LAB ON A CHIP
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2024
·Vol. 24
·期 8
·2272-2279
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IF 6.1
Huilan Hu,Jiaqi Tian,Rui Shu,Huihui Liu,Shaochi Wang,Xuechi Yin,Jianlong Wang,Daohong Zhang
A highly sensitive lateral flow immunoassay (LFIA) is developed for the enzyme-catalyzed and double-reading determination of clenbuterol (CLE), in which a new type of probe was adopted through the direct electrostatic adsorption of ultra-small copper–gold bimetallic enzyme mimics (USCGs) and monoclonal antibodies. In the assay, based on the peroxidase activity of USCG, the chromogenic substrate TMB–H2O2 was introduced to trigger its color development, and the results were compared with those before catalysis. The detection sensitivity after catalysis is 0.03 ng mL−1 under optimal circumstances, which is 6-fold better than that of the traditional Au NPs-based LFIA and 2-fold greater than that before catalysis. This approach was successfully applied to the detection of CLE in milk, pork and mutton samples with an optimum assay time of 7 min and best catalytic time of 80 s, after which satisfactory recoveries of 98.53–117.79% were obtained. Cu–Au nanoparticles as a signal tag and the use of their nanozyme properties are the first applications in the field of LFIA. This work can be a promising exhibition for the application of a cheaper substitute for HRP, ultra-small bimetallic enzyme mimics, in LFIAs.
研究领域:
生物化学
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分析化学
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食品科学
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PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 2
·1-9
·
IF 5.833
Li Wenjun,Chen Siyu,Yang Yue,Song Yiju,Ma Chaoyun,Qiao Xiuwen,Hong Chenglin
A GOx/HRP@ZIF-90 nanomaterial is proposed by coating GOx and HRP in ZIF-90 using a bio-simulated mineralization method to improve the tolerance of the enzyme to the external environment. In the detection process, the ZIF-90 is turned on under mild conditions by the competitive reaction of ATP with Zn 2+ and imidazole-2-carboxaldehyde (2-ICA), and the electrical signal of the system is amplified by the enzyme cascade reaction of GOx and HRP. Finally, based on the signal amplification strategy of the competitive reaction between Zn 2+ and ATP to construct a “signal on” mode, electrochemical immunosensor of GOx-HRP enzyme–linked cascade reaction was prepared. The proposed electrochemical immunosensor shows an excellent analytical performance when detecting CA-125, with good selectivity and stability, with a detection range of 0.1 pg mL −1 –40 ng mL −1 and a detection limit of 0.05 pg mL −1 . The test has been performed using chronoamperometry under a constant voltage of −0.4 V. The immunosensor also shows an excellent performance when analysing human blood samples. The recovery of the immunosensor is 97.94–101.8%, with a relative standard deviation of 3.7–6.1%. The proposed sensor provides a novel idea for clinical use of GOx and HRP enzymes and a new method for the clinical detection of tumor markers. Graphical abstract
研究领域:
生物医学工程
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分析化学
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肿瘤标志物检测
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PubMed
高影响因子
ANALYST
·
2020
·Vol. 145
·期 18
·6171-6179
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IF 3.978
Yingqi Pei,Jing Zhang,Kang Wu,Anping Deng,Jianguo Li
A rapid and highly sensitive flow injection chemiluminescence immunoassay was developed to detect ractopamine residues in pork products. Palladium nanocubes with excellent catalytic performance for a traditional luminol–PIP–H2O2 chemiluminescence system were used as effective carriers to connect a ractopamine antibody and horseradish peroxidase (HRP). Double amplification of a chemiluminescence signal was realized because of palladium nanocubes and HRP. Carboxyl-modified resin beads were used as suitable materials to load ractopamine-coating antigens due to their good biocompatibility and large specific surface area. Based on the principle of competitive immunity, ractopamine standard solution would compete with antigen loaded on carboxyl resin beads for limited binding sites on a ractopamine antibody. Thus, the chemiluminescence intensity of an immunosensor has a linear negative correlation with the logarithm value of ractopamine concentration. Under optimal experimental conditions, the detection range of ractopamine was 0.005–1000 ng mL−1, and the limit of detection (LOD) was 1.7 pg mL−1 (S/N = 3). The proposed immunoassay possessed acceptable accuracy, high specificity and reproducibility and RAC was examined in pork and pig feed with satisfactory results, which would provide a better prospect for the detection of small molecules in food and environment analysis.
