ELECTROPHORESIS最新文献

{"title":"Optimized iCIEF Fractionation Enables Deep Profiling of Antibody Drug Conjugate Charge Heterogeneity","authors":"Eunju Jang,&nbsp;Qunying Zhang,&nbsp;Paul M. Thomas","doi":"10.1002/elps.70112","DOIUrl":"10.1002/elps.70112","url":null,"abstract":"<p>Antibody drug conjugates (ADCs) are a novel class of therapeutic drugs that combine selectivity of an antibody moiety with the cell killing potency of cytotoxic agent. ADCs are complex molecules, and their charge variants are one of the important critical quality attributes (CQAs) that need to be monitored throughout the drug development process. Imaged capillary isoelectric focusing (iCIEF) is considered a gold standard in charge variant analysis due to its short analysis time and ease of use. For comprehensive analysis, iCIEF fractionation is a newly developed technique that can be used to enable LC–MS characterization of charge variants. Here, we describe method development for an iCIEF fractionation system, as well as further characterization using LC–MS. Fractionation conditions were optimized to provide stabilized mobilization after separation. Fractions were collected with the optimized method, and native SEC–MS results showed that fractionation enabled detection of species previously undetectable. Further mass spectrometric analysis revealed the major posttranslational modifications (PTMs) and drug load distributions for both light and heavy chains. The results indicate that iCIEF fraction collection enables a deeper understanding of ADC charge variants and will be further useful for studying other complex biomolecules.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 7","pages":"675-683"},"PeriodicalIF":2.6,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70112","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148137032","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":"Validated RP-HPLC–UV Method for the Quantitative Analysis and Stability Evaluation of Amoxicillin in Paediatric Gummy Tablets","authors":"Samuele Bonafè,&nbsp;Anna Imbriano,&nbsp;Cinzia Pagano,&nbsp;Anna Migni,&nbsp;Elisa Bianconi,&nbsp;Laura Mercolini,&nbsp;Luana Perioli,&nbsp;Roccaldo Sardella","doi":"10.1002/elps.70103","DOIUrl":"10.1002/elps.70103","url":null,"abstract":"<p>The development of products that encounter compliance from paediatric patients represents one of the challenges of pharmaceutical technology that aims to develop formulations able to overcome problems related to palatability and dosing flexibility and, in general, to patient acceptance. In this study, a fully validated reversed-phase high-performance liquid chromatography coupled with ultraviolet detection (RP-HPLC–UV) method was applied for the quantitative determination of amoxicillin (AMX) in an innovative gummy tablet formulation designed for paediatric use. The chromatographic method was optimized and validated <b>largely (but not strictly) in accordance</b> with ICH Q2(R1) guidelines, evaluating specificity, linearity, limit of quantification (LOQ), trueness, precision, robustness and ruggedness. A <b>blank</b> matrix derived from a <b>blank</b> formulation was employed to closely simulate the sample matrix during method validation and quantitative analysis. The method exhibited excellent linearity over the concentration range of 0.0039–0.25 mg/mL (<i>R</i>² = 0.99) with satisfactory trueness (recoveries between 101.3% and 102.4%) and high precision (coefficient of variation [CV] ≤ 1.6%). The LOQ was estimated between 0.0020 and 0.0039 mg/mL. Extraction recovery was <b>also</b> evaluated using penicillin G as a model compound due to its chemical and physicochemical similarity to AMX, yielding a recovery of 97.0% ± 0.1%. Application of the validated method to AMX-loaded gummy tablets confirmed a drug content of 94.7% ± 3.7% of the theoretical value. Stability studies, conducted over 6 months under different storage conditions, revealed a progressive decrease in AMX content within the gummy matrix with more pronounced degradation observed under vacuum at 25°C compared to storage in a dry cabinet at room temperature (RT). Overall, the developed RP-HPLC–UV method proved to be reliable and suitable for routine quality control and stability assessment of AMX-containing gummy tablets, supporting further development of this child-friendly dosage form.