Tumor-derived extracellular vesicles (tdEVs) have been emerging as potential biomarkers for cancer diagnosis because the tdEVs precisely reflect tumor cell alterations with significantly increased production. The proteomic profiling study of tdEVs represents a promising approach in a non-invasive manner to novel biomarker discovery for early detection and targeted therapy of cancer. Previously, we have developed a novel microfluidic chip for rapid and selective isolation of tdEVs. This microfluidic chip enables the selection of two types of EVs by using breast tumor-derived proteins (EpiCAM & CD49f) within two minutes. Using a microfluidic chip capable of selectively isolating tdEVs, the study compared the proteomes of EVs isolated from the serum of 100 breast cancer patients with different subtypes and 30 healthy individuals.