研究领域:
分析化学
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食品科学
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环境分析
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PubMed
高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
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2015
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IF 2.128
Ruirui Liu,Aixue Dong,Xuerong Fan,Qiang Wang,Yuanyuan Yu,Artur Cavaco-Paulo
The horseradish peroxidase (HRP)/hydrogen peroxide system can catalyze the free-radical polymerization of aromatic compounds such as lignins. In this study, to improve the hydrophilicity of lignin-rich jute fabrics, acrylamide (AM) was grafted onto their surface by HRP-mediated oxidative polymerization. The resulting AM-lignin graft polymer in jute fibers, from which the lignin was removed by extracting with dioxane, was characterized by gel permeation chromatography, elemental analysis, matrix-assisted laser desorption/ionization mass spectrometry, nuclear magnetic resonance, scanning electron microscopy, and Fourier transform infrared spectroscopy. The results show that AM was grafted to the jute fiber by HRP . Furthermore, the hydrophilicity of the fabrics was determined in terms of wetting time, balance moisture regain, and static contact angle. Compared to the control sample, the wetting time, balance moisture regain, and static contact angle of the grated fabrics varied from 11.62 min to 7.45 min, from 9.78% to 10.40%, and from 122.30° to 92.84°, respectively, indicating that the hydrophilicity of the raw jute fabric improved. The modification conditions of the HRP-catalyzed AM-grafting reaction were optimized in terms of the nitrogen content of the jute fabrics.
研究领域:
材料科学
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生物化学
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高影响因子
NEW JOURNAL OF CHEMISTRY
·
2019
·Vol. 44
·期 1
·194-199
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IF 3.069
Wenda Wang,Xuerong Fan,Qiang Wang,Jin Xu,Lili Wang,Lipeng Liu,Xueming Bao
In this communication, acrylamide (AM) polymerisation to poly(acrylamide) was initiated by horseradish peroxidase (HRP) and acetylacetone (ACAC) without hydrogen peroxide (H2O2). Electron spin resonance (ESR) spectroscopy showed that free radicals of ACAC and AM were present in the reaction. Gel Permeation Chromatography (GPC), FT-IR, 1H-NMR and relative viscosity indicated that the product was PAM. Furthermore, the enzymatic cycling mechanism of the HRP–ACAC–H2O2 ternary system was also explained.
研究领域:
生物化学
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高分子化学
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高影响因子
MICROCHEMICAL JOURNAL
·
2026
·
IF 5.2
Minghui Wang,Qiaolin Guo,Zhenyong Hu,Yue He,Lai Zhang,Cuifei Chen
An ultrasensitive biosensor has been developed for miR-375 detection. • HRP is embedded into COFs to form stable HRP@COFs co-precipitated nanospheres. • The HRP@COFs preserved enzymatic activity and enhanced catalytic efficiency. • The biosensor has been successfully validated using clinical samples. The dysregulated expression of microRNA (miRNA) in various cancers has attracted considerable attention owing to their crucial roles in tumor initiation and progression. Among them, miR-375 has been recognized as a prospective biomarker for early malignancy detection and therapeutic monitoring. In this study, a highly sensitive electrochemical biosensor has been developed based on horseradish peroxidase (HRP)-embedded covalent organic framework (HRP@COFs) co-precipitated nanospheres for the detection of miR-375. The HRP@COFs nanocomposite effectively preserves the enzymatic activity of HRP while providing the structural stability and high loading capacity of COFs, thereby enhancing catalytic efficiency and signal amplification. In the presence of target miR-375, the azide-modified H1 hairpin probes on the HRP@COFs surface are activated, leading to the formation of a rigid H1-H2 duplex structure through competitive hybridization with H2 hairpins. The resulting H2-H1-HRP@COFs complexes are subsequently anchored onto the electrode surface via click chemistry. Upon the addition of H 2 O 2 , HRP catalyzes aniline polymerization and deposition, resulting in a significant electrochemical response. This biosensor achieves ultrasensitive detection of miR-375 with a detection limit as low as 230 aM. It has been successfully validated using clinical samples, demonstrating great potential for reliable breast cancer diagnosis. This work highlights the synergistic advantages of integrating HRP with COFs, providing a robust and efficient platform for microRNA detection and expanding the applications of COFs-based materials in biosensing and clinical diagnostics. Download: Download high-res image (151KB) Download: Download full-size image
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高影响因子
Journal of Environmental Chemical Engineering
·
2023
·
IF 7.7
Siao Li,Weisong Yan,Changyi Yang,Yanshu Zhu,Ying Liu,Yaohua Gu,Zhiqiang Wu,Keren Shi,Huiqin Yao
Acridine , as a nitrogen heterocyclic pollutant, poses a huge ecological and human health hazard, and its efficient degradation is one of the top tasks. A novel photo-enzyme-coupled catalytic system was constructed by coupling horseradish peroxidase (HRP) with graphite carbon nitride (g-C 3 N 4 ) through a simple inorganic hybridization process. The obtained g-C 3 N 4 /HRP nanoflowers not only increase the enzyme loading capacity and enzyme activity , but also promote the efficient separation of carriers. More importantly, the H 2 O 2 generated by g-C 3 N 4 directly participates in the redox cycle reaction of HRP, inducing photo-enzyme synergistic catalysis. Therefore, compared to HRP nanoflowers and g-C 3 N 4 nanoflowers, g-C 3 N 4 /HRP nanoflowers exhibited higher catalytic efficiency for the degradation of acridine. Thus, the photo-enzyme coupled catalyst proposed in this study provides a promising platform for the degradation of acridine pollutants by combining enzymes with photocatalysts through inorganic hybridization.
