ELECTROPHORESIS最新文献_第4页

Construction of A Novel 5-Dye Fluorescent Multiplex System With 30 Y-STRs for Patrilineal Relationship Prediction. 30个Y-STRs的新型5染料荧光多路系统的构建用于父系关系预测。

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-08-20 DOI: 10.1002/elps.70015

Chaoran Sun, Zhirui Zhang, Xindi Wang, Bo Liu, Chengye Zhou, Yufei Yang, Chuanxu Wang, Sunxi Xu, Chang Wang, Lagabaiyila Zha, Jienan Li, Haibo Luo, Feng Song

{"title":"Construction of A Novel 5-Dye Fluorescent Multiplex System With 30 Y-STRs for Patrilineal Relationship Prediction.","authors":"Chaoran Sun, Zhirui Zhang, Xindi Wang, Bo Liu, Chengye Zhou, Yufei Yang, Chuanxu Wang, Sunxi Xu, Chang Wang, Lagabaiyila Zha, Jienan Li, Haibo Luo, Feng Song","doi":"10.1002/elps.70015","DOIUrl":"https://doi.org/10.1002/elps.70015","url":null,"abstract":"Nowadays, Y chromosome short tandem repeats (Y-STRs) are widely used in forensic medicine practice, which has great significance for ancestry tracing, male lineage evolution, and male paternal relatives. Rapidly mutating Y-STRs (RM Y-STRs) have been shown to have greater potential to distinguish males from males in the patrilineal line. Therefore, a novel 5-dye fluorescent multiplex system with 30 Y-STRs was developed and optimized to screen out more RM Y-STRs and fasting mutating Y-STRs (FM Y-STRs). New primers were designed, and composite system construction was carried out. A series of experiments were conducted following the guidelines of the Scientific Working Group on DNA Analysis Methods (SWGDAM), including polymerase chain reaction (PCR) amplification conditions, sensitivity, stability, species specificity, mixture and degraded sample studies, mutation analysis, and population studies. The results suggested that changing PCR amplification conditions in a reasonable range hardly affected the genotyping. The system showed excellent sensitivity and stability in sensitivity and stability studies. Even with UV-C exposure for up to 96 h, the system performed well in male blood samples and semen-vaginal secretion mixtures. Mutation analysis was performed on 582 father-son pairs, and 4 RM Y-STRs and 7 FM Y-STRs were identified, with mutation rates ranging from 1.72 × 10-3 to 20.62 × 10-3. Furthermore, on the basis of mutation rate analysis, eight machine learning methods were used to construct and compare patrilineal relationship prediction models, inferring the relationship by predicting the number of meiosis. Overall, the multiplex system displays favorable performance and has a greater potential for application in forensic science practice.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144947064","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}

引用次数: 0

Editorial Board: Electrophoresis 11–12'25 编辑委员会：电泳11-12 '25

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-08-20 DOI: 10.1002/elps.70012

引用次数: 0

Epigenetic Age Signatures in Postmortem Rib Samples. 死后肋骨样本的表观遗传年龄特征。

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-08-13 DOI: 10.1002/elps.70014

Siqi Chen, Changquan Zhang, Dan Wen, Chudong Wang, Xuan Tang, Xue Li, Xiaoyi Fu, Jienan Li, Xin Jin, Haibo Luo, Feng Song, Ying Liu, Lagabaiyila Zha

