Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX, 79409, USA.
Biomed Microdevices. 2021 Apr 28;23(2):28. doi: 10.1007/s10544-021-00566-z.
Microfluidic, flow cytometry, and immunomagnetic methods for cancer cell isolation have heavily relied on the Epithelial Cellular Adhesion Molecule (EpCAM) for affinity separation. While EpCAM has been used extensively for circulating tumor cell isolation, it cannot be used to isolate non-epithelial cells. The human transferrin receptor (CD71) can also be used for cancer cell isolation and has the advantage that as an affinity target it can separate virtually any cancer cell type, regardless of disease origin. However, direct comparison of the capture ability of EpCAM and CD71 has not been reported previously. In this work, cell capture with both EpCAM and CD71 were studied using a novel higher-throughput herringbone cell separation microfluidic device. Five separation chip models were designed and the one with the highest capture efficiency (average 90 ± 10%) was chosen to compare antigen targets for cell capture. Multiple cancer cell lines including CCRF-CEM, PC-3 and MDA-MB-231 were tested for cell capture performance using both ligands (anti-CD71 and anti-EpCAM) in the optimized chip design. PC-3 and MDA-MB-231 cells were spiked into blood at concentrations ranging from 0.5%-10%. PC-3 cells were separated by anti-CD71 and anti-EpCAM with 32-37% and 31-50% capture purity respectively, while MDA-MB-231 were separated with 35-53% and 33-56% capture purity using anti-CD71 and anti-EpCAM for all concentrations. The enrichment factor for the lowest concentrations of cells in blood ranged from 66-74X. The resulting enrichment of cancer cells shows that anti-CD71 was found to be statistically similar to anti-EpCAM for epithelial cancer cells, while anti-CD71 can be further used for non-epithelial cells, where anti-EpCAM cannot be used.
微流控、流式细胞术和免疫磁珠方法在分离癌细胞时严重依赖于上皮细胞黏附分子（EpCAM）进行亲和分离。虽然 EpCAM 已被广泛用于循环肿瘤细胞的分离，但它不能用于分离非上皮细胞。人转铁蛋白受体（CD71）也可用于癌细胞分离，其优点是作为亲和靶标，它可以分离几乎任何类型的癌细胞，而与疾病起源无关。然而，以前没有报道过 EpCAM 和 CD71 的捕获能力的直接比较。在这项工作中，使用新型高通量人字形细胞分离微流控装置研究了 EpCAM 和 CD71 的细胞捕获。设计了五种分离芯片模型，并选择了捕获效率最高（平均 90±10%）的芯片模型来比较细胞捕获的抗原靶标。使用两种配体（抗-CD71 和抗-EpCAM）在优化的芯片设计中测试了包括 CCRF-CEM、PC-3 和 MDA-MB-231 在内的多种癌细胞系的细胞捕获性能。将 PC-3 和 MDA-MB-231 细胞以 0.5%-10%的浓度范围混入血液中。PC-3 细胞分别通过抗-CD71 和抗-EpCAM 以 32-37%和 31-50%的捕获纯度分离，而 MDA-MB-231 细胞以 35-53%和 33-56%的捕获纯度使用抗-CD71 和抗-EpCAM 分离所有浓度。血液中最低细胞浓度的富集因子范围为 66-74X。癌细胞的富集表明，抗-CD71 与上皮癌细胞的抗-EpCAM 在统计学上相似，而抗-CD71 可进一步用于上皮细胞无法使用的非上皮细胞。
