生物分子利用共價鍵修飾在基材上已經廣泛運用於生醫感測器、生醫材料等，隨著生物科技的進步與生醫感測器等的蓬勃發展，共價鍵修飾獲得了越來越多的關注，其步驟包含：(1)在基材上修飾自組性單分子膜，(2)利用交聯劑，如：碳二亞胺，與自組性單分子膜之官能基反應形成活化層，(3)活化層再與欲修飾於基材表面之生物辨識分子反應，如抗體、DNA等，以共價鍵—肽鍵連接。然而目前的修飾仍是依據經驗法則，不同文獻所使用的修飾條件更是大相逕庭，缺乏基礎理論作為修飾的依據。因此本研究致力於探討生物分子修飾之動態過程，以提供修飾的依據，進而改善生醫感測器。表面增顯紅外光技術結合衰弱式全反射(ATR-SEIRAS)的分析由於具有高靈敏度、表面選擇性與低背景溶液干擾等優點，逐漸受到重視，已成功應用於修飾過程的界面量測。本實驗利用衰弱式全反射表面增顯紅外光即時現址觀測分子層的修飾，探討11-巰基烷酸做為自組性單分子膜修飾於金表面，利用EDC/NHS活化其表面，並使11-巰基烷酸與氨基二戊鐵(Aminoferrocene)以肽鍵連接，藉SEIRAS的觀測，而得到較佳的修飾參數。

Covalent binding has been wildly used in biosensors and biomaterials. With the vigorous development of biosensors, covalent immobilization is getting more and more attention. The process of forming covalent bonds on biosensors including: (1) forming a self-assembly monolayer (SAM), (2) activating the SAM with crosslinkers to form an active layer, and then (3) covalent attachment of biomolecules. However, the immobilization parameters are still based on experience, and vary from one study to another. In this study, 11-mercaptoundecanoic acid (MUA) was used to form SAM on gold surface, activating the surface through EDC/NHS, and then formed peptide with aminoferrocene (AmFc). Known for high sensitivity, low background interference, and in-situ observation, attenuated total reflection surface enhanced infrared adsorption spectroscopy (ATR-SEIRAS) was used to investigate the immobilization process and then to find a relative well optimized parameters.

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