Analytical Methods
·
2024
·
IF 2.7
Dumei Ma,Yiting Luo,Xiaoya Zhang,Zehu Xie,Yinghua Yan,Chuan-Fan Ding
Exosomes have gained recognition as valuable reservoirs of biomarkers, holding immense potential for early cancer detection. Consequently, there is a pressing need for the development of an economical and highly sensitive exosome detection methodology. In this work, we present a fluorescence method for breast cancer-derived exosome detection based on Cu-triggered click reaction of azide-modified CD63 aptamer and alkyne functionalized Pdots. The detection threshold for the exosomes obtained from the breast cancer serum was determined to be 6.09 × 107 particles per μL, while the measurable range spanned from 6.50 × 107 to 1.30 × 109 particles per μL. The employed methodology achieved notable success in accurately distinguishing breast cancer patients from healthy individuals through serum analysis. The application of this method showcases the significant potential for early exosome analysis in the clinical diagnosis of breast cancer patients.
研究领域:
医学
·
生物技术
·
肿瘤学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Jing Huang,Tucan Chen,Yunong Zhao,Dandan Li,Qing Huang,Lina Cao,Jingqiu Chen,Dongjuan Chen,Liang Hu,Huan Liu
Small extracellular vesicles (sEVs), primarily referring to exosomes, have emerged as potent tools for investigating pathological mechanisms and facilitating precise diagnostics owing to their ability to transfer informative molecules between cells and their accessibility from diverse biological fluids. Despite the multitude of biomedical applications, achieving enhanced specificity and sensitivity in detecting trace sEV samples continues to pose a significant challenge. Considering the potential of electrochemical sensing in efficient molecular detection, this study proposes a viable strategy for specifically recognizing sEVs by targeting their membrane proteins. Additionally, it demonstrates sensitive and quantitative detection of sEVs based on the unique performance of PbS colloidal quantum dots (CQDs) nanomaterials. The good immobilization of target proteins on the chemically modified electrode is achieved through the synergistic effect of the charge–discharge mechanism and strong suspension bonds of PbS CQDs. The results obtained from the analysis of differential pulse voltammetry demonstrate rapid and highly sensitive electrochemical responses. The CD63 antibody-modified sensor can achieve highly sensitive detection of sEVs, providing a wide linear detection range (10 2 ∼ 10 8 particles mL −1 ) and a low detection limit (19 particles mL −1 ). The established method is ultimately employed to differentiate the sEVs originating from distinct tumor cells, thereby demonstrating its potential in membrane-biomarker-based diagnostics. Consequently, the presented electrochemical sensor demonstrates remarkable efficiency in quantifying EVs with exceptional specificity and sensitivity, thereby offering extensive prospects for applications in EV analysis and point-of-care tests.
研究领域:
生物医学
·
纳米技术
·
诊断学
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产品关联
DOI
高影响因子
ACS Sensors
·
2020
·
IF 7.333
Chenyun Wang,Cancan Wang,Dan Jin,Yi Yu,Fan Yang,Yulin Zhang,Qunfeng Yao,Guo-Jun Zhang
In this study, we report a gold nanoparticle (AuNP)-amplified surface acoustic wave (SAW) sensor for exosome detection with high sensitivity. The SAW chip was self-assembled with mercapto acetic acid to generate carboxylic groups via the Au–S bond. Anti-CD63 was then anchored onto the chip by pretreatment with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide,1-hydroxypyrrolidine-2,5-dione (NHS). Due to the existence of a membrane protein, CD63, on the exosome surface, exosomes could be bound onto the antibody-immobilized SAW chip. To amplify the detection signal, both the biotin-conjugated epithelial cell adhesion molecule (EpCAM) antibody as a secondary antibody and AuNP-labeled streptavidin were applied onto the exosome-bound SAW chip, resulting in AuNP assembly on the chip through biotin–avidin recognition. The sensor was capable of detecting 1.1 × 103 particles/mL exosomes, which was about 2 orders of magnitude higher than those detected by the strategy without using signal amplification. The sensor also achieved a satisfactory specificity and could detect the low-abundance exosomes directly in blood samples from cancer patients with minimal disturbance. This makes the SAW sensor useful for early diagnosis of cancer.
