Glucose Oxidase Research Articles

Glucose Oxidase Patterned Meta‐Chemical Surface for Sensing Glucose Using Dip‐Pen Nanolithography

AbstractNumerous studies have been done to advance the development of efficient, simple, and cheap sensors, especially for glucose, which are important in the pharmaceutical and food industries. The most common sensors and biosensors used today are based on electrochemistry. The novelty of this study is the use of dip‐pen nanolithography (DPN) in order to pattern a mixture of glucose oxidase with PMMA as nanoclusters and develop meta‐chemical surface (MCS), which leads to highly efficient working electrodes with increased detection sensitivity compared to the today‐known glucose sensors ( M was detected in the current study). Our novel and important results support the feasibility of using DPN to develop sensors on various electrode surfaces patterned with different active species, especially in systems requiring target substance detection over large concentration ranges.

An injectable selenite-containing hydrogel for synergistic tumor therapy by triggering ROS/RNS generation and disrupting NADPH homeostasis

The anti-cancer effect of selenite (SeO32−) is limited by its short half-life and narrow therapeutic window. Peroxynitrite (ONOO−), the most active reactive nitrogen species (RNS) derived from nitric oxide (NO) reacting with superoxide anion (O2•−), is still not fully exploited in the combination therapy against cancers. Herein, an injectable hydrogel is developed to trigger intracellular reactive oxygen species (ROS)/RNS bursts and disrupt nicotinamide adenine dinucleotide phosphate (NADPH) homeostasis via an innovative SeO32−-based strategy. Calcium selenite/L-Arginine (L-Arg) nanospheres (SL NPs) are synthesized as the donor of NO and SeO32−. The injectable Alg@SLGA sol composed of SL NPs, glucose oxidase (GOx), 6-aminonicotinamide (6-AN, an inhibitor of the pentose phosphate pathway (PPP)), and sodium alginate (Alg) can convert to gel in a physiological environment. Peritumoral injection of Alg@SLGA hydrogel is performed for safe and targeted treatment. The loaded GOx catalyzes glucose oxidation to produce acidic H2O2, initiating the release of NO and SeO32− from Alg@SLGA. SeO32− reacts with intracellular GSH and H2O2 to generate •OH and O2•−, which further interacts with NO to form ONOO−. The SeO32− and 6-AN synergistically trigger intracellular NADPH exhaustion by promoting cystine metabolism-mediated NADPH consumption and inhibiting NADPH generation, respectively. Moreover, Alg@SLGA effectively induces immunogenic cell death (ICD) of tumor cells, further provoking systemic antitumor immune responses. As a consequence, the injectable Alg@SLGA hydrogel achieves enhanced ROS/RNS-mediated combination therapy by synergistically disrupting NADPH homeostasis and improving antitumor immunity.

Bilosomes and Biloparticles for the Delivery of Lipophilic Drugs: A Preliminary Study.

In this study, bile acid-based vesicles and nanoparticles (i.e., bilosomes and biloparticles) are studied to improve the water solubility of lipophilic drugs. Ursodeoxycholic acid, sodium cholate, sodium taurocholate and budesonide were used as bile acids and model drugs, respectively. Bilosomes and biloparticles were prepared following standard protocols with minor changes, after a preformulation study. The obtained systems showed good encapsulation efficiency and dimensional stability. Particularly, for biloparticles, the increase in encapsulation efficiency followed the order ursodeoxycholic acid < sodium cholate < sodium taurocholate. The in vitro release of budesonide from both bilosytems was performed by means of dialysis using either a nylon membrane or a portion of Wistar rat small intestine and two receiving solutions (i.e., simulated gastric and intestinal fluids). Both in gastric and intestinal fluid, budesonide was released from bilosystems more slowly than the reference solution, while biloparticles showed a significant improvement in the passage of budesonide into aqueous solution. Immunofluorescence experiments indicated that ursodeoxycholic acid bilosomes containing budesonide are effective in reducing the inflammatory response induced by glucose oxidase stimuli and counteract ox-inflammatory damage within intestinal cells.

