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1.
Biosens Bioelectron ; 254: 116218, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38518559

RESUMO

Biodetection for non-invasive diagnostics of fluids, especially urine, remains a challenge to scientists due to low target concentrations. And biological complexes of the detection target may contain contaminants that also interfere with any assay. Dengue non-structural 1 protein (Dengue NS1) is an important biomarker for dengue hemorrhagic fever and dengue shock syndrome. Here, we developed an Au-decorated nanowire platform and applied it with a sandwich fluorophore-linked immunosorbent well plate assay (FLISA) to detect Dengue NS1 in urine. For the platform, we fabricated zinc oxide (ZnO) nanowires to provide a high surface area and then coated them with gold nanoparticles (ZnO/Au nanowires) to simply modify the Dengue NS1 antibody and enhance the fluorescence intensity. Our platform employs a sandwich FLISA that exhibits high sensitivity, specifically detecting Dengue NS1 with a limit of detection (LOD) of 1.35 pg/mL. This LOD was 4500-fold lower than the LOD of a commercially available kit for Dengue NS1 enzyme-linked immunosorbent assay. We believe that our ZnO/Au nanowire platform has the potential to revolutionize the field of non-invasive diagnostics for dengue.


Assuntos
Técnicas Biossensoriais , Vírus da Dengue , Dengue , Nanopartículas Metálicas , Nanofios , Óxido de Zinco , Humanos , Dengue/diagnóstico , Ouro , Sensibilidade e Especificidade , Proteínas não Estruturais Virais , Antígenos Virais , Ensaio de Imunoadsorção Enzimática , Imunoadsorventes , Anticorpos Antivirais
2.
Sci Adv ; 9(27): eade6958, 2023 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-37418532

RESUMO

Cancer cell-derived extracellular vesicles (EVs) have unique protein profiles, making them promising targets as disease biomarkers. High-grade serous ovarian carcinoma (HGSOC) is the deadly subtype of epithelial ovarian cancer, and we aimed to identify HGSOC-specific membrane proteins. Small EVs (sEVs) and medium/large EVs (m/lEVs) from cell lines or patient serum and ascites were analyzed by LC-MS/MS, revealing that both EV subtypes had unique proteomic characteristics. Multivalidation steps identified FRα, Claudin-3, and TACSTD2 as HGSOC-specific sEV proteins, but m/lEV-associated candidates were not identified. In addition, for using a simple-to-use microfluidic device for EV isolation, polyketone-coated nanowires (pNWs) were developed, which efficiently purify sEVs from biofluids. Multiplexed array assays of sEVs isolated by pNW showed specific detectability in cancer patients and predicted clinical status. In summary, the HGSOC-specific marker detection by pNW are a promising platform as clinical biomarkers, and these insights provide detailed proteomic aspects of diverse EVs in HGSOC patients.


Assuntos
Vesículas Extracelulares , Nanofios , Neoplasias Ovarianas , Feminino , Humanos , Proteômica , Cromatografia Líquida , Espectrometria de Massas em Tandem , Vesículas Extracelulares/metabolismo , Biomarcadores , Proteínas , Neoplasias Ovarianas/metabolismo
3.
ACS Nano ; 17(3): 2235-2244, 2023 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-36655866

RESUMO

Extracellular vesicles (EVs) have promising potential as biomarkers for early cancer diagnosis. The EVs have been widely studied as biological cargo containing essential biological information not only from inside vesicles such as nucleic acids and proteins but also from outside vesicles such as membrane proteins and glycolipids. Although various methods have been developed to isolate EVs with high yields such as captures based on density, size, and immunoaffinity, different measurement systems are needed to analyze EVs after isolation, and a platform that enables all-in-one analysis of EVs from capture to detection in multiple samples is desired. Since a nanowire-based approach has shown an effective capability for capturing EVs via surface charge interaction compared to other conventional methods, here, we upgraded the conventional well plate assay to an all-in-one nanowire-integrated well plate assay system (i.e., a nanowire assay system) that enables charge-based EV capture and EV analysis of membrane proteins. We applied the nanowire assay system to analyze EVs from brain tumor organoids in which tumor environments, including vascular formations, were reconstructed, and we found that the membrane protein expression ratio of CD31/CD63 was 1.42-fold higher in the tumor organoid-derived EVs with a p-value less than 0.05. Furthermore, this ratio for urine samples from glioblastoma patients was 2.25-fold higher than that from noncancer subjects with a p-value less than 0.05 as well. Our results demonstrated that the conventional well plate method integrated with the nanowire-based EV capture approach allows users not only to capture EVs effectively but also to analyze them in one assay system. We anticipate that the all-in-one nanowire assay system will be a powerful tool for elucidating EV-mediated tumor-microenvironment crosstalk.


Assuntos
Neoplasias Encefálicas , Vesículas Extracelulares , Nanofios , Humanos , Vesículas Extracelulares/metabolismo , Biomarcadores/metabolismo , Neoplasias Encefálicas/diagnóstico , Proteínas de Membrana/metabolismo , Microambiente Tumoral
4.
J Nanosci Nanotechnol ; 18(10): 6786-6790, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-29954494

RESUMO

Graphene has received tremendous interest owing to its excellence excellent in electrical conductivity and very high specific surface area. In this work, density functional theory (DFT) is used in order to investigate optical and electrical properties of functionalized graphene which interacted with cypermethrin pesticide molecules. The structures of graphene and pesticide were designed and optimized with M06-2x/6-31G(d,p) method of calculation. Graphene was functionalized by carboxyl, amine and hydroxyl groups to investigate the cypermethrin molecules. The binding energy, band spectra, optical properties and electron distribution of complexes were analyzed. The results reveal that the functionalized graphene with hydroxyl group can highly improve the interaction between graphene and cypermethrin with the lowest binding energy.

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