Rapid
antigen tests are currently used for population screening
of COVID-19. However, they lack sensitivity and utilize antibodies
as receptors, which can only function in narrow temperature and pH
ranges. Consequently, molecularly imprinted polymer nanoparticles
(nanoMIPs) are synthetized with a fast (2 h) and scalable process
using merely a tiny SARS-CoV-2 fragment (∼10 amino acids).
The nanoMIPs rival the affinity of SARS-CoV-2 antibodies under standard
testing conditions and surpass them at elevated temperatures or in
acidic media. Therefore, nanoMIP sensors possess clear advantages
over antibody-based assays as they can function in various challenging
media. A thermal assay is developed with nanoMIPs electrografted onto
screen-printed electrodes to accurately quantify SARS-CoV-2 antigens.
Heat transfer-based measurements demonstrate superior detection limits
compared to commercial rapid antigen tests and most antigen tests
from the literature for both the alpha (∼9.9 fg mL
–1
) and delta (∼6.1 fg mL
–1
) variants of the
spike protein. A prototype assay is developed, which can rapidly (∼15
min) validate clinical patient samples with excellent sensitivity
and specificity. The straightforward epitope imprinting method and
high robustness of nanoMIPs produce a SARS-CoV-2 sensor with significant
commercial potential for population screening, in addition to the
possibility of measurements in diagnostically challenging environments.
There has been a rapid increase in the fabrication of various kinds of edible nanoparticles for oral delivery of bioactive agents, such as those constructed from proteins, carbohydrates, lipids, and/or minerals. It is currently difficult to compare the relative advantages and disadvantages of different kinds of nanoparticle-based delivery systems because researchers use different analytical instruments and protocols to characterize them. In this paper, we briefly review the various analytical methods available for characterizing the properties of edible nanoparticles, such as composition, morphology, size, charge, physical state, and stability. This information is then used to propose a number of standardized protocols for characterizing nanoparticle properties, for evaluating their stability to environmental stresses, and for predicting their biological fate. Implementation of these protocols would facilitate comparison of the performance of nanoparticles under standardized conditions, which would facilitate the rational selection of nanoparticle-based delivery systems for different applications in the food, health care, and pharmaceutical industries.
We
demonstrate that a novel functionalized interface, where molecularly
imprinted polymer nanoparticles (nanoMIPs) are attached to screen-printed
graphite electrodes (SPEs), can be utilized for the thermal detection
of the cardiac biomarker troponin I (cTnI). The ultrasensitive detection
of the unique protein cTnI can be utilized for the early diagnosis
of myocardial infraction (i.e., heart attacks), resulting in considerably
lower patient mortality and morbidity. Our developed platform presents
an innovative route to develop accurate, low-cost, and disposable
sensors for the diagnosis of cardiovascular diseases, specifically
myocardial infraction. A reproducible and advantageous solid-phase
approach was utilized to synthesize high-affinity nanoMIPs (average
size = 71 nm) for cTnI, which served as synthetic receptors in a thermal
sensing platform. To assess the performance and commercial potential
of the sensor platform, various approaches were used to immobilize
nanoMIPs onto thermocouples or SPEs: dip coating, drop casting, and
a covalent approach relying on electrografting with an organic coupling
reaction. Characterization of the nanoMIP-functionalized surfaces
was performed with electrochemical impedance spectroscopy, atomic
force microscopy, and scanning electron microscopy. Measurements from
an in-house designed thermal setup revealed that covalent functionalization
of nanoMIPs onto SPEs led to the most reproducible sensing capabilities.
The proof of application was provided by measuring buffered solutions
spiked with cTnI, which demonstrated that through monitoring changes
in heat transfer at the solid–liquid interface, we can measure
concentrations as low as 10 pg L–1, resulting in
the most sensitive test of this type. Furthermore, preliminary data
are presented for a prototype platform, which can detect cTnI with
shorter measurement times and smaller sample volumes. The excellent
sensor performance, versatility of the nanoMIPs, and reproducible
and low-cost nature of the SPEs demonstrate that this sensor platform
technology has a clear commercial route with high potential to contribute
to sustainable healthcare.
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