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MICROCHIMICA ACTA · 2024 ·Vol. 191 ·期 11 ·1-9 · IF 5.3

MOF-mediated cascade reaction system for ultrasensitive detection of alkaline phosphatase activity and organophosphorus pesticides

Zhang Shengyuan,Yang Ying,Yang Dezhi,Yang Yaling

A metal–organic-framework (MOF) fluorescent sensor is reported based on NH 2 -MIL-101(Fe) propelled pesticide and alkaline phosphatase (ALP) catalytic reaction. Different from previous reports, a cascade reaction system combined with MOF structural changes to generate fluorescence was employed. The rationale is that ALP can hydrolyze L-ascorbic acid 2-phosphate (AAP) into L-ascorbic acid (AA), which can reduce Fe 3+ to decompose structurally NH 2 -MIL-101(Fe), resulting in 2-aminoterephthalic acid (NH 2 -BDC) with intense fluorescence. The  fluorescence can be decreased to different degrees due to inhibition of organophosphate pesticides (OPPs) on the activity of ALP. By taking chlorpyrifos (CPY) as the model compound of an OPP pesticide and adding ALP and CPY into the NH 2 -MIL-101(Fe) framework, the resulting cascade reaction fluorescence sensors exhibit a good sensitivity for CPY and ALP sensing. The working ranges are  0.02–2 μg/L and 0.2–20 mU/mL with detection limits (LOD) of 5.31 ng/L and 0.05 mU/mL, respectively. The proposed sensor has been actually applied to satisfactory detection of CPY and ALP in food and serum samples. This fluorescence-based assay may extend the application of MOF-based biosensors. Graphical Abstract

研究领域: 分析化学 · 环境科学 · 生物化学 · 材料科学
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Journal of Huazhong University of Science and Technology-Medical Sciences · 2012 ·Vol. 32 ·期 4 ·563-570 · IF 0.385

Synthesis and characterization of UPPE-PLGA-rhBMP2 scaffolds for bone regeneration

Tian Zhichao,Zhu Yuanli,Qiu Jinjun,Guan Hanfeng,Li Liangyu,Zheng Shouchao,Dong Xuehai,Xiao Jun

A novel unsaturated polyphosphoester (UPPE) was devised in our previous research, which is a kind of promising scaffold for improving bone regeneration. However, the polymerization process of UPPE scaffolds was unfavorable, which may adversely affect the bioactivity of osteoinductive molecules added if necessary, such as recombinant human bone morphogenetic protein-2 (rhBMP2). The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2 (UPB) and to investigate the bioactivity of rhBMP2 in this scaffold. Furthermore, the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro . A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres, and then the microspheres were added to UPPE for synthesizing UPB scaffold. The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy. The cumulative release of UPB scaffolds was detected by using ELISA. The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells (bMSCs) seeded on the surface of UPB scaffolds. The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates (ALP) activity in bMSCs seeded. The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2. The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h. bMSCs attached and grew on the surface of soaked UPB scaffolds, exerting well biocompatibility. The ALP activity of bMSCs seeded was significantly enhanced, indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds. It was concluded that UPB scaffolds have low cytotoxicity, good biocompatibility and preserve bioactivity of rhBMP2. UPB scaffolds are promising in improving bone regeneration.

研究领域: 材料科学 · 生物医学工程 · 组织工程
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ANALYTICA CHIMICA ACTA · 2024 · IF 5.7

Pyrroloquinoline quinone exhibiting peroxidase-mimicking property for colorimetric assays

