Electrochemical immunosensor AuNPs/NG-PANI/ITO-PET for the determination of BDNF in depressed mice serum.

A novel electrochemical immunosensor was developed for highly sensitive detection of brain-derived neurotrophic factor (BDNF), a well-known depression marker. The immunosensor was fabricated by modifying indium tin oxide-coated polyethylene terephthalate (ITO-PET) with N-doped graphene-polyaniline (NG-PANI) and gold nanoparticles (AuNPs) to enhance the conductivity and protein loading capacity. Subsequently, BDNF was immobilized onto the electrode surface via gold-sulfur bonds, followed by the attachment of biotinylated antibody (Biotin-Ab) and horseradish peroxidase-avidin (HRP-Avidin) to create the final immunosensor (HRP-Avidin-Biotin-Ab-BDNF-AuNPs/NG-PANI/ITO-PET). The proposed immunosensor exhibited a linear range of determination (0.781-400 pg/mL) with a low limit of detection (LOD) of 0.261 pg/mL (S/N = 3) and excellent reproducibility (RSD = 1.4%) and stability (92.7%, RSD = 3.1%). Additionally, the immunosensor demonstrated good anti-interference performance and good recovery (98.1-107%). To evaluate the practical utility of the immunosensor, BDNF levels were quantified in the serum of mice with depression induced by chronic unpredictable mild stress (CUMS). The results indicated that the serum BDNF levels were significantly decreased in the depression model group compared with the control group, highlighting the potential of this immunosensor for clinical detection of BDNF in depression diagnosis and treatment.

Ultrasensitive cortisol electrochemical immunosensor amplifying by Au single-atom nanozymes and HRP enzymes.

Cortisol, a corticosteroid hormone as a primary stress hormone response to internal and external stress, has been regarded as a gold standard reliable biomarker to evaluate human mental stress. The double enzymes strategy, using nanozyme and enzyme amplifying the electrochemical signal, has been widely used to improve the performance of electrochemical biosensors. An ultra-sensitive electrochemical cortisol sensor based on Au single-atom nanozymes had been fabricated through HRP labeled anti-cortisol antibody binding with Au by Au-S bond. Based on the high catalytic activity of Au single-atom nanozymes and the high selectivity of HRP-labeled anti-cortisol antibodies, the cortisol electrochemical sensor-based Au single-atom nanozymes had an excellent response to cortisol, such as high electrochemical activity, high sensitivity, high selectivity, and wide linear range (0.15-300 ng mL-1) and low detection (0.48 pg mL-1) through the four-parameter logistic model with 95% confidence. The electrochemical cortisol sensor was used to determine the cortisol concentration of human saliva at different times.

Detection of depression marker ASS1 in urine by gold nanoparticles based dual epitope-peptides imprinted sensor.

Trace detection of argininosuccinate synthetase 1 (ASS1), a depression marker, in urine samples is difficult to achieve. In this work, a dual-epitope-peptides imprinted sensor for ASS1 detection in urine was constructed based on the high selectivity and sensitivity of the "epitope imprinting approach". First, two cysteine-modified epitope-peptides were immobilized onto gold nanoparticles (AuNPs) deposited on a flexible electrode (ITO-PET) by gold-sulfur bonds (Au-S), then a controlled electropolymerization of dopamine was carried out to imprint the epitope peptides. After removing epitope-peptides, the dual-epitope-peptides imprinted sensor (MIP/AuNPs/ITO-PET) which with multiple binding sites for ASS1 was obtained. Compared with single epitope-peptide, dual-epitope-peptides imprinted sensor had higher sensitivity, which presented a linear range from 0.15 to 6000 pg ml-1 with a low limit of detection (LOD = 0.106 pg mL-1, S/N = 3). It had good reproducibility (RSD = 1.74%), repeatability (RSD = 3.60%), stability (RSD = 2.98%), and good selectivity, and the sensor had good recovery (92.4%-99.0%) in urine samples. This is the first highly sensitive and selective electrochemical assay for the depression marker ASS1 in urine, which is expected to provide help for the non-invasive and objective diagnosis of depression.

Direct electrochemistry and electrocatalysis of lobetyolin via magnetic functionalized reduced graphene oxide film fabricated electrochemical sensor.

