REVIEW
Nanoenzymes for Biomolecules Detection
Submitted by: Submitted to:
Monisha Singh Prof. Pramod Gware
200010540005 Department of Biotechnology
VIII Semester AEC, Agra.
Review: Nanoenzymes for Biomolecules Detection
Abstract
Nanoenzymes have emerged as promising tools for ultrasensitive biomolecule detection due
to their unique catalytic properties and high surface-to-volume ratio. In this review, we
explore recent advances in the field of nanoenzymes for biomolecule detection, focusing on
their synthesis methods, applications, and challenges. Drawing insights from graphene-based
nanocomposites, affibody-functionalized beads, chromogenic enzyme substrates, and
template-assisted plasmonic nanogap shells, we discuss the potential of nanoenzymes to
revolutionize biomolecular sensing platforms and address critical issues such as sensitivity,
selectivity, and scalability.
Introduction:
The rapid advancement of nanotechnology has lined the way for innovative approaches in
biosensing and diagnostics, particularly through the development of nanoenzymes for
biomolecule detection. The detection of biomolecules, including proteins, nucleic acids, and
small molecules, is essential for various fields such as healthcare, environmental monitoring,
and biotechnology. Nanoenzymes, engineered nanomaterials with enzyme-like catalytic
properties, offer unique advantages over traditional enzymes, including enhanced stability
and catalytic activity. Nanoenzymes represent a versatile and promising class of
nanomaterials for biomolecule detection, offering enhanced sensitivity, specificity, scalability
and multiplexing capabilities. Conventional detection methods often suffer from limitations
such as low sensitivity, long assay times, and complex instrumentation. Nanoenzymes,
synthetic enzymes with nanoscale dimensions, offer a promising solution to overcome these
challenges by leveraging their catalytic activity and biocompatibility with targeted
biomolecules. These advancements in biosensing technology, facilitate early disease
detection, personalized medicine, and biomedical research
Synthesis Methods for Nanoenzymes:
1. Graphene-based Nanocomposites:
Graphene-based nanocomposites are promising materials for nanoenzyme synthesis
due to their high surface area, excellent electrical conductivity, and chemical stability.
Krishnan et al. (2019) reviewed various synthesis methods for graphene-based
nanocomposites, including chemical vapor deposition (CVD) and solution-phase
synthesis. CVD involves the growth of graphene on a metal substrate by exposing it
to carbon-containing precursors at high temperatures. Solution-phase synthesis
methods, on the other hand, involve the reduction of graphene oxide or graphene
oxide derivatives in solution to obtain graphene-based nanocomposites with desired
properties.
2. Affibody-functionalized Beads:
Affibody-functionalized beads are utilized for highly sensitive detection of cancer
cell-derived exosomes. The synthesis of these beads involves functionalizing
commercially available beads with affibody molecules, which are small protein
scaffolds engineered to bind with high specificity to target biomolecules such as
exosomes. Sayyadi et al. (2021) discuss the Evaluation of the EGFR Expression in
A549 Cells Using immunofluorescence staining of A549 cells with anti-EGFR
Affibody using fluorescent microscopy then Characterized Human Lung Cancer Cell-
Derived Exosomes by isolating A549 cell-derived exosomes from the cell culture.
Also discuss the synthesis and application of affibody-functionalized beads for
exosome detection, highlighting their potential for early cancer diagnosis.
3. Chromogenic Enzyme Substrates:
Na et al. (2021) utilize chromogenic enzyme substrates for signal amplification in
multiplexed detection of biomolecules using time-of-flight secondary ion mass
spectrometry (ToF-SIMS), which is a sensitive surface analytical tool. The synthesis
of chromogenic enzyme substrates involves conjugating enzyme substrates with
chromogenic moieties that undergo a color change upon enzymatic cleavage. This
allows for visual or spectrophotometric detection of enzymatic activity, thereby
amplifying the signal for biomolecule detection in complex samples.
4. Template-Assisted Plasmonic Nanogap Shells:
Kang et al. (2021) developed template-assisted plasmonic nanogap shells for highly
enhanced detection of cancer biomarkers based on surface-enhanced Raman
scattering (SERS). The synthesis of these nanogap shells involves templated
electrodeposition of metallic nanoparticles within a nanoporous membrane, followed
by removal of the template to create nanoscale gaps between the nanoparticles. This
results in localized surface plasmon resonance (LSPR) effects, which significantly
enhance the sensitivity of biomolecule detection.
5. Nanozyme-Based Theranostics:
Sisakhtnezhad et al. discussed the biomedical applications of MnO2 nanomaterials as
nanoenzyme-based theranostics. MnO2 nanoenzymes possess intrinsic catalytic
activity, allowing for the detection of biomolecules through enzymatic reactions.
MnO2-NEs also show scavenging properties against reactive oxygen species (ROS) in
a range of pathological conditions. In addition, due to the decomposition of MnO 2-
Nanoenzymes in the tumor microenvironment (TME) and the production of Mn2+,
they can act as a contrast agent for improving clinical imaging diagnostics. This
multifunctional platform holds great potential for precision medicine and personalized
healthcare applications.
These synthesis methods offer precise control over the size, shape, and surface properties of
nanoenzymes, enabling tailored design and optimization for specific biomolecule detection
applications.
Applications of Nanoenzymes in Biomolecule Detection:
1. Electrochemical Biosensors:
Electrochemical biosensors nanoenzymes are extensively employed in
electrochemical biosensors for the detection of various biomolecules, including
proteins, nucleic acids, and small molecules. Graphene-based nanocomposites, for
instance, have been utilized as electrode materials in electrochemical biosensors due
to their high conductivity and large surface area. These nanoenzymes improve the
sensitivity and specificity of these biosensors, enabling rapid and accurate detection of
biomolecular targets.
