Se pu = Chinese journal of chromatography最新文献

{"title":"[Lipidomics analysis of glycine-induced bacterial outer membrane vesicles].","authors":"Jing-Yuan Song, Xiu-Lei Qi, Huai-Zhong Guo, Liang-Hai Hu","doi":"10.3724/SP.J.1123.2024.10017","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.10017","url":null,"abstract":"<p><p>Outer membrane vesicles (OMVs) are nanoparticles with double-phospholipid membrane structures that are secreted by gram-negative bacteria and carry a variety of bioactive substances from parental bacterial cells; consequently, OMVs serve as disease markers. Moreover, bacterial OMVs are potential anticancer- and antibacterial-drug carriers. While the addition of glycine during bacterial culturing promotes the secretion of bacterial OMVs, glycine-induced differences in the lipid compositions of such OMVs have not yet been reported. In this study, the key plasma membrane components of bacterial OMVs before and after glycine induction were analyzed using lipidomics. Bacterial OMVs were captured from bacterial-culture supernatants using an amphiphilic dendritic polymeric supramolecular probe. Two sets of enriched bacterial OMVs were characterized and their enrichment efficiencies determined, after which the numbers and purities of the OMVs within the samples were determined using a nanofluidic assay. Lipids were extracted using the methyl <i>tert</i>-butyl ether lipid-extraction method when consistent numbers were recorded. The lipid compositions of the bacterial OMVs before and after glycine induction were analyzed using an ultra-performance liquid chromatography-ion mobility spectrometry-quadrupole time-of-flight mass spectrometer (UPLC-IMS-QTOF-MS) and MS-DIAL software. Differential lipid species in the key plasma membranes of the bacterial OMVs following glycine induction were recorded along with their corresponding amounts. Detection was accomplished in positive-ion scanning mode using an ACQUITY UPLC BEH C18 column following UPLC-MS injection, and MS<sup>E</sup> mass-spectrometry data-acquisition mode. The lipid components in the two groups were determined by combining mass-spectrometric and software-analysis data, which revealed that the addition of glycine to the <i>E. coli</i> Nissle 1917 culture led to two-to-three-times higher concentrations of OMVs than observed for the untreated group under the same culturing and enrichment conditions. Particle numbers measured for the same volume revealed one-order-of-magnitude more bacterial OMVs after induction than before, with the treated group exhibiting slightly larger particles (on average); however, these particles were better dispersed and less likely to aggregate. The identified lipid components were categorized to determine the amount of each lipid type. Differentially expressed lipids were subsequently screened according to experimental conditions; significantly different expression levels were observed following glycine induction, with 820 lipids identified among the 10165 components detected. The lipid classes were ranked in order of quantity as: glycerolipids (GL), fatty acids (FA), sphingolipids (SP), glycerophospholipids (GP), saccharolipids (SL), and sterol lipids (ST), among which 463 GL lipid fractions (56.4% of all characterized lipids) were recorded. The ST lipid fraction cont","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"547-555"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059986/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144049371","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Isolation and proteomics analysis of cerebrospinal fluid exosome subtypes].","authors":"Xiao-Fei Chen, Wei Liu, Wen-Jia Zhang, Yan-Peng Li, Zhi-Hua Wang, Ming-Xia Gao, Xiang-Min Zhang","doi":"10.3724/SP.J.1123.2024.10014","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.10014","url":null,"abstract":"<p><p>Exosomes are small extracellular vesicles 30-200 nm in diameter that contain many bioactive macromolecules, including proteins, lipids, and nucleic acids; consequently, they play important roles in many physiological and pathological processes and are classified into various property-dependent subtypes. Research into exosome heterogeneity helps broaden our understanding of the physiological and pathological mechanisms associated with exosomes. Exosomes exist in many human biological fluids, with those derived from cerebrospinal fluid (CSF) regarded as potential disease biomarkers. Despite this, few studies have focused on their proteomics, and little research into CSF-derived exosome subtypes has been reported. Traumatic brain injury (TBI) is a major public health issue characterized by a large number of patients and complex pathological processes. While a comprehensive understanding of the pathophysiological processes that underpin TBI is essential for developing therapeutic interventions, proteomic studies into CSF-derived exosomes in patients with TBI