ELECTROPHORESIS最新文献

{"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}

{"title":"Numerical Simulation Noise and Its Significant Impact on the Dielectrophoretic Motion of Particles Near the Electrode Edges.","authors":"Aaditya Venkatesha Babu Bangaru, Stuart J Williams","doi":"10.1002/elps.8115","DOIUrl":"https://doi.org/10.1002/elps.8115","url":null,"abstract":"<p><p>Dielectrophoresis (DEP) has been extensively researched over the years for filtration, separation, detection, and collection of micro/nano/bioparticles. Numerical models have historically been employed to predict particle trajectories in three-dimensional (3D) DEP systems, but a common issue arises due to inherent noise near the edges of electrodes due to electric potential discontinuity, specifically when calculating electric field and gradient of electric field-squared, <math> <semantics> <mrow><mrow><mo>∇</mo> <mo>|</mo> <mi>E</mi></mrow> <msup><mo>|</mo> <mn>2</mn></msup> </mrow> <annotation>$nabla |E{|^2}$</annotation></semantics> </math> . This noise can be reduced to a certain extent with a finer mesh density but results near the electrode edge still have significant error. Realizing the importance of particle-electrode edge interactions prevalent in positive DEP systems, analytical solutions given by Sun et al. was incorporated to demonstrate an improved 3D model of interdigitated electrodes. The results of electric field and gradient of electric field-squared of the numerical model and the improved analytical 3D model were compared, within a simulation space of 50 µm height, 10 µm width, and 50 µm length with interdigitated electrodes of the same width and gap of 10 µm. The DEP particle trajectory error due to the noise was quantified for different particle sizes at various heights above the electrode edge. For example, at 5 V_rms, a trapped 500 nm particles exhibited a velocity error of 10⁴ µm/s (it should have been zero).</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":"143448547","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":"Enhanced Particle Trap: Design and Simulation of Pillar-Based Contactless Dielectrophoresis Microfluidic Devices","authors":"Peyman Torky Harchegani,&nbsp;Mohsen Mashhadi Keshtiban,&nbsp;Mahdi Moghimi Zand,&nbsp;Zahra Azizi","doi":"10.1002/elps.202400110","DOIUrl":"10.1002/elps.202400110","url":null,"abstract":"<div>\u0000 \u0000 <p>Contactless and conventional dielectrophoresis (DEP) microfluidic devices are extensively utilized in lab-on-a-chip applications, particularly for cell isolation and analysis. Nonetheless, these devices typically operate at low throughput and require high applied voltages, posing limitations for microfluidic cell isolation and separation. Addressing these challenges, this study explores the utilization of diverse micro-pillar geometries within the microfluidic device to augment THP-1 cell trapping efficiency numerically using FEM modeling. Furthermore, the simulations examine the influence of pillar gap and quantity on cell trapping efficiency in a contactless DEP device. Notably, elliptical pillars demonstrate superior cell trapping efficiency at elevated flow rates compared to alternative configurations, making the microchip more amenable for high-throughput cell separation, trapping, and isolation applications. Remarkably, employing elliptical pillars in a contactless DEP microfluidic chip yields nearly 100% cell trapping efficiency at higher flow rates. Ellipse configuration showed 122% higher cell trap efficiency at the maximum flowrate compare to the previous study with circular configuration. Additionally, it is observed that reducing the gap between pillars correlates with enhanced cell trapping efficiency. Simulation outcomes indicate that employing two rows of elliptical pillars with a 40-µm gap achieves optimal performance. The findings of this investigation underscore the importance of pillars in contactless DEP devices and provide valuable insights for future designs of such microfluidic devices.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"232-239"},"PeriodicalIF":3.0,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448542","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":"Determination of Withanolides and Withanosides in Ashwagandha Based Products Using HPLC-Drift-Tube-Ion-Mobility Quadrupole Time-of-Flight Mass Spectrometry.","authors":"Laura Zellner, Markus Wierer, Markus Himmelsbach, Benjamin Schatzmann, Bernhard Thalhamer, Christian W Klampfl","doi":"10.1002/elps.202400188","DOIUrl":"https://doi.org/10.1002/elps.202400188","url":null,"abstract":"<p><p>A total of 19 products based on extracts from Withania somnifera (L.) Dunal, better known by its more common name ashwagandha, and five products based on ashwagandha root powder were analyzed with respect to their content in the biologically relevant substances belonging to the group of withanolides and withanosides. Using HPLC coupled to drift-tube ion-mobility quadrupole time-of-flight mass spectrometry (DT-IM-QTOF-MS), 19 withanolides and withanosides could be tentatively identified. The comparison of the results from the quantitative analysis with the information on the product labels showed that the percentage of withanolides and withanosides deviated from the stated specifications by at least a factor of two and at most a factor of 35.