石墨烯氧化物薄膜於表面電漿子共振生物感測器之研發

Abstract

表面電漿子共振(Surface plasmon resonance, SPR)生物感測器為利用金膜表面與生物分子交互作用所造成的微量折射率變化，進而達到免標記且高靈敏度之檢測技術，為目前光學式生物感測技術中的重要方法之一。

本論文將石墨烯氧化物(Graphene oxide, GO)鍵結於黃金薄膜上，所得到的薄膜稱為石墨烯氧化物薄膜，並透過表面電漿子共振生物感測技術，將此薄膜應用於生物感測上。石墨烯氧化物薄膜的製作首先將胱胺二鹽酸(Cystamine, Cys)作為連接劑，並透過分子自組裝單層膜(Self-assembled monolayer, SAM)技術，將胱胺鍵結於金膜上，形成胱胺薄膜。接著將石墨烯氧化物以共價鍵結的方式固定於胱胺薄膜表面，形成石墨烯氧化物薄膜(GO sheets)。論文中利用本實驗室之SPR量測系統並透過牛血清白蛋白與肺結核桿菌等生物分子進行檢測，並比較石墨烯氧化物薄膜與傳統金膜之靈敏度、檢測極限以及動力學分析之差異。

本論文發展之石墨烯氧化物薄膜表面電漿子共振生物感測器，與傳統金膜比較後，石墨烯氧化物薄膜之靈敏度約可提升12倍，檢測極限與動力學分析後之結果亦皆優於傳統金膜。最重要的是，石墨烯氧化物薄膜可直接透過官能基與生物分子鍵結，不必再透過抗原抗體的交互作用來執行，因此可有效地降低檢測成本。未來本系統除了可供醫學與疾病檢測外，亦可應用於製藥工業、環境檢測、農業科技等領域之分析應用，以造福人群。The advantage of surface plasmon resonance (SPR) biosensors includes high sensitivity, label-free, and real-time detection. These advantage leading SPR biosensors to identified as one of the most important optical detection methods for biomolecules.

The goal of this thesis is to develop graphene oxide (GO) sheets SPR biosensor that used GO as sensing layer for biomolecules detection. First, we used cystamine as linker layer, forming Cys film by self-assembled monolayer (SAM) technique. Then, GO solution was immobilized on the surface of Cys film through covalent attachment, forming GO sheets. The GO sheets SPR biosensor was used to detect the dynamic interactions of biomolecules and antibody-antigen. For experimental verification of this system, bovine serum albumin (BSA) and LAMP DNA products were used to demonstrate by GO sheets and conventional Au films. We also compared these two films by sensitivity, detection limits and kinetic analysis.

The sensitivity of GO sheets that can be determinedby this thesis is about 12-fold higher than that obtained with the sensor based on conventional Au films. The detection limits and the results of kinetic analysis of GO sheets were also better than conventional Au films. Most important of all, GO sheets have the ability for directly detect biomolecules, skipped the immunization interactions, leading to a low cost experiment. We anticipate that graphene oxide sheet SPR biosensors will not only enable for possible applications for biomedicine and diseases but also have wide spectrum of application in pharmaceutical industry, environmental monitoring, and agriculture in the near future.