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The BCSCs were found to have characteristics of CD44+/CD24-/low epithelial surface antigen (ESA) and ALDH+ with flow cytometry. The phage was enriched to 200-fold after three rounds of screening for MDA-MB-231 BCSCs. The positive phages were sequenced; then a polypeptide named M58 was synthesized according to sequencing results. Polypeptide M58 has a specific affinity to MDA-MB-231 BCSCs in vivo and in vitro. Conclusion Specific polypeptides binding to MDA-MB-231 BCSCs were screened out by PD screening method, which laid a theoretical f