are limited. Herein, we designed a tandem size-exclusion chromatography protocol for isolating and profiling the proteins of CSF-derived exosome subtypes from patients with TBI using nanoscale liquid chromatography and trapped-ion mobility spectrometry time-of-flight mass spectrometry (nanoLC-TIMS-TOF-MS). We first centrifuged the collected CSF to remove cells and cell debris, after which it was concentrated by ultrafiltration to increase the exosome concentration and remove small proteins and peptides. A mini-size exclusion chromatography (Mini-SEC) column was then used to separate the exosomes from large amounts of interfering proteins, after which high performance liquid-SEC (HPL-SEC) was used to further separate exosomes according to size. The entire extracellular-vesicle-subset separation and purification process takes approximately 1 h for a single CSF sample. Four differently sized exosome subtypes were successfully isolated and are referred to as S1, S2, S3, and S4 in order of descending size. The S1 subtype exhibited the highest exosome purity according to the particle-to-protein ratio. Multiple characterization methods, including transmission electron microscopy (TEM), Western blotting (WB), and nanoparticle tracking analysis (NTA), confirmed that the exosome subtypes had been successfully acquired. NanoLC-TIMS-TOF-MS, combined with database searching were then used to characterize the proteins. A total of 739 proteins were identified, of which 79% and 72% matched all proteins and the top 100 proteins in the Vesiclepedia database, respectively. Moreover, gene ontology analysis revealed that the identified proteins are mainly located in extracellular exosomes, and that the isolated exosome subtypes are closely related to multiple biological processes, including cell signaling, coagulation, and immune responses. Hierarchical cluster analysis revealed that samples from the same exoso","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"518-528"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059988/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144049459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Progress and prospect of separation and analysis of single-cell and single-particle exosomes].","authors":"Ai-Xiang Bu, Guang-Yao Wu, Liang-Hai Hu","doi":"10.3724/SP.J.1123.2024.11001","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.11001","url":null,"abstract":"<p><p>Exosomes are nanoscale vesicles secreted by cells and are encapsulated in lipid bilayers. They play crucial roles in cell communication and are involved in a variety of physiological and pathological processes, including immune regulation, angiogenesis, and tumor initiation and metastasis. Exosomes carry a variety of biomolecules from maternal cells and are therefore important vehicles for discovering disease markers. Traditional detection methods only provide average cell-population information for a given sample and cannot establish clear relationships between the biological functions of exosomes and subtype owing to the significant heterogeneity associated with exosomes from different cell subsets. Therefore, characterizing exosomes at the single-cell and single-particle levels requires exosome specificities to be further explored and the characteristics of various exosome subtypes to be distinguished. Commonly used single-particle exosome characterization technologies include flow cytometry, super-resolution microscopy, atomic force microscopy, surface-enhanced Raman spectroscopy, proximity barcoding assay and MS. In this paper, we summarize recent advances in the separation and characterization of single-cell exosomes based on microfluidics and provide future applications prospects for emerging technologies (such as Olink proteomics, click chemistry, and molecular imprinting) for studying single-cell and single-particle exosomes.</p>","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"399-412"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059993/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144056768","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Exosome separation and enrichment technologies and their applications in disease diagnosis and treatment].","authors":"Guo-Shan Hou, Hui-Ming Yuan, Zhen Liang, Li-Hua Zhang, Yu-Kui Zhang","doi":"10.3724/SP.J.1123.2024.09007","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.09007","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Exosomes are nanoscale vesicles wrapped in lipid bilayers that are secreted by cells and carry a variety of proteins, lipids, nucleic acids, and metabolites. Exosomes are widely present in various bodily fluids and mediate intercellular communication. They participate in a variety of physiological and pathological processes, including immune regulation, angiogenesis, tumorigenesis, and metastasis, and have significant clinical diagnosis and treatment potential. Exosomes