An orthogonal barcoding enabled smart nanodevice for highly efficient isolation and proteomic profiling of tumor-derived extracellular vesicles

exosomal protein extraction and digestion within the nanopores of the MOSF-tag. This integrated strategy not only streamlines the process by eliminating complex steps and minimizing sample loss but also significantly enhances protein identification efficiency. Compared to traditional methods for T-EVs isolation and protein digestion, the smart nanodevice has demonstrated a remarkable improvement in the detection of exosomal proteins and specific proteins from the cell culture medium. As a proof of concept, we applied this strategy to serum samples from prostate cancer (PCa) patients, confirming its efficacy. A total of 832 proteins were identified, with 211 showing differential expression between patients and healthy controls. Among these, 113 proteins were significantly upregulated in the PCa group. These uniquely expressed proteins are likely associated with PCa development, invasion, and metastasis, highlighting their potential as biomarkers for the early diagnosis and prognosis of PCa in the future. This innovative approach not only advances the field of T-EVs research but also opens new avenues for the discovery of clinically relevant biomarkers in cancer.