Blackwell Sci., 1998.
advances in nanorobotic manipulation inside scanning electron microscopes, [179] D.A. Armbruster, T. Pry, Limit of blank, limit of detection and limit of
Microsystems Nanoeng 2 (2016) 1–16. quantitation, Clin. Biochem. Rev. 29 (2008) S49.

17
B. Purohit et al. Sensors International 1 (2020) 100040

[180] D.W. Tholen, K. Linnet, M. Kondratovich, D.A. Armbruster, P.E. Garret, R.L. Jones, [204] Y. Huang, J. Ren, X. Qu, Nanozymes: classification, catalytic mechanisms, activity
M.H. Kroll, R.M. Lequin, T.J. Pankratz, G.A. Scassellati, H. Schimmel, J. Tsai, regulation, and applications, Chem. Rev. 119 (2019) 4357–4412, https://doi.org/
Protocols for Determination of Limits of Detection and Limits of Quantitation; 10.1021/acs.chemrev.8b00672.
Approved Guideline Global Consensus Standardization for Health Technologies, [205] K. Wang, C. Wu, F. Wang, M. Liao, G. Jiang, Bimetallic nanoparticles decorated
2004. www.nccls.org. hollow nanoporous carbon framework as nanozyme biosensor for highly sensitive
[181] A. Vallee-Belisle, F. Ricci, K.W. Plaxco, Engineering biosensors with extended, electrochemical sensing of uric acid, Biosens. Bioelectron. 150 (2020) 111869,
narrowed, or arbitrarily edited dynamic range, J. Am. Chem. Soc. 134 (2012) https://doi.org/10.1016/j.bios.2019.111869.
2876–2879, https://doi.org/10.1021/ja209850j. [206] D. Zhang, Q. Liu, Biosensors and bioelectronics on smartphone for portable
[182] W.J. Peveler, M. Yazdani, V.M. Rotello, Selectivity and specificity: pros and cons in biochemical detection, Biosens. Bioelectron. 75 (2016) 273–284, https://doi.org/
sensing, ACS Sens. 1 (2016) 1282–1285, https://doi.org/10.1021/ 10.1016/j.bios.2015.08.037.
acssensors.6b00564. [207] S. Kanchi, M.I. Sabela, P.S. Mdluli, K. Bisetty, Biosensors and Bioelectronics
[183] L. Soleymani, F. Li, Mechanistic challenges and advantages of biosensor Smartphone based bioanalytical and diagnosis applications : a review, Biosens.
miniaturization into the nanoscale, ACS Sens. 2 (2017) 458–467, https://doi.org/ Bioelectron. 102 (2018) 136–149, https://doi.org/10.1016/j.bios.2017.11.021.
10.1021/acssensors.7b00069. [208] J. Jiang, X. Wang, R. Chao, Y. Ren, C. Hu, Z. Xu, G.L. Liu, Smartphone based
[184] X. Guo, S. Liu, M. Yang, H. Du, F. Qu, Dual signal amplification portable bacteria pre-concentrating microfluidic sensor and impedance sensing
photoelectrochemical biosensor for highly sensitive human epidermal growth system, Sensor. Actuator. B Chem. 193 (2014) 653–659.
factor receptor-2 detection, Biosens. Bioelectron. 139 (2019) 111312, https:// [209] D. Zhang, J. Jiang, J. Chen, Q. Zhang, Y. Lu, Y. Yao, S. Li, G. Logan Liu, Q. Liu,
doi.org/10.1016/j.bios.2019.05.017. Smartphone-based portable biosensing system using impedance measurement
[185] I.S. Kucherenko, O.O. Soldatkin, S.V. Dzyadevych, A.P. Soldatkin, Electrochemical with printed electrodes for 2,4,6-trinitrotoluene (TNT) detection, Biosens.
biosensors based on multienzyme systems: main groups, advantages and Bioelectron. 70 (2015) 81–88, https://doi.org/10.1016/j.bios.2015.03.004.
