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Acta Biomaterialia · 2023 · IF 9.7

Glucose-responsive enzymatic biomimetic nanodots for H2O2 self-supplied catalytic photothermal/chemodynamic anticancer therapy

Yinghui Xu,Jiayi Bian,Xin Liu,Zhengzheng Qian,Minghao Sun,Cheng Zhang,Ruiyang Pan,Qitong Li,Changrui Sun,Bin Lin,Kun Peng,Nan Lu,Xikuang Yao,Wenpei Fan

Photothermal therapy (PTT) combined with chemodynamic therapy (CDT) presents an appealing complementary anti-tumor strategy, wherein PTT accelerates the production of reactive oxygen species (ROS) in CDT and CDT eliminates residual tumor tissues that survive from PTT treatment. However, nanomaterials utilized in PTT/CDT are limited by non-specific damage to the entire organism. Herein, a glucose-responsive enzymatic Fe@HRP-ABTS/GOx nanodot is judiciously designed for tumor-specific PTT/CDT via a simple and clean protein-templated biomimetic mineralization synthesis. By oxidizing glucose in tumor cells, glucose oxidase (GOx) activates glucose-responsive tumor therapy and increases the concentration of H 2 O 2 at the tumor site. More importantly, the self-supplied peroxide hydrogen (H 2 O 2 ) can convert ABTS (2,2′-Hydrazine-bis(3-ethylbenzothiazoline-6-sulfonic acid) diamine salt) into oxidized ABTS (oxABTS) through horseradish peroxidase (HRP) catalysis for PTT and photoacoustic (PA) imaging. Furthermore, the Fe 2+ arising from the reduction of Fe 3+ by overexpressed GSH reacts with H 2 O 2 to generate intensely reactive •OH through the Fenton reaction, concurrently depleting GSH and inducing efficient tumor CDT. The in vitro and in vivo experiments demonstrate superior cancer cell killing and tumor eradication effect of Fe@HRP-ABTS/GOx nanodot under near-infrared (NIR) laser irradiation. Collectively, the nanodots provide mutually reinforcing catalytic PTT/CDT anti-tumor strategies for treating liver cancer and potentially other malignancies. Statement of significance Combinatorial antitumor therapy with nanomedicines presents great prospects for development. However, the limitation of non-specific damage to normal tissues hinders its further clinical application. In this work, we fabricated tumor-selective biomimetic Fe@HRP-ABTS/GOx nanodots for H 2 O 2 self-supplied catalytic photothermal/chemodynamic therapy of tumors. The biomimetic synthesis strategy provides the nanodots with enzymatic activity in response to glucose to produce H 2 O 2 . The self-supplied H 2 O 2 initiates photothermal therapy with oxidized ABTS and enhances chemodynamic therapy through simultaneous •OH generation and GSH depletion. Our work provides a new paradigm for developing tumor-selective catalytic nanomedicines and will guide further clinical translation of the enzymatic biomimetic synthesis strategy.

研究领域: 纳米技术 · 癌症治疗 · 生物医学工程 · 材料科学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Analytical Methods · 2021 ·Vol. 13 ·期 9 ·1164-1171 · IF 2.896

An ultrasensitive colorimetric assay based on a multi-amplification strategy employing Pt/IrO2@SA@HRP nanoflowers for the detection of progesterone in saliva samples

Minglei Lu,Qiyi He,Yingying Zhong,Junkang Pan,Zhiting Lao,Mingxia Lin,Tiantian Wang,Xiping Cui,Jinlong Ding,Suqing Zhao

Progesterone (P4) belongs to a factor that affects stress response and is a potential carcinogen, and saliva levels are expected to be a standard measurement for clinical diagnosis. In this study, a new type of nanoflower with both recognition functionality and catalytic substrate ability was prepared by copper phosphate, Pt/IrO2 nanocomposites (Pt/IrO2 NPs), streptavidin (SA) and horseradish peroxidase (HRP) via a one-pot co-precipitation strategy. Due to the enhanced catalytic activity and stability of Pt/IrO2@SA@HRP nanoflowers, we developed a powerful and sensitive multiple-catalysis ELISA to monitor progesterone in saliva. Multiple-catalysis ELISA based on a specific antibody and Pt/IrO2@SA@HRP nanoflowers exhibited a linear interval range from 0.217 ng mL−1 to 7.934 ng mL−1. The median inhibitory concentration (IC50) for progesterone is 1.311 ng mL−1 and the limit of detection (LOD = IC10) is 0.076 ng mL−1 in the proposed method. Satisfactory recoveries were in a range of 79.6–107% with an acceptable coefficient of variation (below 10.6%). Results of the multiple-catalysis ELISA and LC-MS/MS had a good coincidence. Our result unraveled that multiple-catalysis ELISA is a potentially serviceable tool for the detection of progesterone in saliva.

