Circulating Tumor Cell Detection In Epithelial Ovarian Cancer Using Dual-Component Antibodies Targeting EpCAM And FR[alpha]

Purpose: Circulating tumor cell (CTC) detection methods based on epithelial cell adhesion molecule (EpCAM) have low detection rates in epithelial ovarian cancer (EOC). Meanwhile, folate receptor alpha (FR[alpha]) has high expression in EOC cells. We explored the feasibility of combining FR[alpha] and EpCAM as CTC capture targets in EOC. Patients and methods: EpCAM and FR[alpha] antibodies were linked to magnetic nanospheres (MNs) using the principle of carbodiimide chemistry. Blood samples from healthy donor spiked with A2780 ovarian cancer cells were used for detecting the capture rate. Ninety-five blood samples from 30 patients with EOC were used for comparing the positive rate of detection when using anti-EpCAM-MNs alone with that when using combination of anti-EpCAM-MNs and anti-FR[alpha]-MNs. Samples from 28 patients initially diagnosed with EOC and 20 patients with ovarian benign disease were used for evaluating the sensitivity and specificity of combination of anti-EpCAM-MNs and anti-FR[alpha]-MNs. Results: Regression analysis between the number of recovered and that of spiked A2780 cells revealed [y.sub.EpCAM] = 0.535x ([R.sup.2] = 0.99), [y.sub.FR[alpha]] = 0.901x ([R.sub.2] = 0.99), and [y.sub.EpCAM+FR[alpha]] = 0.928x ([R.sup.2] = 0.99). In mixtures of A2780 and MCF7 cells, the capture rate was 92% using the combination of anti-EpCAM-MNs and anti-FR[alpha]-MNs, exceeding the rate when using anti-EpCAM-MNs or anti-FR[alpha]-MNs alone by approximately 20% (P < 0.01). The combination of anti-EpCAM-MNs and anti-FR[alpha]-MNs showed a significantly increased positive rate of CTC detection in EOC patients compared with anti-EpCAM-MNs alone ([chi square] = 14.45, P < 0.001). Sensitivity values were 0.536 and 0.75 and specificity values were 0.9 and 0.85 when using anti-EpCAM-MNs alone and when using the combination of anti-EpCAM-MNs and anti-FR[alpha]-MNs, respectively. Conclusion: The combination of FR[alpha] and EpCAM is feasible as a CTC capture target of CTC detection in patients with EOC. Keywords: circulating tumor cells, ovarian cancer, epithelial cell adhesion molecule, folate receptor alpha