limitations – a review, Anal. Chim. Acta 1111 (2020) 114–131, https://doi.org/ [210] P. Chandra, Miniaturized label-free smartphone assisted electrochemical sensing
10.1016/j.aca.2020.03.034. approach for personalized COVID-19 diagnosis, Sensors Int 1 (2020) 100019,
[186] J. Xi, C. Xie, Y. Zhang, L. Wang, J. Xiao, X. Duan, J. Ren, F. Xiao, S. Wang, Pd https://doi.org/10.1016/j.sintl.2020.100019.
nanoparticles decorated N-doped graphene quantum dots@N-doped carbon [211] B. Purohit, A. Kumar, K. Mahato, P. Chandra, Smartphone-assisted personalized
hollow nanospheres with high electrochemical sensing performance in cancer diagnostic devices and wearable sensors, Curr. Opin. Biomed. Eng. 13 (2020) 42–50.
detection, ACS Appl. Mater. Interfaces 8 (2016) 22563–22573, https://doi.org/ 
[212] J. Kim, A.S. Campbell, B.E.-F. de Avila, J. Wang, Wearable biosensors for
10.1021/acsami.6b05561. healthcare monitoring, Nat. Biotechnol. 37 (2019) 1, https://doi.org/10.1038/
[187] S. Emaminejad, W. Gao, E. Wu, Z.A. Davies, H. Yin Yin Nyein, S. Challa, S.P. Ryan, s41587-019-0045-y.
H.M. Fahad, K. Chen, Z. Shahpar, S. Talebi, C. Milla, A. Javey, R.W. Davis, [213] W. Gao, S. Emaminejad, H.Y.Y. Nyein, S. Challa, K. Chen, A. Peck, H.M. Fahad,
Autonomous sweat extraction and analysis applied to cystic fibrosis and glucose H. Ota, H. Shiraki, D. Kiriya, D.-H. Lien, G.A. Brooks, R.W. Davis, A. Javey, Fully
monitoring using a fully integrated wearable platform, Proc. Natl. Acad. Sci. Unit. integrated wearable sensor arrays for multiplexed in situ perspiration analysis,
States Am. 114 (2017) 4625–4630, https://doi.org/10.1073/pnas.1701740114. Nature 529 (2016) 509–514, https://doi.org/10.1038/nature16521.
[188] E. Bakker, Can calibration-free sensors Be realized? ACS Sens. 1 (2016) 838–841, [214] M. Elsherif, M.U. Hassan, A.K. Yetisen, H. Butt, Wearable contact lens biosensors
https://doi.org/10.1021/acssensors.6b00247. for, ACS Nano 12 (2018) 5452–5462, https://doi.org/10.1021/acsnano.8b00829.
[189] J. Visentin, L. Minder, J.H. Lee, J.L. Taupin, C. Di Primo, Calibration free [215] W. Jia, A.J. Bandodkar, G. Valde, J.R. Windmiller, Z. Yang, J. Ram, G. Chan,
concentration analysis by surface plasmon resonance in a capture mode, Talanta J. Wang, Electrochemical Tattoo Biosensors for Real-Time Noninvasive Lactate
148 (2016) 478–485, https://doi.org/10.1016/j.talanta.2015.11.025. Monitoring in Human Perspiration, 2013, https://doi.org/10.1021/ac401573r.
[190] H. Li, P. Dauphin-Ducharme, G. Ortega, K.W. Plaxco, Calibration-free [216] A.J. Bandodkar, W. Jia, C. Yardımcı, X. Wang, J. Ramirez, J. Wang, Tattoo-based
electrochemical biosensors supporting accurate molecular measurements directly noninvasive glucose monitoring: a proof-of-concept study, Anal. Chem. 87 (2015)
in undiluted whole blood, J. Am. Chem. Soc. 139 (2017) 11207–11213. 394–398, https://doi.org/10.1021/ac504300n.
