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A simple and non-invasive method for detecting endometrial cancer in women with abnormal uterine bleeding is required. For this purpose, we prepared immuno-magnetic beads conjugated with anti-human EpCAM rat monoclonal antibody (mAb) for isolating exfoliated endometrial cells including endometrial cancer cells in vaginal discharge. The affinities of the anti-human EpCAM rat mAbs were analyzed by flow cytometry and immunocytochemistry and then magnetic beads were conjugated with the mAbs. The rate of retrieval of endometrial cells using the immuno-magnetic beads was calculated. Endometrial cells were isolated using the immuno-magnetic beads from the vaginal discharges of 22 patients with endometrial cancer and 16 non-malignant controls. The isolated cells were stained using endometrial cancer specific-mAbs and analyzed by flow cytometry and imaging cytometry. The immuno-magnetic beads conjugated with high-affinity mAb (clone 1456) appeared to have very low auto-fluorescence. Sufficient enrichment of Ep-CAMpositive cells using immuno-magnetic beads was observed in both simulation and clinical samples. The overall sensitivities of flow cytometry and imaging cytometry to detect endometrial cancer cells were 72.7% and 45.5%, respectively. Meanwhile, the overall specificities of flow cytometry and imaging cytometry for healthy controls were 75.0% and 81.3%, respectively. Our immuno-magnetic beads have very low auto-fluorescence, so they could be useful for fluorescent analysis, such as fluorescent immunochemical staining. In the future, these novel immuno-magnetic beads could be used for cytological study.

Usefulness of Immuno-Magnetic Beads Conjugated with Anti-EpCAM Antibody for Detecting Endometrial Cancer Cells