研究领域:
环境科学
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生物化学
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高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
·
2014
·
IF 2.745
Teng Zhang,Xiao-Ling Xu,Yi-Ning Jin,Jian Wu,Zhi-Kang Xu
Carbon nanotubes are widely applied in electronics, which offers great potential for enhancing the activity of redox enzyme. In this work, polyimide (PI) was blended with multiwalled carbon nanotubes (MWCNTs) and then electrospun into nanofibers for the immobilization of horseradish peroxidase (HRP), a redox enzyme. SEM and TEM were used to characterize the surface morphology of the PI/MWCNTs nanofibers and to analyze the protruding parts induced by the blending of MWCNT. The results indicate that, compared to the virgin PI nanofibers, the blending of MWCNTs increases the retention activity of immobilized HRP from 2.38% to 12.50%. Circular dichroism and UV–vis molecular absorption spectrometry were used to explore the interactions between HRP and MWCNTs . It seems that MWCNTs increase the enzyme activity by the facilitation of electron transfer among the catalytic intermediates “HRP I”, “HRP II” and MWCNTs.
研究领域:
生物化学
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纳米生物技术
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酶工程
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生物材料
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分析化学
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2024
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IF 5.7
Wenrui Hu,Hang Ao,Ziyi Lv,Wencheng Xiao,Wei Li,Jianping Lei,Jie Wu,Huangxian Ju
Background Chemiluminescence (CL) bioassay is one of the most advanced and used detection method in clinical diagnosis and biomedical research because of the advantages of low background, easy operation, and wide-field imaging without a light source or microscope. The luminol/hydrogen peroxide/horseradish peroxidase (luminol/H 2 O 2 /HRP) system is the most popular CL system, but its application in high-throughput imaging detection is challenged due to its low luminescence efficiency and flash-type emission which is difficult in ensuring the reproducibility and consistency of detection results. Results We reported a glow-type CL system of luminol@CD/H 2 O 2 /HRP by using a supramolecular enhancer of cyclodextrin (CD). This luminol@CD/H 2 O 2 /HRP system exhibited a luminescence lifetime of 41 min for sensitive and accurate imaging analysis. The long-lasting CL emission was attributed to the formation of a 1:1 host–guest complex between luminol and CD, which could stabilize the emitter and effectively reduce nonradiative relaxation. The formation of luminol@CD complex was determined through NMR experiments and theoretical analysis. Under optimum conditions, the luminol@CD/H 2 O 2 /HRP system showed higher sensitivity and much better precision than classical luminol/H 2 O 2 /HRP system for imaging detection of HRP. Especially, this glow-type luminol@CD/H 2 O 2 /HRP system realized CL imaging of microwell arrays on microfluidic chips. In addition, the luminol@CD/H 2 O 2 /HRP system was successfully applied for point-of-care detection of 17 β- estradiol based on a competitive mechanism of host–guest recognition. Significance An efficient CL system is crucial for obtaining reproducible and consistent results for accurate detection. Our luminol@CD/H 2 O 2 /HRP system emitted strong and persistent luminescence, resulting in reliability and efficiency at both CL macroscopic and microscopic imaging detection. We expected the luminol@CD/H 2 O 2 /HRP CL system to be applied in various detection fields.
