Analytical biochemistry最新文献_第7页

In-vivo implementation of the HPTLC methodology to rat plasma for the concurrent determination of a novel combination therapy for the management of COVID-19 (favipiravir and nitazoxanide) 在体内实施大鼠血浆HPTLC方法，以同时确定一种用于治疗COVID-19的新型联合疗法（favipiravir和nitazoxanide）。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-19 DOI: 10.1016/j.ab.2025.115779

Fadwa H. Edrees , Maha M. Abdelrahman , Amal B. Ahmed

The 2019 coronavirus outbreak has prompted scientists to investigate pharmaceuticals to prevent the spread of the disease. Favipiravir (FAV) has received Food and Drug Administration FDA approval for the treatment of various viral infections with notable efficacy in clinical trials for COVID-19. Nitazoxanide (NTZ) is a broad-spectrum antiparasitic and antiviral agent used for the treatment of parasitic illnesses. Recently, the antiviral medications FAV and NTZ have been utilized therapeutically as early antiviral combination therapy for COVID-19, highlighting their safety, efficacy, and immunomodulatory effects. Despite several clinical studies on both FAV and NTZ, no analytical method has been developed for their concurrent detection. The objective of this study is to develop a TLC-densitometric method for their assessment and application to rat plasma. FAV, NTZ, and tizoxanide (the active metabolite of nitazoxanide (TZM)) were simultaneously determined using an HPTLC method embracing sulfasalazine as an internal standard (IS). Silica gel 60 F254 used as the stationary phase for chromatographic separation, the mobile phase consisted of chloroform-methanol-formic acid (9.5:0.5:0.2, v/v/v), with UV detection at 230 nm demonstrating linearity within the range of 0.5–5 μg/band. The proposed methodology has been shown to effectively measure the analyzed elements in both pure form and in-vivo rat plasma.

2019年冠状病毒的爆发促使科学家们研究药物来防止疾病的传播。Favipiravir （FAV）已获得美国食品和药物管理局（FDA）批准，用于治疗各种病毒感染，在COVID-19的临床试验中疗效显著。Nitazoxanide （NTZ）是一种广谱抗寄生虫和抗病毒药物，用于治疗寄生虫病。最近，抗病毒药物FAV和NTZ作为COVID-19的早期抗病毒联合治疗被用于治疗，突出了它们的安全性、有效性和免疫调节作用。尽管有几项关于FAV和NTZ的临床研究，但没有一种分析方法可以同时检测它们。本研究的目的是建立一种薄层色谱密度测定方法，以评估其在大鼠血浆中的应用。采用以柳氮磺胺为内标（IS）的高效液相色谱法同时测定FAV、NTZ和替唑昔尼特（硝唑昔尼特的活性代谢物TZM）。以硅胶60f254为固定相进行色谱分离，流动相为氯仿-甲醇-甲酸（9.5:0.5:0.2,v/v/v），在230 nm处紫外检测，在0.5 ~ 5 μg/波段范围内呈线性关系。所提出的方法已被证明可以有效地测量纯形式和体内大鼠血浆中的分析元素。

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引用次数: 0

Rapid, flexible fabrication of a microfluidic electrochemical chip nucleic acid target for selective, label-free detection of influenza virus DNA using catalytic redox-recycling 利用催化氧化还原循环快速、灵活地制备用于流感病毒DNA选择性、无标记检测的微流控电化学芯片核酸靶标。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-16 DOI: 10.1016/j.ab.2025.115771

