Anne B. Ries, Maximilian N. Merkel, Kristina Coßmann, Marina Paul, Robin Grunwald, Daniel Klemmer, Franziska Hübner, Sabine Eggensperger, Frederik T. Weiß
{"title":"Universal Study Design for Instrument Changes in Pharmaceutical Release Analytics","authors":"Anne B. Ries, Maximilian N. Merkel, Kristina Coßmann, Marina Paul, Robin Grunwald, Daniel Klemmer, Franziska Hübner, Sabine Eggensperger, Frederik T. Weiß","doi":"10.1002/elps.70004","DOIUrl":"10.1002/elps.70004","url":null,"abstract":"<p>In pharmaceutical quality control (QC), analytical methods need to maintain release analytics over decades. Over a product's lifecycle, vendors may update instrument hardware and/or software, or a switch between vendors may become necessary upon discontinuation of an instrument. Both situations pose a challenge to pharmaceutical QC.We designed an efficient instrument comparability study to gain a comprehensive understanding of potential performance differences between instruments and therefore rationalize the risk assessment and decision process for a path forward. The results may either point out whether a full or partial re-validation is necessary or whether a science-based update can be pursued based on the data generated in the study. The study design is universally applicable to a substantial range of release analytical methods. In a straightforward setup of two experiments with the new instrument, a statistically meaningful data set is generated for comparison with available historical or validation data of the original instrument. In a Good Manufacturing Practice (GMP) environment, we implemented the study design in a benchmark study comparing the ICE3 and Maurice C imaged capillary isoelectric focusing (icIEF) instruments. The core-study confirmed equal or better performance of the Maurice C in all parameters and serves as a basis for seamless continuation of release icIEF measurements on Maurice C.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 11-12","pages":"669-678"},"PeriodicalIF":2.5,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144495222","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}
Timothy Blanc, Hermann Wätzig, Cari Sänger - van de Griend
{"title":"Peak Integration of Electropherograms in GMP and Research Labs: Navigating Increased Scrutiny Amid Data Integrity Audits and Inspections","authors":"Timothy Blanc, Hermann Wätzig, Cari Sänger - van de Griend","doi":"10.1002/elps.70002","DOIUrl":"10.1002/elps.70002","url":null,"abstract":"<p>Capillary electrophoresis (CE) often offers superior separation relative to chromatography for macromolecules like monoclonal antibodies (mAbs), a major pharmaceutical class. However, electropherogram baselines pose challenges that traditional chromatography algorithms cannot address, requiring complex integration processes. Integration in good manufacturing practice (GMP) laboratories is critically important and has become a focus of data integrity-centric regulatory inspections. Electropherogram integration challenges, the increased use of CE, data systems developed for chromatograms rather than electropherograms, and the increased regulatory scrutiny call for a resolution. This necessity also extends to R&D, clinical, and academic labs. This review examines authoritative sources such as pharmacopoeias, World Health Organization (WHO), Parenteral Drug Association (PDA), and scientific literature. However, none address electropherogram integration. These sources concur that companies should develop integration policies and SOPs. Training programs must ensure analysts are proficient in integration techniques and reviewers are appropriately qualified to assess integrations. Integration parameters must be captured, including slope sensitivity, smoothing factors, and timed events like peak start and stop and baseline correction. Analytical procedures (APs) should include illustrations that define proper integration. Although automatic integration is preferred for its efficiency and objectivity, it is not always accurate. Therefore, manual integration should be permitted under specific conditions, with all settings and iterations documented, justified, and reviewed. Industry collaboration is proposed to create practical integration guidelines specifically for CE.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 11-12","pages":"653-668"},"PeriodicalIF":2.5,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144332651","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}
Emmanuelle Lipka, Roberto Dallocchio, Barbara Sechi, Mikheil Rukhaia, Giorgi Jibuti, Bezhan Chankvetadze, Victor Mamane, Paola Peluso
