| Summary: | 碩士 === 輔仁大學 === 生命科學系碩士班 === 93 === Proteomics is a new knowledge of biological total proteome research. Within the proteomic technologies, the combination of Two-dimensional electrophoresis（2-DE） and mass spectrometry（MS）is the most popular skill.Using this method makes scientists able to separate and identify thousands of proteins simultaneously. If this high throughput operation platform can be practically applied on clinical diagnosis such as the identification of tumor markers, it will be a great achievement to the initial stage diagnosis and treatment of human disease. Non-invasive sample collections are prioritized in the clinical diagnosis of cancerous disease. Among all source of sample, blood is the best choice because of it`s consistent nature, which are not easily affected by exterior factors such as diet. However, the composition of proteins in blood are extremely uneven. Albumin and immunoglobulin（IgG）consist up to 80% and 10% of total protein in blood respectively. In contrast, most important tumor markers exists in much smaller proportions. That cannot be easily identified. To identify these tumor markers, it is necessary to remove those higt abundant proteins in blood sample in order to enrich the minor proteins. Then finding the meaningful target markers from human plasma makes possible to work. Therefore, to develop a high resolution and high repeatable protein separation technology is the main key of seroproteome analyzation. The research purpose of this thesis is to develope the seroproteomics operation platform by using the Parkinson`s disease（PD）patient plasma as an experiment material. In this study, identification of the PD disease specific protein by the seroproteomics operation platform are optimistically expected which can offer to clinical diagnosis consult. The success development of this platform may possibly apply on all the plasma-based clinical diagnosis. In this thesis, we had a initial result：Firstly, We successfully depleted albumin with Cibacorn blue sepharose, depleted IgG with protein A and established pH 3-10NL, pH 4-7 plasma 2-DE platform SOP. Secondly, base on the well-developed plasma 2-DE SOP , we have processed the plasma group（Normal and PD）from Cardinal-Tien Hospital. In a depletion of albumin and IgG situation, we found a specific target protein tumor marker after MALDI-Tof identification. On the other hand, we also purified the plasma high abundant protein as an antigen to immuno BALB/c mice for monoclonal antibody produced. Now we successfully gain the transferring, albumin, antitrypsin and ApoA-1`s monoclonal antibody. These monoclonal antibody will be used for removed plasma high abundant protein and enriched minor protein for achievement tumor marker identification purpose
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