13 July 2004 Separation of breast cancer cells from peripherally circulating blood using antibodies fixed in microchannels
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Bio-Micro Electro Mechanical System (Bio-MEMS) technology was applied to the problem of early breast cancer detection and diagnosis. A micro-device is being developed to identify and specifically collect tumor cells of low abundance (1 tumor cell among 107 normal blood cells) from circulating whole blood. By immobilizing anti-EpCAM (Epithelial Cell Adhesion Molecule) antibodies on polymer micro-channel walls by chemically modifying the surface of the PMMA, breast cancer cells from the MCF-7 cell line, which over-express EpCAM, were selected from a sample volume by the strong binding affinity between the antibody and antigen. To validate the capture of the breast cancer cells, three fluorochrome markers, each identified by a separate color, were used to reliably identify the cancer cells. The cancer cells were defined by DAPI+ (blue), CD45- and the FITC-cell membrane linker+ (green). White blood cells, which may interfere in the detection of the cancer cells, were identified by DAPI+ (blue), CD45+ (red), and the FITC-cell membrane linker+ (green). EpCAM/anti-EpCAM binding models from the literature were used to estimate an optimal velocity, 2mm/sec, for maximizing the number of cells binding and the critical binding force. At higher velocities, shear forces (> 0.48 dyne) will break existing bonds and prevent the formation of new ones. This detection micro-device can be assembled with other lab-on-a-chip components for follow-up gene and protein analysis.
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Juan Feng, Juan Feng, Steven A Soper, Steven A Soper, Robin L McCarley, Robin L McCarley, Michael C. Murphy, Michael C. Murphy, "Separation of breast cancer cells from peripherally circulating blood using antibodies fixed in microchannels", Proc. SPIE 5312, Lasers in Surgery: Advanced Characterization, Therapeutics, and Systems XIV, (13 July 2004); doi: 10.1117/12.529748; https://doi.org/10.1117/12.529748

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