{"title":"Epigenetic Age Signatures in Postmortem Rib Samples.","authors":"Siqi Chen, Changquan Zhang, Dan Wen, Chudong Wang, Xuan Tang, Xue Li, Xiaoyi Fu, Jienan Li, Xin Jin, Haibo Luo, Feng Song, Ying Liu, Lagabaiyila Zha","doi":"10.1002/elps.70014","DOIUrl":"https://doi.org/10.1002/elps.70014","url":null,"abstract":"Skeletal remains, often partially or completely decomposed, are among the most common biological forensic samples found at crime scenes. Analyzing these incomplete specimens to estimate the age of the deceased is crucial. Previous studies on DNA methylation-based age prediction in bones have not evaluated differences across skeletal elements or clarified how bone type influences prediction accuracy. This study focuses on postmortem rib samples-a common forensic specimen-to develop a DNA methylation-based age prediction model specific to ribs. DNA methylation levels at eight CpG sites within the ELOVL2, FHL2, KLF14, and FAM123C genes were analyzed using pyrosequencing in 81 postmortem rib samples and 112 postmortem blood samples, with 50 individuals providing both sample types simultaneously. The rib-derived age prediction model exhibited an R2 value of 0.908, whereas the blood model achieved an R2 value of 0.927. For the rib model, the mean absolute deviation (MAD) of the training set was 4.813 years, and the MAD of the testing set was 5.084 years. The blood model showed slightly higher accuracy in predicting the age of the same individuals. Notably, cross-tissue application of models led to significant prediction bias, emphasizing the necessity of tissue-specific calibration for methylation-based age estimation. Exploratory analysis of postmortem sternum, rib, and frontal bone samples from 12 individuals revealed no statistically significant differences in methylation levels or age estimates across bone types. However, broader generalizability of the rib model to these skeletal elements requires validation in larger, independent cohorts. This work establishes a robust age prediction framework for rib samples, highlights the critical role of tissue specificity in epigenetic forensic models, and provides preliminary evidence for potential cross-bone applicability.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144834459","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}

引用次数: 0

Label-Free Sorting of Human Mesenchymal Stem Cells Using Insulating Dielectrophoresis. 人间充质干细胞绝缘介质电泳的无标记分选。

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-24 DOI: 10.1002/elps.70001

Zuri A Rashad, Kiara L Lacy, Emmanuel Egun, Jazmine S Moore, Tayloria N G Adams

{"title":"Label-Free Sorting of Human Mesenchymal Stem Cells Using Insulating Dielectrophoresis.","authors":"Zuri A Rashad, Kiara L Lacy, Emmanuel Egun, Jazmine S Moore, Tayloria N G Adams","doi":"10.1002/elps.70001","DOIUrl":"https://doi.org/10.1002/elps.70001","url":null,"abstract":"Human mesenchymal stem cells (hMSCs) are a multipotent yet heterogeneous cell population with immunosuppressive and regenerative properties, making them highly promising for stem cell therapies targeting metabolic diseases. However, the inherent heterogeneity of hMSCs presents challenges for producing consistent therapeutic outcomes, emphasizing the need to isolate functionally distinct subpopulations. In this study, we employed insulating dielectrophoresis (DEP) via a trap-and-release sorting strategy to generate and characterize subpopulations of adipose tissue (AT)-derived hMSCs. Voltage and frequency parameters were systematically tuned, revealing that higher voltages increased the percentage of trapped cells, while higher frequencies had less impact. Sorted cells underwent a 14-day adipogenic differentiation process, assessed by Oil Red O staining. Our results demonstrated that untrapped cell populations generated at lower voltage and frequency thresholds exhibited enhanced adipogenic differentiation compared to unsorted controls. These findings suggest that DEP can be leveraged to isolate progenitor cells within hMSC populations, enabling the production of homogeneous cell subsets with targeted functional potential. This work highlights the utility of insulating DEP for addressing hMSC heterogeneity and advancing the development of stem cell therapies.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":"e70001"},"PeriodicalIF":3.0,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144706831","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}

引用次数: 0

Investigating Weak Polypeptide-Cyclodextrin Interactions in Biologic Formulation Development Using Affinity Capillary Electrophoresis and Flow-Induced Dispersion Analysis. 利用亲和毛细管电泳和流动诱导分散分析研究生物制剂开发中的弱多肽-环糊精相互作用。

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-24 DOI: 10.1002/elps.70000

Yunxiao Zhu, Sharadvi Thati, Megan Mccallum, Rao Mantri, William Ying, Neil Mathias, Mark Bolgar, Wenkui Lan

