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Identification of Podoplanin Aptamers by SELEX for Protein Detection and Inhibition of Platelet Aggregation Stimulated by C-Type Lectin-like Receptor 2. 用 SELEX 法鉴定 Podoplanin 短链肽,用于蛋白质检测和抑制由 C 型凝集素样受体 2 刺激的血小板聚集。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-27 DOI: 10.3390/bios14100464
Hui-Ju Tsai, Kai-Wen Cheng, Jou-Chen Li, Tsai-Xiang Ruan, Ting-Hsin Chang, Jin-Ru Wang, Ching-Ping Tseng

Tumor cell-induced platelet aggregation (TCIPA) is a mechanism for the protection of tumor cells in the bloodstream and the promotion of tumor progression and metastases. The platelet C-type lectin-like receptor 2 (CLEC-2) can bind podoplanin (PDPN) on a cancer cell surface to facilitate TCIPA. Selective blockage of PDPN-mediated platelet-tumor cell interaction is a plausible strategy for inhibiting metastases. In this study, we aimed to screen for aptamers, which are the single-stranded DNA oligonucleotides that form a specific three-dimensional structure, bind to specific molecular targets with high affinity and specificity, bind to PDPN, and interfere with PDPN/CLEC-2 interactions. The systematic evolution of ligands by exponential enrichment (SELEX) was employed to enrich aptamers that recognize PDPN. The initial characterization of ssDNA pools enriched by SELEX revealed a PDPN aptamer designated as A1 displaying parallel-type G-quadruplexes and long stem-and-loop structures and binding PDPN with a material with a dissociation constant (Kd) of 1.3 ± 1.2 nM. The A1 aptamer recognized both the native and denatured form of PDPN. Notably, the A1 aptamer was able to quantitatively detect PDPN proteins in Western blot analysis. The A1 aptamer could interfere with the interaction between PDPN and CLEC-2 and inhibit PDPN-induced platelet aggregation in a concentration-dependent manner. These findings indicated that the A1 aptamer is a candidate for the development of biosensors in detecting the levels of PDPN expression. The action by A1 aptamer could result in the prevention of tumor cell metastases, and if so, could become an effective pharmacological agent in treating cancer patients.

肿瘤细胞诱导的血小板聚集(TCIPA)是血液中保护肿瘤细胞、促进肿瘤进展和转移的一种机制。血小板 C 型凝集素样受体 2(CLEC-2)能与癌细胞表面的 podoplanin(PDPN)结合,从而促进 TCIPA。选择性阻断 PDPN 介导的血小板-肿瘤细胞相互作用是抑制转移的一种可行策略。在这项研究中,我们的目标是筛选出能形成特定三维结构、以高亲和力和特异性与特定分子靶标结合、与 PDPN 结合并干扰 PDPN/CLEC-2 相互作用的单链 DNA 寡核苷酸配体。通过指数富集配体的系统进化(SELEX)来富集识别 PDPN 的适配体。通过对 SELEX 富集的 ssDNA 池进行初步表征,发现了一种被命名为 A1 的 PDPN 类似物,它具有平行型 G-四重链和长茎环结构,能与 PDPN 结合,其解离常数(Kd)为 1.3 ± 1.2 nM。A1 合体既能识别原生形式的 PDPN,也能识别变性形式的 PDPN。值得注意的是,A1 合体能够在 Western 印迹分析中定量检测 PDPN 蛋白。A1 合体能干扰 PDPN 与 CLEC-2 之间的相互作用,并以浓度依赖性的方式抑制 PDPN 诱导的血小板聚集。这些研究结果表明,A1 拟合物是开发检测 PDPN 表达水平的生物传感器的候选材料。A1 aptamer 的作用可以防止肿瘤细胞转移,因此可以成为治疗癌症患者的有效药剂。
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引用次数: 0
Correction: Hu et al. Ultrasensitive Silicon Nanowire Biosensor with Modulated Threshold Voltages and Ultra-Small Diameter for Early Kidney Failure Biomarker Cystatin C. Biosensors 2023, 13, 645. 更正:Hu 等人,用于早期肾衰竭生物标志物胱抑素 C 的具有调制阈值电压和超小直径的超灵敏硅纳米线生物传感器,生物传感器 2023,13,645。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-26 DOI: 10.3390/bios14100461
Jiawei Hu, Yinglu Li, Xufang Zhang, Yanrong Wang, Jing Zhang, Jiang Yan, Junjie Li, Zhaohao Zhang, Huaxiang Yin, Qianhui Wei, Qifeng Jiang, Shuhua Wei, Qingzhu Zhang

In the original publication [...].

