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\begin{document}

\title{Biosensors And Bioelectronics Template}


\author[ ]{safaryan}

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\date{}

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\selectlanguage{english}
\begin{abstract}
This paper describes a new method for preparation of chromogen free
oxidase based optical biosensors. Oxidases enzymes were deposited on the
titania film consisting of uniformly distributed nanoparticles reacting
with hydrogen peroxide thus providing formation of yellow color due to
complex Ti (IV)-H\textsubscript{2}O\textsubscript{2}. Using ink jet
printing and specially developed titania ink we have created a universal
platform for the oxidase based biosensors and successfully tested it on
glucose and cholesterol detection. The paper fully discusses new
biomaterials as well as biosensor creation technology and describes the
mechanism of action, study on stability and working concentration limits
and adjusting composition of the inks for good printability and scale-up
production.%
\end{abstract}%



\section*{Introduction}\label{introduction}

Recent years, low-cost, simple and automated biomaterials production
methods are widely recognized in enzymatic biosensor
development\textsuperscript{1,2}. In most cases, this requires the
application of sensitive liquid components containing enzymes on the
substrate surface\textsuperscript{3}. For example, such methods include
additive technology that can accurately control the amount of components
applied to a substrate\textsuperscript{4}. At the moment, there are
different additive methods, ranging from microdispensing to
printing\textsuperscript{5}. Inkjet printing is a perspective direction
for material science development as it allows to apply different
reagents from active substances solutions up to complex sol-gel systems
with unique properties in the ink form\textsuperscript{6}. A distinctive
feature of this method is the possibility of a high-precision deposition
of variety materials, the creation of ordered layers and complex
three-dimensional structures of an active substance\textsuperscript{7}.

A relatively new direction in the field of inkjet printing is the
producing of biomaterials and biosensors\textsuperscript{8,9}. The
possibilities of accurate dosing and positioning along with the
autonomous manufacturing largely determine the advantages of a printing
method for the production of high-precision diagnostic
biosensors\textsuperscript{10}. There is a great variety of types of
biomolecules\textsuperscript{11}, bio-inks\textsuperscript{12} and based
on them biosensors\textsuperscript{13}. The simplest and most widely
recognized biosensing method is still a qualitative enzymatic reaction
based on oxidases due to their high selectivity and
sensitivity\textsuperscript{14}. Oxidases catalyze the
oxidation-reduction reactions\textsuperscript{15}. They specifically
react with the substrate and transfer the hydrogen from substrate to
atmospheric oxygen to form water or hydrogen peroxide. There are many
oxidases meaningful for biochemical analysis and biosensors, such as
Glucose oxidase, Lactate oxidase, Urate oxidase, Cholesterol oxidase,
Glycerol-3-phosphate oxidase, Choline oxidase, Alcohol oxidase and
others\textsuperscript{14}.

In the widely used method, the additional reaction is used to visualize
the substrate oxidation products. In this reaction a chromogenic
substrate (dye) and peroxidase are used for detection the formed
peroxide\textsuperscript{16}. Peroxidase transfers oxygen from the
peroxide to the dye molecule. As a result, the oxidized form of the
chromogen acquires or changes the color. However, many chromogenic dyes
are not stable in the light\textsuperscript{17}, they are readily
oxidized in the air and require additional components to protect against
undesirable oxidation at storage conditions\textsuperscript{18}. This
multicomponent system overloads and complicates the sensor, requires
careful calculation of optimal concentrations and ratios of each
component for the reproducibility of the results, and increases the
costs of creating a biosensor.

In addition to the use of chromogenic dyes, colorimetric biosensors
based on nanoparticles were reported\textsuperscript{19}. These assays
usually use noble metals nanoparticles\textsuperscript{20} such as gold
or silver. The detection is based on the principle of particles
aggregation or dispersion\textsuperscript{21}. However, most of these
nanoparticles are used to DNA analysis\textsuperscript{22}, only a few
of them being adapted to the enzymatic reaction. For example, the use of
redox ceria nanoparticles as colorimetric probes in bioassay were
reported\textsuperscript{23}. The method is based on changes in the
physicochemical properties of cerium oxide nanoparticles operating as
chromogenic indicator.

Along with ceria nanoparticles, the color qualitative reaction to
titanium with a solution of hydrogen peroxide is stated to
exist\textsuperscript{24}. This method is highly sensitive which allows
the detection of titanium trace amounts, due to the yellow complex Ti
(IV) -H\textsubscript{2}O\textsubscript{2} formation with the
tetravalent titanium and hydrogen peroxide\textsuperscript{25}. In this
work we have used titanium dioxide (titania) for the detection of
hydrogen peroxide. Since hydrogen peroxide is formed in substrate
oxidation reactions, oxidase enzymes are used for glucose and
cholesterol determination.

In this work we for the first time propose an approach to obtain
biosensors based on redox nanostructured material applied by desktop
inkjet printing.

Using ink jet printing and special titania ink we have created a
universal substrate for the oxidase biosensors and successfully tested
it on glucose and cholesterol. The application of inkjet printing
titania substrate as a colorimetric component achieves high
repeatability with improved surface properties for subsequent layer
deposition. The minimum concentration of peroxide that is possible to
detect using this method is 0.4 mM.

\par\null

\section*{Results}\label{results}

This section is only included in papers that rely on primary research.
This section catalogues the results of the experiment. The results
should be presented in a clear and unbiased way. Most results sections
will contain \href{http://authorea.com}{links}~as well as
citations~\hyperref[csl:1]{(Einstein, 1916)}~and equations such as~\(e^{i\pi}+1=0\)\\

\section*{Conclusion}\label{auto-label-section-853974}

The conclusion should reinforce the major claims or interpretation in a
way that is not mere summary. The writer should try to indicate the
significance of the major claim/interpretation beyond the scope of the
paper but within the parameters of the field. The writer might also
present complications the study illustrates or suggest further research
the study indicates is necessary.

\selectlanguage{english}
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\section*{References}\sloppy
\phantomsection
\label{csl:1}Einstein, A., 1916. {Näherungsweise Integration der Feldgleichungen der Gravitation}, in: Albert Einstein: Akademie-Vorträge. Wiley-Blackwell, pp. 99–108. \url{https://doi.org/10.1002/3527608958.ch7}

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