{"title":"Investigating Weak Polypeptide-Cyclodextrin Interactions in Biologic Formulation Development Using Affinity Capillary Electrophoresis and Flow-Induced Dispersion Analysis.","authors":"Yunxiao Zhu, Sharadvi Thati, Megan Mccallum, Rao Mantri, William Ying, Neil Mathias, Mark Bolgar, Wenkui Lan","doi":"10.1002/elps.70000","DOIUrl":"https://doi.org/10.1002/elps.70000","url":null,"abstract":"Understanding protein-excipient interactions is vital for biopharmaceutical formulation, as they influence stability and pharmacokinetics (PK). Cyclodextrins (CDs) are widely used excipients that enhance solubility and stability, but their weak interactions with polypeptides remain poorly characterized. Relaxin (RLX), a potent anti-heart failure polypeptide, was selected due to its PK relevance and in vivo interaction with human serum albumin (HSA). Given RLX's poor solubility, CDs were identified as the most effective solubilizers. However, traditional affinity assays lack the sensitivity to detect weak CD-polypeptide interactions. To overcome this limitation, we employed affinity capillary electrophoresis and flow-induced dispersion analysis (FIDA) to assess RLX's binding with hydroxypropyl-β-cyclodextrin (HP-β-CD) and sulfobutylether-β-cyclodextrin (SBE-β-CD). Our results showed a higher affinity for SBE-β-CD than HP-β-CD, though both interactions were significantly weaker than RLX's binding to HSA. These findings provide key insights into weak CD-polypeptide interactions, supporting SBE-β-CD as an excipient to improve solubility without compromising PK performance. Additionally, the effectiveness of these rapid, nonconventional analytical methods was validated through in vivo PK studies in a cynomolgus monkey model, highlighting their value in excipient-protein binding research.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":"e70000"},"PeriodicalIF":3.0,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144706830","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}

引用次数: 0

Editorial Board: Electrophoresis 9–10'25 编辑委员会：电泳9-10 '25

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-18 DOI: 10.1002/elps.70010

引用次数: 0

Sample Preparation 2024 样品制备2024

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-18 DOI: 10.1002/elps.70003

Carlos D. Garcia, Pavel Kubáň, Petr Kubáň

引用次数: 0

Sialylation Impacts Separation of a Biotherapeutic by Capillary Gel Electrophoresis 唾液化对毛细管凝胶电泳分离生物治疗药物的影响。

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-08 DOI: 10.1002/elps.70006

Jeremy D. Osko, Zhengqi Zhang, Andrew Semple, Karen Bern, Julie C. McIntosh, Xiaoyu Yang, Thomas P. Niedringhaus

{"title":"Sialylation Impacts Separation of a Biotherapeutic by Capillary Gel Electrophoresis","authors":"Jeremy D. Osko, Zhengqi Zhang, Andrew Semple, Karen Bern, Julie C. McIntosh, Xiaoyu Yang, Thomas P. Niedringhaus","doi":"10.1002/elps.70006","DOIUrl":"10.1002/elps.70006","url":null,"abstract":"<div>\u0000 \u0000 Monitoring of critical quality attributes (CQAs) is essential for the development of biotherapeutics. One example of a CQA is molecular fragmentation, which is often analyzed by capillary gel electrophoresis with sodium dodecyl sulfate (SDS). Sialylation is a post-translational modification and form of glycosylation that can impact purity profiles of biotherapeutics, resulting in complex structure elucidation. Here, we studied the heterogeneity of Biotherapeutic 1 as a result of O-linked glycosylation with sialylation. Biotherapeutic 1 displayed a second unidentified peak in SDS–capillary gel electrophoresis (SDS-CGE) under reducing conditions that directly impacted peak integration practices and, therefore, method validation. The two peaks were highly reproducible in SDS-CGE as well as complementary LabChip experiments. The apparent molecular weights were calculated using molecular weight ladders with known protein standards. A combination of ion exchange chromatography (IEX), hydrophilic interaction chromatography mass spectrometry (HILIC-MS), and ultra-high-performance size exclusion chromatography (UP-SEC) were used to identify O-linked glycosylation as responsible for the production of reduced peak 1 and reduced peak 2 in SDS-CGE. Specifically, reduced peak 2 contained sialylation that was not observed in reduced peak 1, resulting in two distinct migration times due to impacts in SDS binding efficacy. Enzymatic removal of the sialic acids simplified the heterogeneity into a single uniform peak (reduced peak 1). This work and methodologies highlight the impact a single O-linked glycan can have on SDS-CGE and is applicable to analyzing future biotherapeutics involving complex structure profiles resulting from sialylation.\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 11-12","pages":"687-693"},"PeriodicalIF":2.5,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590708","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}

