Efficient and Discriminative Isolation of Circulating Cancer Stem Cells and Non-Stem-like Circulating Tumor Cells Using a Click-Handle-Loaded M13 Phage-Based Surface

Circulating tumor cells (CTCs) are crucial for cancer research and clinical applications, with circulating cancer stem cells (cCSCs) being a rare but key subpopulation responsible for metastasis, recurrence, and therapy resistance. Current limitations in efficiently isolating these cells, particularly distinguishing cCSCs from non-stem-like CTCs (nsCTCs), hinder our understanding of cancer progression and precision medicine strategies. Herein, we developed a novel CTC isolation approach that integrates cell metabolic chemical tagging with a <i><u>c</u></i>lick-<i><u>h</u></i>andle-loaded M13 <i><u>ph</u></i>age-based surf<i><u>ace</u></i> (CHPhace). The multivalent nature of flexible M13 nanofibers, featuring thousands of modification sites for click reactions, significantly enhances CTC capture across diverse tumor types. Leveraging the unique slow-cycling characteristic of cCSCs, CHPhace demonstrated selective cCSCs isolation through metabolic labeling and demetabolism processes. The robust performance of CHPhace allows efficient isolation of both cCSCs and nsCTCs from complex blood sample matrices, achieving capture efficiencies exceeding 80%. This approach represents a promising tool for advancing our understanding of cancer progression and enhancing precision in clinical diagnosis and cancer prognosis.