CAS / MDL: 302-97-6
研究领域: 分析化学 · 生物化学 · 纳米技术 · 临床诊断
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Biosensors-Basel · 2024 ·Vol. 14 ·期 3 ·135 · IF 4.9

Simple, Visual, Point-of-Care SARS-CoV-2 Detection Incorporating Recombinase Polymerase Amplification and Target DNA–Protein Crosslinking Enhanced Chemiluminescence

Hui Chen,Zhiyuan Zhuang,Naihan Xu,Ying Feng,Kaixin Fang,Chunyan Tan,Ying Tan

The ongoing COVID-19 pandemic, driven by persistent SARS-CoV-2 transmission, threatens human health worldwide, underscoring the urgent need for an efficient, low-cost, rapid SARS-CoV-2 detection method. Herein, we developed a point-of-care SARS-CoV-2 detection method incorporating recombinase polymerase amplification (RPA) and DNA–protein crosslinking chemiluminescence (DPCL) (RPADPCL). RPADPCL involves the crosslinking of biotinylated double-stranded RPA DNA products with horseradish peroxidase (HRP)-labeled streptavidin (SA-HRP). Modified products are captured using SA-labeled magnetic beads, and then analyzed using a chemiluminescence detector and smartphone after the addition of a chemiluminescent substrate. Under optimal conditions, the RPADPCL limit of detection (LOD) was observed to be 6 copies (within the linear detection range of 1–300 copies) for a plasmid containing the SARS-CoV-2 N gene and 15 copies (within the linear range of 10–500 copies) for in vitro transcribed (IVT) SARS-CoV-2 RNA. The proposed method is convenient, specific, visually intuitive, easy to use, and does not require external excitation. The effective RPADPCL detection of SARS-CoV-2 in complex matrix systems was verified by testing simulated clinical samples containing 10% human saliva or a virus transfer medium (VTM) spiked with a plasmid containing a SARS-CoV-2 N gene sequence or SARS-CoV-2 IVT RNA. Consequently, this method has great potential for detecting targets in clinical samples.

CAS / MDL: 540-38-5 521-31-3
研究领域: 分子诊断 · 病毒学 · 生物医学工程 · 分析化学
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TALANTA · 2022 · IF 6.556

Comparative study of Pd@Pt nanozyme improved colorimetric N-ELISA for the paper-output portable detection of Staphylococcus aureus

Xin Wang,Min Zhang,Xiaoxu Pang,Kunlun Huang,Zhiyi Yao,Xiaohong Mei,Nan Cheng

Staphylococcus aureus is a frequent pathogenic bacterium that has a significant detrimental influence on the health of the human body. Therefore, developing a practical and portable detection platform is critical to ensuring food safety. Nanozymes are a kind of engineered nanomaterials with superior enzyme-like activities, providing infinite possibilities for the development of highly sensitive analytical assays. In this study, mesoporous core−shell [email protected] ( [email protected] ) nanozymes were synthesized and then applied as a signal amplifier in Staphylococcus aureus colorimetric immunoassay . At the same time, a careful comparative study of the catalytic performance with natural horseradish peroxidase (HRP), [email protected] nanozymes and their complexes [email protected] (HRP coupling with [email protected] nanozymes) were firstly performed, as well as clever using a colorimeter to achieve portable signal output. [email protected] bioprobes enable remarkable peroxidase-like catalytic activity, resulting in the highest sensitivity with a limit of detection (LOD) improved from 1 × 10 5 to 1 × 10 3  CFU/mL. In all, this work conducted a valuable comparative study and presented an improved strategy for the portable detection of pathogenic bacteria, which was expected to be used in a wide range of applications in food safety regulation and biomedicals arenas.

