ELECTROPHORESIS最新文献_第6页

Reshaping Capillary Electrophoresis With State-of-the-Art Sample Preparation Materials: Exploring New Horizons. 用最先进的样品制备材料重塑毛细管电泳：探索新视野。

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-30 DOI: 10.1002/elps.202400114

Alaa Bedair, Mahmoud Hamed, Fotouh R Mansour

{"title":"Reshaping Capillary Electrophoresis With State-of-the-Art Sample Preparation Materials: Exploring New Horizons.","authors":"Alaa Bedair, Mahmoud Hamed, Fotouh R Mansour","doi":"10.1002/elps.202400114","DOIUrl":"https://doi.org/10.1002/elps.202400114","url":null,"abstract":"Capillary electrophoresis (CE) is a powerful analysis technique with advantages such as high separation efficiency with resolution factors above 1.5, low sample consumption of less than 10 µL, cost-effectiveness, and eco-friendliness such as reduced solvent use and lower operational costs. However, CE also faces limitations, including limited detection sensitivity for low-concentration samples and interference from complex biological matrices. Prior to performing CE, it is common to utilize sample preparation procedures such as solid-phase microextraction (SPME) and liquid-phase microextraction (LPME) in order to improve the sensitivity and selectivity of the analysis. Recently, there have been advancements in the development of novel materials that have the potential to greatly enhance the performance of SPME and LPME. This review examines various materials and their uses in microextraction when combined with CE. These materials include carbon nanotubes, covalent organic frameworks, metal-organic frameworks, graphene and its derivatives, molecularly imprinted polymers, layered double hydroxides, ionic liquids, and deep eutectic solvents. The utilization of these innovative materials in extraction methods is being examined. Analyte recoveries and detection limits attained for a range of sample matrices are used to assess their effects on extraction selectivity, sensitivity, and efficiency. Exploring new materials for use in sample preparation techniques is important as it enables researchers to address current limitations of CE. The development of novel materials has the potential to greatly enhance extraction selectivity, sensitivity, and efficiency, thereby improving CE performance for complex biological analysis.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":" ","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343835","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

Analytical Methods to Evaluate RNA Circularization Efficiency 评估 RNA 循环效率的分析方法

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-30 DOI: 10.1002/elps.202400067

Yali Sun, Anis H. Khimani, Yanhong Tong, Zhi-xiang Lu

引用次数: 0

Imaged Capillary Isoelectric Focusing Coupled to High-Resolution Mass Spectrometry (icIEF-MS) for Cysteine-Linked Antibody–Drug Conjugate (ADC) Heterogeneity Characterization Under Native Condition 成像毛细管等电聚焦与高分辨率质谱联用技术（icIEF-MS）用于原生态条件下半胱氨酸连接抗体-药物共轭物（ADC）的异质性表征。

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-30 DOI: 10.1002/elps.202400083

Xiaoxi Zhang, Gang Wu, Min Du, Tao Bo, Tong Chen, Tiemin Huang

{"title":"Imaged Capillary Isoelectric Focusing Coupled to High-Resolution Mass Spectrometry (icIEF-MS) for Cysteine-Linked Antibody–Drug Conjugate (ADC) Heterogeneity Characterization Under Native Condition","authors":"Xiaoxi Zhang, Gang Wu, Min Du, Tao Bo, Tong Chen, Tiemin Huang","doi":"10.1002/elps.202400083","DOIUrl":"10.1002/elps.202400083","url":null,"abstract":"<div>\u0000 \u0000 Native mass spectrometry (nMS) is a cutting-edge technique that leverages electrospray ionization MS (ESI-MS) to investigate large biomolecules and their complexes in solution. The goal of nMS is to retain the native structural features and interactions of the analytes during the transition to the gas phase, providing insights into their natural conformations. In biopharmaceutical development, nMS serves as a powerful tool for analyzing complex protein heterogeneity, allowing for the examination of non-covalently bonded assemblies in a state that closely resembles their natural folded form. Herein, we present an imaged capillary isoelectric focusing–MS (icIEF–MS) workflow to characterize cysteine-linked antibody–drug conjugate (ADC) under native conditions. Two ADCs were analyzed: a latest generation cysteine-linked ADC polatuzumab vedotin and the first FDA-approved cysteine-linked ADC brentuximab vedotin. This workflow benefits from a recently developed icIEF system that is MS-friendly and capable of directly coupling to a high-sensitivity MS instrument. Results show that the icIEF separation is influenced by both drug payloads and the post-translational modifications (PTMs), which are then promptly identified by MS. Overall, this native icIEF–MS method demonstrates the potential to understand and control the critical quality attributes (CQAs) that are essential for the safe and effective use of ADCs.\u0000 </div>","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"1915-1926"},"PeriodicalIF":3.0,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343832","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 15–16'24 编辑委员会：电泳 15-16'24

