ELECTROPHORESIS最新文献

{"title":"Development and Validation of the 9-Dye Fluorescent Autosomal and Y Chromosome Multiplex STR Amplification System","authors":"Yu Hao,&nbsp;Zhang Zhe,&nbsp;Cui Yuling,&nbsp;Li Chenjia,&nbsp;Mo Xiaoting,&nbsp;Shang Lei,&nbsp;Song Wen,&nbsp;Jia Lina,&nbsp;Wang Jing,&nbsp;Li Wanshui,&nbsp;Bai Xue","doi":"10.1002/elps.8130","DOIUrl":"10.1002/elps.8130","url":null,"abstract":"<div>\u0000 \u0000 <p>To develop a 9-dye fluorescent autosomal and Y chromosome STR multiplex amplification system and verify its performance. Using the 9-dye fluorescent labeling technology, construct a multiplex amplification system that uses the autosomal and Y chromosome core and preferred loci recommended by GB/T 41009-2021 national standard as candidate loci, then evaluate its accuracy, sensitivity, species specificity, and other system performance index. A 9-dye fluorescent multiplex amplification system containing 70 loci, including Amel, D5S818, D21S11, D7S820, CS1FPO, D2S1338, D19S433, D8S1132, D2S441, VWA, D8S1179, D16S539, D10S1435, PentaD, D3S3045, TPOX, TH01, D22S1045, D18S51, FGA, D19S253, D10S1248, D6S477, D6S1043, D13S317, D12S391, D15S659, PentaE, D1S1656, D3S1358, DYS460, DYS389I, DYS390, DYS533, DYS392, DYS518, DYS557, DYS570, DYS458, DYS437, DYS385, GATA-H4, DYS576, DYS643, DYS508, DYS627, DYS456, DYS391, DYF404S1, DYS438, DYS448, DYF387S1, DYS596, DYS549, DYS399S1, DYS393, DYS635, DYS439, DYS19, DYS444, DYS449, DYS481, DYS527a/b, and DYS447 was successfully constructed. The various loci in this system are well balanced, with a sensitivity of 0.0625 ng and a certain ability to detect mixture samples. This system can obtain rich locus information through a single test and meet the needs of routine testing cases and DNA database construction. It has a good practical significance in forensic medicine.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 7-8","pages":"404-412"},"PeriodicalIF":3.0,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143566304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiscale Formulation for Assessment of Electroosmotic Flow in Paper-Based Microfluidics.","authors":"Joselynne C Salazar Bove, Sebastian Toro, Pablo A Kler","doi":"10.1002/elps.202400228","DOIUrl":"https://doi.org/10.1002/elps.202400228","url":null,"abstract":"<p><p>In this work, multiscale techniques to model the pressure driven and electroosmotic flows in porous materials with paper-like microstructures are studied and applied. The multiscale technique is based on the definition of a representative volume element (RVE) of the material, where the microstructure is built from connected channels, where the fluid moves inside the void of the porous material. For fluid flow, the velocity is solved under incompressible flow conditions in the Stokes regime at the microscale level, while the homogeneous Darcy problem is solved at the macroscale level. Similarly, for electroosmotic flow, the velocity and pressure are also solved at the microscale under incompressible flow conditions in the Stokes regime. However, in this case a Helmholtz-Smoluchowsky term is considered at the surface of the solid microstructure. Such term is calculated by solving the electric field via the charge conservation equation. Consequently, the electroosmotic velocity is included in the fluid dynamic problem as a boundary condition, significantly reducing the computational demand. Afterward, once the homogenized velocity field of the microscale problem is obtained, an effective pressure-based permeability and an effective electroosmotic permeability are estimated at the macroscale. To validate the results, a comparison is made with experimental data and other numerical studies reported in the literature for common papers used in microfluidics, such as Whatman <math><semantics><mo>#</mo> <annotation>$#$</annotation></semantics> </math> 1 and Munktel 00A, but also through comparisons with direct numerical simulations. Finally, we propose a microcell structure for representing such papers for matching fluid flow and electrical properties. With such topology, electroosmotic and mixed fluid flow are solved in order to demonstrate the capabilities of the multiscale technique for representing different phenomena involved in paper-based microfluidics. With these microcells will be also possible to predict other physicochemical phenomena which are important for paper-based microfluidics such as capillary imbibition or scalar dispersion, among others.