Hosna Ehzari , Masoud Amiri , Rahman Hallaj , Marzieh Sadeghi

H5N1 flu is a highly virulent and variable subtype of influenza with significant epidemic and pandemic potential. In this study, we introduce a novel, maskless, and rapid manufacturing process for a microfluidic chip integrated with electrodes for the quantitative detection of H5N1-DNA sequences. This detection leverages a catalytic redox-recycling signal via a novel Fe₃O₄@TMU-8 nanocomposite, which facilitates the turnover of the oxidation state of [Ru(NH₃)₆]³⁺, thereby amplifying the electrochemical signal output. The positively charged [Ru(NH₃)₆]³⁺ molecule associates with the phosphate backbone of the nucleic acids in H5N1-DNA. Changes in the aptasensor's redox-recycling signal, due to the hybridization of DNA sequences with [Ru(NH₃)₆]³⁺, were used as the electrochemical sensing response. Under optimal conditions, the signal exhibited a linear relationship with H5N1-DNA concentration, ranging from 1 fM to 1 nM, with a detection limit of 0.16 fM. This report details the fabrication of the microfluidic device using Poly(methyl methacrylate) (PMMA) sheet substrates. A laser system was employed to generate microfluidic patterns directly on the PMMA sheet. This biosensing device demonstrated long-term stability and good reproducibility, making it suitable for the quantitative assay of H5N1-DNA sequences. The results from food sample analyses further confirmed the applicability and effectiveness of the resulting biosensor.

H5N1流感是一种高毒力和可变的流感亚型，具有显著的流行和大流行潜力。在这项研究中，我们介绍了一种新的、无掩模的、快速制造的微流控芯片集成电极，用于定量检测H5N1-DNA序列。这种检测利用了一种新型Fe₃O₄@TMU-8纳米复合材料的催化氧化还原回收信号，促进了[Ru（NH₃）₆]³⁺氧化态的转换，从而放大了电化学信号输出。带正电的[Ru（NH₃）₆]³+分子与H5N1-DNA中核酸的磷酸主链结合。由于DNA序列与[Ru（NH₃）₆]³⁺杂交，偶联传感器的氧化还原回收信号发生了变化，这被用作电化学传感响应。在最佳条件下，该信号与H5N1-DNA浓度呈线性关系，范围为1 fM ~ 1 nM，检出限为0.16 fM。本报告详细介绍了使用聚甲基丙烯酸甲酯（PMMA）片基板的微流控装置的制造。采用激光系统直接在PMMA薄片上产生微流控图案。该生物传感装置具有长期稳定性和良好的重复性，适用于H5N1-DNA序列的定量分析。食品样品分析的结果进一步证实了该生物传感器的适用性和有效性。

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引用次数: 0

Innovative determination of Let-7a for lung cancer diagnosis using a duplex specific nuclease (DSN)-based electrochemical biosensor 基于双特异性核酸酶（DSN）的电化学生物传感器对肺癌诊断中Let-7a的创新测定。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-13 DOI: 10.1016/j.ab.2025.115770

Jianhao Xu, Haifeng Li

In this study, we emphasize the importance of identifying Let-7a, a microRNA that is key in diagnosing and predicting lung cancer outcomes. Let-7a′s function as a biomarker is essential, as it affects tumor suppression and controls cell differentiation and growth. We developed a novel device, an electrochemical biosensor based on Duplex Specific Nuclease (DSN), that is designed for the accurate detection of Let-7a. This biosensor has a very low detection limit of 3.9 aM, showing its high sensitivity. The design is easy to use, requiring little training to operate. Its small size and portability enable the possibility of bedside and home use, which is a significant improvement for personalized healthcare. This versatility of the biosensor is very promising, as it can be applied to other disease biomarkers besides lung cancer. We expect this technology to improve disease diagnosis and patient recovery tracking, playing an important role in the future of medical diagnostics. The use of such sensitive and specific biosensors can transform the way diseases are managed, providing timely and precise information on disease progression and treatment effectiveness. The potential for this technology to help advance personalized medicine and patient care is huge, creating new opportunities in medical diagnostics and therapy monitoring.

在这项研究中，我们强调了鉴定Let-7a的重要性，这是诊断和预测肺癌预后的关键microRNA。Let-7a作为一种生物标志物的功能是必不可少的，因为它影响肿瘤抑制和控制细胞分化和生长。我们开发了一种新的装置，一种基于双工特异性核酸酶（DSN）的电化学生物传感器，用于精确检测Let-7a。该生物传感器的检测限很低，仅为3.9 aM，具有很高的灵敏度。该设计易于使用，几乎不需要培训就可以操作。它的小尺寸和便携性使床边和家庭使用成为可能，这是个性化医疗保健的重大改进。这种生物传感器的多功能性非常有前景，因为它可以应用于肺癌以外的其他疾病生物标志物。我们期望这项技术能够改善疾病诊断和患者康复跟踪，在未来的医疗诊断中发挥重要作用。使用这种敏感和特异的生物传感器可以改变疾病管理的方式，提供关于疾病进展和治疗效果的及时和精确的信息。这项技术帮助推进个性化医疗和患者护理的潜力是巨大的，在医疗诊断和治疗监测方面创造了新的机会。

