金奈米粒子親水化及與DNA一對一鍵結之探討; Study of One to One Linkage between Water Soluble Gold Nanoparticle and DNA

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Title:	金奈米粒子親水化及與DNA一對一鍵結之探討;Study of One to One Linkage between Water Soluble Gold Nanoparticle and DNA
Authors:	吳冠毅;Kuan-yi Wu
Contributors:	化學工程與材料工程研究所
Keywords:	金奈米粒子;親水化;一對一鍵結;DNA;gold nanoparticle
Date:	2007-07-17
Issue Date:	2009-09-21 12:28:45 (UTC+8)
Publisher:	國立中央大學圖書館
Abstract:	本研究主要的目的是合成一種能夠穩定的懸浮於高鹽高溫的水溶液中並且尺寸分布均一的金奈米粒子，然後利用這種水相金奈米粒子發展出一個可和DNA進行之ㄧ對一接合的方法。我們使用還原劑硼氫化鈉(NaBH4)將氯化金(HAuCl4)在甲苯中還原成粒徑7nm的金奈米粒子，再藉由兩性高分子Poly(maleic anhydride alt-1-tetradecene)修飾金奈米粒子表面使其親水化。結果顯示改質後的回收率可達到85.3%，另外在鹽穩定性方面在NaCl濃度低於200mM穩定度都可達到95%以上，即使在500mM的NaCl下也都有很好的穩定度。熱穩定性的部分，當溫度升高至80℃也不會產生聚集而沉澱。在進行一對一接合反應時，先利用胺基修飾的DNA能經由活化劑(EDC)活化和羧基修飾的矽膠體(silica gel)產生鍵結，再加入互補DNA進行雜合反應(Hybridization)，隨後將親水改質後的金奈米粒子鍵結至互補DNA末端，最後靠著去雜合反應(Dehybridization)，即可獲得金奈米粒子與DNA之一對一接合。透過毛細電泳(Capillary Electrophoresis)與穿透式電子顯微鏡(TEM)等儀器鑑定合成反應所得到的產物，藉此判斷此方法合成的一對一接合之成功性。 The main purposes of this study are to synthesize gold nanoparticles, to modify the surface of the gold nanoparticles from organic to aqueous solution by Poly(maleic anhydride alt-1-tetradecene)(PMATD), and to develop a method to conjugate one ssDNA onto this water soluble gold nanoparticles. In the synthesis, HAuCl4 was reduced by NaBH4 to form 7nm Au nanoparticle. Then using PMATD to modify Au nanoparticle, we found that the recovery of modification is 85.3%. The salt endurability of water soluble Au nanoparticles can reach 95% when the NaCl concentration is below 200mM. Even if in the 500mM NaCl, the salt endurability can keep 87%. The Au nanoparticles won’t aggregate when the temperature increase to 80℃ In one-to-one synthesis, the COOH-silica gel can be activated by N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide(EDC) and then conjugated with NH2-DNA (＋). After conjugation, we added complementary NH2-DNA (－) to hybrid the ssDNA which onto the silica gel. Using EDC to activate the water soluble Au nanoparticles to conjugate with NH2-DNA (－). The last step, we adjusted the temperature to dehybride the dsDNA. We can get Au-DNA one-to-one conjugation. Detail the production by capillary electrophoresis(CE) and transmission electron microscopy(TEM) to confirm the Au-DNA structure.
Appears in Collections:	[National Central University Department of Chemical & Materials Engineering] Electronic Thesis & Dissertation

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