Developing a convenient detection method for dopamine holds a significant incentive due to its high clinical significance. Herein, we synthesize crystalline MnOOH nanowires (MNWs) via a simple solvothermal treatment of KMnO4 and demonstrate that they possess excellent oxidase-like activity owing to the presence of pure Mn3+ sites on the MNWs. Particularly, MNWs catalyze the rapid oxidation of dopamine into aminochromes, which show a vivid brown color. The dopamine oxidase-like activity of MNWs follows the typical Michaelis–Mentenkinetics with excellent storage stability. Based on the affirmative catalytic features, a paper dipstick incorporating MNWs in the detection zone is constructed for the one-step colorimetric detection of target dopamine. By immersing the dipstick into the sample solution for 30 min, the sample spontaneously moves to the detection zone due to capillary force, yielding a brown color proportional to the amount of dopamine, which is quantified from an image acquired using a smartphone. With the MNW-containing solution-based assay and MNW-incorporated paper dipstick, dopamine is successfully determined with high selectivity, sensitivity, and detection precision when using spiked human serum and pharmaceutical dopamine injection samples, respectively. Successful analytical values such as the dynamic linear ranges of 3–60 μM and 0.05–7 mM are achieved with the solution-based assay and paper dipstick, respectively, along with excellent detection accuracy (95–99%) and precision (1.0–3.1%). Hence, we developed a simple and efficient nanozyme-based paper dipstick biosensor for dopamine that can be used in point-of-care testing environments.
Amino acid arrays comprising bioluminescent amino acid auxotrophic Escherichia coli are effective systems to quantitatively determine multiple amino acids. However, there is a need to develop a method for convenient long-term preservation of the array to enable its practical applications. Here, we reported a potential strategy to efficiently maintain cell viability within the portable array. The method involves immobilization of cells within agarose gel supplemented with an appropriate cryoprotectant in individual wells of a 96-well plate, followed by storage under freezing conditions. Six cryoprotectants, namely dimethyl sulfoxide, glycerol, ethylene glycol, polyethylene glycol, sucrose, and trehalose, were tested in the methionine (Met) auxotroph-based array. Carbohydrate-type cryoprotectants (glycerol, sucrose, and trehalose) efficiently preserved the linearity of determination of Met concentration. In particular, the array with 5% trehalose exhibited the best performance. The Met array with 5% trehalose could determine Met concentration with high linearity (R2 value = approximately 0.99) even after storage at −20 °C for up to 3 months. The clinical utilities of the Met and Leu array, preserved at −20 °C for 3 months, were also verified by successfully quantifying Met and Leu in spiked blood serum samples for the diagnosis of the corresponding metabolic diseases. This long-term preservation protocol enables the development of a ready-to-use bioluminescent E. coli-based amino acid array to quantify multiple amino acids and can replace the currently used laborious analytical methods.
The accurate and simultaneous detection of neurotransmitters, such as dopamine (DA) and epinephrine (EP), is of paramount importance in clinical diagnostic fields. Herein, we developed cerium–molybdenum disulfide nanoflowers (Ce-MoS2 NFs) using a simple one-pot hydrothermal method and demonstrated that they are highly conductive and exhibit significant peroxidase-mimicking activity, which was applied for the simultaneous electrochemical detection of DA and EP. Ce-MoS2 NFs showed a unique structure, comprising MoS2 NFs with divalent Ce ions. This structural design imparted a significantly enlarged surface area of 220.5 m2 g−1 with abundant active sites as well as enhanced redox properties, facilitating electron transfer and peroxidase-like catalytic action compared with bare MoS2 NFs without Ce incorporation. Based on these beneficial features, Ce-MoS2 NFs were incorporated onto a screen-printed electrode (Ce-MoS2 NFs/SPE), enabling the electrochemical detection of H2O2 based on their peroxidase-like activity. Ce-MoS2 NFs/SPE biosensors also showed distinct electrocatalytic oxidation characteristics for DA and EP, consequently yielding the highly selective, sensitive, and simultaneous detection of target DA and EP. Dynamic linear ranges for both DA and EP were determined to be 0.05~100 μM, with detection limits (S/N = 3) of 28 nM and 44 nM, respectively. This study shows the potential of hierarchically structured Ce-incorporated MoS2 NFs to enhance the detection performances of electrochemical biosensors, thus enabling extensive applications in healthcare, diagnostics, and environmental monitoring.
Nucleic acid aptamer-based research has focused on achieving the highest performance for bioassays. However, there are limitations in evaluating the affinity for the target analytes in these nucleic acid aptamer-based bioassays. In this study, we mainly propose graphene oxide (GO)-based electrical and optical analyses to efficiently evaluate the affinity between an aptamer and its target. We found that an aptamer-coupled GO-based chip with an electrical resistance induced by a field-effect transistor, with aptamers as low as 100 pM, can detect the target, thrombin, at yields as low as 250 pM within five minutes. In the optical approach, the fluorescent dye-linked aptamer, as low as 100 nM, was efficiently used with GO, enabling the sensitive detection of thrombin at yields as low as 5 nM. The cantilever type of mechanical analysis also demonstrated the intuitive aptamer–thrombin reaction in the signal using dBm units. Finally, a comparison of electrical and optical sensors’ characteristics was introduced in the attachment and detachment of aptamer to propose an efficient analysis that can be utilized for various aptamer-based research fields.