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 7","pages":"605-612"},"PeriodicalIF":2.6,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70103","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147812318","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":"Integrated Purification and Desalting of Plasmid DNA Using Monolith Columns","authors":"Rok Miklavčič,&nbsp;Tadej Sever,&nbsp;Polona Skrt,&nbsp;Tina Vodopivec Seravalli,&nbsp;Aleš Štrancar,&nbsp;Urh Černigoj","doi":"10.1002/elps.70104","DOIUrl":"10.1002/elps.70104","url":null,"abstract":"<div>\u0000 \u0000 <p>Efficient purification and desalting of nucleic acids are essential unit operations in downstream processing (DSP). Conventional methods offer good solutions for either desalting (e.g., tangential flow filtration, dialysis and size exclusion) or purification (e.g., chromatography and precipitation), but integrating both into a single step on preparative scale has remained a big challenge. Here, we present a novel chromatographic method using a multimodal anion-exchanging (AEX) Convective Interaction Media (CIM) Swiper column that enables binding of nucleic acids at slightly acidic pH and elution at neutral pH with low salt concentration. Swiper desalting capability was first demonstrated for purified plasmid DNA (pDNA) samples from two standard pDNA DSP elution buffers: Tris-EDTA (TE), 0.9 M NaCl, pH 7.2 (AEX) and TE, 0.9 M ammonium sulphate (AS) and pH 7.2 (hydrophobic interaction chromatography). A scalable desalting process of enriched and partially purified pDNA samples was developed; with desalting capacity of &gt;1 mg pDNA per mL of column and pDNA recovery of &gt;90%, achieved with a simple TE, pH 7.2 elution buffer. Building from here, a single-step DSP combining purification and desalting of pDNA directly from clarified bacterial lysate was evaluated. Dynamic binding capacity (DBC) for pDNA from lysate using 4 mL CIM Swiper column was 1.0 mg/mL with the elution recovery of 90%. RNA, proteins and endotoxins in the pDNA elution fractions were below or close to the FDA-specified guidelines for pDNA purity, whereas host cell DNA and open circular (OC) pDNA were not efficiently depleted. Although the demonstrated process does not reach the specified product quality parameters, we pave the way to simplifying pDNA DSP flow by demonstrating an integration of two previously separate process steps at preparative scale, reducing footprint and costs.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 7","pages":"644-654"},"PeriodicalIF":2.6,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147835227","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 Simulation Study of Inhibition Assay With Theophylline on the Enzymatic Hydrolysis of p-Nitrophenyl Phosphate With Alkaline Phosphatase in a Continuous Mode of Electrophoretically Mediated Microanalysis","authors":"Toshio Takayanagi,&nbsp;Asuka Nomoto,&nbsp;Hitoshi Mizuguchi","doi":"10.1002/elps.70108","DOIUrl":"10.1002/elps.70108","url":null,"abstract":"<p>Inhibition assays in a continuous mode of electrophoretically mediated microanalysis (EMMA) were simulated using a MATLAB matrix. The enzymatic hydrolysis of <i>p</i>-nitrophenyl phosphate (NPP) with alkaline phosphatase (ALP) was adopted as a model enzymatic reaction, where theophylline (THE) was used as a model inhibitor. In the simulation, the substrate NPP and the inhibitor THE were tandemly injected into a separation capillary filled with a separation buffer containing ALP. The formation rate of the product <i>p</i>-nitrophenol (NP) was calculated using a Michaelis–Menten constant, the inhibition constant, and the reagent concentrations. A plateau response of the product was reproduced in the simulation based on the continuous enzymatic reaction and the continuous resolution of the product in EMMA. In addition to the plateau response of the product, a depressed plateau was also simulated as a consequence of the inhibition reaction. This depressed plateau was caused by the overlapping between the injected zones of the substrate and the inhibitor. The simulated electropherograms including the inhibition-induced depression were consistent with the experimental data. The Michaelis–Menten constant, the inhibition constant, and the inhibition type were also reproduced from the simulated plateau responses. Factors affecting the plateau depression were further investigated through simulations by varying the injected zone lengths and experimental conditions. This study successfully demonstrated the utility of the simulation approach for designing inhibition assays of enzymatic reactions in a continuous mode of EMMA.