2. Fluorescent Biosensors:
Nanoenzymes have also set up operations in fluorescent biosensors, where they serve
as catalytic labels for signal amplification. By conjugating fluorescent dyes to
nanoenzymes, researchers can get highly sensitive detection of biomolecules through
fluorescence amplification. Graphene-based nanocomposites, with their unique
optical properties, suggest excellent platforms for developing fluorescent biosensors
with improved detection limits and signal-to-noise ratios.
3. Exosome Detection:
Affibody-functionalized beads are used for the highly sensitive detection of exosomes
derived from cancer cells. Exosomes are extracellular vesicles released by cells, and
their detection holds great promise for cancer diagnosis and monitoring. Affibody-
functionalized beads selectively capture exosomes expressing specific surface
markers, enabling the detection of cancer biomarkers with high sensitivity and
specificity. This approach offers potential applications in early cancer diagnosis and
personalized medicine.
4. Multiplexed Detection using Surface Mass Spectrometry:
Nanoenzymes, such as chromogenic enzyme substrates, are utilized for amplification
of signal in multiplexed biomolecules detection using surface mass spectrometry. By
coupling enzyme-catalyzed reactions with mass spectrometry detection, researchers
can achieve simultaneous detection of multiple biomolecular targets with high
sensitivity and specificity. This approach allows for rapid and quantitative analysis of
complex biological samples, making it suitable for various diagnostic and research
applications.
5. Enhanced Detection of Cancer Biomarkers:
Template-assisted plasmonic nanogap shells are employed for highly enhanced
detection of cancer biomarkers. These nanostructures exhibit localized surface
plasmon resonance (LSPR) effects, which significantly improve the sensitivity of
biomolecule detection. By functionalizing the nanogap shells with specific ligands or
antibodies, researchers can selectively capture cancer biomarkers from complex
biological samples, enabling early cancer diagnosis and monitoring with improved
sensitivity and accuracy.
Challenges and Future Perspectives in Nanoenzyme-Based Biomolecule
Detection:
One of the primary challenges in the field of nanoenzyme-based biomolecule detection is
optimizing synthesis methods and standardizing protocols. Different synthesis techniques and
conditions can yield nanoenzymes with varying properties, affecting their catalytic activity
and performance in detection assays. Standardizing synthesis protocols and optimizing
parameters such as size, shape, and surface chemistry will be crucial for ensuring
reproducibility and reliability across different studies and applications. Another challenge is
assessing the biocompatibility and potential toxicity of nanoenzymes, especially for in vivo
applications. While nanoenzymes offer exciting opportunities for sensitive and selective
biomolecule detection, their interaction with biological systems must be thoroughly evaluated
to ensure safety and minimize adverse effects. Future research should focus on developing
robust methods for biocompatibility assessment and toxicity profiling to facilitate the
translation of nanoenzyme-based detection technologies into clinical and biomedical settings.
Improving the sensitivity and specificity of nanoenzyme-based detection assays is crucial for
detecting biomolecules at low concentrations and discriminating between closely related
analytes. Strategies such as surface functionalization, enzyme engineering, and signal
amplification techniques can enhance the performance of nanoenzyme-based biosensors.
Future research efforts should focus on developing novel approaches to enhance sensitivity
and specificity while minimizing background noise and nonspecific binding.
Nanoenzyme-based detection technologies hold great potential for multiplexed analysis,
allowing simultaneous detection of multiple biomarkers in complex samples. However,
integrating nanoenzymes into multiplexed detection platforms presents several challenges,
including cross-reactivity, signal crosstalk, and assay interference. Developing innovative
assay formats, signal processing algorithms, and data analysis techniques will be essential for
realizing the full potential of nanoenzyme-based multiplexed detection platforms.
In addition to detection, there is growing interest in exploring the theranostic potential of
nanoenzymes, particularly for targeted drug delivery and imaging applications. However,
challenges such as achieving precise control over drug loading and release kinetics, as well as
optimizing targeting efficiency and biocompatibility, need to be addressed. Future research
should focus on developing multifunctional nanoenzyme-based theranostic platforms capable
of real-time detection, imaging, and therapy for personalized medicine applications.
Conclusion
The review paper delves into the advancements and applications of nanoenzymes for
biomolecule detection, drawing insights from recent studies on graphene-based
nanocomposites, affibody-functionalized beads, chromogenic enzyme substrates, and
template-assisted plasmonic nanogap shells. Nanoenzymes offer distinctive catalytic
properties and high surface-to-volume ratios, making them promising candidates for
ultrasensitive biomolecule detection in various fields, including healthcare, environmental
monitoring, and biotechnology.
Despite the progress made in nanoenzyme synthesis and functionalization, several challenges
remain to be addressed. These include optimizing synthesis methods for enhanced stability
and reproducibility, assessing biocompatibility and toxicity concerns, integrating
nanoenzymes with emerging detection platforms, and improving sensitivity and selectivity
for biomolecule detection. In addition, scalability and commercialization hurdles must be
overcome to realize the full potential of nanoenzyme-based detection technologies for
practical applications.
Moving forward, interdisciplinary collaboration, innovative research approaches, and
concerted efforts from academia, industry, and regulatory agencies are essential to address
these challenges and release the transformative potential of nanoenzymes in biomolecule
detection. By overcoming these hurdles, nanoenzyme-based detection technologies can
revolutionize biomolecular sensing platforms, enabling early disease diagnosis, personalized
medicine, and environmental monitoring with exceptional sensitivity and specificity.
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