RESUMO
Given the importance of hydrogen peroxide (H2O2) in many biological processes and its wide application in various industries, the demand for sensitive, accurate, and economical H2O2 sensors is high. In this study, we used Fenton reaction-stimulated fluorescence quenching of N-acetyl-L-cysteine-protected gold nanoclusters (NAC-AuNCs) as a reporter system for the determination of H2O2. After the experimental conditions were optimized, the sensing platform enabled the analysis of H2O2 with a limit of detection (LOD) as low as 0.027 µM. As the glucose oxidase cascade leads to the generation of H2O2 and catalase catalyzes the decomposition of H2O2, these two biocatalytic procedures can be probed by the Fenton reaction-mediated quenching of NAC-AuNCs. The LOD for glucose was found to be 0.18 µM, and the linear range was 0.39-27.22 µM. The LOD for catalase was 0.002 U mL(-1), and the linear range was 0.01-0.3 U mL(-1). Moreover, the proposed sensing methods were successfully applied for human serum glucose detection and the non-invasive determination of catalase activity in human saliva, demonstrating their great potential for practical applications.
Assuntos
Acetilcisteína/química , Técnicas Biossensoriais/métodos , Glicemia/análise , Catalase/análise , Peróxido de Hidrogênio/análise , Ferro/química , Nanopartículas Metálicas/química , Glucose/análise , Humanos , Peróxido de Hidrogênio/química , Limite de Detecção , Modelos Moleculares , Saliva/química , Saliva/enzimologiaRESUMO
Protein-supported nanoparticles have a great significance in scientific and nanotechnology research because of their "green" process, low cost-in-use, good biocompatibility, and some interesting properties. Ruthenium oxide nanoparticles (RuO2NPs) have been considered to be an important member in nanotechnology research. However, the biosynthetic approach of RuO2NPs is relatively few compared to those of other nanoparticles. To address this challenge, this work presented a new way for RuO2NP synthesis (BSA-RuO2NPs) supported by bovine serum albumin (BSA). BSA-RuO2NPs are confirmed to exert peroxidase-like activity, electrocatalytic activity, in vitro salt resistance (2 M NaCl), and biocompatibility. Results indicate that BSA-RuO2NPs have higher affinity binding for 3,3',5,5'-tetramethylbenzidine or H2O2 than bare RuO2NPs. Moreover, BSA turns out to be a crucial factor in promoting the stability of RuO2NPs. Taking the advantages of these improved properties, we established colorimetric (linear range from 2 to 800 µM, a limit of detection of 1.8 µM) and electrochemical (linear range from 0.4 to 3850 µM, a limit of detection of 0.18 µM) biosensors for monitoring in situ H2O2 secretion from living MCF-7 cells. Herein, this work offers a new biosynthesis strategy to obtain BSA-RuO2NPs and sheds light on the sensitive biosensors to monitor the H2O2 secreted from living cells for promising applications in the fields of nanotechnology, biology, biosensors, and medicine.
Assuntos
Materiais Biocompatíveis/química , Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/análise , Nanopartículas Metálicas/química , Compostos de Rutênio/química , Soroalbumina Bovina/química , Cloreto de Sódio/química , Animais , Benzidinas/química , Catálise , Bovinos , Colorimetria , Técnicas Eletroquímicas , Eletrodos , Humanos , Limite de Detecção , Células MCF-7RESUMO
In this study, a novel biomolecule immobilization approach has been proposed to the synthesis of multi-functional core-shell glucose oxidase-Au-polydopamine-Fe3O4 magnetic bionanoparticles (GOx-Au-PDA-Fe3O4 MBNPs) using the one-pot chemical polymerization method. Then, a high performance biosensor has been constructed by effectively attaching the proposed GOx-Au-PDA-Fe3O4 MBNPs to the surface of the magnetic glassy carbon electrode. Scanning electron microscope, energy dispersive x-ray spectrometer, UV-vis spectroscopy, and electrochemical methods were used to characterize the GOx-Au-PDA-Fe3O4 MBNPs. The resultant GOx-Au-PDA-Fe3O4 MBNPs not only have the magnetism of Fe3O4 nanoparticles which makes them easily manipulated by an external magnetic field, but also have the excellent biocompatibility of PDA to maintain the native structure of the GOx, and good conductivity of Au nanoparticles which can facilitate the direct electrochemistry of GOx in the biofilm. Hence, the present GOx-Au-PDA-Fe3O4 biofilm displays good linear amperometric response to glucose concentration ranging from 0.02 to 1.875 mM. This efficient biomolecule immobilization platform is recommended for the preparation of many other MBNPs with interesting properties and application potentials in many fields, such as biosensing, biocatalysis, biofuel cells, and bioaffinity separation.
Assuntos
Técnicas Biossensoriais/métodos , Glucose Oxidase/química , Glucose/isolamento & purificação , Nanopartículas de Magnetita/química , Quitosana/química , Enzimas Imobilizadas , Compostos Férricos/química , Ouro/química , Indóis/química , Polímeros/químicaRESUMO
In this work, the magnetic core-shell Fe(3)O(4)@Au nanoparticles attached to the surface of a magnetic glassy carbon electrode (MGCE) were applied to the immobilization/adsorption of myoglobin (Mb) for fabricating Mb/Fe(3)O(4)@Au biofilm. The morphology, structure, and electrochemistry of the nanocomposite were characterized by transmission electron microscope, UV-vis spectroscopy, electrochemical impedance spectroscopy, and cyclic voltammetry, respectively. The resultant Fe(3)O(4)@Au NPs not only have the magnetism of Fe(3)O(4) NPs that make them easily manipulated by an external magnetic field, but also have the good conductivity and excellent biocompatibility of Au layer which can maintain the bioactivity and facilitate the direct electrochemistry of Mb in the biofilm. The modified electrode based on this Mb/Fe(3)O(4)@Au biofilm displayed good electrocatalytic activity to the reduction of H(2)O(2) with a linear range from 1.28 to 283 microM. The proposed method simplified the immobilization methodology of proteins and showed potential application for fabricating novel biosensors and bioelectronic devices.