ELECTROPHORESIS最新文献

{"title":"A Separation Buffers Platform Set to Facilitate CZE Charge Heterogeneity Method Development for Monoclonal Antibodies.","authors":"Joanne Baxter, Kevin Waltham, Lori Fitton, Cari E Sänger-van de Griend","doi":"10.1002/elps.70089","DOIUrl":"https://doi.org/10.1002/elps.70089","url":null,"abstract":"<p><p>Monoclonal antibodies (mAbs) are complex therapeutic proteins exhibiting heterogeneity due to post-translational modifications, making charge variant analysis essential for defining critical quality attributes. Although ion-exchange chromatography and capillary isoelectric focusing are established techniques, they require extensive optimisation, whereas the widely adopted ε-aminocaproic acid-based capillary zone electrophoresis (CZE) (eACA-CZE) method provides a simpler, robust platform. However, its performance can be limited for mAbs whose charge profiles or isoelectric point (pI) values fall outside the method's optimal range. To expand CZE capabilities while maintaining a platform approach, we developed a series of background electrolytes (BGEs) spanning a range of pH values, polyamine concentrations and buffering capacities. These BGEs were formulated for chemical compatibility, long-term stability and fixed-component composition to ensure consistent pH and reproducible currents. Using a pH 5.7 BGE as the starting point, key components were optimised across the full buffer set. A multivariate approach showed that combining these buffers enhanced resolution, resolving additional impurity peaks in 9 of 10 mAbs. This work establishes a flexible toolkit for screening and optimising charge-variant resolution across diverse mAbs.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147503407","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 3'26","authors":"","doi":"10.1002/elps.70090","DOIUrl":"https://doi.org/10.1002/elps.70090","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 3","pages":"229-231"},"PeriodicalIF":2.5,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70090","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147567937","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":"Concentration Approaches for mRNA and mRNA–LNP Formulations: Enabling mRNA Integrity Quantification in Low-Concentration Formulations","authors":"Shang-Yin Wu,&nbsp;Yun-Jie He,&nbsp;Zhi-Jun Guo,&nbsp;Min Wang,&nbsp;Chang-Yun Xiong,&nbsp;Tingjun Hou,&nbsp;Bin Di,&nbsp;Wei-Jie Fang","doi":"10.1002/elps.70076","DOIUrl":"10.1002/elps.70076","url":null,"abstract":"<div>\u0000 \u0000 <p>mRNA-based vaccines and self-amplifying mRNA (saRNA) have gained growing attention for disease prevention and treatment, but precise detection of low-concentration RNA (including mRNA–lipid nanoparticles, mRNA–LNPs) stability and integrity remains challenging—limiting quality control of RNA-based therapeutics. Capillary electrophoresis (CE)-based instruments show potential, yet suitable concentration strategies for low-abundance, labile RNAs are lacking. This study established two distinct concentration methods (freeze-drying and ultrafiltration) applicable to mRNA, mRNA–LNPs, and saRNA formulations, enabling the conversion of dilute solutions to high-concentration preparations while preserving the structural and molecular integrity of the target nucleic acids. Subsequent stability evaluations and conventional high-performance liquid chromatography analyses of the concentrated products verified their superior storage stability and strong practical applicability. This article aims to reduce the difficulty of RNA integrity assessment and improve the accuracy of detecting various low-concentration RNA samples that may arise in the future.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 3","pages":"261-272"},"PeriodicalIF":2.5,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146084931","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 Monoethylene Glycol in Gas Condensate Samples by Microchip Micellar Electrokinetic Chromatography Integrated With Capacitively Coupled Contactless Conductivity Detection","authors":"Maurício M. L. Pereira,&nbsp;Kariolanda C. A. Rezende,&nbsp;Iris Medeiros Junior,&nbsp;Bruno Charles do Couto,&nbsp;Rogerio M. Carvalho,&nbsp;Claudimir L. do Lago,&nbsp;Wendell K. T. Coltro","doi":"10.1002/elps.70070","DOIUrl":"10.1002/elps.70070","url":null,"abstract":"<p>This study describes the use of microchip micellar electrokinetic chromatography (MEKC) integrated with capacitively coupled contactless conductivity detection (C⁴D) for the determination of monoethylene glycol (MEG) in gas condensate samples. The samples were subjected to a liquid–liquid extraction step and then analyzed by chip-based MEKC-C⁴D. For this purpose, sodium dodecyl sulfate (SDS) was used as a surfactant at a concentration of 30 mmol L⁻¹ added in 50 mmol L⁻¹ phosphate (pH = 9.0). Samples were introduced into microchannels through floating injection mode by applying a voltage of 600 V during 10 s. Separations were performed under an electric field of 82 V cm⁻¹ and monitored by C⁴D measurements recorded applying a 1200-kHz frequency sinusoidal wave with 20-V_pp excitation voltage. The proposed methodology employing MEKC-C⁴D revealed a linear behavior (r² ≥ 0.99) in the MEG concentration range between 150–450 µmol L⁻¹ and LOD equal to 33 µmol L⁻¹. Three gas condensate samples were then analyzed, and the achieved MEG concentration values ranged from 173 to 213 µmol L⁻¹. Recovery experiments provided values between 89 and 102%. Based on the results reported in this study, MEKC-C⁴D devices have demonstrated to be a promising and ecological analytical tool for MEG analysis with huge potential for in-field applications.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 3","pages":"235-242"},"PeriodicalIF":2.5,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13005714/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145997649","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":"Editorial–Dielectrophoresis 2025","authors":"Federica Caselli,&nbsp;Georg R. Pesch","doi":"10.1002/elps.70066","DOIUrl":"10.1002/elps.70066","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 3","pages":"232-234"},"PeriodicalIF":2.5,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146141336","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":"Enantioselective Determination of Amphetamine-Type Stimulants in Environmental Waters Using SPE and CE–MS/MS","authors":"Pol Clivillé-Cabré,&nbsp;Francesc Borrull,&nbsp;Carme Aguilar,&nbsp;Marta Calull","doi":"10.1002/elps.70074","DOIUrl":"10.1002/elps.70074","url":null,"abstract":"<p>The growing concern over drug abuse, particularly involving amphetamine-type substances (ATS), has led to their monitoring in environmental waters. These chiral compounds, commonly found in river water and wastewater treatment plant (WWTP) effluents, require analytical methods capable of enantiomeric discrimination, as their enantiomers can exhibit different pharmacological and environmental behaviours. A method based on capillary electrophoresis (CE) coupled with tandem mass spectrometry (MS/MS) was developed using a dual cyclodextrin (CD) system consisting of 0.1% 2-hydroxypropyl-β-CD and 0.1% γ-CD in the background electrolyte (BGE), which enabled baseline resolution of the target enantiomers of the ATS under study. Samples were pretreated with solid-phase extraction using a mixed-mode cation exchange sorbent, ExtraBond SCX. Samples of 100 mL for WWTP influent and 250 mL for river water and WWTP effluent were extracted and then eluted with 5 mL of 5% NH₄OH in methanol. Recoveries ranged between 40% and 67% for all amphetamines studied, with detection limits between 0.1 and 0.8 µg/L. Analysis of environmental samples from the Ebre River and WWTPs in Reus and Tarragona (Catalonia, Spain) confirmed the presence of some of the target compounds. Both enantiomers of 3,4-methylenedioxymethamphetamine (MDMA) were determined in WWTP influents and effluents, whereas <i>R</i>-amphetamine was quantified in an influent sample. No target compounds were detected in the analysed river water samples. These findings demonstrate the potential of the developed chiral CE–MS/MS method for robust enantiomeric profiling in environmental waters, attributed to efficient separation based on differences in effective mobility (<i>µ</i>_eff), electroosmotic flow (<i>µ</i>_eo) and selective interactions with CD chiral selectors. The method showed moderate adherence to green analytical principles, achieving an AGREE score of 0.47. Its environmental advantages include the use of CE, minimal solvent consumption and non-toxic chiral selectors, offering a more suitable alternative to more traditional LC-based methods.