具生物活性中孔洞二氧化矽之製備及其在三維蛋白質生物晶片之應用

Preparation of Bioactive mesoporous silica and its application on 3-D Protein Biochips

陳韻竹(Yun-Chu Chen)

指導教授 ： 陳玉惠

中原大學/理學院/奈米科技碩士學位學程/碩士(2009年)

https://doi.org/10.6840/cycu200900861

摘要

本研究利用離子熔液為溶劑兼模板使用溶膠-凝膠法，將四乙氧基矽烷分別與帶有環氧基、胺基或二胺基之矽烷類共同水解與縮合，製備出三種具有不同生物活性官能基之中孔洞二氧化矽，經修飾後之中孔洞二氧化矽具與生物分子鍵結之能力，並將其於載玻片上製作出具有三維立體結構之蛋白質晶片，利用其生物活性抓取蛋白質分子進行三明治免疫分析，可用於檢測疾病感染等蛋白質分子間之變化。本研究針對可作為一種發炎感染指標之抗原分子-白血球間質素-6 進行檢測，探討所製備之三維立體結構之蛋白質晶片對此抗原分子之偵測靈敏度。
所製備具生物活性之中孔洞二氧化矽材料，利用FTIR、TGA、固態29Si-NMR 鑑定其結構性質，並由BET、SEM 與TEM 鑑定可知，此材料為具高比表面積之孔洞性材料。生物晶片掃描儀分析顯示，具生物活性之中孔洞二氧化矽為低背景值之蛋白質晶片基材。抗體固定化效果則顯示，三維蛋白質晶片比平板式晶片可抓取較多之抗體樣品，而三種具不同官能基之三維蛋白質晶片中，以胺基三維蛋白質晶片(3D AP chip)之固定化效果最佳。於偵測白血球間質素-6 之三明治免疫分析結果顯示，胺基三維蛋白質晶片確實具提昇提高靈敏度之效果，且可偵測較低濃度之白血球間質素-6。以上結果顯示，本研究發展之胺基三維蛋白質晶片為高偵測靈敏度與低偵測極限之蛋白質晶片。

關鍵字

中孔洞二氧化矽 ； 三維蛋白質生物晶片

並列摘要

In this study, the mesoporous silica aerogel with bioactive functional groups was prepared through the successive sol-gel reactions of the tetraethyl orthosilicate (TEOS) with three different silane reagents: 3-Glycidoxypropyl trimethoxysilane (GLYMO), (3-Aminopropyl)trimethoxysilane (APTS), and N-[3-(trimethoxysilyl) propyl]-1,2-ethanediamine (DAMO) , separately. An ionic liquid was utilized as a pore template and extractable solvent for the aerogel product. The as-prepared aerogel was loaded in arrayed dots on glass microscopy slide to perform as a protein biochip.
The as-prepared aerogel was characterized using FTIR, TGA, and 29Si solid-state NMR to verify its three-dimensional (3D) network structure. The results of BET, SEM, and TEM demonstrated that it is a porous material with large internal surface area. In addition, the microarray scanner images of the as-prepared 3D biochips exhibited clean fluorescent-excitation background, facilitating analyses for protein analytes. The human Interleukin-6 (IL-6) served as the target immune antigen and was demonstrated to receive a higher capturing efficiency by the as-prepared 3D APTS biochips than the traditional planar slides. Furthermore, a lower detection limit of the human IL-6 was also obtained through a sandwichimmunoassay on the 3D biochips prepared in this study than the plain-glass slides.

並列關鍵字

protein biochip ； mesoporous silica ； bioactive

參考文獻

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