{"title":"Insights Into the Enantioseparation of Polyhalogenated 4,4′-Bipyridines With a Cellulose Tris(3,5-Dimethylphenylcarbamate)-Based Chiral Column by Using Supercritical Fluid Chromatography","authors":"Emmanuelle Lipka, Roberto Dallocchio, Barbara Sechi, Mikheil Rukhaia, Giorgi Jibuti, Bezhan Chankvetadze, Victor Mamane, Paola Peluso","doi":"10.1002/elps.8156","DOIUrl":"10.1002/elps.8156","url":null,"abstract":"<p>In the last decade, by integrating experimental and computational analyses, it was demonstrated that halogen bond (HaB) may contribute to binding and enantiorecognition mechanisms underlying the HPLC enantioseparation of halogenated chiral analytes by using cellulose <i>tris</i>(3,5-dimethylphenylcarbamate) (CDMPC)-based chiral columns and <i>n</i>-hexane-based mixtures as mobile phases. When used as a pivotal component of the mobile phase in supercritical fluid chromatography (SFC), carbon dioxide is often considered as an <i>n</i>-hexane-like nonpolar solvent because of its low dielectric constant and zero molecular dipole moment. On the other hand, carbon dioxide may also serve as hydrogen bond (HB) and HaB acceptor due to the presence of nonbonding electrons on the two oxygen atoms, interacting with analyte enantiomers, chiral selectors, and co-solvents. On this basis, we report herein the results of a study aiming at evaluating the impact of using carbon dioxide in SFC in place of <i>n</i>-hexane in HPLC on halogen-dependent enantioseparations by using atropisomeric halogenated 4,4′-bipyridines as analytes and Lux Cellulose-1 as CDMPC-based chiral column. The experimental investigation was complemented by a computational study performed using (a) quantum mechanics (QM) calculations to map and quantify noncovalent interactions possibly underlying the contact of the analytes with carbon dioxide and with the distinctive pendant groups of the CDMPC and (b) molecular dynamics (MD) simulations to visualize noncovalent interactions acting in the analyte <b>1</b>/CDMPC chromatographic system in different media. The use of MD simulations to model enantioseparations performed in carbon dioxide-based media was not reported in the literature so far.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 11-12","pages":"702-715"},"PeriodicalIF":2.5,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/elps.8156","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191619","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":"Quantifying Critical Quality Attributes of Protein Therapeutics by Sodium Dodecyl Sulfate-Capillary Gel Electrophoresis With Native Fluorescence Detection.","authors":"Zaifang Zhu","doi":"10.1002/elps.8154","DOIUrl":"10.1002/elps.8154","url":null,"abstract":"<p><p>[08-08-2025]: This article published in Early View in error and has been taken down temporarily. The article will republish shortly.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191620","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}
Cemil Aydoğan, Zeynep Günyel, Sarah Alharthi, Hakiye Aslan, İbrahim Y Erdoğan, Ziad El Rassi
{"title":"Rapid Separation and Analysis of Exosomes in Milk Sample by on-Line Nano-Liquid Chromatography.","authors":"Cemil Aydoğan, Zeynep Günyel, Sarah Alharthi, Hakiye Aslan, İbrahim Y Erdoğan, Ziad El Rassi","doi":"10.1002/elps.8155","DOIUrl":"https://doi.org/10.1002/elps.8155","url":null,"abstract":"<p><p>Exosomes are very small vesicles of 30-150 nm average particle size and hold great potential in new therapeutic applications. The aim of this study is to develop a new method for the isolation and analysis of exosomes in milk via hydrophobic interaction chromatography (HIC), including salting-out process in nano-liquid chromatography (nano-LC). On the basis of this approach, a trap column combined with graphene oxide (GO)-based monolithic nano-column was used for on-line analysis of exosomes in nano-LC. The monolith was prepared by an in situ polymerization of butyl methacrylate (BMA), ethylene glycol dimethacrylate (EDMA), and methacryloyl graphene oxide nanoparticles (MGONPs). The final solution was introduced into a fused silica capillary with a 50 µm i.d. for polymerization. After preparation, the column was further modified with dimethyloctadecylchlorosilane (DODCS) to increase its hydrophobicity. The characterization of monolith was performed using scanning electron microscopy (SEM) and chromatographic examination. The final monolith was applied for the isolation and analysis of exosomes in milk via HIC-nano-LC. Nanoparticle tracking analysis (NTA), SEM, and Fourier transform-infrared (FT-IR) for the tandem characterization of milk exosomes were used, whereas step gradient elution was employed for HIC. The results demonstrated good ability to isolate exosomes from milk with three dilution factors, and a loading capacity of 7.3 ± 02 × 10<sup>11</sup> exosomes could be obtained using the on-line nano-LC system. The developed method holds many advantages and may be adapted for the isolation of exosomes from a diverse range of media.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144119225","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}
Saurabh K. Maurya, Santanu Saha, Partha P. Gopmandal
