新型態中孔洞材料之合成與應用：磷酸化蛋白質純化與去氧核醣核酸藥物載體之設計

Synthesis and application of a new type mesoporous silica: phosphopeptide purification and DNA drug carrier

劉依欣(Yi-hsin Liu)

指導教授 ： 謝發坤

國立中央大學/理學院/化學系/碩士(2014年)

摘要

本篇論文分為兩個部分:
第一部分，本實驗室成功在具有羧酸官能基的中孔洞材料表面，進一步修飾可調節環狀大小之雙胺官能基，合成新型環狀中孔洞材料，命名為XC-CAR-10，X代表雙端胺基化合物中間的碳數。此環狀中孔洞材料可利用大環分子效應，提升對金屬的吸附效果，以鑭系元素中的鉺金屬為例，其吸附效果可達174 μg/mg，並藉由鉺金屬和磷酸化蛋白質間的螯合作用，溶液中的磷酸化蛋白質會與中孔洞材料中的鉺金屬產生鍵結，利用固定化金屬離子層析法(IMAC)，達到使磷酸化蛋白質具有增量純化的成果，回收率可高達百分之30以上。
第二部分，以表面修飾胺基的中孔洞材料AM-SBA-15發展出新型藥物載體，將孔洞材料內部吸附螢光物質，且因為胺基在中性環境下帶正電荷因此可以吸附負電荷的 DNA雙股螺旋，因此我們期望將DNA作為藥物載體之蓋子，保護孔洞內部的螢光物質不被釋放，但若加入可以使DNA斷裂之切割酶，則可以控制藥物載體使螢光物質不受DNA的保護進而被釋放出來。

關鍵字

中孔洞材料 ； 藥物載體 ； 磷酸化蛋白 ； 純化

並列摘要

The first part of my research is about annular rings with controllable sizes are successfully created on the surface of COOH-functionalized mesoporous silica (namely, XC-CAR-10, where X represents the number of carbons in the diamines used) for the adsorption of metal ions. Particularly, excellent adsorption ability of 3C-CAR-10 towards Er3+ (ca. 174 μg/mg) was achieved.
Reversible phosphorylation of proteins is a common theme in the regulation of important cellular functions such as growth, metabolism, and differentiation. Although, the number of cellular phosphoproteins is relatively high, the phosphorylated residues themselves are generally of low abundance due to the sub-stoichiometric nature. Additionally, phosphopeptides with phosphate groups are able to form stable complexes with the lanthanide ions by the formation of precipitation. In this study, thus, we reported an annular mesoporous adapted with trivalent lanthanide ions Er3+ on for the applications of phosphopeptide enrichment and cleaning-up.
The second part of my research is about a new style drug carrier with designed DNA as a cap to control drug release. We synthesized mesoporous silicas with an amine group on the surface, named AM-SBA15. Due to owning positive charge of AM-SBA15, it can be formation of positive-negative interaction between AM-SBA15 and DNA phosphate groups at neutral condition. Thus, we designed a short length DNA with 17 EcoRV cutting cites as caps on the surface of AM-SBA15 mesoporous silica. According to investigation results, DNA shows a good capability for covering pores on the surface of AM-SBA15 so as to not allow fluorescein released. Remarkably, the fluorescein will be released upon EcoRV added because restriction enzyme of EcoRV digests DNA into smaller fragments. Finally, we develop a novel platform as a drug carrier and release system. Drugs will be released when composites are triggered by specific restriction enzyme.

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