Ning Xia,Fengli Gao,Gang Liu,Yong Chang,Lin Liu

Background Small molecule mimics offer the advantages of easy preparation, good thermodynamic stability, and reproducible catalytic activity. However, most of the reported organic artificial mimics face challenges including low catalytic activity, oxidative self-destruction, and auto-aggregation into inactive dimers. Therefore, novel organic mimics with high catalytic activity as well as good thermal and environmental stability are highly desirable. Results We found that pyrroloquinoline quinone (PQQ), a nutritionally important growth factor, displayed significant peroxidase-like activity to catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxidized TMB (oxTMB) by H 2 O 2 . Its catalytic activity surpassed that of other organic small molecules reported previously, including fluorescein derivatives and adenosine/guanosine triphosphate. Based on the reduction of oxTMB by the enzymatically generated ascorbic acid (AA), the PQQ/TMB-based system was used to monitor alkaline phosphatase (ALP) activity with ascorbic acid 2−phosphate (AAP) as the substrate. This sensing system allowed for the determination of ALP with a detection limit of 1 mU/mL. Furthermore, we discovered that PQQ can coordinate with Cu 2+ to form metal-organic nanoparticles. The building blocks of two active cofactors endowed the nanoparticles with impressive functionalities for biocatalytic applications. With Cu-PQQ nanoparticles as signal labels, colorimetric immunoassays of prostate specific antigen (PSA) were achieved with a lowest detectable concentration down to 0.1 pg/mL. Significance This work is valuable for the design of novel biosensors by utilizing PQQ as an artificial enzyme because of its high catalytic activity, good stability, ease of storage, and simple chemical structure.

研究领域: 生物化学 · 分析化学 · 纳米技术 · 生物传感器
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LAB ON A CHIP · 2016 ·Vol. 17 ·期 1 ·120-127 · IF 5.586

Microfluidic PDMS on paper (POP) devices

Jin-Wen Shangguan,Yu Liu,Jian-Bin Pan,Bi-Yi Xu,Jing-Juan Xu,Hong-Yuan Chen

In this paper, we propose a generalized concept of microfluidic polydimethylsiloxane (PDMS) on paper (POP) devices, which combines well the merits of paper chips and PDMS chips. First, we optimized the conditions for accurate PDMS spatial patterning on paper, based on screen printing and a high temperature enabled superfast curing technique, which enables PDMS patterning to an accuracy of tens of microns in less than ten seconds. This, in turn, makes it available for seamless, reversible and reliable integration of the resulting paper layer with other PDMS channel structures. The integrated POP devices allow for both porous paper and smooth channels to be spatially defined on the devices, greatly extending the flexibility for designers to be able to construct powerful functional structures. To demonstrate the versatility of this design, a prototype POP device for the colorimetric analysis of liver function markers, serum protein, alkaline phosphatase (ALP) and aspartate aminotransferase (AST), was constructed. On this POP device, quantitative sample loading, mixing and multiplex analysis have all been realized.

CAS / MDL: 328-50-7
研究领域: 微流控技术 · 生物医学工程 · 分析化学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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MICROCHIMICA ACTA · 2024 ·Vol. 191 ·期 7 ·1-8 · IF 5.3

Atomically Co-dispersed nitrogen-doped carbon for sensitive electrochemical immunoassay of breast cancer biomarker CA15-3

Han Lei,Cai Shuanglong,Chen Xiaogeng

The development of an ultrasensitive and precise measurement of a breast cancer biomarker (cancer antigen 15-3; CA15-3) in complex human serum is essential for the early diagnosis of cancer in groups of healthy populations and the treatment of patients. However, currently available testing technologies suffer from insufficient sensitivity toward CA15-3, which severely limits early large-scale screening of breast cancer patients. We report a versatile electrochemical immunoassay method based on atomically cobalt-dispersed nitrogen-doped carbon (Co-NC)–modified disposable screen-printed carbon electrode (SPCE) with alkaline phosphatase (ALP) and its metabolite, ascorbic acid 2-phosphate (AAP), as the electrochemical labeling and redox signaling unit for sensitive detection of low-abundance CA15-3. During electrochemical detection by differential pulse voltammetry (DPV), it was found that the Co-NC-SPCE electrode did not have a current signal response to the AAP substrate; however, it had an extremely favorable response current to ascorbic acid (AA). Based on the above principle, the target CA15-3-triggered immunoassay enriched ALP-catalyzed AAP produces a large amount of AA, resulting in a significant change in the system current signal, thereby realizing the highly sensitive detection of CA15-3. Under the optimal AAP substrate concentration and ALP catalysis time, the Co-NC-SPCE-based electrochemical immunoassay demonstrated a good DPV current for CA15-3 in the assay interval of 1.0 mU/mL to 10,000 mU/mL, with a calculated limit of detection of 0.38 mU/mL. Since Co-NC-SPCE has an excellent DPV current response to AA and employs split-type scheme, the constructed electrochemical immunoassay has the merits of high preciseness and anti-interference, and its clinical diagnostic results are comparable to those of commercial kits. Graphical abstract