Hierarchically decorated magnetic nanoparticles amplify the oxidative stress and promote the chemodynamic/magnetic hyperthermia/immune therapy

Magnetic nanoparticles (MNPs) are promising in tumor treatments due to their capacity for magnetic hyperthermia therapy (MHT), chemodynamic therapy (CDT), and immuno-related therapies, but still suffer from unsatisfactory tumor inhibition in the clinic. Insufficient hydrogen peroxide supply, glutathione-induced resistance, and high-density extracellular matrix (ECM) are the barriers. Herein, we hierarchically decorated MNPs with disulfide bonds (S-S), dendritic L-arginine (R), and glucose oxidase (GOx) to form a nanosystem (MNPs-SS-R-GOx). Its outer GOx layer not only enhanced the H2O2 supply to produce .OH by Fenton reaction, but also generated stronger oxidants (ONOO−) together with the interfaced R layer. The inner S-S layer consumed glutathione to interdict its reaction with oxidants, thus enhancing CDT effects. Importantly, the generated ONOO− tripled the MMP-9 expression to induce ECM degradation, enabling much deeper penetration of MNPs and benefiting CDT, MHT, and immunotherapy. Finally, the MNPs-SS-R-GOx demonstrated a remarkable 91.7% tumor inhibition in vivo. Statement of significanceMagnetic nanoparticles (MNPs) are a promising tumor therapeutic agent but with limited effectiveness. Our hierarchical MNP design features disulfide bonds (S-S), dendritic L-arginine (R), and glucose oxidase (GOx), which boosts H2O2 supply for ·OH generation in Fenton reactions, produces potent ONOO−, and enhances chemodynamic therapy via glutathione consumption. Moreover, the ONOO− facilitates the upregulation of matrix metalloprotein expression beneficial for extracellular matrix degradation, which in turn enhances the penetration of MNPs and benefits the antitumor CDT/MHT/immuno-related therapy. In vivo experiments have demonstrated an impressive 91.7% inhibition of tumor growth. This hierarchical design offers groundbreaking insights for further advancements in MNP-based tumor therapy. Its implications extend to a broader audience, encompassing those interested in material science, biology, oncology, and beyond.

Constructing a novel pH-mediated magnetic relaxation sensor via the Mn(II)/Mn(IV) conversion

Herein, we demonstrate a new concept of chemical sensing for the construction of pH-mediated magnetic sensors. The strategy is based on the notion that the oxidation of paramagnetic Mn2+ ions to insoluble Mn(IV)O(OH)2 by dissolved oxygen under alkaline conditions can lead to a significant decrease in the transverse relaxation rate (1/T2) of water protons. Crucially, the conversion of Mn2+ ions to MnO(OH)2 correlates with the amount of NaOH, that is, the reaction is pH-dependent. Thus, by combining this reaction with enzymatic reactions that cause pH changes, we can construct pH-mediated chemical sensors based on magnetic relaxation principles. In this work, we selected glucose and urea as model substrates to verify the feasibility of this strategy, since the oxidation of glucose by glucose oxidase and the hydrolysis of urea by urease lead to a notable decrease and increase of pH, respectively, thereby causing a change in the 1/T2 of the solution. Experimental results showed that the linear detection range was 20–2000 µM with a detection limit of 10 µM for glucose, and 50–1000 µM with a detection limit of 20 µM for urea, validating the proof of concept and potential broad applicability of this strategy.

Biodegradable hollow mesoporous bimetallic nanoreactors to boost chemodynamic therapy

As an endogenous catalytic treatment, chemodynamic therapy (CDT) was attracting considerable attention, but the weak catalytic efficiency of Fenton agents and the non-degradation of nanocarriers severely limited its development. In this work, a biodegradable bimetallic nanoreactor was developed for boosting CDT, in which Fe-doped hollow mesoporous manganese dioxide (HMnO2) was selected as nanocarrier, and the Fe/HMnO2@DOX-GOD@HA nanoprobe was constructed by loading doxorubicin (DOX) and modifying glucose oxidase (GOD) and hyaluronic acid (HA). The glutathione (GSH) responsive degradation of HMnO2 promoted the release of DOX, by which the release rate significantly increased to 96.6%. Moreover, by the GSH depletion, the reduction of Mn2+/Fe2+ achieved strong bimetallic Fenton efficiency, and the hydroxyl radicals (·OH) generation was further enhanced using the self-supplying H2O2 of GOD. Through the active targeting recognition of HA, the bimetallic nanoreactor significantly enriched the tumor accumulation, by which the enhanced antitumor efficacy was realized. Thus, this work developed biodegradable bimetallic nanoreactor by consuming GSH and self-supplying H2O2, and provided a new paradigm for enhancing CDT.