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 3","pages":"250-260"},"PeriodicalIF":2.5,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13005715/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146040604","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":"Determination of AAV Capsid Protein Ratios by Sodium Dodecyl Sulfate Capillary Electrophoresis (CE-SDS)—UV Absorption Method","authors":"Yiran Li,&nbsp;Yufei Sun,&nbsp;Xiang Li,&nbsp;Hua Bi,&nbsp;Dening Pei,&nbsp;Zhihao Fu,&nbsp;Youxue Ding,&nbsp;Yue Pan,&nbsp;Xi Qin,&nbsp;Chenggang Liang,&nbsp;Lan Wang","doi":"10.1002/elps.70073","DOIUrl":"10.1002/elps.70073","url":null,"abstract":"<p>Recombinant adeno-associated viruses (AAVs) have become a leading platform for in vivo gene therapy. The AAV capsid is assembled from 60 individual protein subunits derived from three overlapping viral proteins (VPs): VP1, VP2, and VP3. The VP's ratio distribution affects the capsid trafficking and transgene expression. Quantitative analysis of the ratio is important in monitoring the potency of the AAVs. Sodium dodecyl sulfate capillary electrophoresis (CE-SDS) offers automated and quantitative determination of the VP ratio distribution, and the method has been widely used in the application. The CE-SDS method described was designed to ensure the consistent determination of AAV VP ratios across various sample concentrations. UV-absorption detection was chosen for this purpose as it negated the need for any labeling in the process. To compensate for the lower sensitivity associated with UV-absorption detection, methanol precipitation was employed during sample preparation, and a stacking injection technique was optimized for the samples in the study under CE conditions. These two techniques were found to be highly compatible. The method demonstrated robust linearity for AAV serotype 9 concentrations and exhibited good multiday precision. The VP ratios were successfully determined in AAV9 samples across a concentration range of 6.15 × 10¹² to 1.85 × 10¹³ GC/mL. The relative standard deviation (RSD%) for VP ratio determination within this concentration range was maintained below 7% for VP3, VP2, and VP1. Additionally, this method was further applied to other AAV serotypes, including AAV2 and AAV8, illustrating its versatility and broad applicability.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 3","pages":"243-249"},"PeriodicalIF":2.5,"publicationDate":"2026-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13005713/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146164708","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":"A Continuum Modeling Approach to Nanoparticle Polarizability Characterization Using Fluorescence Intensity Profiles.","authors":"Mens Wout, Wood Jeffery A, Liu Chengxun, Lagae Liesbet, Willems Kherim","doi":"10.1002/elps.70087","DOIUrl":"https://doi.org/10.1002/elps.70087","url":null,"abstract":"<p><p>The use of dielectrophoresis (DEP) for the manipulation of electrically polarizable particles has received a lot of interest in the past decades. Theories relying on the Clausius-Mossotti (CM) factor describe the DEP behavior of macroscale particles with great accuracy. However, under nanoscale conditions, these classical CM factor theories can break down. Therefore, experimental characterization of particle polarizabilities is of utmost importance. We present an integrated experimental-computational methodology that allows the quantification of effective particle polarizabilities from DEP experiments. The methodology is based on the comparison of experimental and simulated concentration profiles of fluorescent nanoparticles captured by a set of electrodes. We obtain effective particle polarizabilities for 52 and 105 nm particles that agree well with values predicted by CM theory. The considered particles hence serve as a calibration model for our approach, demonstrating that realistic particle polarizabilities can be obtained and paving the way for polarizability quantification for particles where existing theories are inadequate.