研究领域: 分析化学 · 生物化学 · 电化学 · 癌症诊断
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BIOMEDICINE & PHARMACOTHERAPY · 2024 · IF 6.9

The bone-protective benefits of kaempferol combined with metformin by regulation of osteogenesis-angiogenesis coupling in OVX rats

Zhongyuan Zhang,Wenshu Xu,Zhenhua Zhang,Xiaoxue Chen,Hui Jin,Ningning Jiang,Hui Xu

This study was to investigate the potential mechanisms of treatment with metformin (Met) combined with kaempferol (Kae) against postmenopausal osteoporosis. Experiments were conducted in both ovariectomy (OVX)-induced osteoporosis rats and in vitro using RAW264.7 cells, MC3T3-E1 cells, and HUVECs. Results demonstrated the therapeutic effect of Met combined with Kae on osteoporosis. In vivo, Kae alone and in combination with Met treatments enhanced tibial trabecular microstructure, bone mineral density (BMD), and mechanical properties in OVX rats without causing hepatotoxicity and nephrotoxicity. It also reduced bone resorption markers (CTX-1 and TRAP) and increased the bone formation marker (PINP) level in the serum of OVX rats. The expression of bone resorption marker TRAP was reduced, while bone formation markers Runx2 and ALP were enhanced in the bone tissue of OVX rats. Furthermore, Met combined with Kae also promoted the expression of angiogenesis-related markers CD31 and VEGF in OVX rats. In vitro, MC3T3-E1s cells treated with Met combined with Kae showed higher expression of ALP, Runx2, and VEGF. Interestingly, the treatment did not directly promote HUVECs migration and angiogenesis, but enhanced osteoblast-mediated angiogenesis by upregulating VEGF levels. Additionally, Met combined with Kae treatment promoted VEGF secretion in MC3T3-E1, and activated the Notch intracelluar pathway by upregulating HES1 and HEY1 in HUVECs. Meantime, their stimulation on CD31 expression were inhibited by DAPT, a Notch signaling inhibitor. Overall, this study demonstrates the positive effects of Met combined with Kae on osteoporotic rats by promoting osteogenesis-angiogenesis coupling, suggesting their potential application in postmenopausal osteoporosis.

研究领域: 药理学 · 骨科 · 分子生物学 · 细胞生物学
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TALANTA · 2018 · IF 4.244

Construction of liquid crystal droplet-based sensing platform for sensitive detection of organophosphate pesticide

Lele Zhou,Qiongzheng Hu,Qi Kang,Li Yu

Alkaline phosphatase (ALP), as an essential disease biomarker, widely distributes in various tissues, and plays a significant role in diagnosis and treatment of some diseases (e.g. cancers and liver dysfunction). Dichlorvos (DDVP), as one of typical organophosphate pesticide , could be hydrolyzed by ALP. In this work, we constructed a simple but robust 4-cyano-4′-pentylbiphenyl (5CB) droplet sensing platform for sensitive and convenient detection of DDVP based on its hydrolysis by ALP. The optical responses of liquid crystals (LCs) originated from controlling orientations of 5CB by using an ALP cleavable surfactant, sodium monododecyl phosphate (SMP). A dark crossed optical image of 5CB was observed due to formation of the SMP monolayer at the aqueous/LC droplet interface. Subsequently, 5CB turned to bright fan-shaped optical appearance upon addition of ALP and SMP mixture. Interestingly, LCs remained dark crossed optical images in contact with the pre-incubated mixture of DDVP and ALP. The detection limit of DDVP reached ~0.1 ng/mL. Isothermal titration calorimetry (ITC) and humam ALP ELISA kit provided strong evidence for the detection mechanism. This strategy was also used to evaluate whether the maximum pesticide residue limits of tomatoes conform to the relevant national standard requirement or not. Our study holds promise for real-time and label-free detection of pesticides with high sensitivity by employing portable LC-based sensing platform.