Characterization of glucose Oxidase from Penicillium chrysogenum MDC 8358: Prospects for application in food industry

Backgrounds: Food enzymes play a crucial role in enhancing specific food attributes, such as improving texture, eliminating toxins and allergens, producing carbohydrates, and enhancing flavor and appearance characteristics. Glucose oxidase (GOX) catalyzes the oxidation of glucose to produce gluconic acid and hydrogen peroxide, making it applicable in various practical scenarios. This enzyme could be utilized to eliminate glucose, thereby facilitating the creation of functional food suitable for individuals with diabetes and glucose intolerance.Our main goal was to characterize GOX from a Penicillium chrysogenum strain isolated from Armenian ecosystems.Methods:GOX of P. chrysogenum MDC 8358 was obtained by surface fermentation using glucose as a carbon source. The enzyme was purified 230-fold to homogeneity using a three-step purification scheme consisting of ion exchange, adsorption, and gel filtration chromatography. Results:The specific activity of the purified enzyme reached 271.2 U/mg. GOX from P. chrysogenum MDC 8358 has shown to be dimeric with a molecular weight of 135.5 kDa, consisting of two equal subunits with a molecular weight of 70 kDa. Isoelectric focusing has revealed a pH of 4.6. The enzyme has displayed a temperature optimum of 40 °C, and a pH optimum of 6.8, being more thermostable at acidic pH. Characterized GOX was highly specific for β-D-glucose and has shown only minor activities to mannose, xylose, and galactose. Keywords:Penicillium chrysogenum MDC 8358, surface fermentation, glucose oxidase, purification, characterization, functional food preservation

An enzymatic glucose biosensor using the BESOI MOSFET

A reconfigurable transistor glucose biosensor was fabricated using the BESOI MOSFET structure. Glucose oxidase enzyme was immobilized on the sensitive regions of the device to specifically detect glucose. The experimental measurements showed a positive linear relationship between the sensitivity and the glucose solution concentration, in the measured range (0–200 mM). The maximum value of 0.52 was obtained for programming gate voltage VGB = 15 V. Next, numerical simulations based on the previous experimental results enabled the study of the channel thickness influence on the sensitivity. An optimal channel thickness value of 10 nm was obtained, in which the generated charges due to the glucose oxidation reaction most impact the drain current level of this device.

Photosynthetic and Self-Draining Biohybrid Dressing for Accelerating Healing of Diabetic Wound.

Wound healing is a well-orchestrated progress associated with angiogenesis, epithelialization, inflammatory status, and infection control, whereas these processes are seriously disturbed in diabetic wounds. In this study, a biohybrid dressing integrating the inherent ability of Bromeliad leaf (photosynthesis and self-draining) with the therapeutic effect of artificial materials (glucose-degrading and ROS-scavenging) is presented. The dressing consists of double-layered structures as follows: 1) Outer layer, a Bromeliad leaf substrate full of alginate hydrogel-immobilized glucose oxidase (GOx@Alg@Bromeliad substrate, abbreviated as BGA), can generate oxygen to guarantee the GOx-catalyzed glucose oxidation by photosynthesis, reducing local hyperglycemia to stabilize hypoxia inducible factor-1 alpha (HIF-1α) for angiogenesis and producing hydrogen peroxide for killing bacteria on the surface of wound tissue. The sophisticated structure of the leaf drains excessive exudate away via transpiration-mimicking, preventing skin maceration and impeding bacterial growth. 2) Inner layer, microneedles containing catalase (CAT-HA MNs, abbreviated as CHM), reduces excessive oxidative stress in the tissue to promote the proliferation of fibroblasts and inhibits proinflammatory polarization of macrophages, improving re-epithelialization of diabetic wounds. Together, the biohybrid dressing (BGA-CHM, abbreviated as BCHM) can enhance angiogenesis, strengthen re-epithelialization, alleviate chronic inflammation, and suppress bacterial infection, providing a promising strategy for diabetic wound therapy.