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 7","pages":"595-604"},"PeriodicalIF":2.6,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70108","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147940582","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 Novel HILIC-UPLC-DAD-MS/MS Method for the Analysis of Mycosporine-Like Amino Acids and Their Quantification in Diverse Algae","authors":"Armin Oberosler,&nbsp;Anastasiia Fedorova,&nbsp;Ignacio Zegri,&nbsp;Fabian Hammerle,&nbsp;Michael Zwerger,&nbsp;Thomas Werner,&nbsp;Markus Ganzera","doi":"10.1002/elps.70110","DOIUrl":"10.1002/elps.70110","url":null,"abstract":"<p>Mycosporine-like amino acids (MAAs) are photoprotective compounds primarily produced by marine organisms, especially red algae. Chemically, MAAs and their precursors are low-molecular-weight natural products that absorb ultraviolet radiation in the range of 270–360 nm without generating free radicals. Owing to these properties, MAAs have attracted considerable interest for potential pharmaceutical and cosmetic applications as natural sunscreen agents. In this study, the first hydrophilic interaction liquid chromatography-ultra-performance liquid chromatography (HILIC-UPLC) method, coupled with both diode array detector (DAD) and tandem mass spectrometry (MS/MS), was developed for the determination of 11 MAAs and 2 MAA precursors in various algal species. As column, a YMC-Triart Diol-HILIC (particle size: 1.9 µm) was used, and the separation of the MAAs realized in under 5 min. The validation of the method was carried out following International Council for Harmonisation (ICH) guidelines, demonstrating linearity, selectivity, precision, and accuracy. The analysis of extracts prepared from well-known species, such as <i>Porphyra</i> sp. and <i>Chondrus crispus</i>, confirmed practical applicability. Demonstrating feasibility of MS/MS-based quantification of co-eluting compounds, the method represents an appealing alternative that surpasses established procedures not only in terms of analysis time but also by providing a substantially increased informative value.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 7","pages":"620-628"},"PeriodicalIF":2.6,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70110","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148005072","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":"Development and Evaluation of a Novel Microhaplotype Panel for DNA Mixture and Kinship Analysis","authors":"Haoyan Jiang,&nbsp;Jiaming Xue,&nbsp;Mengyu Tan,&nbsp;Mengna Wu,&nbsp;Qiushuo Wu,&nbsp;Guihong Liu,&nbsp;Yazi Zheng,&nbsp;Yuanyuan Xiao,&nbsp;Shanshan Wei,&nbsp;Tangleixin Li,&nbsp;Meili Lv,&nbsp;Shengqiu Qu,&nbsp;Weibo Liang,&nbsp;Lin Zhang","doi":"10.1002/elps.70111","DOIUrl":"10.1002/elps.70111","url":null,"abstract":"<div>\u0000 \u0000 <p>Microhaplotypes (MHs), together with the development of next-generation sequencing (NGS), hold great promise for forensic genetics, particularly for mixture interpretation and kinship analysis. In this study, we performed genome-wide screening for new MH markers composed of two or more variants (InDels or single nucleotide polymorphism [SNP]) within 300 bp, based on the Chinese Southern Han (CHS) population from the 1000 Genomes Project (1KG). We developed a 31-plex MH panel specifically for the Chinese Han population, consisting of six MHs and 25 compound markers. The panel demonstrated high polymorphism, with an average effective number of alleles (<i>A</i>_e) of 5.10. The combined power of discrimination (CPD) and cumulative probability of exclusion (CPE) reached 1–1.58 × 10⁻²⁹ and 1–1.24 × 10⁻¹³, respectively. Subsequently, allele detection rates and unique allele detection rates of minor contributors were evaluated in two- to four-person DNA mixtures with varying input levels and contributor ratios. The panel maintained stable detection performance under low-template or moderately unbalanced conditions, with significant declines observed only when both factors were present. The panel also demonstrated great capacity for close relatives (first-degree and second-degree relatives). Parent–child and full-sibling pairs were completely distinguished from unrelated individuals, with consistently strong classification performance in both simulated datasets and real pedigrees. The mean log₁₀ likelihood ratio (LR) values for second-degree relative pairs provided moderate to strong evidential support, whereas discrimination of third- to fifth-degree relative pairs remained limited. In conclusion, these findings indicate that the panel is a valuable supplementary tool for forensic DNA analysis, particularly for mixture deconvolution and kinship inference.