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引用次数: 0

Microplate spectrophotometric method for regioselective lipase screening using structured triglycerides with punicic acid as probe 结构酸甘油三酯筛选区域选择性脂肪酶的微孔板法：从念珠菌异构体中发现具有sn-2区域选择性的Lip4脂肪酶。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-11 DOI: 10.1016/j.ab.2025.115769

Enrique Ordaz , Osvaldo Gómez-Secundino , Hiram Y. Guerrero-Elias , M. Angeles Camacho-Ruiz , Ruben Espinosa-Salgado , Antonio Escobedo-Reyes , Juan C. Mateos-Díaz , Jorge A. Rodríguez

In this study, we propose a continuous assay that provides a high-throughput, efficient method for screening the regioselectivity of lipases at the sn-1,3 and sn-2 positions on triacylglycerols (TAGs). This assay measures the specific hydrolysis rates at the primary and secondary positions of TAGs derivates containing oleic (O) and punicic (P) acids. The method is based on the absorbance ratio of released punicic acid from the hydrolysis of sn-POP (sn-1,3 regiospecific lipases) and sn-OPO (sn-2 regiospecific lipases). The method was validated using pure lipases with known and unknown regioselectivity. Unexpectedly, we found that recombinant Lipase 4 from Candida rugosa (rCRLip4) exhibited significant sn-2 regioselectivity, indicating greater regioselectivity than recombinant lipase A from Candida antarctica (rCALA). In silico analysis and molecular docking studies were conducted to elucidate the main structural differences between CRLip4 and the non-regioselective isoform CRLip1. This continuous regioselective lipase assay on TAGs is versatile and can be used to screen for sn-1,3, sn-2 or non-regioselective lipases. It holds significant potential applications in the biocatalytic production of structured lipids and other industrial processes where regioselectivity is crucial.

在本研究中，我们提出了一种连续测定法，它提供了一种高通量、高效率的方法来筛选三酰甘油（TAGs）上 Sn-1,3 和 sn-2 位上的脂肪酶的区域选择性。该测定法可测量含有油酸（O）和布酸（P）的 TAG 衍生物的主位和次位的特定水解率。该方法基于 sn-POP（sn-1,3 区域特异性脂肪酶）和 sn-OPO（sn-2 区域特异性脂肪酶）水解释放出的布匿酸的吸光度比值。我们使用已知和未知区域选择性的纯脂肪酶对该方法进行了验证。出乎意料的是，我们发现重组粗毛念珠菌脂肪酶 4（rCRLip4）表现出显著的 sn-2 区域选择性，表明其区域选择性高于重组南极念珠菌脂肪酶 A（rCALA）。为阐明 CRLip4 与非区域选择性异构体 CRLip1 之间的主要结构差异，进行了硅学分析和分子对接研究。这种针对 TAG 的连续区域选择性脂肪酶测定方法用途广泛，可用于筛选 sn-1、3、sn-2 或非区域选择性脂肪酶。它在生物催化生产结构脂和其他对区域选择性至关重要的工业过程中具有重要的潜在应用价值。

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引用次数: 0

A highly sensitive creatine kinase detection in human serum using 11-mercaptoundecanoic acid modified ITO-PET electrodes 使用11-巯基十四酸修饰ITO-PET电极检测人血清中高灵敏度肌酸激酶。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-10 DOI: 10.1016/j.ab.2025.115768