在最初的出版物中 [......] 。
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引用次数: 0
Harnessing CRISPR/Cas Systems for DNA and RNA Detection: Principles, Techniques, and Challenges. 利用 CRISPR/Cas 系统检测 DNA 和 RNA:原理、技术和挑战。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-26 DOI: 10.3390/bios14100460
Heyjin Son

The emergence of CRISPR/Cas systems has revolutionized the field of molecular diagnostics with their high specificity and sensitivity. This review provides a comprehensive overview of the principles and recent advancements in harnessing CRISPR/Cas systems for detecting DNA and RNA. Beginning with an exploration of the molecular mechanisms of key Cas proteins underpinning CRISPR/Cas systems, the review navigates the detection of both pathogenic and non-pathogenic nucleic acids, emphasizing the pivotal role of CRISPR in identifying diverse genetic materials. The discussion extends to the integration of CRISPR/Cas systems with various signal-readout techniques, including fluorescence, electrochemical, and colorimetric, as well as imaging and biosensing methods, highlighting their advantages and limitations in practical applications. Furthermore, a critical analysis of challenges in the field, such as target amplification, multiplexing, and quantitative detection, underscores areas requiring further refinement. Finally, the review concludes with insights into the future directions of CRISPR-based nucleic acid detection, emphasizing the potential of these systems to continue driving innovation in diagnostics, with broad implications for research, clinical practice, and biotechnology.

CRISPR/Cas 系统以其高特异性和高灵敏度彻底改变了分子诊断领域。本综述全面概述了利用 CRISPR/Cas 系统检测 DNA 和 RNA 的原理和最新进展。本综述从探讨支持 CRISPR/Cas 系统的关键 Cas 蛋白的分子机制开始,探讨了病原体和非病原体核酸的检测,强调了 CRISPR 在识别各种遗传物质方面的关键作用。讨论延伸到 CRISPR/Cas 系统与各种信号读出技术(包括荧光、电化学和比色法)以及成像和生物传感方法的整合,强调了它们在实际应用中的优势和局限性。此外,还对目标放大、多路复用和定量检测等领域的挑战进行了批判性分析,强调了需要进一步完善的领域。最后,综述对基于 CRISPR 的核酸检测的未来发展方向进行了深入分析,强调了这些系统继续推动诊断创新的潜力,对研究、临床实践和生物技术具有广泛的影响。
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引用次数: 0
Microfluidic Detection Platform for Determination of Ractopamine in Food. 用于检测食品中莱克多巴胺的微流控检测平台
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-26 DOI: 10.3390/bios14100462
Cheng-Xue Yu, Kuan-Hsun Huang, To-Lin Chen, Chan-Chiung Liu, Lung-Ming Fu

A novel microfluidic ractopamine (RAC) detection platform consisting of a microfluidic RAC chip and a smart analysis device is proposed for the determination of RAC concentration in meat samples. This technology utilizes gold nanoparticles (AuNPs) modified with glutamic acid (GLU) and polyethyleneimine (PEI) to measure RAC concentration in food products. When RAC is present, AuNPs aggregate through hydrogen bonding, causing noticeable changes in their optical properties, which are detected using a self-built UV-visible micro-spectrophotometer. Within the range of 5 to 80 ppb, a linear relationship exists between the absorbance ratio (A693nm/A518nm) (Y) and RAC concentration (X), expressed as Y = 0.0054X + 0.4690, with a high coefficient of determination (R2 = 0.9943). This method exhibits a detection limit of 1.0 ppb and achieves results within 3 min. The practical utility of this microfluidic assay is exemplified through the evaluation of RAC concentrations in 50 commercially available meat samples. The variance between concentrations measured using this platform and those determined via liquid chromatography-tandem mass spectrometry (LC-MS/MS) is less than 8.33%. These results underscore the viability of the microfluidic detection platform as a rapid and cost-effective solution for ensuring food safety and regulatory compliance within the livestock industry.

