Abstract
This paper proposes a microfluidic device for screening molecules such as aptamers, antibodies, proteins, etc. for target cell-specific binding molecules. The discovery of cancer cell-specific binding molecules was the goal of this study. Its functions include filtering non-target cell-binding molecules, trapping molecules on the surface of target cells, washing away unbound molecules, and collecting target cell-specific binding molecules from target cells. These functions were effectively implemented by using our previously developed micro pillar arrays for cell homogeneous dispersion and pneumatic microvalves for tall microchannels. The device was also equipped with serially connected filter chambers in which non-target cells were cultured to reduce the molecules binding to non-target cells as much as possible. We evaluated the performance of the device using cancer cell lines (N87 cells as target cells and HeLa cells as non-target cells) and two fluorescent dye-labeled antibodies: Anti-human epidermal growth factor receptor 2 (anti-HER2) antibody that binds to target cells and anti-integrin antibody that binds to non-target cells. The results showed that the device could reduce anti-integrin antibodies to the detection limit of fluorescent measurement and collect anti-HER2 antibodies from the target cells.
Highlights
Anti-cancer drugs are widely used for cancer treatment
Cancer cell-specific binding molecules such as cyclic arginine-glycine-aspartic acid tripeptide that binds to malignant brain tumor cells in glioma [4], an aptamer that binds to ovarian cancer cells [5], and a protein that binds to the protein disulfide isomerase, which is highly expressed on the surface of tumor cells [6] have been reported
To screen for cancer cell-specific binding molecules using cancer cells, conventional screening procedures involve the following steps: (Step 1) normal cells and the molecular library are mixed to filter out molecules that bind to normal cells; (Step 2) unbound molecules and target cancer cells are mixed to capture target cancer cell-specific binding molecules; (Step 3) washing of the target cancer cells; (Step 4) collecting bound molecules; and (Step 5) amplifying collected molecules
Summary
Anti-cancer drugs are widely used for cancer treatment. Conventional anti-cancer drugs damage cancer cells and harm normal cells [1]. To screen for cancer cell-specific binding molecules using cancer cells, conventional screening procedures involve the following steps: (Step 1) normal cells and the molecular library are mixed to filter out molecules that bind to normal cells; (Step 2) unbound molecules and target cancer cells are mixed to capture target cancer cell-specific binding molecules; (Step 3) washing of the target cancer cells; (Step 4) collecting bound molecules; and (Step 5) amplifying collected molecules. Cancer cell-specific binding molecules are condensed by repeating steps 1 to 5 In addition to these complicated steps, this screening procedure requires precise manual operations, which are laborious and time-consuming. To conduct screening without human errors and decrease the screening time, automation with precise manipulation is required
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