ZIF-8 encapsulated-enzymes integrated nanozyme cascade biocatalysis platform for the colorimetric sensing of glucose and lactose in milk

Cascade biocatalytic reactions have a wide range of applications, especially in the filed of food analysis. Herein, a multi-enzyme composite (ZGGPC) was prepared by in-situ synthesis of Zeolite imidazole framework-8 (ZIF-8) on Prussian blue (PB) modified carbon cloth (CC). The composite encapsulated both glucose oxidase and β-galactosidase simultaneously during the synthesis process. CC and ZIF-8 showed high loading capacity for PB and natural enzymes, respectively. And ZIF-8 also displayed excellent tolerance in protecting enzyme activity under extreme conditions. Based on the cascade biocatalysis, ZGGPC was used to detect glucose and lactose by colorimetric method with detection limits of 1.2 μM and 1.7 mM, respectively. Benefiting from the merits of low cost, easy preparation, and good stability, the sensing system was used to successfully determine glucose and lactose in different milk samples. The present cascade biocatalysis system is hopeful to develop simple and efficient sensing platforms for food analysis.

Gestational Diabetes Mellitus among women attending antenatal clinic in a Tertiary Hospital in Northern Nigeria

Introduction: Women with gestational diabetes mellitus (GDM) are at an increased risk of adverse perinatal and maternal morbidities and mortalities. Hence, early detection and management of this condition is vital to ensure a good outcome for both mother and baby. Aim: The aim of the study is to determine the prevalence and associated risk factors for GDM among antenatal attendees in Federal Medical Centre Gusau (FMCG), Zamfara state, Nigeria. Methods: A descriptive cross-sectional study was carried out. Screening was done between 24 and 28-weeks gestational age with a 75g oral glucose tolerance test (OGTT). Plasma glucose measurements was performed using the glucose oxidase method. GDM was diagnosed according to the 2013 WHO diagnostic criteria. Data were collected using a proforma and analyzed using the Statistical Package for Social Sciences (SPSS) Version 26. Descriptive data were presented in frequency tables. Statistical testing using Chi-square, and multivariate analysis (Odd ratio and CI) were carried out with a level of significance set as P&lt; 0.05. Results: One hundred and thirty-six women between the gestational ages of 24-28 weeks were recruited. The mean age of the participants was 25.9±5.4 years. The prevalence of GDM was found to be 16.9%. Fasting blood glucose alone correctly diagnosed 87% of the patients. Previous history of macrosomic babies (≥4kg) and history of GDM in previous pregnancies were significantly associated with GDM. Conclusion: The prevalence of GDM is high among antenatal attendees in Federal Medical center, Gusau.

A multivalent polyphenol–metal-nanoplatform for cascade amplified chemo-chemodynamic therapy

Chemodynamic therapy (CDT), as an emerging therapeutic strategy, kills cancer cells by converting intracellular hydrogen peroxide (H2O2) into cytotoxic oxidizing hydroxyl radicals (⋅OH). However, the therapeutic efficiency of CDT is compromised due to the insufficient endogenous H2O2 and metal catalysts in tumor cells. The use of multivalent polyphenols with multiple hydroxyl functions provides a facile yet robust means for efficient CDT augmentation. For this purpose, we reported herein the construction of polyphenol–metal nanoparticles (NPs) via a phenol–metal coordination strategy. The uniqueness of this study is the preparation of only one polymer construct with multivalency that can afford various supramolecular interactions for simultaneous “one-pot” loading of different therapeutic species, i.e., doxorubicin (DOX), glucose oxidases (GOD), and Fe3+ and further co-self-assembly into a stabilized nanomedicine for cascade amplified chemo-chemodynamic therapy. Specifically, the tumor intracellular acidic pH-triggered DOX release could serve for chemotherapy as well as enhance the intracellular H2O2 level. Together with the extra H2O2 and gluconic acid produced by the GOD-triggered glucose consumption, DOX@POAD-Fe@GOD NPs promoted Fe3+participation in the Fe-mediated Fenton reaction for cascade amplified chemo-chemodynamic therapy. Notably, this formulation displayed a greater anti-tumor effect with a tumor inhibition ratio 1.6-fold higher than that of free DOX in a BALB/c mice model bearing 4T1 tumors. Overall, the multivalent polyphenol–metal nanoplatform developed herein integrates chemotherapy, starvation therapy, and CDT for synergistic enhanced anticancer efficiency, which shows great potential for clinical translations. Statement of significanceChemodynamic therapy (CDT) generally suffers from compromised therapeutic efficiency due to insufficient endogenous H2O2 and metal catalysts in tumor cells. To develop a facile yet robust strategy for efficient CDT augmentation, we reported herein construction of a multivalent polyphenol–metal nanoplatform, DOX@POAD-Fe@GOD nanoparticles (NPs) via a phenol–metal coordination strategy. This nanoplatform integrates multiple supramolecular dynamic interactions not only for simultaneously safe encapsulation of doxorubicin (DOX), Fe3+, and glucose oxidases (GOD), but also for cascade amplified chemo-chemodynamic therapy. Specifically, the intracellular acidic pH-triggered dissociation of DOX@POAD-Fe@GOD NPs promoted the release of Fe3+, DOX, and GOD for significantly increased ROS levels that can accelerate Fenton reactions for cascaded chemotherapy, starvation therapy, and CDT with amplified antitumor efficiency in vivo.