{"title":"Comparative Study on the Electrophoresis of Soft and Semisoft Nanoparticles","authors":"Saurabh K. Maurya, Santanu Saha, Partha P. Gopmandal","doi":"10.1002/elps.8149","DOIUrl":"10.1002/elps.8149","url":null,"abstract":"<div>\u0000 \u0000 <p>There is abundant literature that deals with the electrophoresis of biocolloids and environmental entities. Most of these nanoparticles can be classified as soft particles, which are core–shell structured in nature. Most existing studies assume the inner core of the soft particles to be rigid in nature. However, there are various core–shell structured nanoparticles for which the inner core is semisoft in nature, which allows ion penetration but restricts fluid flow. In addition, the distribution of the peripheral layer of the soft (soft particle with rigid inner core) or semisoft (soft particle with semisoft inner core) particle is not necessarily uniform. In the present article, we consider the electrophoresis of soft and semisoft particles with the diffuse distribution of monomers across the peripheral shell layer. The mathematical model adopted here is based on the Poisson–Boltzmann equation for the electric double-layer potential and Darcy–Brinkman extended Stokes equation for fluid flow. The study is carried out considering a weak electric field assumption, which allows us to linearize the set of equations using perturbation analysis. A finite difference–based method is adopted to solve the perturbed set of equations and thus to calculate the electrophoretic mobility. The results are presented to indicate the difference in electrophoretic mobility of soft and semisoft particles under similar electrostatic conditions. We have further indicated the dependence of pertinent parameters on the electrophoretic mobility of soft or semisoft nanoparticles.</p></div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 11-12","pages":"752-761"},"PeriodicalIF":2.5,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143997865","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":"Using Electroosmotic Pumps to Control the Flow Velocity in Cotton Thread-Based µTADs.","authors":"Xionghui Li, Haonan Li, Xuanying Liang, Zejingqiu Chen, Muyang Zhang, Qinghao He, Jie Zhou, Zitao Feng, Yeqian Liu, Xinyi Chen, Huilin Chen, Zitong Ye, Ziwei Huang, Xingwei Zhang, Huiru Zhang, Lok Ting Chu, Weijin Guo","doi":"10.1002/elps.8153","DOIUrl":"https://doi.org/10.1002/elps.8153","url":null,"abstract":"<p><p>Flow velocity control is of great interest for passive microfluidic devices that are used in point-of-care diagnostics. Various methods have been developed for the flow velocity control of microfluidic paper-based analytical devices (µPADs), whereas fewer attempts have been made for microfluidic thread-based analytical devices (µTADs). In this research, we attempt to control the flow velocity in cotton thread-based µTADs with electroosmotic pumping. Utilizing electroosmotic pumps, the flow velocity in the cotton thread-based µTADs can be decreased or increased by 13% and 106%, respectively. Moreover, the dynamic control of the flow velocity in the cotton thread-based µTADs is achieved by adjusting the real-time magnitude and direction of the voltage. Furthermore, we demonstrate that electroosmotic pumps can be used to overcome the hydrophobic valves in the cotton thread-based µTADs. We show that the delivery sequence of different liquid samples for a three-branch µTAD can be controlled. Finally, we show the potential application in lithium detection with a colorimetric assay. This method for flow velocity control shows promise for customizing the flow velocity and reaction time of cotton thread-based µTADs, and this method can potentially increase the sensitivity of detection.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143992450","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":"Analysis of Single Nuclei in a Microfluidic Cytometer Towards Metaphase Enrichment.","authors":"Cristian Brandi, Adele De Ninno, Filippo Ruggiero, Valentina Mussi, Mauro Nanni, Federica Caselli","doi":"10.1002/elps.8152","DOIUrl":"https://doi.org/10.1002/elps.8152","url":null,"abstract":"<p><p>Identifying analyzable metaphase chromosomes is crucial for karyotyping, a common procedure used by clinicians to diagnose genetic disorders and some forms of cancer. This task is often laborious and time-consuming, making it essential to develop automated, efficient, and reliable methods to assist clinical technicians. In this work, an original label-free microfluidic approach to identify potential metaphases is developed that uses impedance-based detection of individual flowing nuclei and machine-learning-based processing of synchronized high-speed videos. Specifically, impedance signals are used to identify nucleus-containing frames, which are then processed to extract the contour of each nucleus. Feature extraction is then performed, and both unsupervised and supervised classification approaches are implemented to identify potential metaphases from those features. The proposed framework is tested on K562 cells, and the highest classification accuracy is obtained with the supervised approach coupled with a feature selection procedure and the Synthetic Minority Over-sampling Technique (SMOTE). Overall, this study encourages future developments aimed at integrating a sorting functionality in the device, thus achieving an effective microfluidic system for metaphase enrichment.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143973171","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}
