CyToCap, ENS Cachan-Institut d'Alembert, 61 Avenue du Président Wilson, 94235 Cachan Cedex, France.
Biomed Microdevices. 2011 Feb;13(1):1-9. doi: 10.1007/s10544-010-9465-7.
We have recently reported the analytical performance of an immunosensor comprising one mm-scale parallel plate laminar flow chamber and applied to capture MCF7 breast cancer cells (Ehrhart et al., Biosens. Bioelectr. 24, 467, 2008). Herein we present a new multiplex immunosensor embodying four parallel plate laminar flow chambers that fit onto a standard, functionalized, microscopy glass slide. The four surfaces are coated with long alkyl chain spacers of 21-aminohenicosyl trichlorosilane (AHTS) and then grafted with a monoclonal anti-human epithelial cell adhesion molecule (EpCAM) antibody specific of target cells to immobilize. We first demonstrate a significantly (P < 0.01) improved capacity of each of the four flow chambers of the multiplex immunosensor to capture MCF7 cells compared to the previous single chamber device. Second, in addition to an increase of cell immobilization, the multiplex device offers a versatile tool easily grafted with various purified antibodies onto the four surfaces. Third, we obtained high cell capture rate and efficiency of various numbers of MCF7 cells spiked in buffer containing an equal number of background leukocytes. And fourth, we demonstrate isolation efficiency of circulating tumor cells (CTCs) from peripheral blood drawn from a small cohort of patients with localized or metastatic breast cancer. This new multiplex immunosensor could be tested for its potential to capture different subpopulations of CTCs.
我们最近报道了一种由一个毫米级平行板层流室组成的免疫传感器的分析性能，并将其应用于捕获 MCF7 乳腺癌细胞（Ehrhart 等人，Biosens. Bioelectr. 24, 467, 2008）。在此，我们提出了一种新的多路复用免疫传感器，该传感器包含四个平行板层流室，可安装在标准的功能化显微镜载玻片上。四个表面都涂有长链烷基间隔物 21-氨基二十一烷三氯硅烷（AHTS），然后用针对靶细胞的单克隆抗人上皮细胞黏附分子（EpCAM）抗体接枝以固定化。我们首先证明了多路复用免疫传感器的四个流室中的每一个都具有明显（P < 0.01）提高的捕获 MCF7 细胞的能力，与以前的单腔装置相比。其次，除了增加细胞固定化外，多路复用设备还提供了一种多功能工具，可轻松地将各种纯化抗体接枝到四个表面上。第三，我们获得了在含有相同数量背景白细胞的缓冲液中添加不同数量 MCF7 细胞时的高细胞捕获率和效率。第四，我们证明了从局部或转移性乳腺癌患者的小队列中抽取的外周血中循环肿瘤细胞（CTC）的分离效率。这种新的多路复用免疫传感器可以测试其捕获不同 CTC 亚群的潜力。