</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":"143448539","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":"Characterizing Recent PDMS Changes in Electrokinetic-Based Microfluidic Devices' Performance and Manufacturing for Cell Sorting Applications.","authors":"Alexandra R Hyler, Dean E Thomas, Kyle S Kinskie, Kyle M Brown, Josie L Duncan, Jaka Cemazar, Jeff Schultz, Simeon Brown, Farhad Shiri, Steven A Soper, Nathan S Swami, Rafael V Davalos","doi":"10.1002/elps.8113","DOIUrl":"https://doi.org/10.1002/elps.8113","url":null,"abstract":"<p><p>Understanding cells from complex biological samples is vital to understanding cellular biology and medical applications. One evolving tool for cell sorting is the use of microfluidic devices to achieve higher precision and remove the need for labeling cell subpopulations. However, few microfluidic devices have been translated commercially beyond academic research often due to challenges in larger scale fabrication. Here, we initially investigated a compelling label-free microfluidic device with complex geometries to perform contactless dielectrophoresis (cDEP) for applications in enriching cell subpopulations in oncology, neurology, stem cells, and sample preparation. We began scaling the manufacturing of cDEP devices using Dow Sylgard 184, more commonly referred to as PDMS (polydimethylsiloxane). However, we began observing a new, dynamic bubble formation phenomenon which had significant impacts on device performance. Within just 5 min of exposure at typical experimental values, cell death was nearly 100%. Variables related to manufacturing, environment, equipment, personnel, raw materials sourcing, lithography methods and experimental conditions/parameters were systematically evaluated to find the root cause of the exacerbated bubble formation observed. Further, alternate polymers were sourced for manufacturing and experimental performance comparisons. All variables investigated failed to solve the significant decline in device performance and increase in cell death. Upon completing chemical analysis in this work, we conclude that the decline in device performance was a direct result of changes to the expected PDMS properties and composition. Despite these challenges, our robust quality control combined with experimental protocols to remove bubbles from the cDEP devices achieved consistent experimental performance including 2-3 h run times and >90% cell viability after sorting. These new PDMS behaviors will need to continue to be monitored and controlled to ensure consistency in experimentation, application and commercialization feasibility for a wide variety of microfluidic device designs and applications.</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":"143448532","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":"3D-Printed Microfluidic-Based Cell Culture System With Analysis to Investigate Macrophage Activation.","authors":"Major A Selemani, Giraso Keza Monia Kabandana, Chengpeng Chen, R Scott Martin","doi":"10.1002/elps.8109","DOIUrl":"https://doi.org/10.1002/elps.8109","url":null,"abstract":"<p><p>In this paper, we describe the development of 3D-printed microfluidic cell culture devices that can be coupled with a circulation system to study the dynamics of both intracellular and extracellular (release) processes. Key to this approach is the ability to quantitate key analytes on a minutes timescale with either on-line (in this study, quantitating nitric oxide production using an amperometric flow cell) or off-line (in this work, quantitating intracellular itaconate production with LC/MS) analytical measurements. To demonstrate the usefulness of this approach, we chose to study macrophage polarization as a function of the extracellular matrix (silk) fiber size, a major area of research in tissue engineering. It was found that the use of larger fibers (1280 nm vs. smaller 512 nm fibers) led to increases in the production of both nitric oxide and itaconate. These findings set the foundation for future research for the creation of finely tuned microfluidic 3D cell culture approaches in areas where flow and the extracellular matrix play a significant role in barrier transport and where integrated analysis of key markers is needed.