CAS / MDL: 50-81-7
研究领域: 分析化学 · 生物技术 · 食品安全 · 生物传感器 · 纳米技术
✓ 已验证 产品关联 DOI PubMed 高影响因子
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FOOD CHEMISTRY · 2024 · IF 8.5

CRISPR/HCR-powered ratiometric fluorescence aptasensor for ochratoxin A detection

Xiaokun Wang,Hao Ding,Yinghui Sun,Yanling Ma,Guoqing Wang,Jiadong Chen,Jaebum Choo,Lingxin Chen

To address the need for highly sensitive and reliable detection of trace ochratoxin A (OTA) in food matrices, we developed a ratiometric fluorescent aptasensor by integrating CRISPR/Cas12a, hybridization chain reaction (HCR), and horseradish peroxidase (HRP)-induced inner filter effect (IFE). The mechanism involves OTA releasing an activator that initiates CRISPR/Cas12a trans-cleavage, blocking HCR assembly. This reduces HRP levels, limiting the conversion of o -phenylenediamine (OPD) to fluorescent 2,3-diaminophenazine (DAP) (emitting at 562 nm) while maintaining strong emission from 2-amino terephthalic acid (BDC-NH 2 ) at 426 nm. The F 426 /F 562 ratio serves as a “signal-on” indicator, enabling sensitive OTA detection over 0.1 pM to 10 nM, with a detection limit of 0.0417 pM. The method exhibits excellent reproducibility, with intra-day and inter-day relative standard deviations (RSDs) of 1.91 %–3.87 % and 1.79 %, respectively, along with recovery rates of 90.1 %–110.6 % in real samples. These advantages highlight its significant potential for CRISPR/Cas-based OTA detection.

研究领域: 食品安全 · 分子生物学 · 分析化学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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LWT-FOOD SCIENCE AND TECHNOLOGY · 2021 · IF 4.952

Effects of dielectric barrier discharge cold plasma on the activity, structure and conformation of horseradish peroxidase (HRP) and on the activity of litchi peroxidase (POD)

Yijie Wang,Zichong Ye,Jiahui Li,Yan Zhang,Yingxi Guo,Jun-Hu Cheng

Litchi is one kind of the precious tropical fruits, but it is easily affected by enzymatic browning resulting in great loss to farmers. In the present study, dielectric barrier discharge cold plasma was used to inactivate the litchi peroxidase (POD), one of the enzymes responsible for browning. Results indicated that after 10 min treatment, the residual activity of the POD extracted from pericarp decreased to 47.16%. The effect of cold plasma on POD structure was studied by using horseradish peroxidase (HRP) as a model enzyme. The HRP activity was decreased by about 85% after 180 s of treatment. UV and fluorescence spectral results suggested that the secondary and tertiary structures of HRP were changed through plasma treatment, hydrophobic aromatic amino acids and hemes were exposed and modified. Circular dichroism (CD) spectra indicated that after exposed to plasma for 180 s, the content of α-helix in HRP were decreased from 21.41% to 8.53%, and the β-sheet content raised from 23.29% to 32.11%. The presented study concluded that cold plasma is capable of reducing the activity of litchi POD, which will provide a new thought for prolonging shelf life of easy-browning fruits.

研究领域: 食品科学 · 生物化学
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ANALYST · 2022 ·Vol. 147 ·期 14 ·3219-3224 · IF 5.227

Portable and sensitive detection of cancer cells via a handheld luminometer

Jing Lei,Lu Shi,Wei Liu,Baoxin Li,Yan Jin

Portable detection of cancer cells is important for early-stage diagnostic applications and prognosis of cancer. Herein, a simple and sensitive chemiluminescence method was proposed for portable detection of cancer cells via a handheld luminometer. It is based on the cancer cell triggered cyclic strand displacement reaction (SDR) for signal amplification. Cancer cells, CCRF-CEM, bind to their aptamer and release the trigger DNA (TDNA) of SDR. The TDNA initiates the cyclic SDR between magnetic bead modified hairpin DNA 1 (MB-H1) and HRP-tagged hairpin DNA 2 (HRP-H2), resulting in an enrichment of HRP onto the surface of magnetic beads by forming H1/H2 duplexes. HRP catalyzed luminol/H2O2/PIP solution to produce a strong chemiluminescence (CL) signal. CCRF-CEM cells were sensitively detected by combining magnetic enrichment with the signal amplification of SDR. The CL intensity showed an excellent linear relationship with the number of CCRF-CEM cells in the range of 100 to 5 × 104 cells per mL. The detection limit was as low as 85 cells per mL. Therefore, it offers a sensitive, cheap and portable method for the chemiluminescence detection of cancer cells and provides a new option for the early diagnosis of cancers.