Pauline Zimmer, Oleh Andreiev, Marion Costella, Emmanuelle Laurenceau, Jean-François Bryche, Jean-Pierre Cloarec, Michael Canva, Marie Frénéa-Robin, Julien Marchalot
{"title":"Electrohydrodynamic Vortex Imaging: A New Tool for Understanding Mass Transfer in Surface-Based Biosensors.","authors":"Pauline Zimmer, Oleh Andreiev, Marion Costella, Emmanuelle Laurenceau, Jean-François Bryche, Jean-Pierre Cloarec, Michael Canva, Marie Frénéa-Robin, Julien Marchalot","doi":"10.1002/elps.8137","DOIUrl":"https://doi.org/10.1002/elps.8137","url":null,"abstract":"<p><p>Surface-based biosensor performance is generally limited by mass transfer, especially when detecting low-concentrated species. To address this, dielectrophoresis (DEP) and alternating current electroosmosis (ACEO) can be combined to enhance mass transfer, increasing the target concentration near the sensor. This article presents a method for real-time direct imaging of electrohydrodynamic (EHD) effects on a microparticle suspension within a microfluidic chamber enclosed by two opposing electrodes. This top-bottom configuration was poorly studied in the literature for ACEO. The system presented thereby allows measurements of fluid flow profiles perpendicular to the electrode surface. The velocity of fluorescent latex microsphere tracers was measured as a function of signal frequency, potential, and electrolyte conductivity. This setup enables direct observation of vortices and particle-depleted areas, offering a valuable tool for selecting optimal input parameters-such as electric field, conductivity, and electrode dimensions-to efficiently concentrate microparticles near the sensor. Additionally, a numerical model developed in COMSOL and adapted for this top-bottom configuration enhances understanding of key parameters influencing EHD phenomena.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143997931","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}
Debmalya Roy, Aniruddha Guha, James Yates, Suman Chakraborty
{"title":"Precise Shrinkage of Silicon Nitride Nanopores Via Externally Sourced Hydrocarbons.","authors":"Debmalya Roy, Aniruddha Guha, James Yates, Suman Chakraborty","doi":"10.1002/elps.8150","DOIUrl":"https://doi.org/10.1002/elps.8150","url":null,"abstract":"<p><p>Solid-state nanopores (SSNPs) are progressively gaining importance in biomolecular sensing and ionic circuit applications. Unlocking their full potential, however, requires the development of fabrication techniques that enable precise control over their sizes and shapes. Electron-beam (EB) shrinking provides precise, real-time feedback and is ideally suited to address these requirements. However, it necessitates an initial pore diameter smaller than the membrane thickness for effective shrinking without material addition. Typical focused ion beam (FIB)-drilled pores in silicon nitride membranes often fail to meet these requirements. Alternative efforts towards mitigating these bottlenecks through deploying hydrocarbon-mediated EB shrinkage face challenges due to uncontrolled carbon contamination or a lack thereof in cleaner transmission electron microscope (TEM) chambers. To address these challenges, here we report an alternative approach of high-precision hydrocarbon-mediated EB shrinking with hydrocarbons sourced externally through controlled surface reactions on exposure to ethanol. This provides several decisive advantages, including the reduction of pore diameters much larger than the membrane thickness and controlled shrinking in cleaner environments without contaminations. These measures accelerate nanopore fabrication, improve its predictability by eliminating the dependence on variable carbon contamination in vacuum chambers, and provide high-resolution live feedback during dimension tuning. As a result, our method supports the large-scale production of nanopores with analyte-specific, tuneable dimensions. This capability is particularly imperative for low-noise biomolecular sequencing applications that leverage electrically-modulated transport and sensing over nanoscales. These features could pave the way for the broader application of SSNPs, addressing long-standing challenges in their fabrication and functionalisation that remained unresolved thus far.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143962762","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}