[191] A. Idili, C. Parolo, G. Ortega, K.W. Plaxco, Calibration-free measurement of [217] A.K. Yetisen, R. Moreddu, S. Seifi, N. Jiang, K. Vega, X. Dong, J. Dong, H. Butt,
phenylalanine levels in the blood using an electrochemical aptamer-based sensor M. Jakobi, M. Elsner, A.W. Koch, Dermal tattoo biosensors for colorimetric
suitable for point-of-care applications, ACS Sens. 4 (2019) 3227–3233, https:// metabolite detection, Angew. Chem. Int. Ed. 58 (2019) 10506–10513, https://
doi.org/10.1021/acssensors.9b01703. doi.org/10.1002/anie.201904416.
[192] H. Ren, M.A. Coughlin, T.C. Major, S. Aiello, A. Rojas Pena, R.H. Bartlett, [218] G.-P. Nikoleli, C.G. Siontorou, D.P. Nikolelis, S. Bratakou, S. Karapetis,
M.E. Meyerhoff, Improved in vivo performance of amperometric oxygen (PO2) N. Tzamtzis, Biosensors based on microfluidic devices lab-on-a-chip and
sensing catheters via electrochemical nitric oxide generation/release, Anal. Chem. microfluidic technology, in: Nanotechnol. Biosens., Elsevier, 2018, pp. 375–394.
87 (2015) 8067–8072, https://doi.org/10.1021/acs.analchem.5b01590.  Ríos, M. Zougagh, M. Avila, Miniaturization through lab-on-a-chip: utopia or
[219] A.
[193] M. Pawlak, E. Grygolowicz-Pawlak, G.A. Crespo, G. Mistlberger, E. Bakker, PVC- reality for routine laboratories? A review, Anal. Chim. Acta 740 (2012) 1–11,
based ion-selective electrodes with enhanced biocompatibility by surface https://doi.org/10.1016/j.aca.2012.06.024.
modification with “click” chemistry, Electroanalysis 25 (2013) 1840–1846. [220] L.R. Volpatti, A.K. Yetisen, Commercialization of microfluidic devices, Trends
[194] N. Hui, X. Sun, S. Niu, X. Luo, PEGylated polyaniline nanofibers: antifouling and Biotechnol. 32 (2014) 347–350, https://doi.org/10.1016/j.tibtech.2014.04.010.
conducting biomaterial for electrochemical DNA sensing, ACS Appl. Mater. [221] J. Zhuang, J. Yin, S. Lv, B. Wang, Y. Mu, Advanced “lab-on-a-chip” to detect
Interfaces 9 (2017) 2914–2923, https://doi.org/10.1021/acsami.6b11682. viruses – current challenges and future perspectives, Biosens. Bioelectron. 163
[195] J. Tavakoli, Y. Tang, Hydrogel based sensors for biomedical applications: an (2020) 112291, https://doi.org/10.1016/j.bios.2020.112291.
updated review, Polymers 9 (2017) 364. [222] Y. Sekine, S.B. Kim, Y. Zhang, A.J. Bandodkar, S. Xu, J. Choi, M. Irie, T.R. Ray,
[196] G. Wang, R. Han, X. Su, Y. Li, G. Xu, X. Luo, Zwitterionic peptide anchored to P. Kohli, N. Kozai, T. Sugita, Y. Wu, K. Lee, K.T. Lee, R. Ghaffari, J.A. Rogers,
conducting polymer PEDOT for the development of antifouling and ultrasensitive A fluorometric skin-interfaced microfluidic device and smartphone imaging
electrochemical DNA sensor, Biosens. Bioelectron. 92 (2017) 396–401, https:// module for: in situ quantitative analysis of sweat chemistry, Lab Chip 18 (2018)
doi.org/10.1016/j.bios.2016.10.088. 2178–2186, https://doi.org/10.1039/c8lc00530c.
