ELECTROPHORESIS最新文献

{"title":"Characterization of Hemoglobin Variants by Capillary Electrophoresis, UV-Vis, and FTIR Spectroscopy.","authors":"Julia Werle, Katerina Dunovska, Jakub Podhajsky, Michal Cerny, Jana Cepova, Arli Aditya Parikesit, Geir Bjørklud, Karel Kotaska, Eva Klapkova, Richard Prusa, Egon Werle, Rene Kizek","doi":"10.1002/elps.202400154","DOIUrl":"https://doi.org/10.1002/elps.202400154","url":null,"abstract":"<p><p>Hemoglobinopathies, hereditary disorders affecting the structure or production of hemoglobin, were detected by routine HbA_1c measurements by capillary electrophoresis (CE) at the University Hospital Motol, Prague. The potential of ultraviolet-visible (UV-Vis) and Fourier-transform infrared (FTIR) spectroscopy for the detection and characterization of hemoglobinopathies was investigated. FTIR spectra were recorded with a very high resolution (0.5 cm^-1) with 128 scans. The broad amide I peak, located at 1700-1600 cm^-1, can be formed by superimposition of the conformational structures of hemoglobin. These secondary protein structures were subjected to mathematical analysis. The application of band narrowing techniques, followed by curve fitting and integration processes, provided the basis for the quantitative estimation of protein secondary structure. As a result, unambiguous differences in UV-Vis spectra among patients with presumably normal hemoglobin, an HbC or a hemoglobin S/hemoglobin G (HbS/HbG)-Philadelphia variant could not be demonstrated. However, FTIR spectra indicated slight differences in α-helix, β-turns, β-sheet, or random coil secondary hemoglobin structures for these mutations. In the spectral wavenumber range of 950-850 cm^-1, there were some obvious FTIR differences at specific wavenumbers between patients with normal hemoglobin and those with the HbC variant. Further investigations are needed with a sufficient number of hemoglobin variants to elucidate the potency of FTIR spectroscopy for the characterization of hemoglobinopathies.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142946480","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":"Contents: Electrophoresis 23–24'24","authors":"","doi":"10.1002/elps.202470123","DOIUrl":"https://doi.org/10.1002/elps.202470123","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 23-24","pages":"2073-2074"},"PeriodicalIF":3.0,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143244952","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 Ge.F.I. Collaborative Study: Evaluating Reproducibility and Accuracy of a DNA-Methylation-Based Age-Predictive Assay for Routine Implementation in Forensic Casework","authors":"Martina Onofri,&nbsp;Federica Alessandrini,&nbsp;Serena Aneli,&nbsp;Loredana Buscemi,&nbsp;Elena Chierto,&nbsp;Matteo Fabbri,&nbsp;Paolo Fattorini,&nbsp;Paolo Garofano,&nbsp;Fabiano Gentile,&nbsp;Silvano Presciuttini,&nbsp;Carlo Previderè,&nbsp;Carlo Robino,&nbsp;Simona Severini,&nbsp;Federica Tommolini,&nbsp;Pamela Tozzo,&nbsp;Andrea Verzeletti,&nbsp;Eugenia Carnevali","doi":"10.1002/elps.202400190","DOIUrl":"10.1002/elps.202400190","url":null,"abstract":"<p>The increasing interest in DNA methylation (DNAm) analysis within the forensic scientific community prompted a collaborative project by Ge.F.I. (Genetisti Forensi Italiani). The study evaluated a standardized bisulfite conversion–based Single Base Extension (SBE) protocol for the analysis of the methylation levels at five age-predictive loci (ELOVL2, FHL2, KLF14, C1orf132/MIR29B2C, and TRIM59). The study encompassed three phases: (1) setting up and validating the protocol to ensure consistency and reproducibility; (2) comparing fresh peripheral blood with blood spots; and (3) evaluating sources of intra- and inter-laboratory variability. Samples from 22 Italian volunteers were analyzed by 6 laboratories in replicates for a total of 528 records. From phase I emerged that the choice of genetic sequencer significantly contributed to inter-laboratory data variation, resulting in separate regression analyses performed for each laboratory. In phase II, blood spots were found to be a reliable source for DNAm analysis, despite exhibiting increased experimental variation compared to fresh peripheral blood. In phase III, a strong correlation between the individual's predicted and true ages was observed across different laboratories. Analysis of variance (ANOVA) of the residuals indicated that one-third of the total variance could be attributed to laboratory-specific factors, whereas two-thirds could be attributed to inter-individual biological differences. The leave-one-out cross-validation (LOO-CV) method yielded an overall mean absolute deviation (MAD) value of 4.41 years, with an average 95% confidence interval of 5.24 years. Stepwise regression analysis proved that a restricted model (ELOVL2, C1orf132/MIR29B2C, and TRIM59) produced results virtually indistinguishable from the five-loci model. Additionally, the analysis of samples in replicates greatly improved the fit of the regression model, balancing the slight effects of intra-laboratory variability. In conclusion, the bisulfite conversion–based SBE protocol, combined with replicate analysis and in-lab calibration of a regression-prediction model, proves to be a reliable and easily implementable method for age prediction in forensic laboratories.