Synergistic co-catalytic nanozyme system for highly efficient one-pot colorimetric sensing at neutral pH: Combining molybdenum trioxide and Fe(III)-Modified covalent triazine framework

This study investigates the co-catalytic capabilities of MoO3 nanosheets in enhancing the enzyme-like catalytic activity of a two-dimensional ultrathin Fe(III)-modified covalent triazine framework (Fe-CTF) under neutral pH conditions. The unique physicochemical surface properties and two-dimensional structures of Fe-CTF enable the direct immobilization of native enzymes (glucose oxidase (GOD) and xanthine oxidase (XOD)) through adsorption, eliminating the need for chemical processes. Efficient immobilization of the native enzymes within the Fe-CTF/GOD(XOD) hybrid is achieved through multipoint attachment involving various interactions. The Fe-CTF/MoO3 co-catalytic system exhibits enzyme-mimicking activity at neutral pH and, when combined with the high catalytic activity of the immobilized native enzymes, enables the development of a colorimetric method for glucose detection. This method demonstrates excellent facilitation, rapidity, sensitivity, and selectivity, with a linear detection range of 50–1000 μM and a limit of detection of 8.8 μM for glucose. Furthermore, a straightforward one-pot colorimetric method is established for screening XOD inhibitors. The inhibitory potential of a crude extract derived from Chinese water chestnut peel on XOD activity is evaluated using this method. The findings of this study pave the way for the utilization of nanozyme/native enzyme hybrids in pH-neutral conditions for one-pot colorimetric sensing. This work contributes to the advancement of enzyme-based sensing technologies and holds promise for various applications in biosensing and biomedical research.

Spatiotemporal expression patterns of ZBP1 in the brain of mouse experimental stroke model

Z-DNA binding protein 1 (ZBP1) is a cytosolic nucleic acid sensor, functioning as a critical mediator of inflammation and cell death pathways. Since neuroinflammation could occur in response to damage-associated molecular patterns (DAMPs), ZBP1 might be involved in neuroinflammation after stroke. However, the spatiotemporal expression profile of ZBP1 in the post-stroke brain remains to be elucidated. The aim of this study is to demonstrate the spatiotemporal expression patterns of ZBP1 in the post-stroke brain using a mouse photothrombotic stroke model. Real-time PCR assays showed that ZBP1 is induced on days 3–14 post stroke. ZBP1 immunoreactivity was observed in Iba1-positive microglia/macrophages in peri-infarct regions by immunohistochemistry. ZBP1-positive cells were spread in layers surrounding the infarct core by 7–14 days post stroke. Interestingly, ZBP1 immunoreactivity was also detected in CD206-positive border-associated macrophages (BAMs) in the meninges. Furthermore, ZBP1-expressing cells were positive for antibodies against inflammatory mediators such as Toll-like receptor 4 (TLR4), Toll/IL-1R domain-containing adaptor-inducing IFN-β (TRIF), and receptor-interacting serine/threonine-protein kinase 1 (RIPK1). Morphological analysis with confocal microscopy showed that the co-localization signals of ZBP1 and its adaptor, TRIF, are increased by glucose oxidase (GOx) treatment, which has been reported to induce mitochondrial DNA (mtDNA) release. These results suggest that ZBP1 is induced in peri-infarct microglia/macrophages and may be involved in DAMPs-mediated neuroinflammation involving mtDNA in the post-infarct brain.