研究领域:
生物医学工程
·
纳米技术
·
癌症诊断
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产品关联
DOI
PubMed
高影响因子
ACS Applied Bio Materials
·
2019
·
IF 0
Zaizai Dong,Chuanhao Tang,Zhen Zhang,Wei Zhou,Rong Zhao,Lina Wang,Jiachao Xu,Yayun Wu,Jiang Wu,Xing Zhang,Li Xu,Libo Zhao,Xiaohong Fang
Exosomes, which are 30–150 nm extracellular vesicles containing a variety of biomolecules (i.e., proteins, RNA, DNA, and lipids), have emerged as important analytes in liquid biopsy. Although exosome detection offers a valuable chance to understand disease status in a multidimensional manner, comprehensive analysis of different exosomal biomolecules with limited specimens remains a challenge. Herein, we introduced an aptamer-based PCR (polymerase chain reaction) platform and realized highly sensitive detection of CD63 protein positive exosomes as low as 50 particles/μL. More importantly, simultaneous analysis of exosomal RNA and surface protein is available using this method. In a proof of concept experiment, simultaneous detection of two immunotherapy-related biomarkers, programmed death-ligand 1 (PD-L1) protein and indoleamine 2,3-dioxygenase 1 (IDO1) mRNA, was demonstrated with clinical samples. Our method provides a strategy for the comprehensive analysis of exosomes, with the advantages of high sensitivity and practicability.
研究领域:
生物技术
·
分子生物学
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癌症研究
·
医学诊断
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DOI
PubMed
Advanced Science
·
2024
·
IF 14.3
Jiankun Zang,Yousheng Wu,Xuanlin Su,Kaiwei Cai,Man Ke,Niu He,Huili Zhu,Zefeng Tan,Jielin Zhu,Wensheng He,Min Peng,Shiqing Zhang,Hongcheng Mai,Anding Xu,Dan Lu
Small extracellular vesicles (sEVs) contain abundant circular RNAs (circRNAs) and are involved in cellular processes, particularly hypoxia. However, the process that packaging of circRNAs into neuronal sEVs under hypoxia is unclear. This study revealed the spatial mechanism of the Fused in Sarcoma protein (FUS) that facilitates the loading of functional circRNAs into sEVs in hypoxia neurons. It is found that FUS translocated from the nucleus to the cytoplasm and is more enriched in hypoxic neuronal sEVs than in normal sEVs. Cytoplasmic FUS formed aggregates with the sEVs marker protein CD63 in cytoplasmic stress granules (SGs) under hypoxic stress. Meanwhile, cytoplasmic FUS recruited of functional cytoplasmic circRNAs to SGs. Upon relief of hypoxic stress and degradation of SGs, cytoplasmic FUS is transported with those circRNAs from SGs to sEVs. Validation of FUS knockout dramatically reduced the recruitment of circRNAs from SGs and led to low circRNA loading in sEVs, which is also confirmed by the accumulation of circRNAs in the cytoplasm. Furthermore, it is showed that the FUS Zf_RanBP domain regulates the transport of circRNAs to sEVs by interacting with hypoxic circRNAs in SGs. Overall, these findings have revealed a FUS-mediated transport mechanism of hypoxia-related cytoplasmic circRNAs loaded into sEVs under hypoxic conditions.
研究领域:
神经科学
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分子生物学
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2021
·
IF 13.273
Weiguo Huang,Yunru Yu,Chaoyu Yang,Xiaohui Zhang,Luoran Shang,Yan Zu,Keqing Shi
Exosomes secreted by most cells possese many biological functions and arouse great interest in biomedical research. However, the effective and convenient methods for the capture of exosomes from clinical samples or cultured cells is still scarce. In this work, we developed novel magnetic graphene oxide nanoparticles (MGONs) for the efficient capture of exosomes, in which Fe 3 O 4 @SiO 2 magnetic nanoparticles (NPs) were decorated with GO through dopamine. CD63 aptamers were attached to the surface of MGONs, which could recognize and bound to CD63 on the exosome membrane. The number of exosomes adsorbed onto the MGONs surface was 1.5 times that attached to the Fe 3 O 4 @SiO 2 surface. The capture rate of exosomes reached as high as 89.4% within 15 min at room temperature. Moreover, exosomes can be effectively enriched and isolated under external magnetic field and the capture process did not affect their morphological structure and biological functions. Through hybridizing with a FAM-labelled single-stranded DNA probe, the concentration of captured exosomes could also be recorded by measuring the fluorescence intensity, and the limit of detection of this system reached 2.4 × 10 7 particles/ml. These aptamer-decorated MGONs also showed potential in high-effieient capture of exosomes using cancer patients samples, which demonstrated that the resultant MGONs had significant practical values in early disease diagnosis and the related clinical applications.
