MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Yelan Yao,Tong Liu,Shanshan Zhang,Yue Zhou,Xu Xu,Yunbo Gai,Lu Fang,Yu Cai,Xuesong Ye,Bo Liang
Biocatalytic reactions within organisms are contingent on sophisticated regulatory mechanisms, the precise control and regulation of which are of paramount importance. Magneto-controlled enzyme reaction, as a powerful regulate tool, enables the activation or inhibition of catalytic processes through external magnetic field signals. These reactions are of particular interest for advanced bioelectronic devices with tunable biosensing capabilities and novel bioelectronic applications. Here in, we report an ON/OFF-switchable magneto-controlled electrochemical platform for precise regulation of horseradish peroxidase (HRP) enzymatic reaction immobilized on magnetic beads (MBs) and highly sensitive enzyme detection in both temporal and frequency domain. The ON/OFF switching characteristic ensures sensitive HRP detection with intrinsic noise rejection capability through time-domain signal decoupling, exhibiting a sensitivity of 0.223 uA·ng −1 ·mL −1 , a low detection limit (LOD) of 0.097 ng·mL −1 and a wide linear detection range of 0.1–200 ng·mL −1 . Meanwhile, the frequency-domain electrochemical analysis relies on an innovative periodic magnetic field modulation strategy, successfully isolates weak redox signals from background current by modulating the MBs-HRP periodically. The magnetically modulated redox cycling process generates periodic electrochemical oxidation and reduction signals for subsequent discrete Fourier transform analysis with enhanced detection performance. This specific approach broke the inherent limitations of electrochemical sensors, significantly decreased the LOD of electrochemical amperometric analysis, achieving an exceptional detection limit of 93.5 fg·mL −1 of HRP. This strategy is expected to provide a viable approach for the development of robust, simple, reversible, and highly selective magneto-controlled enzyme reaction systems, providing a universal platform for highly sensitive detection of ultra-low trace biomarkers.
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高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2020
·Vol. 21
·期 24
·9529
·
IF 4.556
Shanxia Jin,Liping Liu,Mengying Fan,Yaru Jia,Ping Zhou
Pollen grain was explored as a new carrier for enzyme immobilization. After being modified with boric acid-functionalized titania, the pollen grain was able to covalently immobilize glycosylated enzymes by boronate affinity interaction under very mild experimental conditions (e.g., pH 7.0, ambient temperature and free of organic solvent). The glucose oxidase and horse radish peroxidase-immobilized pollen grain became a bienzyme system. The pollen grain also worked as an indicator of the cascade reaction by changing its color. A rapid, simple and cost-effective approach for the visual detection of glucose was then developed. When the glucose concentration exceeded 0.5 mM, the color change was observable by the naked eye. The assay of glucose in body fluid samples exhibited its great potential for practical application.Keywords:enzyme immobilization;boronate affinity;pollen grain;glucose detection
研究领域:
生物化学
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分析化学
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生物技术
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高影响因子
RSC Advances
·
2014
·Vol. 4
·期 20
·10273-10279
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IF 3.708
Wei Xu,Su Liu,Jinghua Yu,Min Cui,Jie Li,Yuna Guo,Hongzhi Wang,Jiadong Huang
In this work, we explored an amplification strategy which was based on graphene oxide–polyaniline (GO–PANI) and horseradish peroxidase (HRP) to construct the competitive aptasensor for ultrasensitive detection of oxytetracycline (OTC). In the protocol, the GO–PANI film was immobilized on the surface of the electrodes. Then, gold nanoparticles (AuNPs) were electrodeposited on the electrode surface using a constant potential stripping technique. The selected aptamer which had high affinity and specificity for OTC was used as a capture probe and labeled with HRP. The linear response to OTC concentration of the developed aptasensor was in the range of 4.0 × 10−6 mg L−1 to 1.0 mg L−1. The detection limit (LOD) of 2.3 × 10−6 mg L−1 was obtained (S/N = 3). In addition to good repeatability and stability, the proposed aptamer sensor also showed the advantages of low background current and high sensitivity to examine OTC in real samples.