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 1-2","pages":"76-91"},"PeriodicalIF":3.0,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11773317/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143055888","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":"Isoelectric Focusing Fractionation Method for Signal Enhancement in Detection of Inactivated Biological Agents Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry","authors":"Filip Duša,&nbsp;Jiří Šalplachta,&nbsp;Marie Horká,&nbsp;Kamila Lunerová,&nbsp;Veronika Čermáková,&nbsp;Michal Dřevínek,&nbsp;Oldřich Kubíček","doi":"10.1002/elps.202400052","DOIUrl":"10.1002/elps.202400052","url":null,"abstract":"<p>Timely identification of highly pathogenic bacteria is crucial for efficient mitigation of the connected harmful health effects. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of intact cells enables fast identification of the microorganisms based on their mass spectrometry protein fingerprint profiles. However, the MALDI-TOF MS examination must be preceded by a time-demanding cultivation of the native bacteria to isolate representative cell samples to obtain indicative fingerprints. Isoelectric focusing (IEF) is capable of separating bacterial cells according to their isoelectric point while effectively removing other non-focusing compounds from sample matrix. In this work, we present a divergent-flow IEF chip (DF-IEF chip) fractionation as an alternative way for sample clean-up and concentration of bacterial cells to prepare samples usable for following MALDI-TOF MS analysis without the need of time-demanding cultivation. By means of DF-IEF chip method, we processed four species of highly pathogenic bacteria (<i>Bacillus anthracis</i>, <i>Brucella abortus</i>, <i>Burkholderia mallei</i>, and <i>Yersinia pestis</i>) inactivated with H₂O₂ vapors or by heat treatment at 62.5°C for 24 h. The DF-IEF chip method continually separated and concentrated the inactivated bacterial cells for subsequent detection using MALDI-TOF MS. The content of the inactivated bacteria in the DF-IEF chip fractions was evaluated with the MS analysis, where inactivated <i>Y. pestis</i> was found to be the most efficiently focusing species. Sensitivity analysis showed limits as low as 2 × 10⁵ colony forming units per mL for inactivated <i>B. anthracis</i>.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 3-4","pages":"212-220"},"PeriodicalIF":3.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.202400052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921174","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":"X-phoresis in Micro/Nano Fluidics 2024","authors":"Xiangchun Xuan,&nbsp;Chun Yang","doi":"10.1002/elps.202470121","DOIUrl":"10.1002/elps.202470121","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 23-24","pages":"2075"},"PeriodicalIF":3.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142920629","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 New Empirical Equation to Take Into Account the Influence of Ionic Strength on Electrophoretic Mobility and Determination of Binding Constants of Ester Betulin Derivatives With Sulfated β-Cyclodextrin by Affinity Capillary Electrophoresis.","authors":"Viktoria V Sursyakova, Vladimir A Levdansky, Anatoly I Rubaylo","doi":"10.1002/elps.202400150","DOIUrl":"https://doi.org/10.1002/elps.202400150","url":null,"abstract":"<p><p>An empirical equation relating electrophoretic mobility and ionic strength was proposed. The equation includes a number of parameters that are found using the mobilities of reference ions: two coefficients in the numerator describing the linear relationship of the multiplier in front of the square root of the ionic strength with the product of the ion mobility in the background electrolyte (BGE) without additives by the modulus of the charge number, raised to a certain power, and also the multiplier in the denominator before the square root of the ionic strength. The proposed equation was