In the present study, we fabricated a dual-mode cardiac troponin I (cTnI) biosensor comprised of multi-functional DNA (MF-DNA) on Au nanocrystal (AuNC) using an electrochemical method (EC) and a localized surface plasmon resonance (LSPR) method. To construct a cTnI bioprobe, a DNA 3 way-junction (3WJ) was prepared to introduce multi-functionality. Each DNA 3WJ arm was modified to possess a recognition region (Troponin I detection aptamer), an EC-LSPR signal generation region (methylene blue: MB), and an anchoring region (Thiol group), respectively. After an annealing step, the multi-functional DNA 3WJ was assembled, and its configuration was confirmed by Native-TBM PAGE for subsequent use in biosensor construction. cTnI was also expressed and purified for use in biosensor experiments. To construct an EC-LSPR dual-mode biosensor, AuNCs were prepared on an indium-tin-oxide (ITO) substrate using an electrodeposition method. The prepared multi-functional (MF)-DNA was then immobilized onto AuNCs by covalent bonding. Field emission scanning electron microscope (FE-SEM) and atomic force microscopy (AFM) were used to analyze the surface morphology. LSPR and electrochemical impedance spectroscopy (EIS) experiments were performed to confirm the binding between the target and the bioprobe. The results indicated that cTnI could be effectively detected in the buffer solution and in diluted-human serum. Based on the results of these experiments, the loss on drying (LOD) was determined to be 1.0 pM in HEPES solution and 1.0 pM in 10% diluted human serum. Additionally, the selectivity assay was successfully tested using a number of different proteins. Taken together, the results of our study indicate that the proposed dual-mode biosensor is applicable for use in field-ready cTnI diagnosis systems for emergency situations.

C-reactive protein (CRP) is an acute-phase reactive protein that appears in the bloodstream in response to inflammatory cytokines such as interleukin-6 produced by adipocytes and macrophages during the acute phase of the inflammatory/infectious process. CRP measurement is widely used as a representative acute and chronic inflammatory disease marker. With the development of diagnostic techniques measuring CRP more precisely than before, CRP is being used not only as a traditional biomarker but also as a biomarker for various diseases. The existing commercialized CRP assays are dominated by enzyme-linked immunosorbent assay (ELISA). ELISA has high selectivity and sensitivity, but its limitations include requiring complex analytic processes, long analysis times, and professional manpower. To overcome these problems, nanobiotechnology is able to provide alternative diagnostic tools. By introducing the nanobio hybrid material to the CRP biosensors, CRP can be measured more quickly and accurately, and highly sensitive biosensors can be used as portable devices. In this review, we discuss the recent advancements in electrochemical, electricity, and spectroscopy-based CRP biosensors composed of biomaterial and nanomaterial hybrids.

The outbreak of the influenza virus (H1N1) has symptoms such as coughing, fever, and a sore throat, and due to its high contagious power, it is fatal to humans. To detect H1N1 precisely, the present study proposed an electrochemical biosensor composed of a multifunctional DNA four-way junction (4WJ) and carboxyl molybdenum disulfide (carboxyl-MoS₂) hybrid material. The DNA 4WJ was constructed to have the hemagglutinin aptamer on the head group (recognition part); each of the two arms has four silver ions (signal amplification part), and the tail group has an amine group (anchor). This fabricated multifunctional DNA 4WJ can specifically and selectively bind to hemagglutinin. Moreover, the carboxyl-MoS₂ provides an increase in the sensitivity of this biosensor. Carboxyl-MoS₂ was immobilized using a linker on the electrode, followed by the immobilization of the multifunctional 4WJ on the electrode. The synthesis of carboxyl-MoS₂ was confirmed by field emission scanning electron microscopy (FE-SEM), and the surface of the electrode was confirmed by atomic force microscopy. When H1N1 was placed in the immobilized electrode, the presence of H1N1 was confirmed by electrochemical analysis (cyclic voltammetry, electrochemical impedance spectroscopy). Through selectivity tests, it was also possible to determine whether this sensor responds specifically and selectively to H1N1. We confirmed that the biosensor showed a linear response to H1N1, and that H1N1 could be detected from 100 nM to 10 pM. Finally, clinical tests, in which hemagglutinin was diluted with human serum, showed a similar tendency to those diluted with water. This study showed that the multi-functional DNA 4WJ and carboxyl-MoS₂ hybrid material can be applied to a electrochemical H1N1 biosensor.

