TITLE:
Usefulness of Immuno-Magnetic Beads Conjugated with Anti-EpCAM Antibody for Detecting Endometrial Cancer Cells
AUTHORS:
Yoshikatsu Koga, Satoshi Katayose, Nobuko Onoda, Takahiro Kasamatsu, Tomoyasu Kato, Shunichi Ikeda, Mitsuya Ishikawa, Ken Ishitani, Yasuo Hirai, Hideo Matsui, Yasuhiro Matsumura
KEYWORDS:
Immuno-Magnetic Beads; Auto-Fluorescence; Endometrial Cancer; Cancer Screening
JOURNAL NAME:
Journal of Cancer Therapy,
Vol.4 No.8,
September
17,
2013
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.