研究领域: 生物医学工程 · 分析化学 · 癌症诊断 · 分子生物学
✓ 已验证 DOI PubMed 高影响因子
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FOOD CHEMISTRY · 2021 · IF 7.514

Development of a signal-enhanced LFIA based on tyramine-induced AuNPs aggregation for sensitive detection of danofloxacin

Huanhuan Bao,Meifang Yuan,Chengui Xiao,Daofeng Liu,Weihua Lai

A signal-enhanced LFIA based on tyramine (TYR)-induced AuNPs aggregation has been developed for the sensitive detection of danofloxacin (DAN). In the model, the hydroxyl radical produced by HRP catalyzing H 2 O 2 can trigger the TYR-AuNPs to aggregate on the T or C line for enhancing the detection signal. The linear range of TYR-AuNPs LFIA was 0.25–5 ng mL −1 with the limit of detection (LOD) of 0.032 ng mL −1 , and the LOD was 8-fold lower than that of the traditional AuNPs LFIA (0.26 ng mL −1 ). The TYR-AuNPs LFIA could be used with the naked eyes to qualitatively detect DAN with a cut-off limit of 2.5 ng mL −1 , which was 4-fold lower than that of the traditional AuNPs LFIA (10 ng mL −1 ). The recoveries of TYR-AuNPs LFIA were 86.04–105.14% and 92.41–110.19%, with the coefficient of variation of 1.71–2.05% and 4.42–5.89% in chicken and pork, respectively.

研究领域: 分析化学 · 纳米技术 · 食品安全
✓ 已验证 产品关联 DOI PubMed 高影响因子
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SENSORS AND ACTUATORS B-CHEMICAL · 2019 · IF 6.393

Dual-mode amplified detection of rabies virus oligonucleotide via Y-shaped DNA assembly

Jiaojiao Zhou,Wenjing Wang,Shuojun Li,Axiu Nie,Zhiyong Song,Mohamed F. Foda,Zhicheng Lu,Ling Zhao,Heyou Han

An innovative fluorescent and colorimetric dual-mode strategy based on Y-shaped DNA was proposed for the detection of rabies virus oligonucleotide. The cascade assembly of hairpin probes was initiated by the target, leading to the formation of Y-shaped DNA accompanied by freeing target to catalyze the next assembly process. The fluorescence recovery of the fluorophore FAM was accompanied by opening of hairpin probe. A colorimetric signal was generated by the proximity of glucose oxidase (GOx) and horseradish peroxidase (HRP), which was reflected by enhanced catalytic capability to TMB. Using this method, target DNA can be detected with a wide linear range from 0.1 to 6 nM with a detection limit of 15 pM, and from 0.5 to 60 nM with a detection limit of 60 pM for fluorescent and colorimetric readout, respectively. Moreover, the dual-mode detection strategy exhibited excellent selectivity towards the target compared with interference oligonucleotides. In addition, recoveries ranging from 96.0% to 106% with RSD values varying from 3.2% to 5.4%, and from 91.0% to 102% with RSD values varying from 3.3% to 7.1% for fluorescent and colorimetric studies, respectively, were obtained in diluted cerebrospinal fluid samples. The proposed dual-mode sensor is easy to operate with excellent detection performance. It would be superior to single mode detection and more practicable for quantitative detection of nucleic acid-related biomarkers due to the self-correction capability.

CAS / MDL: 9001-37-0 9003-99-0
研究领域: 生物化学 · 分子生物学 · 分析化学 · 病毒学
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SENSORS AND ACTUATORS B-CHEMICAL · 2013 · IF 3.535

Ultratrace detection of C-reactive protein by a piezoelectric immunosensor based on Fe3O4@SiO2 magnetic capture nanoprobes and HRP-antibody co-immobilized nano gold as signal tags

Jing Zhou,Ning Gan,Tianhua Li,Hankun Zhou,Xing Li,Yuting Cao,Lihong Wang,Weiguo Sang,Futao Hu

