Development of Polymer-based Lab-on-Chip System for Bioanalysis

• Main Authors: Wang-Chou Sung, 宋旺洲
• Other Authors: Shu-Hui Chen
• Format: Others
• Language: zh-TW
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/5pe9m8

博士 === 國立成功大學 === 化學系碩博士班 === 92 === 　　Integration of analytical miniaturized systems is important because it has the potential for the cost reduction, automation and analysis efficiency. The microdevices have been widely applied in the field of bioanalysis, drug screening and chemical/medical detection. By making use of the microelectronic mechanical system (MEMS) technologies, we can miniaturize conventional analytical techniques and integrate them to make total analysis systems in a glass or polymer based microchip, this is what we called lab-on-chip. However, for example, some difficulties like buffer systems compatibility with the MS detection always arise when interfacing chip with mass instrument. Other problems like chip substrate choosing, detection system development and the connection of different reaction system also have to be overcomed. In this dissertation, we built two kinds of polymer-based; poly(dimetyl siloxane) (PDMS) and poly (methyl methacrylate) (PMMA) for integrated analysis of DNA and protein samples. From the results obtained, the integration on the microchip is successful and proceeds high analysis efficiency in both analysis. 　　Fast clinical screening of abnormal chromosomes demands a high-throughput method including DNA sizing and detection of the amplified products. The first study is to explore the use of polymer microchip electrophoresis for the analysis of PCR products of fragile X (CGG)n alleles to facilitate a fast exclusion test of fragile X syndrome (FXS). The PCR bands with more than six CGG-repeats in difference could be clearly distinguished in less 3 mins by microchip electrophoresis with a separation length of 6 cm. Twelve samples from males and females were tested in this study. From the results shown that not only the samples could be finished in a short time, but show correlation between the microchip electrophoresis and the traditional one dimensional (1D) gel electrophoresis. This proves the feasibility of PMMA chip to be a high throughput and fast exclusion DNA analysis tool. 　　For proteomics research, mass analyzer plays an important role in protein identification. Because MS detection could eliminate the dye-labelling procedure and get some information of each chromatographic peak, like analyzes specices and molecule mass. This makes the coupling of microfluidic chip and MS detection more attractive and useful than traditional optical detection in proteomics research. In th other part of the dissteration, an pulled capillary tip was inserted into the end of the PMMA microfluidic channel to be an electrospary interface. This is an easy method was developed to build an ionization interface on the polymer substrate microchip. The whole device could be easily mounted on the commercial MS instrument and offer a high signal quality compared with the commercially available nano electrospray tips. In this research, this polymer-based microdevice would like to generate MS signals for protein identification from the small amounts of protein samples. 　　In the proteomics research, a good separation tool could improve to identify the proteins. Beside, in the convetional protein analysis, the procedures like enzyme digestion and desalting were also used to increase the protein identification precision. In this study, a microfabricated PDMS electrospray microfluidic that integrated AMPS modified separation channel, an injector with trypsin enzyme cartridge and desalting cartride was developed to build the protein identification microfluidic module (PIMM) to do high-speed proteins analysis. Results show that a four peptide mixture could be separated in 2 mins and identified the amino acid sequence by tandem MS. In order to prove the high efficiency of PIMM, two native proteins could be sequential analysis in two hours. This proposed a protein identification microfluidic module (PIMM) can successfully operated in a high efficiency.