研究领域:
纳米技术
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生物医学工程
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分子生物学
·
临床诊断
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产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2020
·Vol. 412
·期 3
·601-609
·
IF 3.637
Zhao Xianxian,Zhang Wenqing,Qiu Xiaopei,Mei Qiang,Luo Yang,Fu Weiling
Numerous studies have shown that exosomes are closely related to the pathogenesis of various diseases, especially cancers. Therefore, a rapid and sensitive method for exosome detection will be of great importance for the diagnosis and prognosis of diseases. We report here a method for exosome detection based on the CD63 aptamer and clustered regular interspaced short palindromic repeats (CRISPR)/Cas12a system. This method consists mainly of exosomal membrane protein recognition based on the CD63 aptamer and signal amplification based on CRISPR/Cas12a. The CD63 aptamer, as an easily adaptable nucleic acid strand, is responsible for the conversion of the amounts of exosomes into nucleic acid detection, whereas CRISPR/Cas12a is responsible for highly specific nucleic acid signal amplification. The detection range of the method was determined as 3 × 10 3 –6 × 10 7 particles per microliter. Additionally, we successfully applied this method to detect exosomes in clinical samples from both healthy individuals and patients with lung cancer, and the results were highly consistent with those obtained by nanoparticle tracking analysis. In general, this method provides a highly sensitive and specific method for the detection of exosomes and offers an avenue toward future exosome-based diagnosis of diseases.
研究领域:
生物标志物研究
·
分子诊断
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癌症生物学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Lu-Shuang Li,Xiao-Hao Chen,Yu Pei,Xiao-Lin Xie,Cong-Ting Wu,Shuang Cai,Jing Li,Wei Gong
Oral squamous cell carcinoma (OSCC) is the most prevalent histological subtype of oral cancer and urgently requires a noninvasive approach for timely detection to improve patients’ prognose. Salivary exosome act as a promising biomarker, while the complex and expensive analytical methods impeded its popularization in clinical applications. In this study, we developed a label-free aptasensor for salivary exosomes that utilizes a “one exosome to multiple carbon dots” signal conversion method along with nano-micro dual-scale signal amplification strategy. SiO 2 microspheres served as substrates, while carbon dots acted as luminophores, with CD63 aptamers functioning as selective recognition elements. Exosomes compete to bind with the aptamers, thereby leading to the release of non-specifically adsorbed carbon dots from the substrates. The concentration of exosomes is proportional to the variation in fluorescence intensity, enabling highly sensitive detection in artificial saliva samples, with a favorable linearity range of 2.5 × 10 2 ∼5 × 10 8 particles/mL and a relatively low LOD of 100 particles/mL. The anti-interference ability is deemed acceptable, and the fabrication process is affordable coupled with a straightforward signal output mode. The aptasensor was applied successfully to distinguish OSCC patients from healthy individuals, suggesting its potential for screening OSCC in physical examination centers and communities.
研究领域:
医学诊断
·
生物传感
·
肿瘤学
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产品关联
DOI
PubMed
高影响因子
Small
·
2024
·
IF 13
Hongyuan Cui,Tianfang Zheng,Nana Qian,Xueqi Fu,Aijun Li,Shu Xing,Xiao-Feng Wang
Exosomes are nanovesicles secreted by cells, which play a crucial role in various pathological processes. Exosomes have shown great promise as tumor biomarkers because of the abundant secretion during tumor formation. The development of a convenient, efficient, and cost-effective method for simultaneously enriching and detecting exosomes is of utmost importance for both basic research and clinical applications. In this study, an aptamer-functionalized magnetic Ti 3 C 2 composite material (Fe 3 O 4 @Ti 3 C 2 @PEI@DSP@aptamer@FAM-ssDNA) is prepared for the simultaneous enrichment and detection of exosomes. CD63 aptamers are utilized to recognize and capture the exosomes, followed by magnetic separation. The exosomes are then released by cleaving the disulfide bonds of DSP. Compared to traditional methods, Fe 3 O 4 @Ti 3 C 2 @PEI@DSP@aptamer@FAM-ssDNA exhibited superior efficiency in enriching exosomes while preserving their structural and functional integrity. Detection of exosome concentration is achieved through the fluorescence quenching of Ti 3 C 2 and the competitive binding between the exosomes and a fluorescently labeled probe. This method exhibited a low detection limit of 4.21 × 10 4 particles mL −1 , a number that is comparable to the state-of-the-art method in the detection of exosomes. The present study demonstrates a method of simultaneous enrichment and detection of exosomes with a high sensitivity, accuracy, specificity, and cost-effectiveness providing significant potential for clinical research and diagnosis.