本研究提出了一种新型微流控莱克多巴胺(RAC)检测平台,该平台由微流控莱克多巴胺芯片和智能分析装置组成,用于测定肉类样品中的莱克多巴胺浓度。该技术利用谷氨酸(GLU)和聚乙烯亚胺(PEI)修饰的金纳米粒子(AuNPs)来检测食品中的莱克多巴胺浓度。当食品中含有 RAC 时,AuNPs 会通过氢键聚合,从而导致其光学特性发生明显变化,并通过自建的紫外可见微分光光度计进行检测。在 5 至 80 ppb 的范围内,吸光度比值(A693nm/A518nm)(Y)与 RAC 浓度(X)之间存在线性关系,即 Y = 0.0054X + 0.4690,且测定系数较高(R2 = 0.9943)。该方法的检测限为 1.0 ppb,可在 3 分钟内得出结果。通过对 50 份市售肉类样品中的 RAC 浓度进行评估,证明了这种微流控检测方法的实用性。使用该平台测量的浓度与液相色谱-串联质谱法(LC-MS/MS)测定的浓度之间的差异小于 8.33%。这些结果凸显了微流控检测平台的可行性,它是一种快速、经济的解决方案,可确保畜牧业的食品安全和合规性。
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引用次数: 0
High Throughput Screening of Transcription Factor LysG for Constructing a Better Lysine Biosensor. 高通量筛选转录因子 LysG 以构建更好的赖氨酸生物传感器
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-25 DOI: 10.3390/bios14100455
Qinggang Li, Haojie Ren, Zhenjiang Liao, Shuchang Xia, Xue Sun

The biosensors based on transcription factors (TFs) are widely used in high throughput screening of metabolic overproducers. The unsatisfactory performances (narrow detection and dynamic ranges) of biosensors limit their practical application and need more improvement. In this study, using the TF LysG (sensing lysine) as an example, a biosensor optimization method was constructed by growth-coupled screening of TF random mutant libraries. The better the performance of the biosensor, the faster the strain grows under screening pressure. A LysGE15D, A54D, and I164V-based biosensors were obtained, which were about 2-fold of the control in the detection and dynamic ranges. A lysine high-producer was screened effectively using the optimized biosensor with the production at 1.51 ± 0.30 g/L in flasks (2.22-fold of the original strain). This study provided a promising strategy for optimizing TF-based biosensors and was of high potential to be applied in the lysine high-producers screening process.

基于转录因子(TFs)的生物传感器被广泛应用于高通量筛选代谢产物。生物传感器的性能(检测范围和动态范围较窄)不尽人意,限制了其实际应用,需要进一步改进。本研究以传感赖氨酸的 TF LysG 为例,通过对 TF 随机突变体库进行生长耦合筛选,构建了一种生物传感器优化方法。生物传感器的性能越好,菌株在筛选压力下的生长速度就越快。获得了基于 LysGE15D、A54D 和 I164V 的生物传感器,其检测和动态范围约为对照的 2 倍。利用优化后的生物传感器有效筛选出了一种赖氨酸高产菌株,其在烧瓶中的产量为 1.51 ± 0.30 g/L(是原始菌株的 2.22 倍)。该研究为优化基于 TF 的生物传感器提供了一种有前景的策略,并极有可能应用于赖氨酸高产菌筛选过程。
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引用次数: 0
Assessment of Microvascular Function Based on Flowmotion Monitored by the Flow-Mediated Skin Fluorescence Technique. 基于流式皮肤荧光技术监测的血流运动评估微血管功能
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-25 DOI: 10.3390/bios14100459
Andrzej Marcinek, Joanna Katarzynska, Katarzyna Cypryk, Agnieszka Los-Stegienta, Jolanta Slowikowska-Hilczer, Renata Walczak-Jedrzejowska, Jacek Zielinski, Jerzy Gebicki

This review summarizes studies dedicated to the assessment of microvascular function based on microcirculatory oscillations monitored by the Flow-Mediated Skin Fluorescence (FMSF) technique. Two approaches are presented. The first approach uses oscillatory parameters measured under normoxic conditions, expressed as flowmotion (FM), vasomotion (VM), and the normoxia oscillatory index (NOI). These parameters have been used for the identification of impaired microcirculatory oscillations associated with intense physical exercise, post-COVID syndrome, psychological stress, and erectile dysfunction. The second approach involves characterization of the microcirculatory response to hypoxia based on the measurement of hypoxia sensitivity (HS). The HS parameter is used to characterize microvascular complications in diabetes, such as diabetic kidney disease and diabetic foot ulcers. Based on research conducted by the authors of this review, the FMSF parameter ranges characterizing microvascular function are presented. The diagnostic approach to assessing microvascular function based on flowmotion monitored by the FMSF technique has a wide range of applications and the potential to be integrated into widespread medical practice.

