ELECTROPHORESIS最新文献

{"title":"From One-to-One Complexation to Multisite Binding: Analytical Transport Coefficients and Mechanistic Diagnostics in Chiral Capillary Electrophoresis.","authors":"S J Manzi, V D Pereyra, F L Ferreyra, P M Centres, M R Gómez","doi":"10.1002/elps.70160","DOIUrl":"https://doi.org/10.1002/elps.70160","url":null,"abstract":"<p><p>We develop a general analytical framework for reaction-drift-diffusion processes under arbitrary multisite complexation schemes. This framework explains the separation of chiral compounds in capillary electrophoresis (CE) experiments involving multiple complexes, thereby extending beyond the scope of classical one-to-one (1:1) models. By diagonalizing the reaction-drift-diffusion operator in Fourier-Laplace space, we obtain closed-form expressions for the effective electrophoretic mobility and diffusion coefficient in an <math><mi>n</mi></math> -site mechanism. For 1:1 binding, the theory recovers the Wren-Rowe equation and exactly reproduces the diffusion coefficient obtained using the generating-function technique. Application to published data for Tyr-Arg-Phe-Phe-NH₂ stereoisomers interacting with DM- <math><mi>β</mi></math> -cyclodextrin shows that a minimal two-site model reproduces the observed curve morphologies. These results provide a direct link between stochastic kinetics, continuum transport, and practical diagnostics for optimizing chiral CE.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149024231","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":"Selected Topics in Detection and Identification of Molecular and Macromolecular Chirality.","authors":"Tamaki Nakano","doi":"10.1002/elps.70148","DOIUrl":"https://doi.org/10.1002/elps.70148","url":null,"abstract":"<p><p>Methods of detection and identification of chiral molecules and macromolecules (polymer molecules) are overviewed, focusing on rather recent topics. In addition to the established detection methods, including rotation and circular dichroism, those based on circularly polarized luminescence, plasmon, and liquid crystalline phase are introduced. Identification of chiral structures is discussed mainly for macromolecules in which the combination of experimental spectral studies and theoretical analyses is an emerging and versatile technique. Although identifying chiral structures is generally more challenging than determining chemical structures, the experiment-theory combination is a powerful tool to assess molecular and macromolecular asymmetry.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149011954","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 Streamlined Single-Step Salting-Out Approach for Chiral and Achiral Pesticide Analysis in Hazelnut-Based Beverages.","authors":"Giulia Bertini, Davide Vona, Nina Felli, Lorenzo Antonelli, Salvatore Fanali, Chiara Fanali, Stefania Carpino, Vincenzo Di Martino, Chiara Cavaliere, Alessandra Gentili","doi":"10.1002/elps.70154","DOIUrl":"https://doi.org/10.1002/elps.70154","url":null,"abstract":"<p><p>The increasing consumption of plant-based beverages (PBBs), driven by ethical and health-related reasons, highlights the need for robust analytical methods assessing their chemical safety. Among these products, hazelnut-based beverages are of particular interest in Italy, where hazelnut production is economically relevant. A rapid one-step extraction procedure was developed for the multiresidue determination of selected chiral and achiral pesticides in PBBs by ultra-high-performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). The method combines matrix precipitation and analyte extraction in a single step, providing a simple and efficient workflow. Validation, carried out in hazelnut beverage, included recovery, precision, linearity, decision limit (CCα), and detection capability (CCβ) evaluation. As no European Union maximum residue levels (MRLs) are available for PBBs, provisional MRLs for hazelnut milk were derived from raw hazelnut MRLs using a recipe-based processing factor, according to Regulation (EC) No 396/2005. The method achieved recoveries of 60%-90%, intra- and inter-day precision generally below 20%, CCα values of 0.001-0.016 µg L^-1, CCβ values of 0.002-0.027 µg L^-1, and good linearity for most analytes (R² > 0.99). The proposed procedure is suitable for pesticide residue analysis in PBBs, such as hazelnut-based beverages, and represents a valuable approach for monitoring emerging food matrices, for which residue data and regulatory limits are still lacking.