Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004, Jiangsu, PR China.
Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004, Jiangsu, PR China.
Anal Chim Acta. 2018 Dec 31;1044:162-173. doi: 10.1016/j.aca.2018.07.066. Epub 2018 Jul 28.
This work demonstrated an effective strategy for the capture, identification and determination of multiple types of circulating tumor cells (CTCs) on functional and biocompatible immunomagnetic nanosphere interfaces (IMNs). The IMNs were achieved by functionalizing superparamagnetic iron oxide nanospheres (FeO) with polymerized ionic liquid (PIL), and then coating with epithelial-cell-adhesion-molecule antibody (anti-EpCAM). The IMNs exhibited outstanding cell capture efficiency (above 95%) and specificity when employed to separate multiple EpCAM-positive tumor cells, due to the abundant carboxyl groups in the structure of PIL, which enhanced the coupling efficiency of MNs with anti-EpCAM by chemical bonding between carboxyls and amines, thereby enabling more target cells adhered onto IMNs. Under the optimized capture conditions, IMNs were shown an excellent cell capture performance in the range of 5-400 cells/mL in three different cases (e.g., PBS, MCF-7 and THP-1 mixed cell suspension, lysed blood). More significantly, our results indicated that with modification of PIL, in addition to the capture efficiency, the cell viability rate of CTCs was also greatly improved (98%) owing to the nontoxic and biocompatible properties of PIL, which realized the proliferation of the rare number CTCs for further molecular characterization. Finally, the IMNs were successfully applied to the isolation and detection of CTCs in cancer patient peripheral blood samples and as low as one CTC in the whole blood was captured and identified by the ICC method.
这项工作展示了一种在功能化和生物相容的免疫磁性纳米球界面(IMNs)上捕获、鉴定和定量多种循环肿瘤细胞(CTCs)的有效策略。通过将聚合离子液体(PIL)功能化超顺磁性氧化铁纳米球(FeO),然后用上皮细胞黏附分子抗体(anti-EpCAM)涂覆,实现了 IMNs。由于 PIL 结构中丰富的羧基,IMNs 表现出出色的细胞捕获效率(超过 95%)和特异性,可用于分离多种 EpCAM 阳性肿瘤细胞,通过羧基和胺之间的化学键合增强了 MNs 与 anti-EpCAM 的偶联效率,从而使更多的靶细胞黏附到 IMNs 上。在优化的捕获条件下,IMNs 在三种不同情况下(例如 PBS、MCF-7 和 THP-1 混合细胞悬浮液、裂解血液)的 5-400 个细胞/mL 的范围内表现出优异的细胞捕获性能。更重要的是,我们的结果表明,通过 PIL 的修饰,除了捕获效率外,CTCs 的细胞活力率也大大提高(98%),这是由于 PIL 的无毒和生物相容性,实现了稀有数量 CTCs 的增殖,以便进一步进行分子特征分析。最后,IMNs 成功地应用于癌症患者外周血样本中 CTCs 的分离和检测,通过 ICC 方法可以捕获和鉴定低至全血中一个 CTC。