Huang Chao, Santana Steven M, Liu He, Bander Neil H, Hawkins Benjamin G, Kirby Brian J
Department of Biomedical Engineering, Cornell University, Ithaca, NY, USA.
Electrophoresis. 2013 Nov;34(20-21):2970-9. doi: 10.1002/elps.201300242. Epub 2013 Oct 9.
The capture of circulating tumor cells (CTCs) from cancer patient blood enables early clinical assessment as well as genetic and pharmacological evaluation of cancer and metastasis. Although there have been many microfluidic immunocapture and electrokinetic techniques developed for isolating rare cancer cells, these techniques are often limited by a capture performance tradeoff between high efficiency and high purity. We present the characterization of shear-dependent cancer cell capture in a novel hybrid DEP-immunocapture system consisting of interdigitated electrodes fabricated in a Hele-Shaw flow cell that was functionalized with a monoclonal antibody, J591, which is highly specific to prostate-specific membrane antigen expressing prostate cancer cells. We measured the positive and negative DEP response of a prostate cancer cell line, LNCaP, as a function of applied electric field frequency, and showed that DEP can control capture performance by promoting or preventing cell interactions with immunocapture surfaces, depending on the sign and magnitude of the applied DEP force, as well as on the local shear stress experienced by cells flowing in the device. This work demonstrates that DEP and immunocapture techniques can work synergistically to improve cell capture performance, and it will aid in the design of future hybrid DEP-immunocapture systems for high-efficiency CTC capture with enhanced purity.
从癌症患者血液中捕获循环肿瘤细胞(CTC)有助于癌症及转移的早期临床评估以及基因和药理学评估。尽管已经开发出许多用于分离罕见癌细胞的微流控免疫捕获和电动技术,但这些技术往往受到高效与高纯度之间捕获性能权衡的限制。我们展示了一种新型混合DEP-免疫捕获系统中剪切依赖性癌细胞捕获的特性,该系统由在Hele-Shaw流动池中制造的叉指电极组成,该流动池用单克隆抗体J591进行功能化,J591对表达前列腺特异性膜抗原的前列腺癌细胞具有高度特异性。我们测量了前列腺癌细胞系LNCaP的正负DEP响应作为施加电场频率的函数,并表明DEP可以通过促进或阻止细胞与免疫捕获表面的相互作用来控制捕获性能,这取决于施加的DEP力的符号和大小,以及装置中流动细胞所经历的局部剪切应力。这项工作表明DEP和免疫捕获技术可以协同工作以提高细胞捕获性能,并且将有助于未来设计用于高效捕获CTC并提高纯度的混合DEP-免疫捕获系统。