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148987934","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":"Enhancing Lentiviral Purification: Combining CIM QA Monolithic Columns With Design of Experiments Approach.","authors":"Luka Bevc, Ana Pavšič Rijavec, Valentina Novak, Sandra Potušek Šuleski, Ivana Petrović Koshmak, Maja Leskovec, Hana Jug, Aleš Štrancar","doi":"10.1002/elps.70159","DOIUrl":"https://doi.org/10.1002/elps.70159","url":null,"abstract":"<p><p>Use of lentiviral vectors (LVs) has been steadily increasing over recent years, culminating with a recent focus on novel in vivo therapies. Despite an increased focus on LV therapies, the methods for their purification have not yet been standardised. Additionally, no major focus has been given to the problem of heterogeneity of LV harvest and removal of particle impurities. As such, novel methods for LV purification and a better understanding of factors influencing it have to be addressed. In order to address these issues, a purification method for LV has been developed using anion exchange chromatography, specifically, using quaternary amine (QA) Convective Interaction Media (CIM) monoliths. Using 96-Well CIM QA monolithic plates and design of experiments (DOE) approach, we optimised downstream process from 20% recovery to 90%. The process was successfully transferred to a 1 mL CIMmultus QA (6 µm) column, where 70% average recovery was achieved. During purification, an average 3.3 log reduction of host cell protein impurities was achieved, as well as significant removal of particle impurities, with 31% reduction in ratio between total and transducing particles in elution, relative to clarified harvest. The CIM QA monolith column also demonstrated a very high binding capacity of 2.9 × 10¹² particles/mL of stationary phase. These conditions not only enhance purification efficiency but also hold significant promise for adaptation in large-scale manufacturing processes.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148982216","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":"Conference Report: MSB 2026-International Symposium on Microscale Separations and Bioanalysis With Special Focus on Gel and Polymer Network-Based Separations.","authors":"Andras Guttman","doi":"10.1002/elps.70157","DOIUrl":"https://doi.org/10.1002/elps.70157","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148967202","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":"Protein-Based High-Performance Liquid Chromatography and Cyclodextrin-Capillary Electrokinetic Chromatography for the Chiral Separation of Azoles","authors":"Sarah Moreau,&nbsp;Amy Dillon,&nbsp;Giacomo Russo,&nbsp;Susanne K. Wiedmer","doi":"10.1002/elps.70124","DOIUrl":"10.1002/elps.70124","url":null,"abstract":"<p>Liquid chromatographic (LC) and capillary electrokinetic chromatography (EKC) methods with ultraviolet/diode array detection were developed for the enantioseparation of five azoles: econazole (ECO), miconazole (MICO), imazalil (IMA), ketoconazole (KET), and penconazole (PEN). The chiral selectors investigated were human serum albumin (HSA) and eight cyclodextrins (CDs), exploited in LC and EKC, respectively. HSA LC offered partial enantioseparation of IMA and, less effectively, of PEN. Interestingly, these azoles share a high degree of molecular similarity, as confirmed by in silico calculations. This may suggest that a single enantiomer achieves preferential access to enantioselective sites of the protein. To address the limited enantioselectivity of HSA for these azoles, CDs were explored as chiral selectors where EKC was preferable to LC due to its superior separation efficiency. The chiral discrimination capabilities of eight CDs were evaluated for these azole derivatives, aiming for fast separations with minimal amounts of CDs. Effective enantioseparation was achieved for ECO, MICO, IMA, and KET, using two of the tested CDs: 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) and heptakis(2,3,6-tri-<i>O</i>-methyl)-β-cyclodextrin (TM-β-CD). Satisfactory resolutions were obtained using a 40 mM phosphate buffer (pH 3.2) with 1 mM HP-β-CD for ECO and MICO and 8 mM TM-β-CD for KET. For IMA, the optimal conditions consisted of a 20 mM phosphate buffer with 3 mM HP-β-CD. None of the tested CDs were effective for the enantiomeric separation of PEN; therefore, a dual CD system was developed for the first time for this compound, employing succinyl-β-CD (Succ-β-CD) in combination with HP-β-CD in a borax buffer under basic conditions (pH 9.4), enabling complete enantioseparation of PEN. The low concentration of CDs required under optimal conditions renders the developed methods highly cost-effective.