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147485097","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 Biomimetic Protein Immobilization Method for Studying Drug-Protein Interactions Based on Affinity Capillary Electrochromatography.","authors":"Jiaping Cui, Yawen Tian, Xinru Fu, Xinhui Jiang, Kai Zhou, Zhining Xia, Yike Huang","doi":"10.1002/elps.70086","DOIUrl":"https://doi.org/10.1002/elps.70086","url":null,"abstract":"<p><p>In complex systems, a universal and effective method for accurately determining the dissociation constant (K_d) of drug-protein interactions (DPI) has not been identified yet. In this study, a novel biomimetic affinity capillary electrochromatography (ACEC) platform was developed. Polydopamine (PDA) was employed not only as a coating material but also as an immobilization agent for human serum albumin (HSA), resulting in the fabrication of the PDA/PDA/HSA@capillary. A series of characterization experiments on the PDA/PDA/HSA@capillary showed that PDA was successfully coated on the inner wall of the capillary, and HSA was also successfully immobilized on the capillary column. The optimal concentration of PDA for preparing the PDA/PDA/HSA@capillary was determined to be 0.8 mg mL^-1, and the optimal concentration for immobilizing HSA was 0.25 mM, respectively. Moreover, the PDA/PDA/HSA@capillary showed good separation effects in the complex systems of different drugs. Furthermore, the electrophoretic performance of individual and mixed samples was compared across different capillary columns. Through interaction studies between proteins and compounds, the K_d value of rutin was 1.09 × 10³ mol L^-1 that of quercitrin was 2.78 × 10³ mol L^-1, and that of quercetin was 7.44 × 10⁴ mol L^- ¹, aligning with results from other established methods. The method was applied to the Sophora japonica extract, and the K_d was consistent with the rutin in the mixed system. Reproducibility studies demonstrated that high separation efficiency was maintained even after 50 consecutive runs. By integrating biomimetic material science with chromatographic innovation, this work overcomes critical bottlenecks in traditional affinity capillary electrophoresis (ACE), providing a universal tool for high-throughput drug screening and structure-guided therapeutic design.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147484954","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":"Electrokinetic Nucleic Acid Amplification (E-NAAMP) Using Paper-PDMS Microfluidics and High-Frequency Joule Heating.","authors":"Jarad Yost, Md Nazibul Islam, Zachary Gagnon","doi":"10.1002/elps.70088","DOIUrl":"https://doi.org/10.1002/elps.70088","url":null,"abstract":"<p><p>We present a novel paper-based nucleic acid amplification (NAA) technique using electrokinetic nucleic acid amplification (E-NAAMP). In E-NAAMP, a high radio frequency (RF) potential is applied across a conductive aqueous sample to induce an Ohmic current and drive the sample temperature to increase by Joule heating. Using this RF approach, we investigate the ability to induce E-NAAMP in pressurized paper-based microfluidic channels. We use the microfluidic pressure-in-paper (µPiP) method to encapsulate synthetic and natural fiber-based paper channels between thin sheets of polydimethylsiloxane (PDMS) with two strips of conductive PDMS that have been infused with carbon black (PDMS-CB) to act as electrodes in contact with the paper channels. A high-frequency (38 MHz) voltage is applied across a conductive NAA sample via the PDMS-CB electrodes to generate Joule heating within the paper structure. Here, we show that µPiP-based E-NAAMP can amplify NAs using the loop-mediated isothermal amplification (LAMP) reaction. We first investigate the pore-scale temperature profile numerically by solving the relevant energy transport equations within digitized paper fiber domains obtained using micro-computed tomography (micro-CT) scans. We compare these temperature-voltage predictions to those measured experimentally and demonstrate good agreement, suggesting that RF Joule heating is a viable method for electrokinetically heating paper-based microfluidic platforms. We next examine the effect of a porous substrate on NAA and demonstrate that the carrier protein, bovine serum albumin (BSA), is required for paper-based NAA reactions by preventing polymerase adsorption within the paper structure. Finally, we show successful NAA in paper using E-NAAMP with multiple paper fiber types while also further demonstrating BSA's necessity for paper E-NAAMP success. Our results demonstrate that paper-based microfluidic NAA using Joule heating is a viable alternative to traditional microfluidic NAA devices by offering substantial heating element miniaturization and decreased fabrication complexity when compared to both traditional and Joule-heated microfluidic NAA devices.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147467439","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}