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-27 DOI: 10.1002/elps.202470092

引用次数: 0

Front Cover: A dual-reference study design for understanding and improving AAV genome size analysis 封面：了解和改进 AAV 基因组大小分析的双重参考研究设计

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-27 DOI: 10.1002/elps.202470091

Yali Sun, Zhi-xiang Lu, Michael Miller, Ying Valcour, Anis H Khimani, Jana Bauer, Michael Salomon, Yanhong Tong

引用次数: 0

Moment analysis method for the determination of permeation kinetics of coumarin at lipid bilayers of liposomes by using capillary electrophoresis 利用毛细管电泳测定香豆素在脂质体双分子层渗透动力学的矩分析方法。

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-27 DOI: 10.1002/elps.202400100

Kanji Miyabe, Mio Sakai, Shunta Inaba

{"title":"Moment analysis method for the determination of permeation kinetics of coumarin at lipid bilayers of liposomes by using capillary electrophoresis","authors":"Kanji Miyabe, Mio Sakai, Shunta Inaba","doi":"10.1002/elps.202400100","DOIUrl":"10.1002/elps.202400100","url":null,"abstract":"A method was developed for studying mass transfer kinetics at lipid bilayers of liposomes. Elution peaks of coumarin were measured by liposome electrokinetic chromatography (LEKC). Four types of phospholipids having different alkyl chains were used for preparing liposomes, which were used as pseudo-stationary phases in LEKC systems. Rate constants of permeation across lipid bilayers of liposomes or of adsorption at lipid membranes were determined by analyzing the first absolute and second central moments of the elution peaks measured by LEKC. The rate constants of permeation or adsorption tend to decrease with an increase in the carbon number of the alkyl chains of phospholipids. It was demonstrated that the moment analysis of elution peak profiles measured by LEKC is effective for determining lipid membrane permeability or adsorption kinetics. Compared with other conventional techniques, the method has some advantages for studying mass transfer kinetics at lipid bilayers. Solute permeation across or solute adsorption at real lipid bilayers of liposomes is analyzed. The principle of the method is the analysis of separation behavior in LEKC, which is different from that of the other ones. It is expected that the method contributes to the kinetic study of mass transfer at lipid bilayers from various perspectives.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"1885-1894"},"PeriodicalIF":3.0,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343833","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

Contents: Electrophoresis 17–18'24 内容：电泳 17-18'24

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-27 DOI: 10.1002/elps.202470093

引用次数: 0

Comparative investigation of exosome extraction from rat bone marrow mesenchymal stem cells using three different methodologies 使用三种不同方法从大鼠骨髓间充质干细胞中提取外泌体的比较研究。

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-23 DOI: 10.1002/elps.202400055

Na Wang, Mingyue Yin, Jiaqi Yu, Jing Zhang, Xueli Pan

{"title":"Comparative investigation of exosome extraction from rat bone marrow mesenchymal stem cells using three different methodologies","authors":"Na Wang, Mingyue Yin, Jiaqi Yu, Jing Zhang, Xueli Pan","doi":"10.1002/elps.202400055","DOIUrl":"10.1002/elps.202400055","url":null,"abstract":"Exosomes have been identified as crucial mediators in numerous physiological and pathological processes, emerging as a focal point of scientific inquiry. This study aims to compare three methods for isolating exosomes from rat bone marrow mesenchymal stem cells: ultracentrifugation (UC), ultrafast separation system (EXODUS), and commercial precipitation kit (EXO-kit). First, the investigation compared exosomal morphology, particle size distribution, and expression of marker proteins. Subsequently, the RNA content, protein concentration, and purity of exosomes were evaluated. Finally, the impact of these exosomes on cellular metabolic viability and migration capacity was assessed. Results indicated that exosomes exhibited spherical or elliptical membrane structures, and most of the exosomes extracted by the three methods were in the range of 30 to 200 nm. UC-extracted exosomes demonstrated the least impurities and clearest background, followed by EXODUS-extracted exosomes, and lastly EXO-kit-extracted exosomes. The EXO-kit-extracted exosomes yielded the highest RNA and protein content, whereas those isolated through UC exhibited superior purity. Furthermore, exosomes extracted from EXODUS and EXO-kit methods effectively enhanced the metabolic viability and migratory ability of osteoblast precursor cells compared to UC-extracted exosomes. In conclusion, each of the three methodologies presents advantages and limitations. Therefore, the selection of an appropriate exosome extraction technique should be based on specific experimental objectives and requirements.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"2045-2053"},"PeriodicalIF":3.0,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142282186","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

Standardization and application of ARMS TaqMan real-time PCR for screening of folate metabolism genes in Han Chinese ARMS TaqMan 实时 PCR 在汉族叶酸代谢基因筛查中的标准化和应用