A novel lobetyolin electrochemical sensor based on a magnetic functionalized reduced graphene oxide/Nafion nanohybrid film has been introduced in this work. The magnetic functionalized reduced graphene oxide was characterized by fourier transform infrared spectroscopy, atomic force microscope, X-ray diffraction, transmission electron microscopy and thermogravimetric analysis. The scanning electron microscopy characterized the morphology and microstructure of the prepared sensors, and the electrochemical effective surface areas of the prepared sensors were also calculated by chronocoulometry method. The electrochemical behavior of lobetyolin on the magnetic functionalized reduced graphene oxide/Nafion nanohybrid modified glassy carbon electrode was investigated by cyclic voltammetry and differential pulse voltammetry in a phosphate buffer solution of pH6.0. The electron-transfer coefficient (α), electron transfer number (n), and electrode reaction rate constant (Κs) were calculated as 0.78, 0.73, and 4.63s-1, respectively. Under the optimized conditions, the sensor based on magnetic functionalized reduced graphene oxide/Nafion showed a linear voltammetric response to the lobetyolin concentration at 1.0×10-7 to 1.0×10-4mol/L with detection limit (S/N=3)of 4.3×10-8mol/L. The proposed sensor also displayed acceptable reproducibility, long-term stability, and high selectivity, and performs well for analysis of lobetyolin in real samples. The voltammetric sensor was successfully applied to detect lobetyolin in Codonopsis pilosula with recovery values in the range of 96.12%-102.66%.

Protections of bovine serum albumin protein from damage on functionalized graphene-based electrodes by flavonoids.

A sensitive electrochemical sensor based on bovine serum albumin (BSA)/poly (diallyldimethylammonium chloride) (PDDA) functionalized graphene nanosheets (PDDA-G) composite film modified glassy carbon electrode (BSA/PDDA-G/GCE) had been developed to investigate the oxidative protein damage and protections of protein from damage by flavonoids. The performance of this sensor was remarkably improved due to excellent electrical conductivity, strong adsorptive ability, and large effective surface area of PDDA-G. The BSA/PDDA-G/GCE displayed the greatest degree of BSA oxidation damage at 40 min incubation time and in the pH 5.0 Fenton reagent system (12.5 mM FeSO4, 50 mM H2O2). The antioxidant activities of four flavonoids had been compared by fabricated sensor based on the relative peak current ratio of SWV, because flavonoids prevented BSA damage caused by Fenton reagent and affected the BSA signal in a solution containing Co(bpy)3(3+). The sensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). UV-vis spectrophotometry and FTIR were also used to investigate the generation of hydroxyl radical and BSA damage, respectively. On the basis of results from electrochemical methods, the order of the antioxidant activities of flavonoids is as follows: (+)-catechin>kaempferol>apigenin>naringenin. A novel, direct SWV analytical method for detection of BSA damage and assessment of the antioxidant activities of four flavonoids was developed and this electrochemical method provided a simple, inexpensive and rapid detection of BSA damage and evaluation of the antioxidant activities of samples.

Synthesis of a biocompatible gelatin functionalized graphene nanosheets and its application for drug delivery.

A simple and environmentally friendly synthetic route for the preparation of gelatin functionalized graphene nanosheets (gelatin-GNS) was reported by using exfoliated graphene oxide as a precursor, in which gelatin acted as not only a reducing reagent but also a functionalization reagent to guarantee good dispersibility and stability of the GNS in distilled water and various physiological solutions. The obtained biocompatible gelatin-GNS attaching methotrexate (MTX) via strong π-π stacking interaction, exhibited a high drug loading capacity of MTX and excellent ability for controlled drug release. The pH-dependent release behavior of MTX from MTX@gelatin-GNS showed that the release amount under acid conditions is much higher than that under neutral conditions, which experienced a gelatin-mediated sustained release process. From the cytotoxicity assay, we can see that the MTX@gelatin-GNS showed remarkable toxicity while the gelatin-GNS showed nontoxic at appropriate concentration, both of them might be taken up by A549 cells through a nonspecific endocytosis process. The prepared nanohybrids system offers a novel formulation that combines the unique properties of a biodegradable material, gelatin, and graphene for biomedical applications. Therefore, the gelatin-GNS with good stability and biocompatibility can be selected as an ideal drug carrier to be applied in biomedicine studies.