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 9","pages":"813-825"},"PeriodicalIF":2.6,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70124","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148301506","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":"Chiral HPLC Separation of Phosphonothrixin and Phosphonate Derivatives and Electronic Circular Dichroism Characterization of the Isolated Enantiomers","authors":"Mirna Maalouf,&nbsp;Maximilian Knab,&nbsp;Leon Gessner,&nbsp;Marc Engelhardt,&nbsp;Stella Friedrich,&nbsp;Harald Gross,&nbsp;Frank Boeckler,&nbsp;Chambers C. Hughes,&nbsp;Michael Lämmerhofer","doi":"10.1002/elps.70128","DOIUrl":"10.1002/elps.70128","url":null,"abstract":"<p>Phosphonothrixin (PPX), a phosphonate natural product known for its potent herbicidal activity, has attracted renewed interest because of the need for alternative herbicides. Its biological activity is confined to the (<i>S</i>)-enantiomer. Although the only reported enantioselective synthesis is complex, labor-intensive, and low yielding, direct chiral liquid chromatographic separation is likewise challenging because of its high polarity and lack of aromatic moieties, which limit interactions with commonly used chiral stationary phases (CSPs). In addition, phosphonic acid-containing compounds tend to chelate metal ions and adsorb to metal surfaces, further complicating chromatographic separation by causing pronounced peak tailing. Here, we describe a reliable and reproducible analytical and preparative method for the direct chiral separation of PPX using a zwitterionic sulfohexyl quinidine and quinine carbamate CSPs, as well as two synthetic phosphonate derivatives using a cellulose tris(3,5-dichlorophenylcarbamate) stationary phase as an alternative approach. The best separation of PPX was achieved with the zwitterionic sulfohexyl quinidine CSP (Rs = 2.4). The separated enantiomers were isolated, and their chemical and stereochemical identities were confirmed by mass spectrometry, optical rotation, and electronic circular dichroism spectroscopy. This approach enabled the successful isolation of enantiomerically pure (<i>S</i>)-(−)- and (<i>R</i>)-(+)-PPX.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 9","pages":"853-865"},"PeriodicalIF":2.6,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70128","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148419616","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":"Time-Resolved Native Fluorescence Imaging of the Migration and Band Broadening of Monoclonal Antibody During Capillary Gel Electrophoresis With Sodium Dodecyl Sulfate","authors":"Hui Xu,&nbsp;Uyen Nguyen,&nbsp;Tom Yang","doi":"10.1002/elps.70131","DOIUrl":"10.1002/elps.70131","url":null,"abstract":"<div>\u0000 \u0000 <p>We developed an imaging-based approach to directly visualize and monitor molecular migration during capillary electrophoresis and applied it to the separation of monoclonal antibodies (mAbs) by capillary gel electrophoresis with sodium dodecyl sulfate (CGE–SDS). Proteins were detected through their intrinsic fluorescence at 350 nm following excitation at 280 nm, enabling real-time observation of migration behavior without additional labeling. Time-resolved imaging allowed direct visualization of both migration and band broadening of mAbs under reducing and non-reducing conditions throughout the separation process. Analysis of electrophoretic mobilities suggests that mAb separation in the gel matrix is largely governed by the Ogston sieving mechanism. Quantification of band broadening revealed that peak dispersion increased with applied electric field strength, as evidenced by the marked reduction in broadening when the separation voltage was removed. Qualitative evaluation of the dispersion behavior suggests that mAb band broadening arises from the combined effects of electromigration dispersion (EMD) and gel-sieving-induced dispersion, with glycosylation-related heterogeneity providing an additional contribution for glycosylated species such as the heavy chain (HC) and intact mAb. These results demonstrate the utility of real-time imaging for elucidating separation mechanisms and dispersion phenomena in CGE–SDS and provide new insights into factors governing the electrophoretic behavior of therapeutic antibodies.