Burçak Demirbakan

The enzyme creatine kinase (CK) is a biomarker that plays an extremely significant role in the early detection of cardiovascular disorders. Serum levels of CK are regularly monitored in patients with heart attacks, one of the most critical cardiovascular illnesses. In this study, a highly sensitive electrochemical immunosensor system was designed for the importance of early diagnosis of CK. This immunosensor system was developed by immobilizing 11- mercaptoundecanoic acid (11-MuA) on disposable indium tin oxide–polyethylene terephthalate (ITO-PET) electrodes. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and single frequency impedance (SFI) techniques were utilized throughout the immobilization process during the construction of the immunosensor. In addition, the proposed CK immunosensor system involves thorough analytical research, which may include linear determination range, repeatability, reproducibility, square wave voltammetry, storage capability, and regeneration. The suggested immunosensor was also characterized using scanning electron microscopy (SEM). The proposed immunosensor system demonstrated a broad dynamic range (0.1 pg/mL – 100 pg/mL), as well as a low limit of detection (LOD) and a low limit of quantification (LOQ) of 0.018 pg/mL and 0.0394 pg/mL, respectively. Finally, the immunosensor was tested on human serum samples, proving that it could be utilized in clinical situations.

肌酸激酶（CK）是一种生物标志物，在心血管疾病的早期检测中起着极其重要的作用。心脏病是最严重的心血管疾病之一，在心脏病患者中，血清CK水平被定期监测。鉴于早期诊断CK的重要性，本研究设计了一种高灵敏度的电化学免疫传感器系统。该免疫传感器系统是通过将11-巯基十四酸（11- mua）固定在一次性氧化铟锡-聚对苯二甲酸乙二醇酯（ITO-PET）电极上开发的。在构建免疫传感器的整个固定过程中，采用了电化学阻抗谱（EIS）、循环伏安法（CV）和单频阻抗（SFI）技术。此外，拟议的CK免疫传感器系统涉及深入的分析研究，其中可能包括线性测定范围，可重复性，再现性，方波伏安法，存储能力和再生。所提出的免疫传感器也用扫描电镜（SEM）进行了表征。该免疫传感器系统具有较宽的动态范围（0.1 pg mL-1 ~ 100 pg mL-1），低检测限（LOD）和低定量限（LOQ）分别为0.018 pg mL-1和0.0394 pg mL-1。最后，对人体血清样本进行了测试，证明该免疫传感器可用于临床。

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引用次数: 0

Green fabricated bimetallic zinc ferrite nanoparticles mitigate oxidative stress-induced pathogenesis 绿色制备的双金属铁酸锌纳米颗粒减轻氧化应激诱导的发病机制。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-08 DOI: 10.1016/j.ab.2025.115767

Shivakumar Venkataramaiah , Manjula M. Venkatappa , Rajesh Rangappa , Chikkappa Udagani , Devaraja Sannaningaiah

Current study evaluates the beneficial role of bio-functionalized zinc ferrite nanoparticles fabricated from an aqueous extract of Decalepis hamiltonii leaves (DHLE.ZnFe₂O₄ NPs) on sodium nitrite (NaNO₂) and Diclofenac (DFC) induced oxidative stress in RBCs and Sprague Dawley male rat models. DHLE.ZnFe₂O₄ NPs were characterized using PXRD, FTIR, SEM-EDAX, HR-TEM and VSM. The data suggests that, DHLE.ZnFe₂O₄ NPs were crystalline, ellipsoidal in shape with an average size of 10.95 nm and super paramagnetic in nature. DHLE.ZnFe₂O₄ NPs exhibited anti-oxidant properties by scavenging DPPH, H₂O₂ and reducing ferric to ferrous ions. Furthermore, DHLE.ZnFe₂O₄ NPs normalized key parameters of oxidative stress such as LPO, PCC, TT and anti-oxidant enzymes (SOD & CAT). Similar to the previous in-vitro results, DHLE.ZnFe₂O₄ NPs restored all the said stress parameters in homogenates of the liver, kidney, pancreas and heart. In addition, DHLE.ZnFe₂O₄ NPs repaired Diclofenac induced tissue damage in the liver, kidney, pancreas and heart by regulating all biochemical parameters. Most importantly, DHLE.ZnFe₂O₄ NPs exhibited anti-inflammatory, anti-diabetic, anti-thrombotic activities and were non-toxic to RBCs. In conclusion, DHLE.ZnFe₂O₄ NPs through its anti-oxidant potential ameliorate oxidative stress induced pathogenesis such as, inflammation, tissue damage, diabetes and thrombosis.

