Droplet-based chip with Ni-Co coated surface for immunoassay of multiple histidine-tagged protein

• Main Authors: Len-Hao Yao, 姚人豪
• Other Authors: Yao-Jen Chang
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/82219853118974198879

碩士 === 中原大學 === 機械工程研究所 === 101 === Abstract In this study, a droplet-based immunoassay chip was developed for performing immunoassays in the microarray format. The printed circuit board (PCB) was used as the substrate on which a microchannel layer was attached. A Ni–Co layer fabricated on the PCB substrate by electrodeposition is used to immobilize the multiplex histidine-tagged proteins. Then, the droplets are generated and manipulated to flow through the microchannel for rapid mixing and, finally, aligned at the downstream position of the microchannel. Thus, the histidine-tagged proteins are immobilized by the Ni–Co layer in microarray format for consequent immunoassay and fluorescence detection. To test this proposed droplet-based immunoassay chip, the tumor suppressor protein p53 and the extracellular-signal-related kinase 1 (ERK1, p40) were used, and the silicon oil was chosen as the carrier fluid. Compared with the mineral oil working as carrier fluid, the experimental result shows that for silicon oil the monodispersed droplets can be generated under a lower flow rate, since silicon oil has a higher viscosity. Moreover, the zigzag mixing channel provided rapid and thorough mixing effect. The detected fluorescence intensity is also proportioned to the concentration of the encapsulated content in a small droplet. This proposed droplet-based immunoassay chip allows the operation sequence of immunoassay being conducted automatically through the manipulation of droplets.