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 9","pages":"826-839"},"PeriodicalIF":2.6,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148419641","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":"High-Throughput Multi-Capillary Electrophoresis","authors":"Jana Drewes,&nbsp;Sarah Meyer,&nbsp;Isobel Roberts,&nbsp;Cleo Fanny,&nbsp;Philip A. Doble,&nbsp;Dayanne Mozaner Bordin,&nbsp;Janice I. McCauley","doi":"10.1002/elps.70133","DOIUrl":"10.1002/elps.70133","url":null,"abstract":"<p>The potential of multi-capillary electrophoresis (CE) for high-throughput analysis of legacy CE applications was investigated by optimisation of the separation of six pharmaceutical compounds on a single-capillary instrument, followed by transference of the method to an eight-capillary BioPhase 8800 system from SCIEX. Multi-CE had superior performance, allowing construction of calibration curves in a single run, and a five-fold reduction of run time per sample, improved linearity, accuracy and limits of detection. Method transfer from the single-capillary system required minor instrumental parameter adjustments using the same background electrolyte. The potential of multi-CE was shown via quantification of caffeine in a variety of energy drinks, demonstrating broad applicability where high-throughput and rapid analyses are desirable.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 9","pages":"919-923"},"PeriodicalIF":2.6,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.70133","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148673087","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":"Production of Protein Hydrolysates and Bioactive Peptides From Lablab purpureus and Macrotyloma uniflorum via Optimized Extraction and Proteolysis Protocols","authors":"Chitra Yadav,&nbsp;Pooja Yadav,&nbsp;Abhishek Joshi,&nbsp;Jaya Arora","doi":"10.1002/elps.70125","DOIUrl":"10.1002/elps.70125","url":null,"abstract":"<div>\u0000 \u0000 <p>The rising global demand for sustainable, plant-based protein sources necessitates the exploration of underutilized legumes with high nutritional potential. This study systematically evaluates and compares five protein extraction techniques—ultrasound-assisted extraction (UAE), alkaline extraction–acid precipitation, phosphate buffer with TCA-acetone, TCA-acetone alone, and micellization—applied to <i>Lablab purpureus</i> and <i>Macrotyloma uniflorum</i>, two resilient and underexploited pulse crops. Protein recovery and yield were quantified for each method to identify the most efficient extraction approach. Among the tested methods, ultrasound-assisted extraction demonstrated superior performance in both legumes, with maximum protein yields of 84.02% for <i>L. purpureus</i> and 78.4% for <i>M. uniflorum</i> under optimized conditions (40% power, 10–15 min). Alkaline-acid precipitation resulted in the highest recovery (88.4%) but comparatively lower yields, indicating significant protein loss during isoelectric precipitation. Traditional precipitation techniques using TCA-acetone and phosphate buffer showed limited effectiveness, while micellization produced the lowest recovery and yield in both species. Importantly, the study reveals species-specific responses to extraction conditions, with <i>L. purpureus</i> consistently exhibiting higher extractability than <i>M. uniflorum</i>, likely due to differences in seed microstructure and protein distribution. These findings underscore the need for pulse-specific optimisation strategies rather than universal extraction protocols. The outcomes pave the way for their application in food, nutraceutical, and feed industries, contributing to sustainable protein innovation and improved utilisation of climate-resilient crops. Future work should focus on the structural, functional, and bioactive characterisation of the extracted proteins to enable targeted formulation in diverse food systems.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"47 9","pages":"881-893"},"PeriodicalIF":2.6,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148276291","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}