An ultra-sensitive multiplexed signal amplified piezoelectric immunosensor for the detection of C-reactive protein (CRP) in serum was developed. Firstly, a sandwich-type immunoreaction was made between the capture probe [silicon dioxide-coated magnetic Fe 3 O 4 nanoparticles (MNPs) labeled with the primary CRP antibody (MNPs-CRP Ab1)], CRP (different concentrations), and signal tag [horseradish peroxidase (HRP) coupled with the HRP linked secondary CRP antibody co-immobilized on gold nanoparticles (AuNPs-HRP/HRP-CRP Ab2)]. The immunocomplex was subsequently exposed to 3-amino-9-ethylcarbazole (AEC) and hydrogen peroxide. MNPs-CRP Ab1 capture probe could not only improve the CRP enrichment to amplify the signal through magnetic separation, but also be readily immobilized on the electrode by an external magnet to prepare the sensor. The signal tag could immobilize vast HRP, which could catalyze and yield more AEC's insoluble oxidation product on piezoelectric crystal surface, also resulting in the amplified change in frequency response (Δ f   =   f   −   f 0 ). Thus the multiplex signal amplification was achieved. The Δ f was proportional to CRP concentration from 0.001 to 100   ng   mL −1 with a low detection limit of 0.3   pg   mL −1 . The optimum conditions were studied in detail. The proposed method could provide a fast, simple, and sensitive way for the clinical analysis of ultratrace CRP in heart diseases.

研究领域: 生物医学工程 · 分析化学 · 临床检验
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FOOD CHEMISTRY · 2024 · IF 8.5

Antioxidant amyloid fibril derived from rice protein hydrolysate as stabilizer towards preparing high-stable emulsion

Xin Qi,Xinyu Lv,Wentao Pan,Mingyue Shen,Yi Chen,Qiang Yu,Jianhua Xie

An antioxidant amyloid fibril was prepared as an emulsifier by fibrillating limited enzymatic hydrolysis-modified rice protein (HRP). The purpose of this study was to investigate the feasibility of using fibrillated HRP to stabilize oil-in-water emulsion. A free radical scavenging assay revealed that the antioxidant activity of fibrillated HRP was 2.09 times higher than that of native rice protein. Fibrillated HRP demonstrated a marked reduction in interfacial tension, increased surface hydrophobicity and contact angle (> 80°), and rapid adsorption to the interface, with 35.34 ± 2.43% interfacial adsorbed protein content. The fibrillated HRP barriers resisted environment stresses such as NaCl, pH variations, long-term storage, while reducing lipid oxidation degree. Additionally, fibrillated HRP-based emulsion was more effective in protecting β-carotene from degradation compared to other samples. These findings provide theoretical support for the development of rice protein-based antioxidant emulsifiers and modification of emulsifying properties of plant proteins.

CAS / MDL: 7235-40-7
研究领域: 食品科学 · 胶体与表面化学 · 蛋白质工程
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Journal of Materials Chemistry B · 2020 ·Vol. 8 ·期 28 ·6139-6147 · IF 5.344

Efficient enzyme-activated therapy based on the different locations of protein and prodrug in nanoMOFs

Fan Wang,Jian Yang,Yongsheng Li,Qixin Zhuang,Jinlou Gu

Enzyme-activated prodrug therapy (EAPT) is an effective cancer treatment strategy able to transport non-toxic prodrugs and subsequently convert them into drugs at specific times and locations. However, due to the limitation of easy biodegradability and the membrane-impermeable characteristic of exogenous enzymes, there is a need to exploit suitable carriers for the effective protection and simultaneous delivery of activating enzymes into cancer cells. Herein, hierarchically porous MOFs were employed for the loading of enzyme and prodrug in a single nanocarrier thanks to their different cavity sizes. The simple loading process allows entrapping of horseradish peroxidase (HRP) and a monocarboxyl-containing indole-3-acetic acid (IAA) prodrug with high loading capacities in different spaces, which keeps the catalytic activity of the enzyme perfectly intact and avoids the premature activation of the prodrug. The encapsulated HRP and IAA exhibit sustained and synchronized release behaviors. Compared to the native HRP enzyme, the current MOF nanocarriers not only facilitate enzyme delivery into cellular lysosomes and subsequent endosomal escape, but also effectively release enzyme and prodrug in the intracellular environment within 48 h. Eventually, HRP and IAA loaded MOF nanocarriers cause significant cell death with a low IC50 of 4.2 mg L−1, while the IAA prodrug alone is non-toxic even at high concentrations. Thus, hierarchically porous MOFs might offer a promising platform for EAPT with a highly consistent spatiotemporal distribution of enzymes and prodrugs in target tissues.

研究领域: 药学 · 纳米技术 · 癌症治疗 · 材料科学
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