ELECTROPHORESIS最新文献

{"title":"Integrated Purification and Desalting of Plasmid DNA Using Monolith Columns.","authors":"Rok Miklavčič, Tadej Sever, Polona Skrt, Tina Vodopivec Seravalli, Aleš Štrancar, Urh Černigoj","doi":"10.1002/elps.70104","DOIUrl":"https://doi.org/10.1002/elps.70104","url":null,"abstract":"<p><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 >1 mg pDNA per mL of column and pDNA recovery of >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>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-07","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":"Contactless Dielectrophoretic Separation of Circulating Tumor Cells From Red Blood Cells and Platelets in a Viscoelastic Medium.","authors":"Agnibha Das, Nirmalendu Biswas, Amitava Dutta","doi":"10.1002/elps.70101","DOIUrl":"https://doi.org/10.1002/elps.70101","url":null,"abstract":"<p><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>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-07","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":"Enantioseparation of Chiral Compounds From Traditional Chinese Medicines on a Chiral Packed Capillary Column With UV Detection.","authors":"Shiyun Hu, Xiao Hu, Kun Chen, Yingying Liu, Yingying Deng, Lishuang Yu, Lu Huang","doi":"10.1002/elps.70105","DOIUrl":"https://doi.org/10.1002/elps.70105","url":null,"abstract":"<p><p>A chiral packed capillary column (PCC) was prepared by a photonic crystal fiber (PCF) as the frit and cellulose tris(3,5-dimethylphenylcarbamate)-coated silica particles (CDMPC@SiO₂) as the chiral stationary phase. This column coupled with UV detection was employed for the enantioseparation of chiral compounds derived from traditional Chinese medicines (TCMs). Baseline enantioseparation was achieved for schisandrin B and tetrahydropalmatine, whereas partial separation was observed for anisodamine and corynoxeine. The reproducibility and stability of the column were confirmed, with relative standard deviation (RSD) values for retention time and resolution ranging from 2.5% to 3.2% (intra-day), 2.7% to 3.5% (inter-day), and 4.1% to 4.5% (inter-column). Then, the column was successfully applied to the direct enantioseparation and quantitative determination of tetrahydropalmatine in the extract of Yanhusuo (Corydalis Rhizoma). These results demonstrate that the proposed chiral CDMPC@SiO₂ PCC is not only reproducible and stable but also can be effectively applied to the enantioseparation and content determination of chiral compounds in complex TCMs. Furthermore, density functional theory (DFT) calculations were conducted to investigate intermolecular interactions between CDMPC and chiral analytes, which provides a deeper understanding of the enantioseparation mechanism.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147835244","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":"Validated RP-HPLC-UV Method for the Quantitative Analysis and Stability Evaluation of Amoxicillin in Paediatric Gummy Tablets.","authors":"Samuele Bonafè, Anna Imbriano, Cinzia Pagano, Anna Migni, Elisa Bianconi, Laura Mercolini, Luana Perioli, Roccaldo Sardella","doi":"10.1002/elps.70103","DOIUrl":"https://doi.org/10.1002/elps.70103","url":null,"abstract":"<p><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 largely (but not strictly) in accordance with ICH Q2(R1) guidelines, evaluating specificity, linearity, limit of quantification (LOQ), trueness, precision, robustness and ruggedness. A blank matrix derived from a blank 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 (R² = 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 also 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":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Capillary Electrophoresis With Amperometric Detection for Neurotransmitter Analysis: Principles, Electrode Materials, Methodologies, and Applications.","authors":"Petr Kubáň, Jiří Volánek, Nguyen Thi Thu Trang, Tran Dai Lam","doi":"10.1002/elps.70099","DOIUrl":"https://doi.org/10.1002/elps.70099","url":null,"abstract":"<p><p>Capillary electrophoresis with amperometric detection has become a powerful tool for the determination of electroactive neurotransmitters due to its high separation efficiency, minimal sample consumption, and excellent detection sensitivity. This review provides the first comprehensive summary specifically on monoamine neurotransmitters and related compounds in biological systems. Fundamental principles of amperometric detection, electrode positioning strategies, and approaches to high-voltage decoupling are described, followed by a critical comparison of electrode materials including carbon fiber, glassy carbon, graphite, noble metals, boron-doped diamond, and chemically modified or multielectrode designs. Their impact on noise, sensitivity, fouling resistance, and ease of integration is highlighted. Particular attention is given to background electrolyte composition, where phosphate buffers are most widely used, while 2-(N-morpholino)ethanesulfonic acid systems have enabled the lowest limits of detection, often reaching the attomole range. Applications span human urine, blood, lymphocytes, brain tissue, microdialysates, invertebrate tissues, and single cells, demonstrating capillary electrophoresis with amperometric detection as a versatile platform for neurochemical analysis. Advances in online preconcentration, dynamic pH junctions, and redox-cycling electrodes have further improved selectivity and sensitivity. Finally, current limitations, such as electrode fouling and interface robustness, are discussed together with emerging trends, including clinical translation and in vivo neuroanalysis.