During the last 30 years, the World Health Organization (WHO) reported a gradual increase in the number of patients with cardiovascular disease (CVD), not only in developed but also in developing countries. In particular, acute myocardial infarction (AMI) is one of the severe CVDs because of the high death rate, damage to the body, and various complications. During these harmful effects, rapid diagnosis of AMI is key for saving patients with CVD in an emergency. The prompt diagnosis and proper treatment of patients with AMI are important to increase the survival rate of these patients. To treat patients with AMI quickly, detection of a CVD biomarker at an ultra-low concentration is essential. Cardiac troponins (cTNs), cardiac myoglobin (cMB), and creatine kinase MB are typical biomarkers for AMI detection. An increase in the levels of those biomarkers in blood implies damage to cardiomyocytes and thus is related to AMI progression. In particular, cTNs are regarded as a gold standard biomarker for AMI diagnosis. The conventional TN detection system for detection of AMI requires long measurement time and is labor-intensive and tedious. Therefore, the demand for sensitive and selective TN detection techniques is increasing at present. To meet this demand, several approaches and methods have been applied to develop a TN detection system based on a nanostructure. In the present review, the authors reviewed recent advances in TN biosensors with a focus on four detection systems: (1) An electrochemical (EC) TN nanobiosensor, (2) field effect transistor (FET)-based TN nanobiosensor, (3) surface plasmon resonance (SPR)-based TN nanobiosensor and (4) surface enhanced Raman spectroscopy (SERS)-based TN nanobiosensor.

Interferon gamma (IFN-γ) is an important cytokine with antiviral, antibacterial, and immunosuppressive properties. It has been used as a biomarker for the early detection of several diseases, including cancer, human immunodeficiency virus (HIV), tuberculosis, and paratuberculosis. In this study, we developed an electrochemical biosensor composed of multifunctional DNA 3WJ to detect IFN-γ level with high sensitivity. Each multifunctional triple-stranded aptamer (MF-3WJ) was designed to have an IFN-γ aptamer sequence, anchoring region (thiol group), and 4C–C (cytosine–cytosine) mismatch sequence (signal generation), which could introduce silver ions. To generate the electrochemical signal, four Ag⁺ ions were intercalated (3wj b-3wj c) in the 4C–C mismatch sequence. MF-3WJ was assembled through the annealing step, and the assembly of MF-3WJ was confirmed by 8% tris–boric–EDTA native polyacrylamide gel electrophoresis. The Au microgap electrode was manufactured to load sample volumes of 5 µL. The reliability of electrochemical biosensor measurement was established by enabling the measurement of seven samples from one Au microgap electrode. MF-3WJ was immobilized on the Au microgap electrode. Then, cyclic voltammetry and electrochemical impedance spectroscopy were performed to confirm the electrochemical properties of MF-3WJ. To test the electrochemical biosensor’s ability to detect IFN-γ, the limit of detection (LOD) and selectivity tests were performed by square wave voltammetry. A linear region was observed in the concentration range of 1 pg/mL–10 ng/mL of IFN-γ. The LOD of the fabricated electrochemical biosensor was 0.67 pg/mL. In addition, for the clinical test, the LOD test was carried out for IFN-γ diluted in 10% human serum samples in the concentration range of 1 pg/mL–10 ng/mL, and the LOD was obtained at 0.42 pg/mL.

With the acceleration of the Fourth Industrial Revolution, the development of information and communications technology requires innovative information storage devices and processing devices with low power and ultrahigh stability. Accordingly, bioelectronic devices have gained considerable attention as a promising alternative to silicon-based devices because of their various applications, including human-body-attached devices, biomaterial-based computation systems, and biomaterial–nanomaterial hybrid-based charge storage devices. Nanomaterial-based charge storage devices have witnessed considerable development owing to their similarity to conventional charge storage devices and their ease of applicability. The introduction of a biomaterial-to-nanomaterial-based system using a combination of biomolecules and nanostructures provides outstanding electrochemical, electrical, and optical properties that can be applied to the fabrication of charge storage devices. Here, we describe the recent advances in charge storage devices containing a biomolecule and nanoparticle heterolayer including (1) electrical resistive charge storage devices, (2) electrochemical biomemory devices, (3) field-effect transistors, and (4) biomemristors. Progress in biomolecule–nanomaterial heterolayer-based charge storage devices will lead to unprecedented opportunities for the integration of information and communications technology, biotechnology, and nanotechnology for the Fourth Industrial Revolution.