One-step, reagent-free construction of nano-enzyme as visual and reusable biosensor for oxidase substrates

The substrates of oxidase are biologically essential substances that are closely associated with human physiological health. However, current biosensing methods suffer from tough recyclability and undesired denaturation of enzyme due to impurity interference. Herein, we have developed a visual and reusable biosensor for detecting substrate using glucose oxidase (GOx) as a model oxidase. GOx was immobilized onto gold nanoparticles (AuNPs) at −20 °C in one step without additional reagents. The resulting nano-enzyme generated coloimetric signals by coupling with horseradish peroxidase (HRP) using TMB as the substrate. Our results demonstrated that the immobilized GOx exhibited satisfactory sensitivity (0.68 μM) for glucose detection and higher inherent stability than free GOx under harsh conditions, enabling reliable detection of glucose in complex fluids (colored beverages and saliva). Furthermore, the nano-enzyme retained 80 % activity even after four cycles of catalytic oxidation. This strategy constructs a universal biosensor for substrates with nano-enzyme which rely only on intrinsic cysteine within the oxidase while avoiding functional handle modification.

A cascaded enzyme system based on the catalase-like activity of Ti3C2Tx MXene nanosheets for the efficient combination cancer therapy

Catalytic therapy with enzymes and nanozymes is a promising approach to cancer treatment. However, it often proves inadequate when used as a standalone treatment. This work presents the catalase (CAT)-like activity of Ti3C2Tx MXene, a kind of two-dimensional (2D) material with photothermal capability. Ti3C2Tx nanosheets were then used to load glucose oxidase (GOx) and the anticancer drug doxorubicin hydrochloride (DOX), and modified with polyethylene glycol (PEG) to obtain an enzyme cascade system, named Ti3C2Tx-GOx/DOX-PEG, for anticancer purposes. Ti3C2Tx-GOx/DOX-PEG exhibited a photothermal conversion efficiency of ∼28.7% at 808 nm, enabling its use for near-infrared light-based photothermal therapy (PTT). In addition, Ti3C2Tx could decompose H2O2 to O2 in cancer cells, thereby enhancing starvation therapy by sensitizing GOx. Furthermore, alleviation of hypoxia could sensitize the chemotherapy of DOX. The decomposition of glucose by GOx could also generate H2O2 for the production of O2 by Ti3C2Tx. Ultimately, the cycling of the enzyme cascade reaction could alleviate tumor hypoxia and enable enhanced chemo/starvation/photothermal combination therapy. The CAT-like behavior of Ti3C2Tx as well as various enzymatic reactions catalyzed by Ti3C2Tx-GOx/DOX-PEG were systematically investigated by intra- and extracellular experiments. Results from both in vitro and in vivo studies demonstrated the effective anticancer capability of Ti3C2Tx-GOx/DOX-PEG. This enzyme cascade system has the potential to enhance material utilization and reduce side effects, thus opening up new opportunities for designing efficient and safe anticancer systems.

A long lifetime and highly sensitive wearable microneedle sensor for the continuous real-time monitoring of glucose in interstitial fluid

The short lifetime and low sensitivity of the current glucose electrochemical sensors are two major issues for implementing continuous real-time monitoring of glucose in vivo. Here we show that a unique microneedle-based glucose monitoring skin patch (termed here MGMSP) can continuously measure glucose in real time in live animals with micromolar sensitivity and over 14 days of service life. This MGMSP employs a glucose oxidase (GOD) and carbon nanotube (CNT) modified hollow syringe as electrochemical sensor for glucose monitoring, an integrated circuit for signal processing and transmission, and the real-time glucose levels are displayed on smartphone via Bluetooth. The designed microneedle device protects the stability of the sensing molecules immobilized within the inner surface of hollow syringe and simultaneously the interior space of hollow syringe substantially increases the amount of immobilized sensing molecules. This microneedle design thus extends the lifetime as well as improves the detection sensitivity. The final MGMSP enables the continuous real-time monitoring of glucose in the interstitial fluid of live rats. This innovative microneedle-based MGMSP could potentially provide the public with high-accuracy continual glucose monitoring.