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147812410","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":"Establishment of a Highly Accurate and Sensitive Age Prediction Model for Multiple Body Fluids: Blood, Saliva, and Semen Using Pyrosequencing.","authors":"Xudong Zhao, Daijing Yu, Jingjing Xu, Yaru Mai, Kewen Zhang, Lijuan Su, Fenglong Yang, Jiangwei Yan","doi":"10.1002/elps.70100","DOIUrl":"https://doi.org/10.1002/elps.70100","url":null,"abstract":"<p><p>DNA methylation-based age prediction has become a reliable method for individual identification. While current models have achieved high accuracy when targeting a single type of biological fluid, crime scenes often contain multiple body fluids. Applying single fluid models to other fluid types may result in significant prediction errors, potentially misleading investigations. Therefore, age prediction models applicable to multiple biological fluids are of critical importance. In this study, we screened three age-related methylation sites (cg05940966, cg10501210, and cg10528482) from blood, saliva, and semen samples by analysis of public databases. These sites are associated with age. We then quantified methylation levels in peripheral blood samples from 101 healthy individuals via pyrosequencing. Based on these data, machine learning algorithms were applied to construct multiple age prediction models. These models were evaluated for their prediction accuracy, applicability to other body fluids, sensitivity, inhibitor tolerance, and utility with aged forensic samples. A multiple linear regression model constructed using the pyrosequencing results displayed the highest prediction accuracy, with mean absolute deviation (MAD) values of 2.717, 3.506, and 4.154 years in blood, saliva, and semen, respectively. Pyrosequencing also demonstrated high sensitivity, with MAD remaining within 4 years, even in trace samples (0.5 ng of unconverted genomic DNA). However, when the concentration of oxidized heme exceeds 1 ng/µL and the concentration of humic acid exceeds 2 ng/µL, pyrosequencing cannot accurately measure the methylation values at the CpG sites. In summary, this study provides an accurate and reliable tool for age prediction based on multiple bodily fluids. From a practical standpoint, this technology enables rapid age estimation without requiring prior identification of fluid type, thereby conserving valuable biological samples at crime scenes. This breakthrough will significantly enhance the efficiency of criminal investigations, strengthen the reliability of biological evidence interpretation in complex scenarios, and provide scientific support for judicial processes.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147812369","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":"Production and Purification of Mesenchymal Stem Cell-Derived Extracellular Vesicles Using Preferential Exclusion Chromatography.","authors":"Katja Vrabec, Andrej Raspor, Darja Božič, Nastja Bizjak Ternik, Ana Železnik, Valentina Novak, Ivana Petrović Koshmak, Maja Leskovec, Aleš Štrancar","doi":"10.1002/elps.70097","DOIUrl":"https://doi.org/10.1002/elps.70097","url":null,"abstract":"<p><p>Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) therapeutics hold the potential to harness the regenerative power of their parental cells, promoting proliferation and reducing inflammation. There is a need for the production and isolation process that can be easily scaled and deliver the desired quality and quantity of EVs. We present a chromatographic method development based on preferential exclusion principle that can be used for EV isolation from adherent and microcarrier-based upstream. The method employs monolithic hydroxyl columns to purify and concentrate MSC-EVs. The heterogeneity in EV size and composition poses challenges in upstream and downstream processes; therefore, process development was guided by PATfix biochromatography analytics, tracking EV-specific surface antigens. The developed purification method enables direct concentration of MSC-EVs from conditioned media while removing 97% of protein impurities and 95% of double-stranded deoxyribonucleic acid (dsDNA)-related impurities in the process.