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Due to their rarity, the analysis of CTCs is challenging and commonly based on immunomagnetic technologies using antibodies against EpCAM. This study used mass cytometry (CyTOF\u00ae) for the identification and characterization of CTCs from longitudinally monitored metastatic breast cancer (mBC) patients. Functional analysis focused on DNA damage responses, particularly the DNA repair pathway of homologous recombination (HR) validated in BC cells from the pleura. Fifty\u2010two blood samples from 13 mBC patients were collected for the enumeration of CTCs using CellSearch\u00ae technology, isolation of CTCs together with peripheral blood mononuclear cells (PBMCs) and of plasma. Cell\u2010free DNA (cfDNA) from plasma was analyzed by shallow genome sequencing to determine tumor fraction (TF) and HR deficiency (HRD). CTC\/PBMC mixtures were phenotyped by CyTOF\u00ae using a panel of 13 antibodies including anti\u2010\u03b3H2AX, 53BP1, and RAD51. 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