RESUMEN
A simple and sensitive AuNP-coated magnetic beads (AMB)-based electrochemical biosensor platform was fabricated for bioassay. In this study, AuNP-conjugated magnetic particles were successfully prepared using biotin-streptavidin conjugation. The morphology and structure of the nanocomplex were characterized by scanning electron microscopy (SEM) with energy-dispersive X-ray analysis (EDX) and UV-visible spectroscopy. Moreover, cyclic voltammetry (CV) was used to investigate the effect of AuNP-MB on alkaline phosphatase (ALP) for electrochemical signal enhancement. An ALP-based electrochemical (EC) immunoassay was performed on the developed AuNP-MB complex with indium tin oxide (ITO) electrodes. Subsequently, the concentration of capture antibodies was well-optimized on the AMB complex via biotin-avidin conjugation. Lastly, the developed AuNP-MB immunoassay platform was verified with extracellular vesicle (EV) detection via immune response by showing the existence of EGFR proteins on glioblastoma multiforme (GBM)-derived EVs (108 particle/mL) spiked in human plasma. Therefore, the signal-enhanced ALP-based EC biosensor on AuNP-MB was favorably utilized as an immunoassay platform, revealing the potential application of biosensors in immunoassays in biological environments.
RESUMEN
MXene nanosheets of type Ti3C2Tx were modified with ß-hydroxybutyrate dehydrogenase and then used as a biosensor for amperometric sensing of ß-hydroxybutyrate. The MXene and the nanocomposite were characterized by X-ray photoelectron spectroscopy, field-emission scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The MXene has a layered structure and proved to be an excellent immobilization matrix providing good compatibility with the enzyme ß-hydroxybutyrate dehydrogenase. The MXene-based biosensor, best operated at a potential of - 0.35 V (vs. Ag/AgCl), displays a wide linear range (0.36 to 17.9 mM), a sensitivity of 0.480 µA mM-1 cm-2, and a low detection limit (45 µM). The biosensor was successfully applied to the determination of ß-hydroxybutyrate in (spiked) real serum samples. Graphical abstract Schematic representation of the synthesis and decoration of Mxene 2D sheets with ß-hydroxybutyrate dehydrogenase for the amperometric determination of ß-hydroxybutyric acid.
Asunto(s)
Ácido 3-Hidroxibutírico/análisis , Compuestos Inorgánicos de Carbono/química , Hidroxibutirato Deshidrogenasa/química , Nanocompuestos/química , Titanio/química , Ácido 3-Hidroxibutírico/metabolismo , Técnicas Biosensibles , Técnicas Electroquímicas , Electrodos , Hidroxibutirato Deshidrogenasa/metabolismo , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
The growing interest in magnetic materials as a universal tool has been shown by an increasing number of scientific publications regarding magnetic materials and its various applications. Substantial progress has been recently made on the synthesis of magnetic iron oxide particles in terms of size, chemical composition, and surface chemistry. In addition, surface layers of polymers, silica, biomolecules, etc., on magnetic particles, can be modified to obtain affinity to target molecules. The developed magnetic iron oxide particles have been significantly utilized for diagnostic applications, such as sample preparations and biosensing platforms, leading to the selectivity and sensitivity against target molecules and the ease of use in the sensing systems. For the process of sample preparations, the magnetic particles do assist in target isolation from biological environments, having non-specific molecules and undesired molecules. Moreover, the magnetic particles can be easily applied for various methods of biosensing devices, such as optical, electrochemical, and magnetic phenomena-based methods, and also any methods combined with microfluidic systems. Here we review the utilization of magnetic materials in the isolation/preconcentration of various molecules and cells, and their use in various techniques for diagnostic biosensors that may greatly contribute to future innovation in point-of-care and high-throughput automation systems.