Edge-generated N-doped carbon-supported dual-metal active sites for enhancing electrochemical immunoassay

Development of high-precision human epidermal growth factor receptor 2 (HER2) assay is essential for the early diagnostic and prevention of breast cancer. In this work, an innovative Fe/Mn bimetallic nanozyme at the edge of N-doped carbon defects (FeMn-NCedge) with abundant active sites was prepared through the hydrothermal synthetic method. FeMn-NCedge nanozyme displayed excellent peroxidase-like activity relative to the H2O2-catalyzed 3,3′,5,5′-tetramethylbenzidine (TMB) system for generation of the oxidized TMB (oxTMB). As a proof-of-concept application, we constructed an electrochemical immunoassay for the detection of HER2 based on the unique merits of FeMn-NCedge. Initially, a sandwiched immunoreaction was carried out in the microtiter plate coated with monoclonal anti-HER2 capture antibodies using glucose oxidase (GOx)-labeled anti-HER2 as detection antibody. The carried GOx could catalyze glucose to produce H2O2, thus resulting in the formation of oxTMB with the assistance of TMB and FeMn-NCedge nanozyme. The produced oxTMB could be determined on the electrode by the chronoamperometry at an applied potential of +10 mV. Experimental results revealed that the steady-state current increased with the increasing HER2 concentration in the sample, and gave a good linear relationship within the dynamic range of 0.01–10 ng/mL at a limit of detection of 5.4 pg/mL HER2. In addition, good reproducibility, high specificity and acceptable accuracy were acquired for the measurement of human serum samples. Importantly, this method can be extended for quantitative monitoring other disease-related proteins by changing the corresponding antibodies.

Utilization of Interfacial Films of Organo‐Modified Carbon Nanotubes as Adsorption Templates for a Number of Biomolecules while Maintaining their Activity

AbstractWe investigated the versatility of a monolayer of organo‐modified single‐walled carbon nanotubes (SWCNTs) on a water surface as a template to maintain activity by adsorbing various biomolecules from the subphase. Organo‐modified SWCNTs with amphiphilic properties formed a Langmuir film on the subphase containing biomolecules, and adsorbed and immobilized biomolecules via electrostatic interactions. In this study, the interactions of trypsin, lysozyme, cytochrome C protein, glucose oxidase sugar chain polymer, deoxyribonucleic acid, and chitosan with SWCNTs at the air/water interface were investigated. The uniqueness of this study is that it shows the possibility of wide‐ranging applications by utilizing an organo‐modified SWCNT interfacial film as an adsorption template for various biomolecules. The expansion behavior of the surface pressure‐area isotherms indicated the existence of interactions between the organo‐modified SWCNTs monolayer on the water surface and the biomolecules. The morphology and spectroscopic properties of the films transferred onto solid substrates clearly indicate the adsorption and immobilization of biomolecular groups. DNA and chitosan were not immobilized on the organo‐modified SWCNTs film template; however, the other four enzyme/protein molecules were adsorbed onto the solid substrate and maintained their fine second‐order structures, even under harsh conditions.

DNA flexible chain modified MOFs as a versatile platform for chemoenzymatic cascade reactions in glucose catalysis

Glucose oxidase (GOD) is widely used in the pharmaceutical industry, fermentation products and glucose biosensors for its essential role in catalyzing the conversion of glucose to gluconic acid and hydrogen peroxide (H2O2). As H2O2 is the by-product and will have a toxic effect on glucose oxidase, so introducing another enzyme that could consume H2O2 to form an enzymatic cascade reaction is a practical solution. However, this decision will lead to extra expenses and complex condition optimization such as the specific mass ratio, temperature and pH to improve the activity, stability and recyclability. Herein, we describe a mild and versatile strategy by anchoring GOD on carboxyl-activated MOF (Cu-TCPP(Fe)) through DNA-directed immobilization (DDI) technology. Robust MOF nanosheets were utilized as not only the carrier for the immobilization of GOD, but also a peroxidase-like catalyst for the decomposition of H2O2 to reduce its harmful impacts. In this work, the immobilized GOD retained 55.78% of its initial activity after being used for 7 times. More than 60% of the immobilized enzyme's catalytic activity was still maintained after 96 h of being stored at 50 ℃. This study provides a new idea for preparing immobilized enzymes with enhanced stability, fast diffusion and high activity, which can be used in fields such as biocatalysis and biotechnology.