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

Yoshikatsu Koga, Satoshi Katayose, Nobuko Onoda, Takahiro Kasamatsu, Tomoyasu Kato, Shunichi Ikeda, Mitsuya Ishikawa, Ken Ishitani, Yasuo Hirai, Hideo Matsui, Yasuhiro Matsumura

Department of Gynecology, National Cancer Center Hospital, Tokyo, Japan.
Department of Obstetrics and Gynecology, Tokyo Women’s Medical University, Tokyo, Japan.
Department of Obstetrics and Gynecology, Tokyo Women’s Medical University, Tokyo, Japan..
Division of Developmental Therapeutics, Research Center for Innovative Oncology, National Cancer Center Hospital East, Kashiwa, Japan.
R & D Department Unit 1, JSR Life Sciences Corporation, Tsukuba, Japan.

DOI: 10.4236/jct.2013.48150                

Abstract

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.

Keywords

Immuno-Magnetic Beads; Auto-Fluorescence; Endometrial Cancer; Cancer Screening

Conflicts of Interest

The authors declare no conflicts of interest.

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