類沸石咪唑骨架材料(ZIF-90)之官能基修飾與 包覆質體DNA的探討

顔嘉儀(Chia-I Yen)

指導教授 ： 謝發坤

國立中央大學/理學院/化學系/碩士(2015年)

摘要

本篇論文包含兩部分，第一部分延續實驗室已畢業的吳哲瑋學長之研究成果，將含有醛官能基的有機金屬骨架材料─ZIF-90─利用後修飾的方式轉換為三種分別帶有羧酸、胺基、硫醇基的不同材料，分別命名為ZIF-90-C、ZIF-90-A、ZIF-90-T。在溫和的反應條件下，修飾後的材料仍保有結晶性和孔洞性質，其中有近80%之微孔體積顯著的保留在修飾後的ZIF-90-C和ZIF-90-A材料上。而ZIF-90-A在純水中因胺基的質子化，其界達電位值提升至30 mV，表示其在水相中的分散穩定度提高，能延緩沉降聚積的速度，加上所有材料修飾前後皆保有溫和的細胞毒性，因此經過轉換官能基後能提升材料在生物方面應用的潛力。
第二部分根據實驗室在2015年成功利用ZIF-90材料包覆蛋白質的經驗，將包覆對象延伸至大分子量的核酸。本部分實驗藉由在ZIF-90水相的合成環境中加入預先混合的乙烯基吡咯烷酮（Polyvinylpyrrolidone, PVP）與質體DNA，使質體在ZIF-90晶體形成時被包覆在材料內，且其結晶性與外觀形態並沒有受影響，並初步利用電泳的方式確認核酸存在材料當中，未來仍需要實驗證實質體的包覆率及核酸序列完整性。期盼得到的複合材料能保護核酸並利用表面易於修飾等特質，有助於做為基因傳輸載體的應用。

關鍵字

機金屬骨架材料 ； 後修飾 ； ZIF-90 ； 乙烯基吡咯烷酮

並列摘要

In the first part of this dissertation, the aldehyde groups of Zeolitic Imidazolate Framework-90 (ZIF-90) were converted into carboxyl, amino, and thiol groups, which were named ZIF-90-C, ZIF-90-A and ZIF-90-T respectively, confirmed by the spectrometric investigations. X-ray diffraction patterns and N2 adsorption isotherm showed that the crystallinity and porosity of the framework were not affected. Remarkably, the micropore volume in ZIF-90-C and ZIF-90-A were retained almost 80% due to the mild reaction condition and the optimal reactant equivalence. According to zeta-potential analysis, the positive charge on surface of ZIF-90-A was significantly enhanced, which implies dispersion stability in water and affinity to slightly negatively charged cell surface. In addition to the surface charge enhancement, the moderate in vitro cytotoxicity of the transformers, as reported by AlamarBlue assay on HEK293 cell line, revealed the functionalization opens a new way for ZIF-90 in bioapplications.
In the second part, on the basis of our previous publication regarding embedded enzyme in MOF crystals, the guest molecules in this study was changing from enzyme to DNA. By adding polyvinylpyrrolidone together with plasmid into the aqueous ZIF-90 synthesis condition, the plasmid could be surrounded by the ZIF-90 framework. The X-ray diffraction patterns and the Scanning Electron Microscope image indicated the crystallinity and the morphology of ZIF-90 were intact after imbedding plasmid. Additionally, agarose gel electrophoresis primarily confirmed the nucleic acid component in acid-digested DNA/ZIF-90 complex material. It is worth mentioning that loading efficiency of plasmid and integrity of the nucleotide sequence need to be further studied. The ease of functionalization for DNA/ZIF-90 complex material is expected to be a benefit to gene delivery.

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