Multifunctional Magnetic Particles for Combined Circulating Tumor Cells Isolation and Cellular Metabolism Detection.
Multifunctional Magnetic Particles for Combined Circulating Tumor Cells Isolation and Cellular Metabolism Detection.
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DOI：
10.1002/adfm.201504184
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发表时间：
2016-06-14
影响因子：
19
通讯作者：
Qian K
中科院分区：
文献类型：
--
作者：
Wu J;Wei X;Gan J;Huang L;Shen T;Lou J;Liu B;Zhang JX;Qian K
我们首次展示了基于多功能磁性颗粒的稀有细胞分离结合下游激光解吸/电离质谱（LDI-MS）来测量富集循环肿瘤细胞（CTC）的代谢。通过探索不同的细胞过程，CTC 代谢的表征在理解肿瘤微环境方面发挥着重要作用。然而，由于检测灵敏度低、样品复杂性高、制备过程繁琐，特别是对于临床研究中的稀有细胞分析，表征细胞代谢仍然具有挑战性。在这里，我们将氧化铁磁性颗粒与表面上的抗 EpCAM 结合，从 PBS 和全血中特异性、高效地富集 CTC，细胞浓度为每毫升 6-100 个细胞。此外，这些亲水性颗粒作为基质，能够对复杂生物混合物中的小代谢物（MW<500 Da）进行灵敏和选择性的 LDI-MS 检测，并且可以进一步与同位素定量相结合，以监测约 50 个 CTC 的选定分子代谢。我们独特的方法将 CTC 的免疫磁性分离和基于 LDI-MS 的代谢分析相结合，这代表了稀有细胞下游代谢物分析的关键一步，以研究 CTC 的生物学特征及其在病理和生理现象中的细胞反应。通过探索不同的细胞过程，循环肿瘤细胞 (CTC) 代谢的表征在理解肿瘤微环境方面发挥着重要作用。在此，开发了多功能磁性颗粒，用于组合稀有细胞分离和下游激光解吸/电离质谱法，以测量富集 CTC 的代谢。
We for the first time demonstrate multi-functional magnetic particles based rare cell isolation combined with the downstream laser desorption/ionization mass spectrometry (LDI-MS) to measure the metabolism of enriched circulating tumor cells (CTCs). The characterization of CTCs metabolism plays a significant role in understanding the tumor microenvironment, through exploring the diverse cellular process. However, characterizing cell metabolism is still challenging due to the low detection sensitivity, high sample complexity, and tedious preparation procedures, particularly for rare cells analysis in clinical study. Here we conjugate ferric oxide magnetic particles with anti-EpCAM on the surface for specific, efficient enrichment of CTCs from PBS and whole blood with cells concentration of 6–100 cells per mL. Moreover, these hydrophilic particles as matrix enable sensitive and selective LDI-MS detection of small metabolites (MW<500 Da) in complex bio-mixtures and can be further coupled with isotopic quantification to monitor selected molecules metabolism of ~50 CTCs. Our unique approach couples the immunomagnetic separation of CTCs and LDI-MS based metabolic analysis, which represents a key step forward for downstream metabolites analysis of rare cells to investigate the biological features of CTCs and their cellular responses in both pathological and physiological phenomena. The characterization of circulating tumor cells (CTCs) metabolism plays a significant role in understanding the tumor microenvironment, through exploring the diverse cellular process. Herein, multi-functional magnetic particles are developed for combined rare cell isolation and downstream laser desorption/ionization mass spectrometry to measure the metabolism of enriched CTCs.