研究领域:
分析化学
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生物传感技术
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环境科学
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Materials Science & Engineering C-Materials for Biological Applications
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2017
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IF 4.164
Buguang Zhou,Min He,Ping Wang,Haitian Fu,Yuanyuan Yu,Qiang Wang,Xuerong Fan
Silk fibroin (SF) as a bioactive protein can offer growth substrates for hydroxyapatite (HAp) deposition. In the current work, graft copolymerization of acrylic acid (AA) onto fibroin chains was carried out using hydrogen peroxide-horseradish peroxidase (H 2 O 2 -HRP) catalytic system, SF-g-polyacrylic acid (PAA) membranes was prepared subsequently, followed by in situ biomimetic mineralization in the Ca/P solutions, aiming at promoting the deposition of HAp and endowing the fibroin-based biocomposite with enhanced bioactivity. Meanwhile, p -hydroxyphenylacetamide (PHAD) and methyl acrylate (MA), as the model compounds of tyrosine residues in SF and vinyl monomer were used to disclose the mechanism of graft copolymerization. The data from FTIR and SEC chromatograms indicated that vinyl monomer was successfully graft copolymerized with SF during H 2 O 2 -HRP treatment. According to the results of XRD, SEM patterns and EDS-Mapping, mineral phases on the surfaces of SF-g-PAA membranes were detected after different cycles of biomimetic mineralization, and the mechanical property of SF-g-PAA/HAp membrane was noticeably improved. Cell viability and adhesion assays revealed that the composite of SF-g-PAA/HAp exhibited acceptable biocompatibility and outstanding adhesion property. The present work provides a novel method for preparation of the fibroin-based biomaterial for bone tissue engineering.
研究领域:
材料科学
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生物材料
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组织工程
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生物医学工程
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高影响因子
ACS Applied Materials & Interfaces
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2019
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IF 8.456
Ding Wang,Yaqin Chai,Yali Yuan,Ruo Yuan
In this work, the array arrangement of cascade enzymes was implemented by alternately and equidistantly anchoring two model enzymes glucose oxidase (GOx) and horseradish peroxidase (HRP) to the vertexes of rigid DNA tetrahedron units in lattice-like nucleic acid scaffold, in which the distance between any adjacent cascade enzymes had been regulated to the optimum for obtaining high enzyme cascade catalytic efficiency. Compared to the enzyme cascade system with no-array arrangement of cascade enzymes, the proposed enzyme cascade system allowed the intermediate H2O2 produced by GOx catalyzing substrate glucose to concurrently and equidistantly diffuse toward the four adjacent HRP enzyme surfaces. In this case, the invalid diffusion effect of intermediate H2O2 between cascade enzymes could be effectively avoided, thereby promoting the enzyme cascade reaction with high catalytic efficiency. The specific catalytic efficiency (kcat/Km) of the cascade enzyme system with array arrangement had been evaluated, which exhibited catalytic efficiency about 3.6 times higher than that of the randomly arranged cascade enzyme system. As a result, this strategy provided a new avenue for constructing a highly efficient enzyme cascade system with ultimate applications in biosynthesis, bioanalysis, and biodiagnostics.
研究领域:
生物化学
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纳米生物学
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酶工程
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高影响因子
SENSORS
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2019
·Vol. 19
·期 3
·640
·
IF 3.031
Meng Li,Jing Wu,Haiping Su,Yan Tu,Yazhuo Shang,Yifan He,Honglai Liu
In this work, three different aqueous solutions containing imidazole-based ILs with different alkyl chain lengths ([Cnmim]Br,n= 2, 6, 12) were adopted as the medium for the synthesis of ionic liquid-polypyrrole (IL-PPy) composites. Herein, the ILs undertook the roles of the pyrrole solvent, the media for emulsion polymerization of PPy and PPy dopants, respectively. The electrochemical performances of the three IL-PPy composites on a glassy carbon electrode (GCE) were investigated by electrochemical experiments, which indicated that [C12mim]Br-PPy (C12-PPy) composites displayed better electrochemical performance due to their larger surface area and firmer immobilization on the GCE. Further, C12-PPy/GCE were decorated with Au microparticles by electrodeposition that can not only increase the conductivity, but also immobilize sufficient biomolecules on the electrode. Then, the obtained C12-PPy-Au/GCE with outstanding electrochemical performance was employed as a horseradish peroxidase (HRP) immobilization platform to fabricate a novel C12-PPy-Au-HRP/GCE biosensor for H2O2detection. The results showed that the prepared C12-PPy-Au-HRP/GCE biosensor exhibited high sensitivity, fast response, and a wide detection range as well as low detection limit towards H2O2. This work not only provides an outstanding biomolecule immobilization matrix for the fabrication of highly sensitive biosensors, but also advances the understanding of the roles of ILs in improving the electrochemical performance of biosensors.Keywords:ionic liquids (ILs);polypyrrole (PPy);gold particles;horseradish peroxidase (HRP);enzyme immobilization;hydrogen peroxide sensor
研究领域:
材料科学
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电化学
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生物传感器
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分析化学
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POLYMER JOURNAL
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2024
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IF 2.3
Wang Wenda,Wang Qiang,Wang Ping
Acrylic acid (AA)/methyl acrylate (MA)-modified starch was prepared via a catalytic process involving horseradish peroxidase (HRP)/acetylacetone (ACAC) as an initiation system. The structural analysis results demonstrated that the grafting of AA and MA onto starch was successful. Furthermore, the impact of the grafting reaction conditions on the grafting effect and the characteristics of the resulting grafted products were also examined. The results demonstrated that AA and MA were successfully grafted onto starch. The optimal conditions for starch grafting with AA/MA were identified as a reaction time of 4 h, an HRP enzyme concentration of 12 U/mL, a total monomer/starch ratio of 4:6, and a molar ratio of AA/MA feeding of 3:7. The viscosity and viscoelasticity of the modified starch slurry increased, and the film-forming property was enhanced. Compared with the original starch film, the modified film markedly increased the flexibility and tensile strength. Furthermore, an increased acrylic acid content maintained the original hydrophilicity of the modified starch film.