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-17 DOI: 10.1002/elps.202400017

Peipei Deng, Xuan Liu, Yuanjing Li, Huanhuan Li, Bangrong Zhao, Shusong Wang, Jing Ma

{"title":"Standardization and application of ARMS TaqMan real-time PCR for screening of folate metabolism genes in Han Chinese","authors":"Peipei Deng, Xuan Liu, Yuanjing Li, Huanhuan Li, Bangrong Zhao, Shusong Wang, Jing Ma","doi":"10.1002/elps.202400017","DOIUrl":"10.1002/elps.202400017","url":null,"abstract":"Folate has antioxidant properties, and low concentration in seminal plasma may be associated with increased DNA damage in sperm. Mutations of the methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR) genes, including MTHFR C677T (rs1801133), MTHFR A1298C (rs1801131), and MTRR A66G (rs1801394), can lead to decreased activity of the encoded folate metabolic enzymes, thereby affecting male reproduction. The current SNP detection methods commonly used in clinical practice have some shortcomings, such as long time-consuming, complex detection steps, or high cost. The purpose of this study was to establish a simple, time-saving, sensitive, accurate, and easy to clinical popularization method for folate metabolism gene detection. We combined ARMS-PCR with TaqMan fluorescent probe to establish an ARMS TaqMan real-time PCR detection method. According to the variation of rs1801131, rs1801133, and rs1801394, two specific primers (one wild type and one mutant) were designed. Mismatched nucleotides were introduced at the penultimate or third position to improve the specificity of the primer. Specific TaqMan probe was introduced to detect PCR products to improve the sensitivity of the method. The results showed that the sensitivity of ARMS TaqMan real-time PCR in SNP genotyping was 1 ng, and the accuracy was 100%. A total of 249 clinical samples were detected by the established method, and the correlation between three SNPs and semen quality was analyzed. We found that individuals carrying the AG + GG genotype of rs1801394 had a lower risk of abnormal semen quality. In conclusion, we developed a highly sensitive, accurate, rapid, and easy to be popularized method for detecting SNPs of rs1801394, rs1801131, and rs1801133. ARMS TaqMan real-time PCR is a reliable SNP genotyping method in folate metabolism genes.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"1995-2004"},"PeriodicalIF":3.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252731","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

Preparation of alkyl microporous organic network-based capillary column for an efficient gas chromatographic separation of position isomers 制备用于高效气相色谱分离位置异构体的烷基微孔有机网络毛细管柱

IF 3 3区生物学

ELECTROPHORESIS Pub Date : 2024-09-17 DOI: 10.1002/elps.202400111

Zhi-Yong Gao, Xi-Jin Li, Yuan-Yuan Cui, Cheng-Xiong Yang

{"title":"Preparation of alkyl microporous organic network-based capillary column for an efficient gas chromatographic separation of position isomers","authors":"Zhi-Yong Gao, Xi-Jin Li, Yuan-Yuan Cui, Cheng-Xiong Yang","doi":"10.1002/elps.202400111","DOIUrl":"10.1002/elps.202400111","url":null,"abstract":"The large surface area, excellent thermal stability and easy modification make microporous organic networks (MONs) good candidates in the field of gas chromatography (GC). Due to the limited species and highly conjugated networks of MONs, their applications are still in infancy and restricted. To accelerate their developments and to enrich their types in GC, here we report the first example of synthesizing alkyl MON and its capillary column for GC separation of position isomers. Linear 1,8-dibromooctane is used as the alkyl monomer instead of traditional aromatic ones to construct novel alkyl MON to decrease the inherent conjugated characteristic of MONs. The alkyl MON exhibits good thermal stability (up to 350°C), large surface area (1173 m2 g−1), and non-polar character, allowing good resolution for alkanes, alkyl benzenes, alcohols, ketones, and diverse position isomers, including dichlorobenzene, trichlorobenzene, bromotoluene, nitrotoluene, methylbenzaldehyde, and ionone with the limits of detection (0.003 mg mL−1) and limits of quantitation of (0.10 mg mL−1). The in situ growth–prepared alkyl MON column demonstrates remarkable duration time and precisions for the retention relative standard deviations, (RSDs%, intra-day, n = 7), 0.06%–0.53% (intra-day, n = 7), and 2.87%–10.59% (column-to-column, n = 3). In addition, the fabricated alkyl MON-coated capillary column offers better resolution than three commercial GC columns for the resolution of methylbenzaldehyde, bromotoluene, and chlorotoluene isomers. This work reveals the practicability for synthesizing alkyl MONs and demonstrates their prospects for position isomers separation.","PeriodicalId":11596,"journal":{"name":"ELECTROPHORESIS","volume":"45 21-22","pages":"1895-1905"},"PeriodicalIF":3.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252730","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