目前的研究评估了生物功能化铁酸锌纳米颗粒的有益作用，这些纳米颗粒是由水提取物制备的。ZnFe2O4 NPs作用于亚硝酸钠（NaNO2）和双氯芬酸（DFC）诱导红细胞和Sprague Dawley雄性大鼠模型氧化应激。DHLE。采用PXRD、FTIR、SEM-EDAX、HR-TEM和VSM对ZnFe2O4纳米粒子进行了表征。数据表明，DHLE。ZnFe2O4 NPs为椭圆形结晶，平均尺寸为10.95 nm，具有超顺磁性。DHLE。ZnFe2O4 NPs通过清除DPPH、H2O2和将铁还原为亚铁离子表现出抗氧化性能。此外,DHLE。ZnFe2O4 NPs规范了氧化应激关键参数LPO、PCC、TT和抗氧化酶（SOD和cat）。与之前的体外实验结果相似，DHLE。ZnFe2O4 NPs在肝脏、肾脏、胰腺和心脏匀浆中恢复了上述所有应力参数。此外，DHLE。ZnFe2O4 NPs通过调节所有生化参数修复双氯芬酸诱导的肝、肾、胰腺和心脏组织损伤。最重要的是，DHLE。ZnFe2O4 NPs具有抗炎、抗糖尿病、抗血栓活性，对红细胞无毒。总之，DHLE。ZnFe2O4 NPs通过其抗氧化潜能改善氧化应激诱导的发病机制，如炎症、组织损伤、糖尿病和血栓形成。

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引用次数: 0

Hydrodynamic characterization of the FtsZ protein from Escherichia coli demonstrates the presence of linear and lateral trimers 来自大肠杆菌的FtsZ蛋白的流体动力学表征表明存在两种类型的三聚体。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-07 DOI: 10.1016/j.ab.2025.115766

Nelson A. Araujo , Marcelo Veloso , Luis Pouchucq

FtsZ is a bacterial protein that plays a crucial role in cytokinesis by forming the Z-ring. This ring acts as a scaffold to recruit other division proteins and guide the synthesis of septal peptidoglycan, which leads to cell constriction. In its native state, the FtsZ protein from Escherichia coli (EcFtsZ) is a multi-oligomer comprising dimers, trimers, tetramers, and hexamers in a dynamic self-association equilibrium depending on its concentration. This study employed classical methods of analytical biochemistry that included native polyacrylamide gel electrophoresis, size-exclusion chromatography, sedimentation through sucrose gradients, and chemical cross-linking with formaldehyde to characterize the EcFtsZ. The dimers, trimers, and tetramers are the most prevalent oligomers of the EcFtsZ protein; however, the trimer has been understudied compared to the dimer. In this study, we characterized uncross-linked trimers by exclusion chromatography and crosslinked trimers by sedimentation. The results of size-exclusion chromatography demonstrated that the uncross-linked trimer of EcFtsZ has a mass of 128.8 kDa and a frictional ratio f/f_o of 1.96, which coincides with the theoretical frictional ratio of 1.80 for a linear trimer. The EcFtsZ protein treated with formaldehyde resulted in a polypeptide band of 128 kDa recognized by anti-FtsZ antibodies and a frictional ratio S_max/S_20,w equal to 1.95, which agrees with the theoretical calculation of the frictional ratio of a lateral trimer. The protein-protein interaction prediction program (PEPPI) identified a contact site between subunits in the C-terminal linker region of the EcFtsZ protein, which has the potential to interfere with the recognition of the C-terminal linker by the ClpX(P) protease.

