Facile PEG-based isolation and classification of cancer extracellular vesicles and particles with label-free surface-enhanced Raman scattering and pattern recognition algorithm.

Extracellular vesicles and particles (EVPs), which contain the same surface proteins as their mother cells, are promising biomarkers for cancer liquid biopsy. However, most of the isolation methods of EVPs are time-consuming and complicated, and hence, sensitive detection and classification methods are required for EVPs. Here, we report a facile polyethylene glycol (PEG)-based method for isolating and classifying EVPs with label-free surface-enhanced Raman scattering (SERS) and pattern recognition algorithm. There are only three steps in the PEG-based isolation method, and it does not require ultracentrifugation, which makes it a low-cost and easy-to-use method. Three types of common male cancer cell lines, namely leukemia (THP-1), prostate cancer (DU-145), and colorectal cancer (COLO-205), and one healthy male blood sample, were utilized to isolate EVPs. To collect the SERS spectra of EVPs, a novel planar nanomaterial, namely amino molybdenum oxide (AMO) nanoflakes, was applied, with the enhancement factor being obtained as 3.2 × 10. Based on the principal component analysis and support vector machine (PCA-SVM) algorithm, cancer and normal EVPs were classified with 97.4% accuracy. However, among the cancer EVPs, the accuracy, precision, and sensitivity were found to be 90.0%, 90.9%, and 83.3% for THP-1; 86.7%, 80.0%, and 92.3% for DU-145; 96.7%, 83.3%, and 100% for COLO-205, respectively. Thus, this work will improve the isolation, detection, and classification of EVPs and promote the development of cancer liquid biopsies.