[197] O.Y.F. Henry, J.L.A. Sanchez, C.K. O'Sullivan, Bipodal PEGylated alkanethiol for [223] B. Bruijns, A. van Asten, R. Tiggelaar, H. Gardeniers, Microfluidic devices for
the enhanced electrochemical detection of genetic markers involved in breast forensic DNA analysis: a review, Biosensors 6 (2016) 41, https://doi.org/10.3390/
cancer, Biosens. Bioelectron. 26 (2010) 1500–1506, https://doi.org/10.1016/ bios6030041.
j.bios.2010.07.095. [224] W.R. Heineman, W.B. Jensen, Leland C. Clark Jr., Biosens. Bioelectron. 21 (2006)
[198] J. Wu, S. Campuzano, C. Halford, D.A. Haake, J. Wang, Ternary surface 1403–1404, https://doi.org/10.1016/j.bios.2005.12.005, 1918–2005.
monolayers for ultrasensitive (Zeptomole) amperometric detection of nucleic acid [225] I. Palchetti, M. Mascini, Biosensor technology: a brief history, in: Sensors and
hybridization without signal amplification, Anal. Chem. 82 (2010) 8830–8837, Microsystems, Springer, 2010, pp. 15–23, https://doi.org/10.1007/978-90-481-
https://doi.org/10.1021/ac101474k. 3606-3_2.
[199] J. Sabate del Río, O.Y.F. Henry, P. Jolly, D.E. Ingber, An antifouling coating that [226] N. Wongkaew, M. Simsek, C. Griesche, A.J. Baeumner, Functional nanomaterials
enables affinity-based electrochemical biosensing in complex biological fluids, and nanostructures enhancing electrochemical biosensors and lab-on-a-chip
Nat. Nanotechnol. 14 (2019) 1143–1149, https://doi.org/10.1038/s41565-019- performances: recent progress, applications, and future perspective, Chem. Rev.
0566-z. 119 (2018) 120–194.
[200] Y. Lin, J. Ren, X. Qu, Catalytically active nanomaterials: a promising candidate for
artificial enzymes, Acc. Chem. Res. 47 (2014) 1097–1105, https://doi.org/
10.1021/ar400250z. List of abbreviations for Table 1 and 2
[201] H. Wei, E. Wang, Nanomaterials with enzyme-like characteristics (nanozymes):
next-generation artificial enzymes, Chem. Soc. Rev. 42 (2013) 6060–6093,
AFM: Atomic force microscopy
https://doi.org/10.1039/c3cs35486e.
BET: Brunauer–Emmett–Teller
[202] J. Wu, X. Wang, Q. Wang, Z. Lou, S. Li, Y. Zhu, L. Qin, H. Wei, Nanomaterials
DCS: Differential centrifugal sedimentation
with enzyme-like characteristics (nanozymes): next-generation artificial enzymes
DLS: Dynamic light scattering
(II), Chem. Soc. Rev. 48 (2019) 1004–1076, https://doi.org/10.1039/
EDX: Energy-dispersive X-ray spectroscopy
c8cs00457a.
EELS: Electron energy loss spectroscopy
[203] L. Gao, J. Zhuang, L. Nie, J. Zhang, Y. Zhang, N. Gu, T. Wang, J. Feng, D. Yang,
EPLS: Ellipticallypolarized light scattering
S. Perrett, X. Yan, Intrinsic peroxidase-like activity of ferromagnetic
EPM: Electrophoretic mobility
nanoparticles, Nat. Nanotechnol. 2 (2007) 577–583, https://doi.org/10.1038/
FMR: Ferromagnetic resonance
nnano.2007.260.
FTIR: Fourier transform infrared spectroscopy

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HRTEM: High-resolution transmission electron microscopy TGA: Thermal gravimetric analysis

ICP-MS: Inductively coupled plasmamass spectrometry TLS: thermal lens spectrometry
MFM: Magnetic force microscopy TOF-SIMS: Time of flight secondary ion mass spectrometry
NMR: Nuclear magnetic resonance SIMS: Secondary ion mass spectrometry
SAED: Selected area electrondiffraction (also, known as electron diffraction (ED)) UV–Vis: Ultra violet-Visible spectroscopy
SAXS: Small angle X-ray scattering VSM: Vibrating sample magnetometry
SIMS: Secondary ion mass spectrometry XAS: X-ray absorption spectroscopy; X-ray absorption spectroscopy
SQUID: Superconducting quantum interference device magnet XPS: X-ray photoelectron spectroscopy
TEM: Transmission electron microscopy XRD: X-ray diffraction
STEM: Scanning transmission electron microscopy XMCD: X-ray magnetic circular dichroism)

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