College of Science and Engineering, Hamad Bin Khalifa University, Doha P.O. Box 34111, Qatar.
Liberal Arts and Science Program, Virginia Commonwealth University, Doha P.O. Box 8095, Qatar.
Sensors (Basel). 2022 Aug 24;22(17):6347. doi: 10.3390/s22176347.
Tumor-derived exosomes may provide biomarkers for cancer treatment. Using sputtering technology, an affinity-based device to capture exosomes was developed using nanoporous substrate (NPG)-coated silicon microscopy. Immunology-based techniques detect and purify exosomes using gold coating with a specific antigen. Inverted fluorescent microscopy was used to detect target exosomes quantitatively utilizing fluorescent nanospheres as the label. We quantified the expression of CD63 surface protein markers on exosomes from conditioned culture media of breast cancer cells. The exosomes that targeted specific proteins with controls were statistically analyzed and compared to those that targeted non-specific proteins. Results from SEM showed that the exosomes were circular, between 30 and 150 nanometers in size. The porous gold substrates captured more exosomes than the nonporous substrates. Nitric acid treatments at different times resulted in a variety of pore sizes. Despite the increase in the size of the pores, the number of exosomes found in the porous gold substrate treated for 10 min nearly doubled compared to the one treated for 5 min. In this work, a fluorescence biosensor was developed to detect breast cancer exosomes using nanoporous gold substrates (NPG). Assay and model exosomes of specific breast cancer cells showed that exosomes exhibit diagnostic surface protein markers, reflecting the protein profile of their parent cells. Furthermore, the specific binding between the exosome surface antibodies and the targets identified the CD63 biomarkers on the exosome, suggesting these markers' diagnostic potential. This study can accelerate exosome research in determining tumor-related exosomes and develop novel cancer diagnostic methods.
肿瘤衍生的外泌体可为癌症治疗提供生物标志物。采用溅射技术，开发了一种基于亲和性的装置，使用涂有纳米多孔基底（NPG）的硅显微镜来捕获外泌体。免疫学法技术使用带有特定抗原的金涂层检测和纯化外泌体。倒置荧光显微镜用于利用荧光纳米球作为标记物定量检测靶向外泌体。我们定量检测了乳腺癌细胞条件培养基中外泌体表面 CD63 蛋白标志物的表达。对靶向特定蛋白质的外泌体与对照组进行了统计学分析，并与靶向非特异性蛋白质的外泌体进行了比较。SEM 结果表明，外泌体呈圆形，大小在 30 到 150 纳米之间。多孔金基底比非多孔基底捕获更多的外泌体。不同时间的硝酸处理会导致各种孔径的出现。尽管孔径增大，但经过 10 分钟处理的多孔金基底中发现的外泌体数量几乎是经过 5 分钟处理的两倍。在这项工作中，开发了一种荧光生物传感器，使用纳米多孔金基底（NPG）来检测乳腺癌外泌体。针对特定乳腺癌细胞的分析和模型外泌体表明，外泌体表现出诊断性表面蛋白标志物，反映了其母细胞的蛋白质谱。此外，外泌体表面抗体与靶标之间的特异性结合鉴定了外泌体上的 CD63 生物标志物，表明这些标志物具有诊断潜力。这项研究可以加速外泌体研究，确定与肿瘤相关的外泌体，并开发新的癌症诊断方法。
