以水膠為基礎結合適體建構非侵入式腫瘤壞死因子-α感測系統用於早期慢性疾病檢測

Assessment of TNF-α Levels with Hydrogel-Based Aptasensor in Noninvasive Technique for Early Chronic Disease Diagnosis

戴妤庭(Yu-Ting Tai)

指導教授 ： 柯富祥

國立交通大學/工學院/奈米科技研究所/碩士(2018年)

摘要

人類預防與診斷疾病一直是很重要的課題，隨著醫療技術的進步，人類平均壽命變長，疾病的提早偵測就更為重要。其中，在全球十大死因裡，慢性疾病占了一半以上，因此，如何提早偵測與診斷慢性疾病是今後主要的研究議題。腫瘤壞死因子-α (TNF-α)是一種與發炎反應有關的細胞因子，本篇研究利用超高分子水膠與特定的適體結合，製造一個可以量測生物標記腫瘤壞死因子-α的生物感測器，來達到慢性疾病的早期診斷。研究中首先利用水相合成方法，以萘乙酸為基礎製備水膠2-Naphthylacetic acid- L-Phenylalanine- L-Phenylalanine (Nap-FF)，並探討其基礎性質與成膠條件。為了能捕捉並偵測與發炎反應有關的細胞因子:腫瘤壞死因子-α，我們利用修飾有胺基的腫瘤壞死因子-α適體 (VR11)與水膠連接，並且觀察並分析再加入我們的目標受體腫瘤壞死因子-α對凝膠化的影響。利用此一特性，我們建構出腫瘤壞死因子-α濃度判定的檢測系統。
此系統的作用機制在於當特定的適體碰到其對應的生物標記時，會產生形變並抓取該目標，而當兩端都接有水膠的腫瘤壞死因子-α適體接上腫瘤壞死因子-α時，產生形變讓兩端的水膠堆疊在一起，使π-π間的相互作用增加，使水膠強度變強，並產生白點。依照成膠的強度與白點造成的穿透率不同為指標，來推斷腫瘤壞死因子-α的濃度。

關鍵字

超高分子水膠 ； 適體 ； 生物感測器 ； 腫瘤壞死因子-α

並列摘要

It is always an important issue on prevention and diagnosis for human diseases. As medical technology advances, life expectancy on humans is longer than before. Therefore, the early detection of diseases is more critical. Among the top 10 causes of death globally, over half of the causes are derived from chronic diseases. Thus, how to detect and diagnose chronic diseases in early stages is a main research topic from now on. Tumor necrosis factor-α (TNF-α), a key member of cytokines, is a meditator of inflammation in immune system. The present study was focused on the fabrication of a biosensor which can detect the biomarker TNF-α through the application of a supramolecular hydrogel and aptamer for early diagnosis of chronic disease. First, we synthesized a novel hydrogel, 2-Naphthylacetic acid- L-Phenylalanine- L-Phenylalanine (Nap-FF), which was based on Napthyl acetic moiety by liquid phase synthesis strategy, and investigated its characteristic and the condition of hydrogelation. To capture and assess the cytokines TNF-α which was related to inflammation, the combination of hydrogel and the TNF-α aptamer (VR11) which was modified by amino group was used. In addition, the effects of gelation were observed and analyzed after the target TNF-α was added. Finally, we constructed a detecting system of TNF-α’s concentration assessment.
In our sensing system, the specificity between the aptamer and its corresponding target plays an important role. When the aptamer VR11 which connected to hydrogel Nap-FF in two terminal sites captured TNF-α, its formation would change and let the hydrogel stacked together. Thus, the π-π interaction would increase, and so would the strength of the hydrogel. Moreover, the white points would be appeared. By means of the strengthening of hydrogelation and the difference of transmittance because of the white points, the concentration of TNF-α could be inferred easily and rapidly.

並列關鍵字

Supramolecular hydrogels ； aptamer ； biosensor ； TNF-α

參考文獻

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