are source-rich, structurally stable, and reflect the states of their parental cells. Therefore, they are expected to serve as novel diagnostic markers for various diseases. In addition, stem-cell-derived exosomes show therapeutic potential and have the advantages of low immunogenicity, high safety and easy storage, and exhibit therapeutic potential for neurodegenerative disorder, cardiovascular disease, and cancer. Furthermore, exosomes are highly biocompatible, have natural homing properties, and are capable of easily penetrating biological barriers, making them excellent drug-delivery carriers. Isolation and enrichment of exosomes is a prerequisite for downstream analysis and application. High-purity, high-yield, and high-throughput exosome-isolation methods are expected to be used in clinical diagnosis and treatment applications. Based on the physicochemical properties of exosomes, including density, size, charge, and surface composition, exosome-isolation methods are mainly divided into density-based (e.g., differential ultracentrifugation, density-gradient ultracentrifugation), size-based (e.g., ultrafiltration, size-exclusion chromatography, field-flow fractionation), polymer-precipitation (e.g., polyethylene-glycol-based precipitation), and chemical affinity (e.g., antibody-based, aptamer-based, and surface-lipid-based lipid probes) methods. Currently, basic research into exosomes and their clinical applications face a number of challenges. Firstly, the complexity and heterogeneity of exosomes and the lack of standardized isolation methods has led to highly variable research results that hinder comparing and reproducing results between different laboratories and clinical settings. Current isolation methods are generally hindered by insufficient purity, low yield, low throughput, and difficulties separating specific subpopulations, which seriously restrict the development of the exosome field. Secondly, exosome-isolation methods that are easy to use in the clinic, have few technical requirements, and are highly efficient and inexpensive are lacking. Commonly used classical methods, such as ultracentrifugation, are time-consuming, labor-intensive, require large sample volumes, and are inappropriate for clinical settings. Methods such as immunoaffinity can be used to isolate exosomes from precious trace samples in clinical practice; however, high costs, low recoveries, and high operating requirements are shortcomings that restrict sample analysis in the clinic. In addit","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"434-445"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059991/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Research advances of liposomes and exosomes in drug delivery and biomarker screening].","authors":"Ya-Ting Su, Xiao-Hong Qian, Wei-Jie Qin","doi":"10.3724/SP.J.1123.2024.08012","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.08012","url":null,"abstract":"<p><p>Vesicles, are categorized as artificial (i.e., liposomes) or natural (i.e., extracellular vesicles (EVs)) and play significant roles in drug-delivery and biomarker-screening applications. Liposomes, as a representative form of artificial vesicle, are spherical lipid structures composed of one or more artificially synthesized phospholipid bilayers. Liposomes are highly biocompatible and bioavailable, very stable, and easily synthesized; hence, they are among the most commonly used and frequently applied nanocarriers in targeted drug-delivery systems (DDS). EVs are natural small membrane-bound vesicles actively secreted by cells and contain a variety of components, including nucleic acids, proteins, and lipids. They also serve as important mediators of intercellular communication. As the smallest EV subtype, with diameters of only 30-100 nm, exosomes contain unique biomolecules that are considered to be the fingerprints of the parent cells. In the pathological state, the content of exosomes will change; consequently, exosomes are potential disease-diagnosis biomarkers. Recent clinical trials have shown that exosomes are ideal nanocarriers in targeted drug-delivery therapies for a variety of diseases. Compared with traditional artificial liposomal carriers, exosomes display unique advantages and provide the DDS field with new possibilities. Liposomes and exosomes are receiving increasing levels of attention in the drug-delivery and biomarker-screening fields. This article introduces techniques for the preparation of liposomes, and the enrichment and separation of exosomes, and delves into research progress on their use in drug-delivery and biomarker-screening applications. Finally, challenges facing the use of liposomes and exosomes in clinical applications are discussed.