FtsZ是一种细菌蛋白，通过形成z环在细胞分裂中起着至关重要的作用。这个环作为一个支架来招募其他分裂蛋白，并指导间隔肽聚糖的合成，从而导致细胞收缩。在其天然状态下，来自大肠杆菌的FtsZ蛋白（EcFtsZ）是一种多低聚物，包括二聚体、三聚体、四聚体和六聚体，根据其浓度处于动态自结合平衡状态。本研究采用经典的分析生物化学方法，包括天然聚丙烯酰胺凝胶电泳，尺寸排除色谱，通过蔗糖梯度沉淀，以及与甲醛的化学交联来表征EcFtsZ。二聚体、三聚体和四聚体是EcFtsZ蛋白最常见的低聚物；然而，与二聚体相比，三聚体的研究还不够充分。在本研究中，我们分别用排除色谱法和沉淀法对非交联三聚体进行了表征。结果表明，EcFtsZ的非交联三聚体质量为128.8 kDa，摩擦比f/fo为1.96，与线性三聚体的理论摩擦比1.80一致。经甲醛处理后，EcFtsZ蛋白的多肽带长度为128 kDa，可被抗ftsz抗体识别，其摩擦比Smax/S20，w = 1.95，与横向三聚体摩擦比的理论计算结果一致。蛋白-蛋白相互作用预测程序（PEPPI）确定了EcFtsZ蛋白c端连接子区域亚基之间的接触位点，该位点有可能干扰ClpX(P)蛋白酶对c端连接子的识别。

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引用次数: 0

Imaged capillary isoelectric focusing and online mass spectrometry for milk whey protein characterization in dairy products 乳制品中乳清蛋白的成像毛细管等电聚焦和在线质谱分析。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-06 DOI: 10.1016/j.ab.2025.115765

She Lin Chan , Teresa Kwok , Niusheng Xu , Tao Bo , Tiemin Huang

Characterizing major bovine milk proteins, including whey and casein, is of significant interest in the dairy industry. The diverse array of protein proteoforms can be different in terms of genetic variation, breed ways, lactation stage, and animal nutritional status. Current routine methods for bovine milk protein profiling are typically based on immunological techniques, infrared spectroscopy, slab gel isoelectric focusing, capillary electrophoresis, and high-performance liquid chromatography. However, there are obvious disadvantages of existing approaches including low throughput, tedious operation, unsatisfactory repeatability, and lack of robust quantitation capability. In this study, we present a novel approach that, for the first time, combines imaged capillary isoelectric focusing with mass spectrometry to separate and characterize whey proteins in milk products. The established method provided a rapid, repeatable, accurate, and simultaneous analysis of α-lactalbumin, β-lactoglobulin A, and β-lactoglobulin B within 10 min for diverse bovine milk samples. The methodology was systematically validated regarding repeatability of pI and peak area, sensitivity, linearity and recovery. The integration of high-resolution mass spectrometry with nano-electrospray ionization and icIEF has been pivotal in accurately identifying intact whey proteins in milk products. This approach has significantly enhanced the precise characterization of protein proteoforms in milk.

表征主要的牛奶蛋白，包括乳清和酪蛋白，是乳品行业的重要兴趣。不同种类的蛋白质在遗传变异、繁殖方式、哺乳阶段和动物营养状况方面可能不同。目前常规的牛乳蛋白分析方法通常是基于免疫学技术、红外光谱、平板凝胶等电聚焦、毛细管电泳和高效液相色谱。然而，现有方法存在通量低、操作繁琐、可重复性差、定量能力不强等明显缺点。在这项研究中，我们提出了一种新的方法，首次将成像毛细管等电聚焦与质谱相结合，以分离和表征乳制品中的乳清蛋白。所建立的方法可在10 min内快速、重复、准确、同时分析多种牛乳样品中的α-乳蛋白、β-乳球蛋白a和β-乳球蛋白B。对该方法的重复性、峰面积、灵敏度、线性度和回收率进行了系统验证。高分辨率质谱与纳米电喷雾电离和icIEF的结合对于准确鉴定乳制品中完整的乳清蛋白至关重要。这种方法大大提高了牛奶中蛋白质的精确表征。

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引用次数: 0

Mesoporous carbon nanospheres-assisted amplified electrochemiluminescence for l-cysteine detection 介孔碳纳米球辅助放大电化学发光检测l -半胱氨酸。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2025-01-02 DOI: 10.1016/j.ab.2025.115764