CAS / MDL: 62-73-7 9000-90-2
研究领域: 分析化学 · 环境科学 · 生物化学 · 食品安全
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

Dual-mode colorimetric–fluorometric immunoassay for foodborne parasitic infection

Yao Yu,Xiaolei Liu,Ning Xu,Lin Li,Yaming Yang,Mingyuan Liu,Lianjing Zhao,Xue Bai

Immunoassays are the primary method used for detection of foodborne parasitic infection. The nanomaterials used in these assays often present challenges such as complex preparation requirements and low stability during long-term storage and transport. Herein, based on cost-effective K 3 [Fe(CN) 6 ]-mediated in-situ generated CuFe Prussian blue analogues (CuFe-PBA) and quinoxaline derivatives with high fluorescence, a dual-mode immunoassay was proposed for foodborne parasitic infection. This platform is relied on the restoration of K 3 [Fe(CN) 6 ] by ascorbic acid (AA). In addition to this process, K 4 [Fe(CN) 6 ]-triggered the generation of CuFe-PBA, and the enhanced fluorogenic reaction of dehydrogenated AA and o -phenylenediamine (OPD) occur sequentially. Without complex probes preparation, the assay enabled the rapid on-site detection of ALP and foodborne parasitic infection through a colorimetric mode accessible via smartphone and offered enhanced sensitivity through a fluorescent mode. The method was used for the detection of ALP, achieving detection limits of 0.38 U/L and 0.03 U/L for colorimetric and fluorescence modes, respectively. Subsequently, the dual-mode immunoassay was applied to Trichinella spiralis infection. The sensitivity and specificity of both the colorimetric and fluorescence modes were 100 % in 950 pig serum samples. This immunosensor provides a universal analysis tool for the accurate, cost-effective and highly sensitive detection of foodborne parasitic infection.

研究领域: 分析化学 · 食品安全 · 生物技术 · 寄生虫学
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SCIENCE OF THE TOTAL ENVIRONMENT · 2024 · IF 8.2

Bioenzyme–nanoenzyme–chromogen all-in-one test strip for convenient and sensitive detection of malathion in water environment

Hanwen Zhang,Jiabin Jin,Xini Wang,Wei Han,Lei Qin,Lin Chen,Xiaoyan Mao,Zhenjiang Liu,Xingwei Xiang

The presence of pesticide residues in aquatic environments poses a significant threat to both aquatic ecosystems and human health. The presence of these residues can result in significant harm to aquatic ecosystems and can negatively impact the health of aquatic organisms. Consequently, this issue requires urgent attention and effective measures to mitigate its impact. However, developing sensitive and rapid detection methods remains a challenge. In this study, an all-in-one test strip, which integrated bioenzymes, nanoenzymes, and a chromogen, was developed in combination with an enzyme labeling instrument for a highly sensitive and convenient sensing of malathion residues. The oxidase activity of heme chloride (Hemin) in the strip can catalyze the oxidation of H 2 O 2 and 3,3′,5,5′-tetramethylbenzidine (TMB) to produce a blue-colored oxide. Simultaneously, the alkaline phosphatase (ALP) present in the strip can break down l -ascorbic acid-2-phosphate to produce ascorbic acid (AA). This AA then acts to reduce the oxidized form of TMB, turning it into a colorless substance and leading to the disappearance of its fluorescent signal. In the presence of a pesticide, the activity of ALP is inhibited and formation of AA is blocked, thereby preventing the reduction of oxidized TMB and producing a colored signal. According to this principle, the integrated test strip detected the target pesticide with high performance as per the optical density value determined via an enzyme marker. The detection limit of the test strip was 0.209 ng/mL with good sensitivity. The method was used for detecting malathion in actual river water samples, and the recoveries were in the range of 93.53 %–96.87 %. The newly devised technique effectively identified malathion in samples of natural water. This research has introduced a novel approach for the precise and convenient surveillance of pesticide remnants. Additionally, these discoveries could inspire the advancement of proficient multi-enzyme detection systems.