</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":"143448530","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":"The Dynamic Behavior of Charged Particles During the Relaxation Process of Electrophoretic Displays","authors":"Sheng Mai,&nbsp;Xuekai Liu,&nbsp;Juncheng Zhao,&nbsp;Zhihang Yu,&nbsp;Jing Jin,&nbsp;Liuyong Shi,&nbsp;Teng Zhou","doi":"10.1002/elps.202400231","DOIUrl":"10.1002/elps.202400231","url":null,"abstract":"<div>\u0000 \u0000 <p>Electrophoretic displays (EPDs) are commonly employed in applications like e-books and electronic price tags due to their benefits of minimal power consumption, excellent contrast, and broad viewing angles. This article establishes a dynamic model for nanoparticles after the removal of the applied electric field. The model combines the Poisson equation, the Navier–Stokes equation, and the Nernst–Planck equation. The Arbitrary Lagrangian–Eulerian method is applied to simulate nanoparticle diffusion motion under varying conditions, such as solution viscosity, particle radius, and reverse micelle radius, after the electric field is removed. The results indicate that after the electric field is removed, high-viscosity solutions exert a stronger hindrance on the particles, resulting in a shorter displacement over the same time period. With equal charge, smaller particle radius exhibits higher surface charge density, allowing them to travel further within the same time frame. Additionally, a smaller reverse micelle radius facilitates the rapid neutralization of surface charge on the particles, thereby limiting their diffusion distance. These findings provide theoretical support for a deeper understanding of the operating mechanism of EPDs.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"221-231"},"PeriodicalIF":3.0,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143406433","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":"Determination of Carbohydrates in Syngonium podophyllum Biomass by Capillary Electrophoresis After Saccharification Process Under Different Sample Preparation Techniques.","authors":"Rafaella Silva Aredes, Daniel Garcez Santos Quattrociocchi, Vinicius Gomes Madriaga, Giovanni Offrede, Maria Clara de Souza Paranhos, Thiago de Melo Lima, Lucas Mattos Duarte, Flávia Ferreira de Carvalho Marques","doi":"10.1002/elps.202400232","DOIUrl":"https://doi.org/10.1002/elps.202400232","url":null,"abstract":"<p><p>To achieve a more sustainable future, it is crucial to conduct studies on lignocellulosic biomass as a source of carbohydrates, ultimately serving as feedstocks for platform molecules that can be converted into biofuels, biopolymers, and chemicals. Urban residue lignocellulosic biomass, exemplified by Syngonium podophyllum, shows promise as an alternative source. This study employed capillary zone electrophoresis with indirect UV detection as an environmentally friendly method for determining xylose and glucose after treating, depolymerizing, and saccharifying Syngonium biomass. Three different hydrolysis methods yielded distinct outcomes. Heating under reflux treatment resulted in a 37 ± 3 mg g^-1 xylose content, whereas ultrasonication significantly increased xylose yield, measuring 110 ± 5 mg g^-1. Interestingly, these two methods did not detect glucose, suggesting they may be less effective at producing detectable monosaccharide levels. On the other hand, microwave-assisted hydrolysis produced the highest xylose content at 290 ± 13 mg g^-1 and glucose at 124 ± 6 mg g^-1. These results highlighted the effectiveness of microwave-assisted hydrolysis in converting carbohydrates from the urban Syngonium biomass, demonstrating its potential in processing lignocellulosic biomass through a cheap and sustainable route.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143406410","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":"Saliva Proteomics Highlights Potential BMI-Related Biomarkers for Forensic and Medical Research","authors":"Shunyi Tian,&nbsp;Qiyan Wang,&nbsp;Xiaolan Huang,&nbsp;Shengjie Zhu,&nbsp;Hongling Zhang,&nbsp;Zheng Ren,&nbsp;Yubo Liu,&nbsp;Meiqing Yang,&nbsp;Jiang Huang,&nbsp;Xiaoye Jin","doi":"10.1002/elps.202400206","DOIUrl":"10.1002/elps.202400206","url":null,"abstract":"<div>\u0000 \u0000 <p>Obesity and underweight pose significant threats to human health as non-communicable diseases. In addition, body shape (like obesity or emaciation) is an important investigative clue in forensic practice. Body mass index (BMI) is a common indicator to reflect body shape of an individual. However, there is a lack of rapid, simple, and effective methods for identifying different BMI individuals. This research aimed to delve into the correlations between salivary proteins and BMI. A total of 418 differential expression proteins (DEPs) were identified through four-dimensional data independent acquisition quantitative proteomics analysis. The Kyoto Encyclopedia of Genes and Genomes analysis revealed that DEPs were primarily involved in oxidative phosphorylation, protein processing in the endoplasmic reticulum, and cholesterol metabolism pathways. Finally, we identified 17 protein markers that were correlated with BMI. Two machine-learning models (random forest and support vector machine) were also built based on these 17 markers. Obtained results demonstrated the efficacy of these 17 protein markers in accurately distinguishing different BMI individuals. In conclusion, our study not only provides potential salivary protein markers for identifying obesity and underweight individuals, it could also present a novel method for inferring BMI of saliva-related samples in forensic case investigation.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"250-257"},"PeriodicalIF":3.0,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143406416","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}