本综述概述了基于流介导皮肤荧光(FMSF)技术监测的微循环振荡对微血管功能进行评估的研究。文中介绍了两种方法。第一种方法使用的是在常氧条件下测量的振荡参数,以流量运动(FM)、血管运动(VM)和常氧振荡指数(NOI)表示。这些参数已被用于识别与剧烈运动、COVID 后综合征、心理压力和勃起功能障碍相关的微循环振荡受损。第二种方法是通过测量缺氧敏感性(HS)来描述微循环对缺氧的反应。HS 参数用于描述糖尿病微血管并发症,如糖尿病肾病和糖尿病足溃疡。根据本综述作者的研究,介绍了表征微血管功能的 FMSF 参数范围。根据 FMSF 技术监测到的血流运动来评估微血管功能的诊断方法具有广泛的应用范围,并有可能被纳入广泛的医疗实践中。
{"title":"Assessment of Microvascular Function Based on Flowmotion Monitored by the Flow-Mediated Skin Fluorescence Technique.","authors":"Andrzej Marcinek, Joanna Katarzynska, Katarzyna Cypryk, Agnieszka Los-Stegienta, Jolanta Slowikowska-Hilczer, Renata Walczak-Jedrzejowska, Jacek Zielinski, Jerzy Gebicki","doi":"10.3390/bios14100459","DOIUrl":"https://doi.org/10.3390/bios14100459","url":null,"abstract":"<p><p>This review summarizes studies dedicated to the assessment of microvascular function based on microcirculatory oscillations monitored by the Flow-Mediated Skin Fluorescence (FMSF) technique. Two approaches are presented. The first approach uses oscillatory parameters measured under normoxic conditions, expressed as flowmotion (FM), vasomotion (VM), and the normoxia oscillatory index (NOI). These parameters have been used for the identification of impaired microcirculatory oscillations associated with intense physical exercise, post-COVID syndrome, psychological stress, and erectile dysfunction. The second approach involves characterization of the microcirculatory response to hypoxia based on the measurement of hypoxia sensitivity (HS). The HS parameter is used to characterize microvascular complications in diabetes, such as diabetic kidney disease and diabetic foot ulcers. Based on research conducted by the authors of this review, the FMSF parameter ranges characterizing microvascular function are presented. The diagnostic approach to assessing microvascular function based on flowmotion monitored by the FMSF technique has a wide range of applications and the potential to be integrated into widespread medical practice.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"14 10","pages":""},"PeriodicalIF":4.9,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11505855/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142510801","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
MNAzyme-Assisted Nucleic Acid Lateral Flow Assay for Cost-Effective, On-Site Mercury Detection. 用于现场汞检测的核酸侧流成本效益分析法(MNAzyme-Assisted Nucleic Acid Lateral Flow Assay)。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-25 DOI: 10.3390/bios14100454
Seok Hyeon Kim, Yujun Kim, Seokjoon Kim, Eun Sung Lee, Byung Seok Cha, Ki Soo Park

Mercury ions (Hg2+) are toxic heavy metals present in the environment that pose significant health risks. An advanced detection system could allow for a prompt response and alleviate serious damage to humans. In this study, we developed a cost-effective, on-site detection method for Hg2+ using a multicomponent nucleic acid enzyme (MNAzyme)-assisted nucleic acid lateral flow assay (NALFA). The MNAzyme, which was engineered to contain thymine-thymine mismatches, is responsive only to the presence of Hg2+ and exerts efficient cleavage activity on substrates that can be captured by the NALFA strip, and thus the proposed system enables the visual detection of Hg2+ in the NALFA strip. Our assay demonstrated sufficient detection sensitivity and specificity to meet the WHO standards, offering a good practical alternative for rapid environmental and public health monitoring.

汞离子(Hg2+)是存在于环境中的有毒重金属,对人体健康构成严重威胁。先进的检测系统可以迅速做出反应,减轻对人类的严重危害。在这项研究中,我们利用多组分核酸酶(MNAzyme)辅助核酸横向流动分析法(NALFA)开发了一种经济有效的现场检测 Hg2+ 的方法。MNA酶被设计成含有胸腺嘧啶-胸腺嘧啶错配,只对Hg2+的存在有反应,并对可被NALFA条带捕获的底物发挥高效的裂解活性,因此所提议的系统可在NALFA条带中实现对Hg2+的肉眼检测。我们的检测方法具有足够的检测灵敏度和特异性,符合世界卫生组织的标准,为环境和公共卫生的快速监测提供了一个很好的实用选择。
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引用次数: 0
NIR-Sensitive Squaraine Dye-Peptide Conjugate for Trypsin Fluorogenic Detection. 用于胰蛋白酶荧光检测的近红外敏感水杨酸染料肽共轭物。
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-25 DOI: 10.3390/bios14100458
Priyanka Balyan, Shekhar Gupta, Sai Kiran Mavileti, Shyam S Pandey, Tamaki Kato

