Isolation of leukemia and breast cancer cells from liquid biopsies and clinical samples at low concentration in a 3D printed cell separation device via transferrin-receptor affinity

Liquid biopsies can be used for early detection, cancer diagnosis, prognosis, monitoring patient treatment, and individualized therapy strategies. Currently, liquid biopsy analysis is limited by the detection methods for identifying and isolating cancer cells. In this work, we developed and validated a 3D printed affinity separation device for cancer cell detection in liquid biopsies with minimal operator intervention using CD71 (transferrin receptor) as the affinity target. Cancer cell isolation was demonstrated with concentration as low as 30 cells per mL. COG-LL-332 patient-derived leukemia cells and MDA-MB-231 breast cancer cells were spiked into lysed blood. Samples with initial cancer cell concentrations as low as 0.05% were separated with a capture purity of 94–98% and an enrichment factor of 98X-1900X for COG-LL-332 cells. The purity of enriched 0.05% cells was 97–99% with an enrichment factor of 99X-1900X for MDA-MB-231 cells. Clinical plasma samples of breast and prostate cancer patients were tested, showing our 3D printed CD71 affinity device could isolate cancer cells from these liquid biopsies. To further show utility, clinical cancer cells were spiked into blood and separated with a capture purity of 93 ± 6% to 94 ± 10% and an enrichment factor of 1860 ± 120 to 1890 ± 190 for breast cancer and a purity of 90 ± 10% to 96 ± 7% with an enrichment factor of 1793 ± 200 to 1920 ± 140 for acute lymphoblastic leukemia samples. The low detection concentration, high enrichment factor, simple fabrication and conjugation, and applicability to multiple cancer types makes this a promising approach for liquid biopsies.