研究领域:
纳米技术
·
生物标志物研究
·
癌症诊断
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MATERIALS & DESIGN
·
2021
·
IF 7.991
Guohao Li,Nanhang Zhu,Jia Cheng,Yujia Zhang,Yue Yu,Xiaolin Zhang,Qiangying Yi,Yao Wu
Enrichment of exosomes with high structural and functional integrity is a critically important step before downstream applications in biological and clinical domains. In this work, dynamic biological interfaces functionalized fructose-responsive immunomagnetic beads (Fr-IMBs) were fabricated by incorporating the dynamic biological interfaces composed of dopamine and 4-carboxyphenylboronic acid with immunomagnetic beads. These fabricated Fr-IMBs were successfully applied to capture the MCF-7 exosomes with a high capture efficacy of 91.0%. After incubation with fructose, the molecular structure of the dopamine-boronic acid complexation can be rearranged and the antibody (anti-CD63) bound exosomes were released competitively, with a yield greater than 79.8%. Those exosomes that underwent both immunomagnetic separation and fructose triggered release remained their good bioactivity to promote MCF-7 cell migration. Better yet, exosomes enriched by Fr-IMBs possessed higher purity and better preservation of tumor-related molecular information of the exosomal proteins than those obtained through the ultracentrifugation approach, as proved by the comparative proteomics. These proposed sugar-responsive immunomagnetic beads may serve as biofriendly candidates in enriching useful biomarkers (e.g., cells, nucleic acids, proteins, etc.) efficiently from complex biological samples with high purity.
研究领域:
生物技术
·
癌症研究
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Min Luo,Yuxin Lou,Yingqi Liang,Jiangang Mei,Di Zhang,Fei Lan,Junfang Zhu,Yuxin Fang,Lei Zheng,Shihua Luo,Ye Zhang
Extracellular vesicles (EVs) contain a wealth of biological information and hold great value for disease diagnosis. In contrast to conventional in vitro detection, in vivo detection has the ability to improve the detection efficiency, reduce external interferences and enable continuous or real-time dynamic monitoring. However, developing a simple and accurate in vivo assay for EVs remains challenging. In this study, we constructed an allosteric aptamer-based flexible microneedle biosensor capable of measuring disease-associated EVs in vivo. Firstly, the aptamer binding to CD63 captures EVs and immobilizes it on the electrode, and then the aptamer binding to EGFR contacts the EVs, which results in a change in redox current, due to the proximity of the reporter molecule to the electrode surface decreases electron efficiency. Continuous monitoring of tumor-derived EVs revealed significant variations in EVs concentrations across different tumor stages, which exhibited a strong correlation with tumor volume. This flexible microneedle sensor offers the advantages of selectivity, biocompatibility, and analytical performance, presenting a promising approach for tumor monitoring in vivo .
研究领域:
生物传感器
·
纳米技术
·
癌症研究
·
分子诊断学
·
生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2020
·
IF 0.936
Wen-Nan FENG,Jin HE,Si-Ying LI,Dan YANG,Hong-Na LI,Guo-Cheng YANG,Qiang FU,Yu-Ping SHAN
In recent years, exosomes as the intercellular vehicles have received extensive attention for their fundamental roles in biosystem and as novel biomarkers for early detection and therapeutic applications. It has been demonstrated that anti-CD63 can effectively target exosomes from cancer cells. In this work, multifunctional nanoelectrode-nanopore nanopipettes (MNNPs) were prepared to detect exosomes secreted from human cervical cancer cells (HeLa cells) at the single-entity level using the nanopore- based resistive pulse sensing technique. Synthetic liposomes were also measured for comparison. By modifying the anti-CD63 on the carbon nanoelectrode surface near the nanopore, the detected current signals were very different from the signals from the non-modified MNNPs. The differences were induced by the specific interactions between anti-CD63 and the membrane protein CD63 at the surface of exosomes. No obvious differences were observed when liposomes were tested. Furthermore, by utilizing micromanipulation technology, the position of the nanopipette tip could be accurately located, which enabled monitoring the secretion of exosomes from individual cells in a real-time manner. In the control experiment, the probe-to-cell distance was optimized and the HeLa cells incubated with exosomes inhibitor (GW4869) were tested by MNNPs. These significantly reduced translocation signals proved that anti-CD63 modified MNNPs were competent for exosomes detection in complex cellular environments.
研究领域:
生物学
·
纳米技术
·
癌症研究
·
生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）