Circulating Tumor Cell Microseparator Based on Lateral Magnetophoresis and Immunomagnetic Nanobeads
Circulating Tumor Cell Microseparator Based on Lateral Magnetophoresis and Immunomagnetic Nanobeads
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DOI：
10.1021/ac303284u
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发表时间：
2013-03-05
影响因子：
7.4
通讯作者：
Han, Ki-Ho
中科院分区：
文献类型：
--
作者：
Kim, Seonyoung;Han, Song-I;Han, Ki-Ho
本文提出了一种循环肿瘤细胞 (CTC) 微分离器，使用带有特异性结合上皮癌细胞的抗上皮细胞粘附分子 (EpCAM) 抗体的免疫磁性纳米珠，从人外周血中分离 CTC。分离是通过横向磁泳进行的，这是由高梯度磁分离技术引起的，涉及镶嵌在微通道底部基板中的铁磁线阵列。实验结果表明，CTC 微分离器以高达 5 mL/h 的流速分离出人外周血中约 90% 的加标 CTC，纯化率约为 97%。 200μL外周血的整个分离过程在15分钟内完成。使用CTC微量分离器分离乳腺癌和肺癌患者外周血中的CTC，并将结果与健康供者的结果进行比较。使用基于荧光的活力测定，观察从癌症患者外周血中分离的 CTC 的活力。此外，通过使用从癌症患者中分离的 CTC 对癌症特异性基因进行逆转录酶聚合酶链式反应 (RT-PCR) 扩增，证实了 CTC 微分离器在后续基因检测中的有用性。
This paper presents a circulating tumor cell (CTC) microseparator for isolation of CTCs from human peripheral blood using immunomagnetic nanobeads with bound antiepithelial cell adhesive molecule (EpCAM) antibodies that specifically bind to epithelial cancer cells. The isolation is performed through lateral magnetophoresis, which is induced by high-gradient magnetic separation technology, involving a ferromagnetic wire array inlaid in the bottom substrate of a microchannel. Experimental results showed that the CTC microseparator isolates about 90% of spiked CTCs in human peripheral blood at a flow rate of up to 5 mL/h and purifies to approximately 97%. The overall isolation procedure was completed within 15 min for 200 mu L of peripheral blood. CTCs from peripheral blood of patients with breast and lung cancers were isolated with the CTC microseparator, and the results were compared with those of healthy donors. Using a fluorescence-based viability assay, the viability of CTCs isolated from peripheral blood of patients with cancer was observed. In addition, the usefulness of the CTC microseparator for subsequent genetic assay was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR) amplification of cancer-specific genes using CTCs isolated from patients with cancer.