開發偵測柯楠因之生物比色法感測器

Abstract

本論文包含兩部分，主要是利用抗癌藥物柯楠因與腺嘌呤具有高度專一性的結合並形成 A2-coralyne-A2 錯合物的特性，設計出針對柯楠因之生化感測器，用以追蹤並定量柯楠因。在第一部分的實驗中，透過人工設計之 poly A 單股 DNA 序列與帶正電之高分子聚合物，於金奈米粒子輔助下檢測分析物柯楠因之含量。當溶液中有分析物存在時，富含腺嘌呤之單股 DNA 序列與柯楠因作用，形成 A2-coralyne-A2 錯合物，使該 DNA 序列無法與帶正電的聚合物作用，進一步導致帶負電之金奈米粒子聚集。當溶液中無分析物時，帶正電的聚合物會與 poly A 單股 DNA 預先作用。此時，金奈米粒子不會聚集，即可利用金奈米粒子之紫外光吸收光譜變化偵測分析物柯楠因。在第二部分實驗中，於金奈米粒子與金包覆四氧化三鐵奈米粒子表面修飾上三磷酸腺苷。當溶液中存在分析物柯楠因時，柯楠因會與粒子表面的三磷酸腺苷作用，導致兩種奈米粒子彼此藉由柯楠因而連接聚集。透過外加磁場，把有作用的粒子從溶液中移除。當溶液中的分析物柯楠因濃度增加時，溶液中金奈米粒子的濃度會因被外加磁場移除而隨之減少，透過偵測金奈米粒子紫外光譜吸收值的改變，即可偵測柯楠因的濃度。本論文所呈現之兩種偵測方法均具有簡單、方便、快速與靈敏度高等優點，可準確定量柯楠因於樣品中之濃度。

It has been demonstrated that coralyne can restrict the leukemia growth in cells and exhibit a noticeable anti-tumor activity, and thus provide a new medicine for the cancer therapy. In this study, two brand new coralyne biosensors were presented. Based on specific interaction between adenine and coralyne, a specific coordination complex of A2-coralyne-A2 formed, which could be used as main reorganization mechanism to detect coralyne. In the first part, a single strain DNA with poly A sequence was used to detect coralyne under the assistance of poly (diallyldimethylammonium) chloride (PDDA) and gold nanoparticles (Au NPs.) Coralyne was first captured by poly A DNA and neutralized its negative charges. After adding positively charged PDDA and Au NPs, PDDA induced the aggregation of Au NPs and the UV-Vis absorbance of Au NPs shifted. The A650/A520 ratio was used to estimate the concentration of coralyne. In the second part, we synthesized the adenosine triphosphate (ATP) stabilized gold nanoparticles (ATP-AuNPs) and ATP stabilized gold coated magnetic nanoparticles (ATP-MNPs), which used for detecting coralyne. In the presence of coralyne, both ATP stabilized nanoparticles aggregated via linking of coralyne. After applying external magnetic field, aggregated Au NPs were removed with MNPs, led to the reducing of UV absorbance. The absorbance difference was corresponding to the coralyne concentration. Both designed biosensors could use to estimate coralyne concentration with high accuracy and good selectivity.