</p>","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"472-486"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059997/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144053917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Efficient capture and proteomics analysis of urinary extracellular vesicles by affinity purification].","authors":"Gui-Yuan Zhang, Zhen Zhan, Wei-Guo Tao, Hao Zhang","doi":"10.3724/SP.J.1123.2024.11013","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.11013","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Liquid biopsy is a promising alternative to traditional tissue biopsies for diagnosing cancer because it offers advantages such as minimal invasiveness, accessibility, and ease of operation. Extracellular vesicles (EVs) are lipid bilayer vesicles that contain proteins, DNA, and RNA and are secreted by cells. Indeed, urinary EVs are important sources of cancer biomarkers. The lipid bilayer protects EV proteins from degradation by enzymes present in bodily fluids. Prostate cancer (PCa) is among the most prevalent malignancies in developed countries and is the second-leading cause of cancer-related mortality in men. Current screening methods commonly used to initially evaluate patients with suspected PCa include serum prostate-specific antigen (PSA) testing and digital rectal examination (DRE), with magnetic resonance imaging (MRI) and transrectal ultrasound often recommended for further assessment. However, both PSA testing and DRE have limited specificities, which results in a substantial number of unnecessary prostate biopsies. Consequently, additional reliable biomarkers need to be urgently discovered for rapidly diagnosing PCa more accurately. Prostate-derived secretions, including those associated with malignancies, are detectable in urine owing to the anatomical proximity of the prostate to the urethra; hence urine is a promising liquid-biopsy medium for discovering PCa biomarkers, which is a topic that has been the focus of extensive research efforts in recent years. However, isolating EVs from biofluids in sufficient yields for proteomics analysis remains challenging. In this study, functional magnetic beads EVlent (extracellular vesicles isoLated efficiently, naturally, and totally) with high-affinity capabilities were developed for selectively enriching EVs from biological fluids.The surfaces of the beads were modified with three antibodies that target CD9, CD63, and CD81, which enables the specific recognition of EV surface proteins. The isolation performance of EVlent was validated by comprehensively characterizing urinary EVs using Western blotting (WB), nanoparticle tracking analysis (NTA), and transmission electron microscopy (TEM). WB revealed prominent bands for EV markers (CD9, TSG101, and HSP70) in EVlent-enriched samples, whereas weaker bands were observed following ultracentrifugation (UC). NTA revealed that the EVs isolated by EVlent are predominantly in the 50-400 nm size range, with a content of 4.1×10&lt;sup&gt;9&lt;/sup&gt; particles/mL, which is significantly higher than the value of 1.8×10&lt;sup&gt;9&lt;/sup&gt; particles/mL obtained by UC. TEM confirmed that the isolated EVs have characteristic elliptical or cup-shaped vesicular structures. These findings demonstrate that EVlent outperforms UC in terms of enrichment efficiency and purity, delivering a separation efficiency of 87.2% compared to the value of 30.3% obtained by UC. We used proteomics to analyze urinary EVs isolated from 15 healthy volunteers and 15 patients with prostate ca","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"508-517"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059984/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Typical strategy and research progress of efficient isolation methods of exosomes based on affinity interaction].","authors":"Hai-Yan Wang, Pei-Juan Xie, Xiao-Qiang Qiao, Li-Yuan Zhang","doi":"10.3724/SP.J.1123.2024.11004","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.11004","url":null,"abstract":"<p><p>Exosomes form a subclass of extracellular vesicle that are secreted by most cells and found in nearly all body fluids, including blood, urine, saliva, amniotic fluid, and milk, as well as in various tissues and intercellular spaces. Exosomes have recently been recognized as crucial intercellular communication mediators, and an increasing number of studies have shown that exosomes are important liquid-biopsy tools that play irreplaceable roles in the diagnosis, prognosis, and treatment of diseases. The ability to isolate high-quality exosomes is a prerequisite for diagnosing and subsequently treating diseases in an accurate and repeatable manner. However, efficiently isolating exosomes from complex biological samples is challenging owing to their relatively low abundances and interference from non-vesicular macromolecules (such as cell debris and proteins). To date, various isolation techniques based on the physical, chemical, and biological characteristics of exosomes have been developed. Indeed, efficient affinity-interaction-based methods have recently overcome the limitations and drawbacks of traditional exosome isolation methods and are widely used in scientific research and clinical applications. This review focuses on exosome isolation and enrichment, and systematically reviews recent research progress on efficient isolation methods based on affinity interactions. Developmental prospects of exosome isolation and enrichment directions are analyzed with the aim of providing a reference for the construction and use of new exosome-isolation strategies.