Trypsin enzyme has gained recognition as a potential biomarker in several tumors, such as colorectal, gastric, and pancreatic cancer, highlighting its importance in disease diagnosis. In response to the demand for rapid, cost-effective, and real-time detection methods, we present an innovative strategy utilizing the design and synthesis of NIR-sensitive dye-peptide conjugate (SQ-3 PC) for the sensitive and selective monitoring of trypsin activity by fluorescence ON/OFF sensing. The current research deals with the design and synthesis of three unsymmetrical squaraine dyes SQ-1, SQ-2, and SQ-3 along with a dye-peptide conjugate SQ-3-PC as a trypsin-specific probe followed by their photophysical characterizations. The absorption spectral investigation conducted on both the dye alone and its corresponding dye-peptide conjugates in water, utilizing SQ-3 and SQ-3 PC respectively, reveals enhanced dye aggregation and pronounced fluorescence quenching compared to observations in DMSO solution. The absorption spectral investigation conducted on dye only and corresponding dye-peptide conjugates in water utilizing SQ-3 and SQ-3 PC, respectively, reveals not only the enhanced dye aggregation but also pronounced fluorescence quenching compared to that observed in the DMSO solution. The trypsin-specific probe SQ-3 PC demonstrated a fluorescence quenching efficiency of 61.8% in water attributed to the combined effect of aggregation-induced quenching (AIQ) and fluorescence resonance energy transfer (FRET). FRET was found to be dominant over AIQ. The trypsin-mediated hydrolysis of SQ-3 PC led to a rapid and efficient recovery of quenched fluorescence (5-fold increase in 30 min). Concentration-dependent changes in the fluorescence at the emission maximum of the dyes reveal that SQ-3 PC works as a trypsin enzyme-specific fluorescence biosensor with linearity up to 30 nM along with the limit of detection and limit of quantification of 1.07 nM and 3.25 nM, respectively.

胰蛋白酶已被认为是结直肠癌、胃癌和胰腺癌等多种肿瘤的潜在生物标记物,这凸显了它在疾病诊断中的重要性。为了满足对快速、经济、实时检测方法的需求,我们提出了一种创新策略,利用设计和合成近红外敏感染料-肽共轭物(SQ-3 PC),通过荧光开/关传感灵敏、选择性地监测胰蛋白酶活性。目前的研究涉及设计和合成三种非对称方碱染料 SQ-1、SQ-2 和 SQ-3,以及作为胰蛋白酶特异性探针的染料-肽共轭物 SQ-3-PC,并对其进行光物理表征。分别利用 SQ-3 和 SQ-3 PC 对单独的染料及其相应的染料-肽共轭物在水中进行的吸收光谱研究表明,与在二甲基亚砜溶液中观察到的结果相比,染料的聚集性增强,荧光淬灭明显。分别利用 SQ-3 和 SQ-3 PC 对水中的染料和相应的染料-肽共轭物进行的吸收光谱研究表明,与在二甲基亚砜溶液中观察到的结果相比,不仅染料聚集增强,而且荧光淬灭也很明显。由于聚集诱导淬灭(AIQ)和荧光共振能量转移(FRET)的共同作用,胰蛋白酶特异性探针 SQ-3 PC 在水中的荧光淬灭效率达到 61.8%。研究发现,FRET 比 AIQ 起主导作用。胰蛋白酶介导的 SQ-3 PC 水解可使淬灭的荧光快速有效地恢复(30 分钟内增加 5 倍)。染料发射最大值处荧光的浓度变化表明,SQ-3 PC 是一种胰蛋白酶特异性荧光生物传感器,其线性关系可达 30 nM,检测限和定量限分别为 1.07 nM 和 3.25 nM。
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引用次数: 0
Recovery and Analysis of Bacterial Membrane Vesicle Nanoparticles from Human Plasma Using Dielectrophoresis. 利用介电泳技术从人体血浆中回收和分析细菌膜泡纳米颗粒
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-25 DOI: 10.3390/bios14100456
Jason P Ware, Delaney K Shea, Shelby L Nicholas, Ella A Stimson, Jessica L Riesterer, Stuart D Ibsen

