Combined use of EpCAM and FRa enables the high-efficiency capture of circulating tumor cells in non-small cell lung cancer

Circulating tumor cells (CTCs) provide a new approach for auxiliary diagnosis, therapeutic effect evaluation, and prognosis prediction for cancer patients. The epithelial cell adhesion molecule (EpCAM)-based separation method (CellSearch) showed good clinical use in multiple types of cancer. Nevertheless, some non-small cell lung cancer (NSCLC) tumor cells have a lower expression of EpCAM and are less frequently detected by CellSearch. Here, we present a highly sensitive immunomagnetic separation method to capture CTCs based on two cell surface markers for NSCLC, EpCAM and Folate receptor alpha (FR alpha). Our method has been demonstrated to be more efficient in capturing NSCLC cells (P < 0.01) by enriching three types of CTCs: EpCAM(+)/FR alpha(-/low), EpCAM(-/low)/FR alpha(+), and EPCAM(+)/FR alpha(+). In 41 NSCLC patients, a significantly higher CTC capture rate (48.78% vs. 73.17%) was obtained, and by using a cutoff value of 0 CTC per 2 ml of blood, the sensitivities were 53.66% and 75.61% and the specificities were 100% and 90% for anti-EpCAM-MNs or a combination of anti-EpCAM-MNs and anti-FR alpha-MNs, respectively. Compared with the tumor-specific LT-PCR based on FR alpha, our method can isolate intact FR alpha(+) CTCs, and it is advantageous for additional CTC-related downstream analysis. Our results provide a new method to increase the CTC capture efficiency of NSCLC.