| Info. | Vol.10 - No.3 (2016.09.20) |
|---|---|
| Title | Preconcentration-enhanced Immunosensing for Whole Human Cancer Cell Lysate based on a Nanofluidic Preconcentrator |
| Authors | Hsuan Franziska Wu1, Tamara G. Amstislavskaya2, Pin-Hsuan Chen3, Ting-Feng Wu4, Yu-Hung Chen1,5,* & Chun-Ping Jen3,* |
| Institutions | 1Department of Medicine, College of Medicine, National Cheng-Kung
University, Tainan, Taiwan, R.O.C. 2Laboratory of Experimental Models of Emotional Pathology,Scientific Research Institute of Physiology and Basic Medicine,Novosibirsk, Russia 3Department of Mechanical Engineering, National Chung Cheng University, Chia Yi, Taiwan, R.O.C. 4Department of Biotechnology, Southern Taiwan University of Science and Technology, Tainan, Taiwan, R.O.C. 5Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng-Kung University, Tainan, Taiwan, R.O.C. *Correspondence and requests for materials should be addressed to C.-P. Jen ( imecpj@ccu.edu.tw) and Y.-H. Chen ( second@mail.ncku.edu.tw) |
| Abstract | Sample preconcentration is an important step that increases the accuracy of subsequent detection,especially for samples with extremely low concentrations. Due to the overlap of electrical double layers in a nanofluidic channel, the concentration polarization effect can be generated by applying an electric field. A nonlinear electrokinetic flow is induced, which results in the fast accumulation of proteins in front of the induced ionic depletion zone, the so-called exclusion-enrichment effect. In this way, a protein sample can be driven by electroosmotic flow and accumulated at a specific location. In the present study, a nanofluidic preconcentrator fabricated with the help of junction gap electric breakdown was integrated with microelectrodes for immunoassay. The preconcentration chip for proteins was fabricated using simple standard soft lithography with a polydimethylsiloxane replica. Human galectin-1 proteins from the cell lysate of T24 cells were concentrated and immunoassayed in the proposed microchip. The capability of the proposed microchip for concentrating multiple proteins from cell lysates and immunoassays after preconcentration was demonstrated. Immunosensing was evaluated by measurements of both fluorescence intensities and impedance, which proved the enhancement of preconcentration for immunoassay. |
| Keyword | Protein, Preconcentration, Exclusion-enrichment effect, Electric breakdown, Cell lysate, Galectin-1, Immunoassay |
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