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143566393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Self-Assembly Pipette Tip Restricted Access Mesoporous Polypyrrole Solid-Phase Extraction Coupled With Capillary Electrophoresis With Diode Array Detection for the Determination of Enalapril in Urine Samples","authors":"Iara Amorim Carvalho,&nbsp;Camilla Fonseca Silva,&nbsp;Keyller Bastos Borges","doi":"10.1002/elps.8126","DOIUrl":"10.1002/elps.8126","url":null,"abstract":"<p>A miniaturized self-assembly pipette tip with restricted access mesoporous polypyrrole solid-phase extraction, combined with capillary electrophoresis with diode array detection (CE-DAD), was developed to rapidly extract and determine enalapril from urine samples. The CE-DAD technique used 50 mmol L⁻¹ phosphate (pH 7) as the background electrolyte, a voltage of 13 kV, a 30 mbar hydrodynamic injection for 4 s, a capillary temperature of 25°C, and a wavelength of 195 nm to achieve a migration time of 6.3 min with satisfactory peak asymmetry and no interfering and/or baseline noise. The factors that influenced the extraction efficiency were evaluated and optimized: 750 µL sample at pH 7.5, 40 mg adsorbent, 250 µL hexane as a washing solvent, and 750 µL acetonitrile as eluent, resulting in recoveries around 74%. Linearity was acceptable in the 100–3000 ng mL⁻¹ range, and the selectivity and accuracy were also suitable. The limits of detection and quantitation were 30 and 50 ng mL⁻¹, respectively. The adsorbent effectively removed 87% of the proteins and may be reused three times. The analytical approach was successfully verified and used to analyze enalapril in urine samples collected from volunteers. Finally, the greenness of the researched technique was assessed using five measures that showed good eco-friendliness.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 7-8","pages":"376-387"},"PeriodicalIF":3.0,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.8126","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143566303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Obituary for Professor Dr. Joachim Klose to be Published in Electrophoresis","authors":"","doi":"10.1002/elps.8118","DOIUrl":"10.1002/elps.8118","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 7-8","pages":"478-479"},"PeriodicalIF":3.0,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143566414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dielectrophoresis Low and High Crossover Frequencies of Cancerous Exosomes.","authors":"Michihiko Nakano, Ryu Nakabayashi, Rie Koyama, Masafumi Inaba, Junya Suehiro","doi":"10.1002/elps.8128","DOIUrl":"https://doi.org/10.1002/elps.8128","url":null,"abstract":"<p><p>Exosomes, which constitute a subclass of extracellular vesicles found in various body fluids, play a crucial role in intercellular communication. Exosomes consist of matter derived from their host, such as nucleic acids, proteins, and membrane proteins. Cancer cell-derived exosomes have cancer-specific molecules and are found in the body fluids. As such, they are highly useful in liquid biopsies for cancer diagnosis. For diagnosis, rapid and simple determination of the sampled exosomes is important. Dielectrophoresis (DEP) manipulation has the potential for use in liquid biopsies. DEP behavior based on variations in exosomes has not been well investigated. In this study, the dielectrophoretic characteristics of exosomes derived from four cancer cell lines were investigated: MCF-7, PC-3, HT-29, and COLO 201. The DEP crossover frequencies of these exosomes were determined by directly observing their DEP behaviors at various suspension medium conductivities. Each exosome type exhibited distinct dielectrophoretic characteristics, with significant differences in the lower crossover frequencies linked to variations in the membrane properties and inner conductivity. A single-shell dielectric model was used to estimate the dielectric and physical parameters of the exosomes, aligning with the experimental results. These findings highlight the potential of DEP for specific exosome separation, enabling rapid and simple exosome-based liquid biopsies for cancer diagnosis.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143566381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simple and Convenient Three-Electrode Layout for Real-Time Electrorotation Measurement Upon Chemical Stimulation.","authors":"Masato Suzuki, Ryoga Yamada, Yuki Imou, Yushi Isozaki, Tomoyuki Yasukawa","doi":"10.1002/elps.8121","DOIUrl":"https://doi.org/10.1002/elps.8121","url":null,"abstract":"<p><p>We developed a novel electrorotation (ROT) device featuring a microwell array with three electrodes. This device allows to monitor the increase in membrane capacitance of cells subjected to chemical stimulation. The microwell array is integrated into the bottom of a fluidic channel and holds rotating cells during stimulation with a solution containing a chemical agent. Positive dielectrophoresis (p-DEP) effectively traps cells in microwells, whereas negative DEP (n-DEP) facilitates the rapid formation of single-cell presence. Alternating current (AC) voltages with a 120° phase shift applied across the three electrodes enable vertical and simultaneous rotation of cells. We observed a peak in rotation rate as a function of applied frequency, with the frequency spectrum shifting to lower frequencies as membrane capacitance increased. A positive correlation was identified between rotation rate and membrane capacitance, so monitoring in the low-frequency range is advantageous. Although n-DEP at lower frequencies risks removing cells from microwells, the continuous monitoring of the ROT rate during chemical stimulation was achieved by regulating the height of the ROT center of cells. We demonstrated the monitoring of membrane capacitance increase induced by Ca²⁺ influx from ionomycin. This simple configuration facilitates statistical analysis of ROT rates without fluorescent labeling, making it suitable for label-free assessments of white blood cells' responses to stimuli.