tested using the mobilities measured in BGEs with the addition of sodium chloride to adjust ionic strength and sulfated β-cyclodextrin (S-β-CD) for 11 anions with charge numbers from -1 to -4. Measuring anion electrophoretic mobility over a wide range of values, from -17 to -76 × 10^-9 m² V^-1 s^-1, was achieved through a combination of conventional separation and separation in a thermostatically controlled part of a capillary. The equation describes experimental data much more accurately (the discrepancy is no more than 1.1%) compared to the previously used simple empirical equation. For the S-β-CD complexes of betulin derivatives, the apparent binding constants determined are equal to 40-60 M^-1.</p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142909331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of Major Chemical Constituents in Jiuwei Decoction Based On Capillary Electrophoresis Coupled With Quadrupole Time-of-Flight Mass Spectrometry","authors":"Jiayu Zhao,&nbsp;Jinhong Li,&nbsp;Hongyu Xue,&nbsp;Qiang Ma","doi":"10.1002/elps.202400204","DOIUrl":"10.1002/elps.202400204","url":null,"abstract":"<div>\u0000 \u0000 <p>An analytical method for identifying the chemical constituents in the Chinese herbal combination Jiuwei decoction was established using capillary electrophoresis coupled with quadrupole time-of-flight mass spectrometry. Nine herbs were extracted with a 60:40 (v/v) ethanol/water solution to prepare the Jiuwei decoction. Electrophoretic separation was carried out using a 50 µm i.d., 125 cm bare fused silica capillary at a constant temperature of 25°C for sample injection. The chemical composition was analyzed and identified in both positive and negative ion modes. In the positive separation mode, a separation voltage of 30 kV was applied, with a 1% aqueous formic acid solution serving as the background electrolyte (BGE), along with a 50:50 (v/v) methanol/water solution containing 0.1% formic acid as the sheath liquid. In the negative separation mode, a separation voltage of −30 kV was employed, utilizing a 50 mM ammonium acetate solution (pH 9.0) as the BGE, combined with a 50:50 (v/v) methanol<i>/</i>water solution containing 5 mM ammonium acetate as the sheath liquid. The MassHunter Qualitative Analysis software (version 10.0) was utilized to identify 52 compounds based on accurate mass, isotope abundance, isotopic patterns, and fragmentation information. The data were compared with the Agilent Personal Compound Database and Library of traditional Chinese medicine (TCM), as well as multiple online databases (TCMSP, ChemSpider, PubChem, PubMed, and Web of Science) and relevant literature. The fragmentation pathways of representative compounds were also characterized. This study identified the chemical composition of Jiuwei decoction, providing a molecular basis for further exploration of its pharmacological effects and presenting a robust methodology for the compositional analysis of complex TCM mixtures.</p>\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 5-6","pages":"322-330"},"PeriodicalIF":3.0,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142881747","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 21–22'24","authors":"","doi":"10.1002/elps.202470112","DOIUrl":"https://doi.org/10.1002/elps.202470112","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"1879-1880"},"PeriodicalIF":3.0,"publicationDate":"2024-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.202470112","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142868793","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":"Front Cover: Development of the SupersonicIEF Method for High-Throughput Charge Variant Analysis","authors":"Will McElroy,&nbsp;Christopher D. Heger","doi":"10.1002/elps.202470111","DOIUrl":"https://doi.org/10.1002/elps.202470111","url":null,"abstract":"<p>DOI: 10.1002/elps.202400117</p><p>The cover image is based on the Article <i>Development of the SupersonicIEF Method for High-Throughput Charge Variant Analysis</i> by Will McElroy and Christopher D. Heger, https://doi.org/10.1002/elps.202400117.\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elps.202470111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142869040","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":"Contents: Electrophoresis 21–22'24","authors":"","doi":"10.1002/elps.202470113","DOIUrl":"https://doi.org/10.1002/elps.202470113","url":null,"abstract":"","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"1881-1883"},"PeriodicalIF":3.0,"publicationDate":"2024-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142869041","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}