Department of Chemistry, University of Kansas, 1567 Irving Hill Road, Lawrence, Kansas 66045, United States.
Center of BioModular Multi-scale Systems for Precision Medicine, University of Kansas, Lawrence, Kansas 66045, United States.
ACS Sens. 2021 May 28;6(5):1831-1839. doi: 10.1021/acssensors.0c02728. Epub 2021 May 3.
Liquid biopsies are becoming popular for managing a variety of diseases due to the minimally invasive nature of their acquisition, thus potentially providing better outcomes for patients. Circulating tumor cells (CTCs) are among the many different biomarkers secured from a liquid biopsy, and a number of efficient platforms for their isolation and enrichment from blood have been reported. However, many of these platforms require manual sample handling, which can generate difficulties when translating CTC assays into the clinic due to potential sample loss, contamination, and the need for highly specialized operators. We report a system modularity chip for the analysis of rare targets (SMART-Chip) composed of three task-specific modules that can fully automate processing of CTCs. The modules were used for affinity selection of the CTCs from peripheral blood with subsequent photorelease, simultaneous counting, and viability determinations of the CTCs and staining/imaging of the CTCs for immunophenotyping. The modules were interconnected to a fluidic motherboard populated with valves, interconnects, pneumatic control channels, and a fluidic network. The SMART-Chip components were made from thermoplastics via microreplication, which lowers the cost of production making it amenable to clinical implementation. The utility of the SMART-Chip was demonstrated by processing blood samples secured from colorectal cancer (CRC) and pancreatic ductal adenocarcinoma (PDAC) patients. We were able to affinity-select EpCAM expressing CTCs with high purity (0-3 white blood cells/mL of blood), enumerate the selected cells, determine their viability, and immunophenotype the cells. The assay could be completed in <4 h, while manual processing required >8 h.
液体活检因其采集的微创性质而在多种疾病的管理中变得越来越流行，因此可能为患者带来更好的结果。循环肿瘤细胞 (CTC) 是从液体活检中获得的许多不同生物标志物之一，已经报道了许多用于从血液中分离和富集它们的高效平台。然而，许多这些平台需要手动处理样本，这可能会由于潜在的样本丢失、污染以及对高度专业化操作人员的需求，在将 CTC 检测转化为临床应用时带来困难。我们报告了一种用于分析稀有靶标的系统模块化芯片 (SMART-Chip)，它由三个特定任务模块组成，可以完全自动化 CTC 的处理。这些模块用于通过亲和性从外周血中选择 CTC，然后进行光释放、同时对 CTC 进行计数和活力测定，以及对 CTC 进行染色/成像以进行免疫表型分析。这些模块与带有阀门、互连、气动控制通道和流体网络的流体母板相互连接。SMART-Chip 组件由热塑性塑料通过微复制制成，这降低了生产成本，使其易于临床实施。通过处理从结直肠癌 (CRC) 和胰腺导管腺癌 (PDAC) 患者获得的血液样本，证明了 SMART-Chip 的实用性。我们能够以高纯度（每毫升血液 0-3 个白细胞）亲和性选择表达 EpCAM 的 CTC，对所选细胞进行计数，确定其活力，并对细胞进行免疫表型分析。该检测可以在<4 小时内完成，而手动处理需要>8 小时。