研究领域: 环境科学 · 分析化学 · 生物技术 · 水质监测
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JOURNAL OF ETHNOPHARMACOLOGY · 2024 · IF 4.8

Desmodium styracifolium (Osb.) Merr. extracts alleviate cholestatic liver disease by FXR pathway

Zhiyuan Zhang,Guoqiang Guan,Zixuan Tang,Weimin Wan,Zhipeng Huang,Yuefeng Wang,Jianzhao Wu,Bo Li,Mingli Zhong,Kefeng Zhang,Lixian Nong,Ya Gao,Houkang Cao

A B S T R A C T Ethnopharmacological relevance Cholestatic liver disease (CLD) is a disease characterized by cholestasis. Farnesoid X receptor (FXR) is a nuclear receptor that maintains homeostasis in bile acid metabolism. Studies have shown that gut microbiota interfered with the FXR pathway. Modulation of FXR to inhibit cholestasis has become a key measure in the treatment of CLD. In traditional folk medicine, Desmodium styracifolium (Osb.) Merr. was used as a primary treatment for gallstones, gonorrhea, jaundice, cholecystitis and other diseases. Modern pharmacological studies had also found that the herb has anti-calculus, anti-inflammatory, antioxidant, diuretic and liver damage. Therefore, we speculated that Desmodium styracifolium (Osb.) Merr. extracts (DME) could alleviate CLD through the FXR pathway and might be associated with the gut microbiota. However, studies of DME alleviating CLD through the FXR pathway have not been reported. Aim of study To study the effect and mechanism of DME in relieving CLD through in vivo and in vitro experiments. Materials and methods First, mice were administrated with alpha-naphthyl isothiocyanate (ANIT) to establish a CLD model in vivo . Meanwhile, HepG2 cells were induced by lithocholic acid (LCA) to establish the CLD model in vitro . To evaluate the therapeutic effect of DME on CLD mice, hematoxylin-eosin (HE) staining, and biochemical indicators were performed. The prototype of the blood components in mice serum was detected by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). 16S rDNA sequencing was used to analyze the gut microbiota. Finally, the protein and mRNA expression of the FXR pathway in mice liver tissues or HepG2 cells were detected by Western blot, qRT-PCR, or immunofluorescence. Results Pathological testing and biochemical indexes showed that DME significantly reduced serum ALT, AST, ALP, TBIL, DBIL, TBA and liver TBA levels, and attenuated liver tissue injury, necrosis and jaundice in CLD mice. In addition, MetagenomeSeq analysis of gut microbiota showed that DME significantly up-regulated the abundance of Parvibacter , down-regulated the abundance of Paenalcaligenes , and regulated bile acid homeostasis. In terms of mRNA expression, DME significantly upregulated the mRNA levels of Nr1h4, Abcb11, Cyp7a1 and Slc10a1. Meanwhile, in terms of protein expression, DME significantly up-regulated the protein expression levels of FXR, BSEP, CYP7A1 and NTCP, which regulated bile acid homeostasis. Finally, the molecular docking results showed that the components of DME, such as Lumichrome, Daidzein and Folic acid, all had good binding ability with FXR, and the surface plasmon resonance (SPR) results also showed that both Lumichrome and Daidzein had a relatively high affinity with FXR. Conclusion DME alleviated CLD through the FXR pathway, and the mechanisms might be associated with the gut microbiota.

