The invention provides a HER2, EGFR, EpCAM and MUC1 multiple antibody immunomagnetic bead and a preparation method thereof. The multiple antibody immunomagnetic bead includes a HER2 immunomagnetic bead, an EGFR immunomagnetic bead and an EpCAM immunomagnetic bead, and a HER2 antibody, an EGFR antibody and an EpCAM antibody are respectively coupled with magnetic microspheres to obtain the HER2 immunomagnetic bead, the EGFR immunomagnetic bead and the EpCAM immunomagnetic bead. The multiple antibody immunomagnetic bead is good in specificity and sensitivity, rapid in magnetic response, short in concentration time and high in capturing efficiency when being used for capturing CTCs (circulating tumor cells). The immunomagnetic bead is stable in nature, small in particle size, good in magnetic response and dispersibility, simple in preparation method and high in practicability.