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143515027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Contents: Electrophoresis 3–4'25","authors":"","doi":"10.1002/elps.202570022","DOIUrl":"https://doi.org/10.1002/elps.202570022","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"131-132"},"PeriodicalIF":3.0,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143496939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Editorial Board: Electrophoresis 3–4'25","authors":"","doi":"10.1002/elps.202570021","DOIUrl":"https://doi.org/10.1002/elps.202570021","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"129-130"},"PeriodicalIF":3.0,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.202570021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143497271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design Guidelines of Free-Flow Counterflow Gradient Focusing Device for Protein Fractionation.","authors":"Matthew Courtney, Oscar Manuel Carreno-Molina, Carolyn L Ren","doi":"10.1002/elps.8106","DOIUrl":"https://doi.org/10.1002/elps.8106","url":null,"abstract":"<p><p>Free-flow counterflow gradient focusing (FF-CGF) is a promising tool for protein fractionation. This work establishes guidelines to construct an FF-CGF system suitable for preparative applications. Numerical and analytical computations are undertaken to investigate design parameters, including device geometry, flow conditions, sample concentration, and applied voltage. This collective information serves as a design tool to guide the fabrication of the device, which, in this case, is constructed from polymethyl methacrylate (PMMA) and double-sided tape. Furthermore, the design tool also supports device operation and provides insights into the expected resolution and throughput. Experimental results, which demonstrated the ability of the designed chip to generate a uniform counterflow gradient while separating small molecules and proteins, validated the accuracy of the design guidelines.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of a cIEF Fractionation Workflow for Offline MS Analysis of Charge Variants of the Monoclonal Antibody Matuzumab","authors":"Antonia Wittmann,&nbsp;Yannick Wilke,&nbsp;Nicolas Grammel,&nbsp;Hermann Wätzig","doi":"10.1002/elps.8108","DOIUrl":"10.1002/elps.8108","url":null,"abstract":"<p>Biological drugs like monoclonal antibodies require careful analysis and characterization to ensure product quality, safety, and efficacy. Charge variants of the molecule are of key interest and are analyzed using imaged capillary isoelectric focusing (icIEF). However, deeper characterization of these variants poses challenges. Two workflows for their characterization exist: an ion-exchange chromatography method for variant collection before mass spectrometry (MS) analysis, which is labor-intensive, and direct coupling of CE to MS, which allows detailed structural characterization but has limitations, for example, due to incompatibilities with ES ionization using high BGE concentrations. This study evaluates a platform that fractionates charge variants for offline MS analysis. The suitability of a procedure in which analytical icIEF methods are converted into preparative cIEF fractionation methods by increasing the sample concentration and adding 20 mM arginine as a cathodic spacer was tested. After chemical mobilization and fraction collection, the identity of the fractions was determined by fluorescence measurement and reinjection of the protein-containing fractions, using the previously developed analytical icIEF method. MS was subsequently performed. The general suitability of the workflow was demonstrated using Matuzumab. Transferring the analytical method from a concentration of 0.2 to 1.2 mg/mL in fractionation yielded an identical number of peaks and visually comparable peak profiles. The preparative separation took 50 min, with an additional 25 min for mobilization and 45 s per fraction collection, totaling approximately 2.5 h. Verification of charge variant isolation was straightforward via analytical icIEF. Following fractionation, MS allowed for the identification of the main peaks. Preliminary results with other antibodies indicated that the concentration range for MS experiments needs further investigation. Future work will aim to optimize sensitivity, selectivity, analysis time, and reproducibility.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"240-249"},"PeriodicalIF":3.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.8108","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}