研究领域: 药理学 · 肝病学 · 分子生物学 · 微生物学
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CHEMICAL ENGINEERING JOURNAL · 2024 · IF 13.3

Increased physiological osteochondral repair via space-specific sequestrating endogenous BMP-2 founctional hydrogel

Yuping Wang,Yiting Lei,Nan Wang,Jun Zhang,Wenguo Cui,Xiaoji Luo

The challenge of achieving graded regeneration of adjacent tissues remains significant in tissue repair and regeneration. For instance, the disruption of the microenvironment caused by “osteogenesis mobilization” often results in fibrosis of the regenerated cartilage, ultimately leading to osteoarthritis. Inspired by the utilization of activated carbon for adsorbing and filtering small particles in air purification and water treatment, we have designed an integrated hydrogel scaffold capable of absorbing the endogenous BMP-2 produced by “osteogenesis mobilization”, confining it to the bone defect area, and preventing its diffusion into the cartilage region, thereby facilitating physiological osteochondral gradient regeneration. The osteogenic layer of this integrated scaffold is NHAP-AHAMA-B2A, which captures the BMP-2 produced by “osteogenesis mobilization” and amplifies its osteogenic effect through the grafted peptide B2A. This upregulation enhances the expression of ALP, OCN, and OPN in MSCs within the osteogenic layer, promoting osteogenic differentiation. The chondrogenic layer, HAMA-B2A, utilizes the peptide B2A to upregulate the expression of COL II and ACAN in MSCs, fostering chondrogenic differentiation. This integrated HAMA-B2A/NHAP-AHAMA-B2A hydrogel scaffold confines the BMP-2 produced by “osteogenesis mobilization” to the bone defect area, prevents its diffusion into the cartilage defect area, and effectively utilizes it to induce stem cell osteogenic differentiation. Simultaneously, it induces stem cell chondrogenic differentiation in the cartilage defect area, achieving transparent cartilage regeneration. In summary, the osteogenic and chondrogenic layers of this integrated scaffold collaborate to achieve a physicochemical gradient regeneration of bone-cartilage.

CAS / MDL: 106797-53-9 760-93-0
研究领域: 组织工程 · 再生医学 · 生物材料科学 · 骨科
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ANALYTICAL CHEMISTRY · 2024 · IF 6.7

Enzymatically Mediated In Situ Generation of Z-Scheme Bi2S3/Bi2MoO6 Heterojunction-Based Organic Photoelectrochemical Transistor for METTL3/METTL14 Detection

Miao Zhang,Haowei Zhang,Yunlei Zhou,Huanshun Yin,Zhengkun Yu,Xinyue Zhang,Shiyun Ai,Minghui Wang

The OPECT biosensing platform, which connects optoelectronics and biological systems, offers significant amplification and more possibilities for research in biological applications. In this work, a homogeneous organic photoelectrochemical transistor (OPECT) biosensor based on a Bi2S3/Bi2MoO6 heterojunction was constructed to detect METTL3/METTL14 protein activity. The METTL3/METTL14 complex enzyme was used to catalyze adenine (A) on an RNA strand to m6A, protecting m6A-RNA from being cleaved by an E. coli toxin (MazF). Alkaline phosphatase (ALP) catalyzed the conversion of Na3SPO3 to H2S through an enzymatic reaction. Due to the adoption of the strategy of no fixation on the electrode, the generated H2S was easy to diffuse to the surface of the ITO electrode. The Bi2S3/Bi2MoO6 heterojunction was formed in situ through a chemical replacement reaction with Bi2MoO6, improving photoelectric conversion efficiency and realizing signal amplification. Based on this “signal on” mode, METTL3/METTL14 exhibited a wide linear range (0.00001–25 ng/μL) between protein concentration and photocurrent intensity with a limit of detection (LOD) of 7.8 fg/μL under optimal experimental conditions. The applicability of the developed method was evaluated by investigating the effect of four plasticizers on the activity of the METTL3/METTL14 protein, and the molecular modeling technique was employed to investigate the interaction between plasticizers and the protein.

研究领域: 生物化学 · 生物传感器 · 光电化学 · 蛋白质活性检测
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