利用真空濕式反應室進行即時就地膠原蛋白固定之研究

On-line Study of Collagen Immobilization Utilizing Wet Cell System

吳奕德(Yi-Te Wu)

指導教授 ： 廖峻德

中原大學/工學院/醫學工程研究所/碩士(2002年)

https://doi.org/10.6840/cycu200200481

摘要

本研究使用順流式微波氮氣電漿對排列完整的自我組裝形成ODT在Au（111）、Ag（111）的單層膜結構反應，使其表面產生極性基團，並隨即以稀釋的氣態丙烯酸單體與該活化之ODT單層膜進行接枝反應。在不明顯破壞硫（單層膜）與金屬鍵結的結構下，形成具丙烯酸之羧基官能基團的電漿處理表面，並與同樣含有羧基官能基分子之MUA單層膜做比較。以特殊設計之真空濕室化學反應室系統分別對前述兩種表面進行膠原蛋白的固定，再以同步輻射軟X-ray區段搭配高解析X光光電子能譜儀，分析各化學結構的變化；並對測得的S 2p、C 1s、N 1s與O 1s能譜做分析。實驗結果顯示：具有羧基之MUA /Au,Ag於真空濕室反應室中及傳統大氣環境下固定膠原蛋白，試片在兩種環境所固定的膠原蛋白結構，在定性及定量分析上無顯著差異，先證實在本真空濕室反應室中進行連續的蛋白質固定程序具體可行。另一方面，將僅電漿處理及隨之以丙烯酸接枝後的兩種ODT表面以相同方式傳送至真空濕室反應室中進行連續的蛋白質固定，經由X光光電子能譜分析後，發現兩者仍可以固定膠原蛋白，但在定量程度上有差異，經推論為：僅電漿處理之ODT/Au,Ag試片受到氧的參與反應，故產生能接著蛋白質分子的氧化基團，其中包括有限量的O=C-OH基。本研究確認可在真空濕室反應室系統完成固定蛋白質程序，以及即時表面結構分析。此結果有助於進一步對生醫應用方面的基礎反應機制的研究。

關鍵字

真空濕室化學反應室系統 ； X光光電子能譜儀 ； 順流式微波電漿 ； 自我組裝單層膜

並列摘要

This study utilized nitrogen downstream microwave plasma to react with the well packed self-assembly monolayers (SAMs) of 1-Octadecanethiol (ODT) adsorbed on Au (111) or Ag (111) substrate. Plasma processing was to create polarizable groups for subsequent grafting with diluted acrylic acid (AAc) monomer. The functional group of AAc, O=C-OH, was purposely bonded on the plasma-treated surface without obvious damage to sulfur/metal bonds. We took SAMs of 11-mercaptoundecanoic acid (MUA) on Au or Ag surface as the reference. Collagen was subsequently immobilized on these two types of surfaces under a custom-made well cell system. Synchrotron soft X-rays combined with high-resolution X-ray Photoelectron Spectroscopy (XPS) were applied to characterize the variations of chemical structures. The S 2p, C 1s, N 1s and O 1s spectra were typically analyzed. Experimental result has demonstrated that using MUA/Au, Ag as the substrates, collagen immobilization process in wet cell system or in atmospheric environment did not have qualitative or quantitative difference. Firstly, our well cell system is feasible to carry out on-line collagen immobilization process. We also compared the plasma-treated ODT surface with that after AAc-grafting. Using similar method to immobilize collagen in wet cell system, both surfaces were capable to have collagen immobilized. Nevertheless, their immobilized quantities were significantly different. The most pronounced processes are the participation of oxygen to the plasma-treated ODT surface, the generated oxidative groups, particularly the limited amount of O=C-OH group, are likely to bind with protein molecules. Overall, this work has completed a combined collagen immobilized process in wet cell system, followed by on-line XPS characterization. Fundamental studies on reactive mechanisms for biomolecules are expectative.

並列關鍵字

Wet Cell System ； Self-Assembly Monolayers ； Downstream Microwave Plasma ； X-ray Photoelectron Spectroscopy.

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