Center for Regenerative Therapeutics and Department of Medicine, Brigham and Women's Hospital, Cambridge, MA 02139, USA.
Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19626-31. doi: 10.1073/pnas.1211234109. Epub 2012 Nov 12.
Capture and isolation of flowing cells and particulates from body fluids has enormous implications in diagnosis, monitoring, and drug testing, yet monovalent adhesion molecules used for this purpose result in inefficient cell capture and difficulty in retrieving the captured cells. Inspired by marine creatures that present long tentacles containing multiple adhesive domains to effectively capture flowing food particulates, we developed a platform approach to capture and isolate cells using a 3D DNA network comprising repeating adhesive aptamer domains that extend over tens of micrometers into the solution. The DNA network was synthesized from a microfluidic surface by rolling circle amplification where critical parameters, including DNA graft density, length, and sequence, could readily be tailored. Using an aptamer that binds to protein tyrosine kinase-7 (PTK7) that is overexpressed on many human cancer cells, we demonstrate that the 3D DNA network significantly enhances the capture efficiency of lymphoblast CCRF-CEM cells over monovalent aptamers and antibodies, yet maintains a high purity of the captured cells. When incorporated in a herringbone microfluidic device, the 3D DNA network not only possessed significantly higher capture efficiency than monovalent aptamers and antibodies, but also outperformed previously reported cell-capture microfluidic devices at high flow rates. This work suggests that 3D DNA networks may have broad implications for detection and isolation of cells and other bioparticles.
从体液中捕获和分离流动细胞和颗粒具有巨大的诊断、监测和药物测试意义,但为此目的而使用的单价黏附分子导致细胞捕获效率低下,并且难以回收捕获的细胞。受能够有效捕获流动食物颗粒的海洋生物的启发,这些生物具有包含多个黏附结构域的长触须,我们开发了一种使用包含重复黏附适体结构域的 3D DNA 网络来捕获和分离细胞的平台方法,这些结构域延伸到数十微米的溶液中。DNA 网络通过滚环扩增从微流控表面合成,其中包括 DNA 接枝密度、长度和序列在内的关键参数可以很容易地进行调整。使用与许多人类癌细胞上过度表达的蛋白酪氨酸激酶-7(PTK7)结合的适体,我们证明 3D DNA 网络显著提高了淋巴母细胞 CCRF-CEM 细胞的捕获效率,超过单价适体和抗体,但保持了捕获细胞的高纯度。当整合到人字形微流控装置中时,3D DNA 网络不仅具有比单价适体和抗体更高的捕获效率,而且在高流速下优于先前报道的细胞捕获微流控装置。这项工作表明,3D DNA 网络可能对细胞和其他生物颗粒的检测和分离具有广泛的意义。