研究领域:
材料科学
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生物催化
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CHEMICAL ENGINEERING JOURNAL
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2025
·
IF 13.2
Xiaozhou Li,Bin Wu,Zhen Gao,Wei Zhuang
To address the challenges of quantitative immobilization and unavoidable spatial hindrance of multiple enzymes in traditional multi-enzyme cascade immobilization, we developed a novel bienzyme immobilization system based on a hierarchically structured MXene@LDHs@Fe₃O₄ (MLF) carrier. The MLF composite, with a high theoretical surface area (∼300 m 2 /g), adjustable interlayer spacing (10–50 nm), and strong magnetization (20 emu/g), provides an ideal support for high-capacity enzyme loading. The system was fabricated through the assembly of MXene nanosheets with layered double hydroxides (LDHs) and magnetic Fe₃O₄ nanoparticles, integrating horseradish peroxidase (HRP) on the MLF core, a polydopamine (PDA) interfacial barrier, and surface-immobilized glucose oxidase (GOx) through melamine-genipin mediated dendritic cross-linking. This spatially organized catalytic architecture enables quantitative enzyme loading while minimizing spatial hindrance, thereby enhancing catalytic efficiency. The co-immobilized bienzyme reactor (GOX@PDA@HRP@MLF) achieved loading capacities of 272.46 mg/g for HRP and 414.33 mg/g for GOx, with an activity recovery of 85.3 % during 2,4-dichlorophenol degradation. The immobilized enzymes retained ∼80 % of their initial activity after five operational cycles. Under optimized conditions, the GOX@PDA@HRP@MLF material achieved a degradation rate of 4.6 for 2,4-dichlorophenol in a continuous flow system, representing a ∼21-fold increase in space-time yield compared to random multi-enzyme immobilization on single-layer resin materials. These results demonstrate that the dendritic cross-linking strategy employed in this study can serve as a universal method for multi-enzyme immobilization, with significant potential for industrial applications.
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TALANTA
·
2025
·
IF 6.1
Yuanyuan Ding,Ping Wang,Na Wang,Liyun Kong,Min Li,Jing Wang,Tao Zhang
The immunoassay methods, stand as the gold standard in biomedicine, are widely used in diagnostics. However, they often encounter inherent limitations in practical applications, such as the poor stability of enzymes and antibodies, and the laborious process of preparing enzyme-antibody conjugates through covalent bonding. Mas-related G-protein-coupled receptor X2 (MRGPRX2) has been identified as a blood-based diagnostic biomarker for chronic spontaneous urticaria (CSU). In this study, a bifunctional enzyme-labeled antibody (anti-MRGPRX2&HRP@ZIF-8) was successfully constructed by integrating horseradish peroxidase (HRP) into ZIF-8 during its synthesis, followed by adsorption of anti-MRGPRX2 onto the surface of HRP@ZIF-8. The resulting anti-MRGPRX2&HRP@ZIF-8 complex not only preserved its antigen-capturing capability but also exhibited improved stability. The established chemiluminescence immunoassay (CLIA) method based on anti-MRGPRX2&HRP@ZIF-8 demonstrated enhanced stability and achieved a lower detection limit (1.035 ng/mL) compared to conventional enzyme-linked immunosorbent assay (ELISA) method. Furthermore, MRGPRX2 was successfully detected in serum samples by the established method, demonstrating its suitability for the diagnosis of CSU in real clinical samples. Consequently, this study presented a promising approach for simplifying the preparation of enzyme-antibody conjugates and highlighted the potential of metal-organic framework (MOF)-based composites in biomedical applications.