研究领域:
分析化学
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生物医学工程
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临床诊断
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2019
·
IF 4.784
Lei Zhang,Jiali Mi,Guannan Hu,Chen Zhang,Haishan Qi
In this study, we developed a smart microfluidic device to prepare biocatalyst HRP-pCBMA. HRP-pCBMA is composed of horseradish peroxidase (HRP) and zwitterionic polymers poly(carboxybetaine methacrylate) (pCBMA), and could be applied to biodegrade bisphenol A (BPA) efficiently. Compared to free HRP , HRP-pCBMA exhibited an obviously enhanced degrading capability for 1 mM BPA with 99.42% degradation efficiency within 20 min, even being superior to 20-fold amount of free HPR. Besides, HRP-pCBMA displayed high stability against the abrupt changes of environmental factors (temperature, pH and organic solvents), and HRP-pCBMA exhibited a relatively high BPA degradation rate of more than 90% even after 10 cycles. The K cat and V max values of HRP-pCBMA were both 7-fold higher than that of free HRP, indicating significant improvement of the catalytic activity. Furthermore, the cytotoxicity assay indicated HRP-pCBMA has excellent biocompatibility. These results demonstrated that HRP-pCBMA possessed great potential in the bioremediation of BPA.
研究领域:
环境科学
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生物化学
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材料科学
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PubMed
高影响因子
TALANTA
·
2026
·
IF 6.7
Xinyi Wang,Minghao Wang,Wenbin Zhang,Dewang Yan,Liubing Kong,Gaofeng Wang
Accurate cortisol monitoring is essential for stress assessment and health management, yet reliable quantification remains challenging because signal fluctuations and matrix effects can compromise the performance of conventional single-mode immunosensors. Herein, we developed a fluorescence-colorimetric (FL-CL) dual-modal immunosensor based on a CuMOF-PEI@AuNPs@HRP (CPA) immunoreceptor for cortisol determination. The integrated sensing architecture generates correlated fluorescence and colorimetric responses from the same recognition event, which are combined through a normalized dual-signal output to improve calibration reliability. Under optimized conditions, the proposed sensor exhibited a detection limit of 0.22 ng/mL and a linear range of 3.1-200 ng/mL, covering physiologically relevant cortisol concentrations in saliva. Compared with single-modal assays and commercial kits, the proposed platform demonstrates enhanced sensitivity, improved correlation, good stability, and batch reproducibility. By integrating signal amplification, dual-modal readout, and simplified immunoreceptor fabrication within a single sensing platform, this work provides a reliable strategy for cortisol monitoring and offers potential for extension to other biomarker detection systems.
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高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 6
·1-9
·
IF 6.232
Wang Huan,Wang Jie,Wang Yuetong,Liu Yuqin,Liu Rui,Wang Xuelin,Tan Hui,Wang Tianfu,Kong Tiantian
A biomimetic antibody is described for colorimetric determination of glycoprotein, and horseradish peroxidase (HRP) is used as model analyte. Use is made of oriented surface imprinted inverse opal hydrogel particles functionalized with phenylboronic acid. The inverse opal hydrogel particles were negatively replicated from silica colloidal crystal beads (SCCBs), so that they were endowed with larger specific surface area than the bulk structure. Benefit from that, there were abundant surface molecularly imprinting sites in the hydrogel particles. Because the imprinting sites match the structure of the template molecules, they can recognize HRP with high selectivity and sensitivity. The recognized glycoprotein was bonded with the phenylboronic acid within the sites. The bonded HRP was determined by colorimetry of 3, 3′, 5, 5′-tetramethylbenzidine (TMB) single-component solution at 450 nm, and it shows a 16.03 imprinting factor under optimized conditions. Alpha-fetoprotein (AFP) was also investigated and demostrated the value of this strategy in practical applications. The results show that the absorbance increases linearly in the 1–50 ng mL −1 AFP concentration range and has a 1.32 ng mL −1 detection limit. The assay of human serum was realized by the standard addition method. This strategy is promising to open new horizons for glycoprotein assay.
Schematic representation of the preparation of oriented boronate affinity–imprinted inverse opal hydrogel particles for glycoprotein assay.
研究领域:
生物化学
·
分析化学
·
材料科学
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PubMed
高影响因子
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