</p>","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"413-423"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059992/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144049821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Exosome separation and analysis based on microfluidics technology and its clinical applications].","authors":"Yu-Hang Xing, Xiang-Shan Ren, Dong-Hao Li, Lu Liu","doi":"10.3724/SP.J.1123.2024.10032","DOIUrl":"10.3724/SP.J.1123.2024.10032","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Exosomes are cell-secreted nanoscale vesicles 30-150 nm in size and encompass a diverse array of biomolecules, including lipids, proteins, and nucleic acids. Exosomes play pivotal roles during the intercellular exchange of materials and information, and are closely associated with the onset and progression of a variety of diseases. Therefore, comprehensively investigating exosomes is very important in terms of disease diagnosis and treatment. However, exosomes are genetically heterogeneous and are composed of different materials. Additionally, exosome-size and packing-specific-biomarker heterogeneities result in biofunction diversity. Moreover, isolating and analyzing exosomes is highly challenging owing to their small sizes and heterogeneities. Accordingly, effective separation methods and analytical techniques for highly specifically and efficiently identifying exosomes are urgently needed in order to better understand their functionalities. While separation and analysis is required to reveal exosome heterogeneity, the former is confronted by three primary challenges. Firstly, exosome heterogeneity (including heterogeneous marker expressions and size heterogeneity that results in heterogeneous functions) results in systems that are very difficult to separate. Secondly, the coexistence of non-vesicular contaminants (lipoprotein nanoparticles, soluble proteins, nucleic acids, etc.) and the complex matrix effects of body fluids also contribute to separation difficulties. Thirdly, enrichment is a highly challenging task owing to low exosome concentrations. Traditional methods, such as ultracentrifugation and size-exclusion chromatography, fall short in terms of their abilities to precisely separate and analyze exosomes. On the other hand, microfluidics has emerged as a robust tool for the efficient analysis of complex biological samples and is characterized by miniaturization, precise control, high throughput, automation, and integration. Firstly, the operability, integrability, and modifiability of a microfluidics system facilitate exosome separation and purification based on surface properties, size, charge, and polarity. Secondly, the use of a microfluidics approach, with its high throughput, low reagent consumption, and multichannel manipulability, greatly facilitates preparing exosomes and enhancing their concentrations. Thirdly, microfluidics ensures that diverse separation methods are compatible with downstream analysis techniques. Exosomes are highly heterogeneous; hence, they are classified by type and subpopulation (according to origin, size, molecular markers, functions, etc.). This paper first discusses microfluidics techniques for separating exosomes and examines various separation strategies grounded in the physicochemical properties of exosomes. We then analyze exosome detection methodologies that use microfluidics platforms and encompass traditional group-exosome analysis techniques and novel single-exosome analysis approaches","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"455-471"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059995/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144034758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Advances in exosome-targeting aptamer-screening techniques].","authors":"Li-Ting Zheng, Ge Yang, Feng Qu","doi":"10.3724/SP.J.1123.2024.10029","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.10029","url":null,"abstract":"<p><p>Exosomes play crucial intercellular-communication roles and regulate various cellular physiological processes. They are considered potential biomarkers for the early diagnosis of cancers and other diseases. Therefore, detecting and isolating exosomes with specific functions has significant clinical implications. Moreover, the development of low-cost, highly sensitive recognition elements for identifying exosomes is essential