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 7","pages":"655-665"},"PeriodicalIF":2.6,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148053387","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":"Fractionation of Milligram Quantities of Gadolinium and Terbium Ions by Annular Free-Flow Isotachophoresis","authors":"Hannah Hallikainen,&nbsp;Xiaofeng Guo,&nbsp;Cornelius Ivory","doi":"10.1002/elps.70094","DOIUrl":"10.1002/elps.70094","url":null,"abstract":"<p>The fractionation of milligram quantities of gadolinium (Gd) and terbium (Tb) ions is demonstrated using annular free-flow isotachophoresis (ITP) with ammonium as the leading ion and hydronium as the terminating ion, and α-hydroxyisobutyric acid (HIBA) and acetate as lanthanide complexing ligands. Buffer conditions were designed using numerical simulations in COMSOL Multiphysics, allowing for rapid screening of candidate buffer systems before laboratory experiments. The leading electrolyte contained 15 mM ammonium acetate, 5 mM acetic acid, and 3.5 mM HIBA at pH 4.9, while the terminating electrolyte consisted of 15 mM glutamic acid, 5 mM acetic acid, and 3.5 mM HIBA at pH 3.1. After 5 h of electrophoresis at 2 kV and at a mass loading of 7.077 mg Gd and 7.074 mg Tb, 84.08% of the Tb mass was recovered in the first 7 fractions removed from the separation column, and 86.51% of the Gd mass was recovered from the remaining 11 fractions. Overall recoveries of Gd and Tb were 95.07% and 99.13%, respectively.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 6","pages":"441-449"},"PeriodicalIF":2.6,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147654042","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":"Contactless Dielectrophoretic Separation of Circulating Tumor Cells From Red Blood Cells and Platelets in a Viscoelastic Medium","authors":"Agnibha Das,&nbsp;Nirmalendu Biswas,&nbsp;Amitava Dutta","doi":"10.1002/elps.70101","DOIUrl":"10.1002/elps.70101","url":null,"abstract":"<div>\u0000 \u0000 <p>A cancerous tumor harbors millions of genetically mutated cells, which after hyperproliferation are picked up and transported to various sites in the human body by the bloodstream or lymphatic system. These circulating tumor cells (CTCs) become the seeds for the subsequent growth of secondary tumors. Hence, CTCs hold information about a tumor that could be the key to cancer diagnosis and treatment. This work focused on the designing and simulating a microfluidic platform using contactless-dielectrophoresis (cDEP) to separate CTCs from platelets and red blood cells (RBCs) in a viscoelastic fluid flowing through a microfluidic channel. Contactless electrodes with alternating potentials have been used in this microfluidic device. Using the finite element-based solver, the separation in a microfluidic device based on size-based segregation has been simulated. The operating conditions have been varied in this work to determine the optimal conditions for segregation. Under optimal conditions of 150 Pa inlet pressure of working fluid, 2.5 V of applied voltage, and 50 kHz of alternating electric field frequency, the cells were effectively segregation yielding the best results. At these conditions, separation efficiency of 98.5% is achieved for platelets, 97.25% for RBCs, and 98.5% for CTCs for a separation period of 100 s. The angle of separation of the particles is suffered by the cells at various applied voltages (2, 2.5, 3, 3.5, and 4 V). The study demonstrates the validity of a size-based segregation device using cDEP for the segregation of tumor cells and the advancement of microscale devices in cancer detection and diagnosis.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 6","pages":"556-580"},"PeriodicalIF":2.6,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147835152","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 Five-Color Multiplex Droplet Digital PCR Assay With Support Vector Regression for Forensic Age Prediction","authors":"Jing Zhang,&nbsp;Chuantong Zhao,&nbsp;Li Jiang,&nbsp;Pu Fan,&nbsp;Caixia Li,&nbsp;Rui Liu,&nbsp;Wenting Zhao","doi":"10.1002/elps.70102","DOIUrl":"10.1002/elps.70102","url":null,"abstract":"<div>\u0000 \u0000 <p>Current forensic methylation analysis faces challenges: Traditional SNaPshot assays often suffer from limited inhibitor tolerance and high reagent costs, whereas massively parallel sequencing (MPS) involves a labor-intensive workflow and long turnaround time. Furthermore, conventional droplet digital PCR (ddPCR) assays typically lack high-order multiplexing capability, and most existing models are restricted to single tissue types. To address this, we validated 98 age-associated CpGs in 1398 Chinese blood methylomes and established a novel 17-plex assay utilizing an advanced 5-color fluorescence ddPCR platform. This assay was applied to 325 saliva, 107 blood, and 91 buccal swab samples. Results showed strong age correlations across tissues (Spearman's |<i>r</i>|: 0.554–0.956). Comparative modeling using support vector regression (SVR) versus multiple linear regression (MLR) demonstrated that SVR significantly outperformed MLR by effectively capturing non-linear methylation changes. The SVR model explained &gt;98% of age variation with mean absolute deviations (MADs) of 2.396 years (saliva), 2.006 years (blood), and 2.044 years (buccal swabs), maintaining robust performance across broad age groups. This five-color multiplex strategy provides a rapid, cross-tissue, and cost-effective analytical method, offering a robust alternative to SNaPshot and MPS workflows for both forensic investigation and potential health-related age estimation.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 6","pages":"489-501"},"PeriodicalIF":2.6,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147940576","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 Comprehensive Numerical Analysis of Dielectric-Patterned Hybrid Induced-Charge Electrokinetics for On-Chip Nanoparticle Enrichment","authors":"Miao Fang,&nbsp;Ye Tao,&nbsp;Yifan Chen,&nbsp;Jingyi Yang,&nbsp;Wen Cao,&nbsp;Wenhui Hou,&nbsp;Kang Xiangli,&nbsp;Weiyu Liu","doi":"10.1002/elps.70096","DOIUrl":"10.1002/elps.70096","url":null,"abstract":"<div>\u0000 \u0000 <p>Nanoparticle enrichment is critical for analytical chemistry applications in microfluidic chemical and biological analyses, enabling sensitive detection of low-abundance species. Among electrokinetic techniques, induced-charge electroosmosis (ICEO) stands out for its nonlinear electric field response, effectiveness in dilute electrolytes, and independence from solution-specific properties—addressing limitations of conventional methods like centrifugation. Here, we present a dielectric-patterned bottom electrode microfluidic device that generates <i>a hybrid ICEO flow field</i> (a synergy of two modes) for efficient nanoparticle enrichment, designed as <i>a high-performance sample pretreatment module for microfluidic analytical systems</i>. Dielectric patterning induces tangential electric fields—indispensable for ICEO flow—creating alternating conductive grooves and dielectric protrusions. The hybrid field comprises low-frequency, strong converging ICEO flows in grooves (drawing particles to groove centers) and high-frequency, weak quadrupolar ICEO flows around dielectrics (enabling supplementary transport). Via numerical simulations coupling electrokinetic, fluid, and mass transport effects, optimal enrichment (concentration factor &gt;3.6, ∼5 µm narrow band) is achieved at 2 kHz and 3 V, driven by converging flows—<i>this performance directly meets the pretreatment requirements for trace nanoparticle analysis by enhancing sample concentration to detectable levels</i>. Voltage modulation enables dual-target capture: groove centers at low voltages (3–6 V) and dielectric corners at high voltages (12–15 V) via ICEO-dielectrophoresis synergy, expanding the adaptability of the system to diverse analytical detection layouts. The hybrid field exhibits strong frequency/conductivity dependence: Converging flow peak frequencies shift to higher values with increasing conductivity, whereas quadrupolar flows always remain robust in the high frequency limit—<i>supporting the analysis of samples with varying matrix conductivities</i>, a key demand in analytical chemistry. This work advances microfluidic nanoparticle manipulation with a frequency-adaptable platform, overcoming unpatterned electrode limitations (narrow frequency range and poor localization). Potential applications include point-of-care diagnostics, lab-on-a-chip analytical systems, and trace environmental/biological nanoparticle detection, all core focuses of analytical chemistry research.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 6","pages":"502-518"},"PeriodicalIF":2.6,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147688512","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}