微陣列蛋白質晶片基材的製作

Substrates preparation for Microarray protein chips

蕭正彥(Cheng-Yen Hsiao)

指導教授 ： 張淑美

國立臺北科技大學/工程學院/有機高分子研究所/碩士(2011年)

摘要

蛋白質微陣列晶片的製作可分為三大部分，共有基材前處理、基材表面化學修飾、固定蛋白質在基材表面。本研究使用分子膜自組裝方法進行表面化學修飾，
使用此方法將矽氧烷修飾在玻璃或矽晶片上，形成末端官能基基材。由於矽氧烷末端官能有親疏水性質的差異，可用接觸角測量儀滴上液滴觀察。另外，我們也使用AFM觀察矽氧烷分子在基材表面成長的情形，並從分子高度推估為多層分子膜，也從roughness來得知末端官能基基材表面的粗糙度。
由於修飾在基材表面的矽氧烷之末端官能基，可與蛋白質中的COOH及NH2官能基進行鍵結，因此可點上Cy5-streptavidin觀察螢光強度，由此結果可判斷蛋白質在自行製作的末端官能基基材固定狀況，與市售玻片的結果比較。另外，本研究也探討在基材上的分子以2D和3D的方式成長在基材表面後，發現以3D成長方式之分子由於spacer較大，能夠鍵結較多的蛋白質，螢光強度比2D方式成長的基材要高。
HSV-1病毒為人體中常見的病毒，因此本研究選用此病毒之抗原，來做抗原-抗體免疫分析，以利日後能夠應用於臨床醫學診斷。

關鍵字

蛋白質微陣列基材 ； Cy5-protein ； 分子膜自組裝

並列摘要

Protein microarray chips is composed of three parts, those are pre-treatment substrates, surface chemical modification, and immobilizing protein on substrate surfaces. In this study, self-assembly monolayers is used for surface chemical modification. By this method to perform silaneization on glass and silicon wafer, forming the terminal group substrates. Due to protein includes COOH and NH2 functional groups, that could bind with silane, dendrimer on substrates, so proteins will immobilize on substrate surface. Finally, we will connected to the substrates, Cy5 in red laser irradiation, it will excite red fluorescence, and then we can use Gene Pix 4000B observing red fluorescence intensity, determining the situation of protein immobilized on substrates, as well as performing quantitative objectives.
This study also explores the substrate molecules to the way 2D and 3D growth of the substrate surface, found that 3D growth mode of the larger molecules as spacer, to bond more protein, fluorescence intensity ratio of 2D growth mode must be high.
HSV-1 virus is a common human virus, so this antigen of HSV-1 is used in this study, performing the antigen - antibody analysis, in order to facilitate the future can be used in clinical diagnosis

並列關鍵字

Microarray protein substrates ； Cy5-protein ； self-assembly

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