for advancing early disease diagnosis and treatment. Nucleic acid aptamers are single-stranded DNA or RNA molecules capable of specifically binding to targets and are produced through the systematic evolution of ligands by exponential enrichment (SELEX) technique. Such aptamers are highly stable, chemically synthesizable, exhibit high affinities and specificities, and are applicable to a broad range of targets, which endow them with unique advantages. Currently, aptamers that target exosomes have been used in a variety of research fields, including cell imaging, drug delivery, and disease diagnosis and treatment. However, selecting aptamers that precisely identify specific exosomes is significantly challenging owing to the complex structures of exosome and their heterogeneity. Consequently, obtaining high-performance aptamers requires efficient screening techniques. This review first summarizes the functions and selection strategies of key targets for exosome-aptamer screening. Furthermore, it outlines the main methods and techniques currently used to screen exosome aptamers, which includes five screening techniques: magnetic bead-SELEX, microfluidic-SELEX, nitrocellulose-SELEX, cell-SELEX, and capillary electrophoresis-SELEX. The separation principles, advantages, limitations, and the latest applications of these techniques are discussed in detail. The review finally addresses current challenges associated with selecting exosome aptamers and provides insight into future research directions.</p>","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"424-433"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059990/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144035966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Isolation and proteomic analysis of bacterial outer membrane vesicle subpopulations].","authors":"Po-Ju Yu, Xun Zou, Yan Wu, Sun-Tao Li, Hua Xiao","doi":"10.3724/SP.J.1123.2024.10028","DOIUrl":"https://doi.org/10.3724/SP.J.1123.2024.10028","url":null,"abstract":"<p><p>Outer membrane vesicles (OMVs) are 20-400 nm in size, membrane-bound, and secreted by gram-negative bacteria. OMVs play important roles in processes such as toxin delivery and immune evasion. Although many studies have revealed the critical roles played by OMVs, their heterogeneity has limited our ability to attain a comprehensive understanding of their protein compositions and functions. Therefore, studying the compositions of heterogeneous OMVs subpopulations and their biological functions is important. Herein, we used ultracentrifugation combined with density-gradient centrifugation and quantitative proteomics to systematically separate, characterize, and comprehensively analyze OMVs secreted by <i>Escherichia coli</i> DH5α and <i>Pseudomonas aeruginosa</i> PAO1. First, crude OMVs extracts from both strains were obtained by ultracentrifugation and subjected to iodixanol density-gradient centrifugation to afford six fractions each. DH5α-OMVs and PAO1-OMVs particle-size distributions were then determined via nanoparticle tracking analysis, with average particle sizes of 131.0-161.0 and 140.0-169.0 nm determined for the two subpopulation, respectively. Vesicles were observed to have classical chattel structures by transmission electron microscopy. OMVs subpopulation distributions in the density-gradated fractions were determined by silver staining and protein immunoblotting, which also identified F1a-F4a and F1b-F5b as the effective DH5α-OMVs and PAO1-OMVs subpopulation fractions, respectively. We then identified 2388 and 905 proteins from the DH5α-OMVs and PAO1-OMVs subpopulation, respectively, and used k-means clustering and gene ontology (GO) enrichment analyses to reveal the heterogeneities of the various density subpopulations in terms of biological functions, such as energy metabolism, material transport and ribosome synthesis. Comparative analysis of the <i>E. coli</i> DH5α-OMVs and <i>P. aeruginosa</i> PAO1-OMVs subpopulations finally revealed that they exhibit different functional characteristics, despite sharing commonalities in their basic OMVs functions. The F1a DH5α-OMVs subpopulation was found to be enriched for functions related to amino-acid metabolism and protein synthesis, while the F2b PAO1-OMVs subpopulation exhibited significant biomolecule synthesis functions. This study revealed that bacterial OMVs subpopulations have distinct biological functions, which in turn provides a new theoretical basis for understanding the pathogenic mechanisms of bacteria and their interactions with the host, thereby expanding their biological applications.</p>","PeriodicalId":101336,"journal":{"name":"Se pu = Chinese journal of chromatography","volume":"43 5","pages":"529-538"},"PeriodicalIF":0.0,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12059987/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143996930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}