Proceedings of the National Academy of Sciences · 2012 · 281 citations · 38 references
EngineeringBioengineeringBiomedical EngineeringBiomaterialsBiosensorsCell AdhesionAdhesion MoleculesBiopolymer NetworksDna NanotechnologyMicrofluidic BiosensorsMicrofluidic Device FabricationMicrofluidicsAptamer ImmobilizationAptamer-based Cell CaptureAptamersMicrofluidic Cell IsolationMicrofluidic Device DesignMicrofluidic Diagnostic DevicesMicrofluidic PlatformsRolling Circle Amplification
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.
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Isolation of rare circulating tumour cells in cancer patients by microchip technology
Sunitha Nagrath, Lecia V. Sequist, Shyamala Maheswaran et al. · Nature · 2007 · 3.5K citations · Full text
Medicine, Cancer Biology, Oncology +15
Detection of Mutations in <i>EGFR</i> in Circulating Lung-Cancer Cells
Shyamala Maheswaran, Lecia V. Sequist, Sunitha Nagrath et al. · New England Journal of Medicine · 2008 · 1.6K citations · Full text