Bacterial membrane vesicle (BMV) nanoparticles are secreted naturally by bacteria throughout their lifecycle and are a rich source of biomarkers from the parent bacteria, but they are currently underutilized for clinical diagnostic applications, such as pathogen identification, due to the time-consuming and low-yield nature of traditional recovery methods required for analysis. The recovery of BMVs is particularly difficult from complex biological fluids. Here, we demonstrate a recovery method that uses dielectrophoretic (DEP) forces generated on electrokinetic microfluidic chips to isolate and analyze BMVs from human plasma. DEP takes advantage of the natural difference in dielectric properties between the BMVs and the surrounding plasma fluid to quickly and consistently collect these particles from as little as 25 µL of plasma. Using DEP and immunofluorescence staining of the LPS biomarker carried on BMVs, we have demonstrated a lower limit of detection of 4.31 × 109 BMVs/mL. The successful isolation of BMVs from human plasma using DEP, and subsequent on-chip immunostaining for biomarkers, enables the development of future assays to identify the presence of specific bacterial species by analyzing BMVs from small amounts of complex body fluid.

细菌膜囊(BMV)纳米粒子是细菌在其整个生命周期中自然分泌的,是母菌生物标志物的丰富来源,但由于传统的分析回收方法耗时长、产量低,目前在病原体鉴定等临床诊断应用中还未得到充分利用。从复杂的生物液体中回收 BMV 尤为困难。在这里,我们展示了一种利用电动微流控芯片上产生的介质电泳(DEP)力从人体血浆中分离和分析 BMV 的回收方法。DEP 利用了 BMV 与周围血浆流体之间介电性质的天然差异,能从 25 µL 的血浆中快速、稳定地收集这些微粒。通过使用 DEP 和对 BMV 上携带的 LPS 生物标记物进行免疫荧光染色,我们证明检测下限为 4.31 × 109 BMVs/mL。利用 DEP 成功地从人体血浆中分离出 BMV,并随后在芯片上对生物标记物进行免疫染色,这使我们能够开发未来的检测方法,通过分析少量复杂体液中的 BMV 来确定特定细菌物种的存在。
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引用次数: 0
Ratiometric Electrochemical Detection of Interleukin-6 Using Electropolymerized Methylene Blue and a Multi-Walled Carbon-Nanotube-Modified Screen-Printed Carbon Electrode. 使用电聚合亚甲基蓝和多壁碳纳米管修饰的丝网印刷碳电极对白细胞介素-6 进行比率电化学检测
IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-25 DOI: 10.3390/bios14100457
Zhuo Liu, Fengyu Liu, Chaofan Wang, Hongjuan Li, Yongqian Xu, Shiguo Sun

Herein, we report a ratio-based electrochemical biosensor for the detection of interleukin-6 (IL-6). We electropolymerized methylene blue (MB) on the surface of screen-printed carbon electrodes; introduced an internal reference signal probe; modified the carboxylate multi-walled carbon nanotubes on the electrode surface to increase the electrochemically active area; and finally linked the amino-modified IL-6 aptamer to the electrode surface through the Schiff base reaction, with bovine serum albumin (BSA) added to mask non-specific adsorption. After adding IL-6 to the samples, the signal of IMB remained almost unchanged, while the signal of I[Fe(CN)6]3-/4- decreased with increasing IL-6 concentration. Thus, a novel ratiometric electrochemical sensor with a linear range of 0.001~1000.0 ng/mL and a low detection limit of 0.54 pg/mL was successfully developed. The sensor had high repeatability, stability, sensitivity, and practicability. It provides a new method for constructing proportional electrochemical sensors and detecting IL-6.

在此,我们报告了一种基于比率的电化学生物传感器,用于检测白细胞介素-6(IL-6)。我们在丝网印刷碳电极表面电聚合亚甲基蓝(MB);引入内部参考信号探针;修饰电极表面的羧基多壁碳纳米管以增加电化学活性面积;最后通过希夫碱反应将氨基修饰的 IL-6 合体连接到电极表面,并加入牛血清白蛋白(BSA)以掩蔽非特异性吸附。在样品中加入 IL-6 后,IMB 的信号几乎保持不变,而 I[Fe(CN)6]3-/4- 的信号则随着 IL-6 浓度的增加而降低。因此,一种线性范围为 0.001~1000.0 纳克/毫升、检测限低至 0.54 皮克/毫升的新型比率电化学传感器研制成功。该传感器具有高重复性、稳定性、灵敏度和实用性。它为构建比例电化学传感器和检测 IL-6 提供了一种新方法。
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引用次数: 0
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