Ziqi Wang, Yahui Ji, Hui Zhang, Gen Liu

Luminol-loaded mesoporous carbon nanospheres (MCs@LU) were utilized to develop a highly sensitive electrochemiluminescence (ECL) sensor for the detection of L-cysteine (L-Cys). L-Cys acted as the coreactant of luminol, and the pore confinement effect of mesoporous carbons (MCs) resulted in a robust ECL signal. Upon optimization, a linear correlation between the ECL intensity and L-Cys concentration was observed over the range of 5.0 × 10⁻¹⁰ mol L⁻¹ to 5.0 × 10⁻⁶ mol L⁻¹. The detection limit, with a signal-to-noise ratio of 3, was determined to be 1.67 × 10⁻¹⁰ mol L⁻¹. Additionally, the ECL sensor exhibited good reproducibility, stability, and selectivity for L-Cys and was successfully applied to the quantification of L-Cys in drug samples.

利用负载鲁米诺的介孔碳纳米球（MCs@LU）开发了一种高灵敏度的电化学发光（ECL）传感器，用于检测l-半胱氨酸（L-Cys）。L-Cys作为鲁米诺的共反应物，介孔碳（MCs）的孔约束效应导致了鲁棒的ECL信号。优化后，ECL强度与L-Cys浓度在5.0 × 10-10 mol·L-1 ~ 5.0 × 10-6 mol·L-1范围内呈线性相关。检测限为1.67 × 10-10 mol·L-1，信噪比为3。此外，ECL传感器对L-Cys具有良好的重现性、稳定性和选择性，并成功应用于药物样品中L-Cys的定量分析。

引用次数: 0

Twice target-recognition mediated exonuclease iii (Exo-iii)-Propelled cascade signal recycling MicroRNA detection system with improved accuracy 二次靶标识别介导的外切酶iii （Exo-iii）推进级联信号循环MicroRNA检测系统的准确性提高。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-12-30 DOI: 10.1016/j.ab.2024.115757

Yuling Jia , Jianhua Yuan , Yanlei Zheng , Yanzhen Huang , Juncai Zhang , Haibin Zhao , Jiefang Zhang

Simple yet specific miRNA detection remains an enormous challenge due to its low abundance in samples and the high similarity among homologous miRNAs. Here, we propose a novel fluorescent approach for miRNA detection with greatly elevated accuracy by utilizing exonuclease-iii (Exo-iii) assisted twice target recognition. The proposed method involves a “Sensing probe” engineered with two loop sections to facilitate dual target miRNA recognition. The collaboration between Exo-iii and miRNA initiates target recycling for signal amplification, resulting in the formation of complete DNAzyme. The intact DNAzyme connects with the “Signal probe” and creates a nicking site within its loop region. The fluorescence signal of the “Signal probe” reemerges, correlating with the quantity of miRNA in the sensing system. The suggested technique demonstrates great selectivity for target miRNA and can readily differentiate sequences with a one-base mismatch, based on dual-target identification. Furthermore, the Exo-iii-assisted signal recycling imparts the approach with great sensitivity and a low detection limit of 548 aM. This method has the potential to be a robust alternative for the detection of miRNAs in real samples due to its high accuracy, simplicity, and resistance to potential fluorescence interferences.

由于样品中的低丰度和同源miRNA之间的高相似性，简单而特异性的miRNA检测仍然是一个巨大的挑战。在这里，我们提出了一种新的荧光检测miRNA的方法，通过利用外切酶-iii （Exo-iii）辅助两次目标识别，大大提高了准确性。所提出的方法涉及一个“传感探针”工程与两个环段，以促进双目标miRNA识别。Exo-iii和miRNA之间的合作启动了信号放大的靶标循环，从而形成完整的DNAzyme。完整的DNAzyme与“信号探针”连接，并在其环路区域内创建一个切口位点。“信号探针”的荧光信号重新出现，与传感系统中miRNA的数量相关。所建议的技术对目标miRNA具有很高的选择性，并且可以很容易地区分基于双靶标鉴定的单碱基错配序列。此外，exo -iii辅助信号回收使该方法具有高灵敏度和548 aM的低检测限。由于该方法具有较高的准确性、简单性和对潜在荧光干扰的抗性，因此有可能成为真实样品中mirna检测的可靠替代方法。

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