引用次数: 0

An Optimized, CE-Compatible, Targeted NGS-Based SSR Genotyping Method Using Primer-Anchored Alignment 基于引物锚定比对的优化、ce兼容、靶向的ngs SSR基因分型方法

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-04 DOI: 10.1002/elps.70007

Kai Liu, Qinghui Meng, Tianlun Zheng, Nan Xie, Fan Zhou

{"title":"An Optimized, CE-Compatible, Targeted NGS-Based SSR Genotyping Method Using Primer-Anchored Alignment","authors":"Kai Liu, Qinghui Meng, Tianlun Zheng, Nan Xie, Fan Zhou","doi":"10.1002/elps.70007","DOIUrl":"10.1002/elps.70007","url":null,"abstract":"<div>\u0000 \u0000 Microsatellites (SSRs) are highly polymorphic DNA sequences widely used in genetic research, including parentage assignment. Traditional SSR analysis relies on capillary electrophoresis (CE), which is time-consuming and has limited capacity. Next-generation sequencing (NGS) offers a high-throughput and cost-effective alternative, but existing NGS-based SSR genotyping methods produce results that are incompatible with CE data, increasing the risk of Mendelian inheritance mismatches. This study presents an optimized, targeted, NGS-based approach for SSR genotyping that prioritizes consistency with CE-based results. We optimized SSRseq, a targeted NGS-based SSR genotyping method, by (1) using primer flanking sequences as anchors for BLAST (Basic Local Alignment Search Tool)-based read alignment to reference SSRs, enabling the utilization of both overlapping and nonoverlapping paired-end reads; (2) inferring motif repeat counts from aligned read lengths, tolerating imperfections within the microsatellite repeat array (MRA); and (3) dynamically adjusting motif definition when discrepancies arose between expected and observed MRAs. We evaluated our optimized SSRseq against the original SSRseq and CE using four 10-plex SSR panels for parentage assignment in Largemouth black bass (Micropterus salmoides). The optimized SSRseq substantially improved parentage assignment accuracy. Multiple combinations of two or more optimized SSRseq panels achieved an assignment rate of 1.000 and an accuracy rate of 0.950, whereas the original SSRseq's highest accuracy was 0.900, requiring all four panels. The optimized method also showed high concordance with CE genotyping at several tested loci. This optimized SSRseq approach provides a robust, efficient, and cost-effective tool, leveraging NGS for accurate SSR genotyping in parentage assignment and other genetic analyses while minimizing Mendelian inheritance mismatches.\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"46 11-12","pages":"716-726"},"PeriodicalIF":2.5,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564696","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}

引用次数: 0

Capillary Electrophoresis Can Detect the Simultaneous Presence of Hairpins and Self-Dimers in Non-Symmetric, Single-Stranded DNA Oligomers 毛细管电泳可以检测非对称单链DNA低聚物中同时存在的发夹和自二聚体。

IF 2.5 3区生物学

ELECTROPHORESIS Pub Date : 2025-07-02 DOI: 10.1002/elps.70005

Earle Stellwagen, Nancy C. Stellwagen

引用次数: 0