细胞外小泡
胞外囊泡
计算生物学
生物标志物
细胞生物学
数字聚合酶链反应
化学
微泡
纳米技术
生物
材料科学
生物化学
小RNA
聚合酶链反应
基因
作者
David E. Reynolds,M. L. Pan,Jingbo Yang,George Galanis,Yoon Ho Roh,Renee Tyler Tan Morales,Shailesh Kumar,Su Jin Heo,Xiaowei Xu,Wei Guo,Jina Ko
标识
DOI:10.1002/advs.202303619
摘要
Extracellular vesicles (EVs) have emerged as a promising source of biomarkers for disease diagnosis. However, current diagnostic methods for EVs present formidable challenges, given the low expression levels of biomarkers carried by EV samples, as well as their complex physical and biological properties. Herein, a highly sensitive double digital assay is developed that allows for the absolute quantification of individual molecules from a single EV. Because the relative abundance of proteins is low for a single EV, tyramide signal amplification (TSA) is integrated to increase the fluorescent signal readout for evaluation. With the integrative microfluidic technology, the technology's ability to compartmentalize single EVs is successfully demonstrated, proving the technology's digital partitioning capacity. Then the device is applied to detect single PD-L1 proteins from single EVs derived from a melanoma cell line and it is discovered that there are ≈2.7 molecules expressed per EV, demonstrating the applicability of the system for profiling important prognostic and diagnostic cancer biomarkers for therapy response, metastatic status, and tumor progression. The ability to accurately quantify protein molecules of rare abundance from individual EVs will shed light on the understanding of EV heterogeneity and discovery of EV subtypes as new biomarkers.
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