Environmental Molecular Sciences Laboratory , Pacific Northwest National Laboratory , Richland , Washington 99354 , United States.
Knight Cancer Institute , Oregon Health & Science University , Portland , Oregon 97239 , United States.
Anal Chem. 2018 Oct 16;90(20):11756-11759. doi: 10.1021/acs.analchem.8b03268. Epub 2018 Oct 5.
Proteome profiling of circulating tumor cells (CTCs) can provide crucial insight into disease progression and the role of CTCs in tumor metastasis. We describe an integrated workflow to measure global protein expression in 1-5 spiked CTCs enriched from whole blood by immunodensity gradient centrifugation. Enriched CTCs were purified and collected by laser capture microdissection, prepared using a recently developed nanodroplet-based processing platform (nanoPOTS), and finally analyzed by ultrasensitive nanoLC-MS/MS. The workflow was capable of identifying an average of 164 and 607 protein groups from samples comprising 1 and 5 LNCaP cells, respectively, that were isolated from human whole blood. A panel of prostate cancer-specific proteins were identified and quantified, which was used to differentiate between spiked CTCs and white blood cells.
循环肿瘤细胞 (CTC) 的蛋白质组分析可为疾病进展和 CTC 在肿瘤转移中的作用提供重要的见解。我们描述了一种集成工作流程，用于通过免疫密度梯度离心从富含全血的 1-5 个掺入 CTC 中测量全局蛋白质表达。通过激光捕获微切割对富集的 CTC 进行纯化和收集，使用最近开发的基于纳米液滴的处理平台 (nanoPOTS) 进行制备，最后通过超灵敏纳升 LC-MS/MS 进行分析。该工作流程能够从从人全血中分离的包含 1 个和 5 个 LNCaP 细胞的样本中分别鉴定出平均 164 个和 607 个蛋白质组。鉴定和定量了一组前列腺癌特异性蛋白，可用于区分掺入的 CTC 和白细胞。