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TALANTA
·
2018
·
IF 4.244
Nuanfei Zhu,Yanmin Zou,Menglu Huang,Shuaibing Dong,Xiangyang Wu,Guoxi Liang,Zhixiang Han,Zhen Zhang
A sensitive and artful colorimetric immunosensor based on horseradish peroxidase (HRP) was designed by labelling metal–organic frameworks (Cu-MOFs) on the second antibody (Cu-MOFs@Ab 2 ) as signal amplification for the detection of trace dibutyl phthalate (DBP). In this system, when Cu-MOFs@Ab 2 was captured by antigen- primary antibody (Ab 1 ) complex, tremendous Cu(II) will be released from Cu-MOFs in the presence of nitric acid (HNO 3 ), and Cu(II) will be further reduced to Cu(I) after the addition of sodium ascorbate (SA), consequently, inhibiting the HRP to catalyse the colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxidized TMB (ox TMB). Under the optimized conditions, the limit of detection (LOD) was 1 μg L −1 , which was almost 60 times lower than that using a conventional ELISA with the same antibody. In addition, our method showed good accuracy and reproducibility (recoveries of 87.73–103.4%; CV values of 1.46–5.95%) through a spike-recovery analysis. The proposed immunosensor indicated great potential for trace DBP determination from environmental and food samples.
研究领域:
环境科学
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食品科学
·
分析化学
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BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Su Liu,Yu Wang,Wei Xu,Xueqi Leng,Hongzhi Wang,Yuna Guo,Jiadong Huang
In this paper, a novel sandwich-type electrochemical aptasensor has been fabricated and applied for sensitive and selective detection of antibiotic oxytetracycline (OTC). This sensor was based on graphene-three dimensional nanostructure gold nanocomposite (GR-3D Au) and aptamer-AuNPs-horseradish peroxidase (aptamer-AuNPs-HRP) nanoprobes as signal amplification . Firstly, GR-3D Au film was modified on glassy carbon electrode only by one-step electrochemical coreduction with graphite oxide (GO) and HAuCl 4 at cathodic potentials, which enhanced the electron transfer and loading capacity of biomolecules . Then the aptamer and HRP modified Au nanoparticles provide high affinity and ultrasensitive electrochemical probe with excellent specificity for OTC. Under the optimized conditions, the peak current was linearly proportional to the concentration of OTC in the range of 5×10 −10 −2×10 −3 g L −1 , with a detection limit of 4.98×10 −10 g L −1 . Additionally, this aptasensor had the advantages in high sensitivity, superb specificity and showed good recovery in synthetic samples. Hence, the developed sandwich-type electrochemical aptasensor might provide a useful and practical tool for OTC determination and related food safety analysis and clinical diagnosis.
研究领域:
分析化学
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食品安全
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生物技术
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电化学
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Journal of Materials Chemistry B
·
2017
·Vol. 5
·期 22
·4052-4059
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IF 4.543
Heping Wang,Fenglong Jiao,Fangyuan Gao,Junjie Huang,Yan Zhao,Yehua Shen,Yangjun Zhang,Xiaohong Qian
Protein glycosylations play important roles in various biological processes and in the disease progression of organisms. The development of specific enrichment materials and strategies before mass spectrometric analysis was a prerequisite to glycoproteomic analysis due to the difficulty caused by substoichiometric levels of glycoproteins. In this work, novel magnetic covalent organic frameworks (denoted as Fe3O4@TpPa-1) were first developed using only a two-step solvothermal reaction and then applied in the hydrophilic enrichment of glycopeptides. Sea urchin-type composites with super-paramagnetic properties were constructed by in situ growth of TpPa-1 covalent organic frameworks on the surface of magnetic nanoparticles. A total of 37 and 22 glycopeptides could be easily detected from IgG and HRP digests, respectively, by hydrophilic enrichment with the newly developed materials. An ultralow detection limit (28 fmol), satisfactory selectivity and high recovery could be achieved using Fe3O4@TpPa-1. The material's excellent enrichment performance was also demonstrated using glycopeptide analysis in real complex samples. Within three independent replicates, 228 glycopeptides corresponding to 114 glycoproteins could be detected from human serum digests, which is better than the performance obtained by commercial HILIC materials. The results suggest that covalent organic frameworks show potential for application in glycoproteomic studies.
研究领域:
分析化学
·
生物化学
·
材料科学
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