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147722175","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":"Metabolomic Signatures of Spontaneous Isolated Superior Mesenteric Artery Dissection: Indole Derivatives and Lysophospholipids as Novel Biomarkers","authors":"Lei Ji,&nbsp;Xun Wang,&nbsp;Yangyan He,&nbsp;Ziheng Wu,&nbsp;Donglin Li,&nbsp;Hongkun Zhang","doi":"10.1002/elps.70078","DOIUrl":"10.1002/elps.70078","url":null,"abstract":"<div>\u0000 \u0000 <p>Spontaneous isolated superior mesenteric artery dissection (SISMAD) is a rare but increasingly recognized vascular disorder that can lead to intestinal dysfunction with severe complications. Currently, no plasma biomarkers, particularly metabolomics-based markers, have been identified for SISMAD. To identify novel plasma biomarkers and potential therapeutic targets for SISMAD and its associated intestinal dysfunction, we analyzed plasma samples from 30 patients with SISMAD and 30 healthy controls. Blood was collected within 72 h of diagnosis. Ultra-high-performance liquid chromatography coupled with orbitrap mass spectrometry was used to profile metabolites, followed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) to analyze the data. A total of 123 and 128 metabolites were significantly changed in the positive and negative ion modes, respectively. Notably, tryptophan-related metabolites, including methyl 2-(1<i>H</i>-indol-3-yl) acetate and indole-3-pyruvic acid (IPA), were significantly reduced in the SISMAD group (<i>p</i> &lt; 0.001), with a strong correlation between the two (Corr. = 0.989, <i>p</i> &lt; 0.001). Pathway analysis revealed that glycerophospholipid metabolism was most enriched, primarily driven by lysophospholipids (LPLs), particularly lysophosphatidylethanolamine (LysoPE), which showed the highest fold change and diagnostic accuracy (area under the curve [AUC] = 0.986). This study identifies significant metabolic alterations in SISMAD, including reductions in indole derivatives (methyl 2-(1<i>H</i>-indol-3-yl) acetate and IPA and several LPLs, such as LysoPE, lysophosphatidylcholine (LysoPC), and lysophosphatidylglycerol (LysoPG). These findings highlight potential diagnostic biomarkers and therapeutic targets for SISMAD and its associated intestinal dysfunction, providing a promising avenue for clinical management and intervention.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 4","pages":"328-338"},"PeriodicalIF":2.5,"publicationDate":"2026-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146164718","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 4'26","authors":"","doi":"10.1002/elps.70095","DOIUrl":"https://doi.org/10.1002/elps.70095","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 4","pages":"273-275"},"PeriodicalIF":2.5,"publicationDate":"2026-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70095","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147686043","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 Genetic Profiling for Cattle Parentage Testing via Next-Generation Sequencing","authors":"Jiaojiao Geng,&nbsp;Jiajun Liu,&nbsp;Zhentang Liu,&nbsp;Zhiyong Liu,&nbsp;Chaobo Bao,&nbsp;Xiaohui Yan,&nbsp;Xiaoyue Wang,&nbsp;Zhanze Wu,&nbsp;Hongyu Sun,&nbsp;Riga Wu","doi":"10.1002/elps.70082","DOIUrl":"10.1002/elps.70082","url":null,"abstract":"<div>\u0000 \u0000 <p>Cattle are frequently involved in civil disputes and offenses. Currently, the paternity testing of cattle is mainly done by analysis of dinucleotide STR (di-STR) or SNPs. However, the di-STR marker usually produces many stutter artifacts that affect the accurate genotyping, and the polymorphic level of SNPs is not high enough. To overcome these limitations, we developed and validated a novel integrated next-generation sequencing (NGS) panel for cattle. It comprises 33 reported di-STRs and 30 newly identified tetranucleotide STRs (tetra-STRs), the mitochondrial displacement loop (D-loop) region, as well as cytochrome b (<i>CYTB</i>) and recombination-activating gene 1 (<i>RAG1</i>). The results showed a high genotyping success rate (99.6% on average) and balanced amplification efficiency across most STR loci. In addition, the panel's average genotyping consistency rate reached 96.4% between blood and hair samples. The total discrimination power and cumulative probability of exclusion for duos of this panel were 1–1.8 × 10⁻¹³ and 1–1.7 × 10⁻⁸, respectively. Application in paternity testing caseworks demonstrates its applicability and reliability for resolving paternity disputes. Overall, this study provided a cattle profiling NGS panel with a substantial advancement over current capillary electrophoresis (CE)-based systems, offering superior accuracy, multiplex capability, and efficiency for forensic genetic testing in cattle.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 4","pages":"339-352"},"PeriodicalIF":2.5,"publicationDate":"2026-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147316759","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}