陳昭岑臺灣大學:化學研究所莊絢婷Chuang, Shiuan-TingShiuan-TingChuang2007-11-262018-07-102007-11-262018-07-102006http://ntur.lib.ntu.edu.tw//handle/246246/52024近年來研究發現,醣類衍生物在生物系統中扮演著重要角色,大多數蛋白質經過醣基的後修飾才具有功能性,除了單醣基的修飾外,寡聚醣基本身的複雜多變的結構,得以使它在複雜生物系統下仍具有專一性,所以目前有許多生物與化學家致力於醣蛋白化學這個領域。由於寡聚醣在合成的困難,所以本研究目的在將寡聚醣基拆解為簡單的單醣基修飾在金奈米粒子,利用多價性提高與蛋白質作用,並且當醣配基與蛋白質結合後會藉由兩者間的多價性作用導致金奈米粒子聚集使得表面電漿共振吸收改變,可而作為偵測工具。以路易士醣配基的受體蛋白翅莢豌豆凝集素(LTL)與L-選擇素為研究對象。 在之前的研究指出Recent research results show that carbohydrates play many biological roles such as information carriers. Oligosaccharides on the glycoproteins or glycolipids display highly specific recognition because of their extremely complicated and various stereoisomers. For example, oligosaccharides Lewis antigens have been shown in mammalian systems to be important recognition determinants in cell–cell interactions. Therefore, many scientists devote their efforts to carbohydrate research. However, the advancement of the field is highly hampered by rather difficult and challenging synthesis of oligosaccharides. To circumvent the synthetic dilemma, we plan to identify the key monosaccharides of oligosaccharides, which are pivotal to the biological functions, and immobilize them on 32-nm gold nanoparticles (GNPs) as multivalent systems to achieve the desired effects. Tetragonolobus purpureas (LTL) and L-selectin, Lewis binding lectins, was chosen to demonstrate our molecular. Changes of surface plasma resonance (SPR) absorption were carefully monitored and used as indication of binding. Previous studies showed that the binding strength of第一章 醣類與外源凝集素…………………………………………1 1.1 醣質科學(Glycosciene)的發展…………………………...…. 1 1.2 醣類對生物系統的重要性……………………………………….1 1.3 複合醣體的分類介紹…………………………………….………3 1.4 醣類與受體間的專一結合……………………………….………4 1.5外源凝集素(Lectins)簡介…………………………….……….6 1.6 外源凝集素的分類……………………………………....………8 第二章 醣類與外源凝集素…………………………….……….…11 2.1路易士醣配基的結構……………………………………………11 2.2第一型路易士醣配基的生物功能與研究……………………....13 2.3 第二型路易士醣配基的生物功能………………………….....14 2.3.1 Lex在胚胎發育過程的角色………………………....14 2.3.2 Lex、sLex與選擇素(Selectins)……………….....16 2.4模擬LeX、sLeX的類似物………………………………………21 第三章 翅莢碗豆凝集素與L-選擇素的偵測與純化……………31 3.1 金奈米粒子的介紹……………………………………………...31 3.1.1金奈米粒子的合成………………………………...…..32 3.1.2金奈米粒子在生物感測器上的應用………….………33 3.2以金奈米粒子修飾混合單醣基偵測與純化翅莢豌豆凝集素....36 3.2.1翅莢豌豆凝集素與Lex……………...……………..….36 3.2.2 醣類衍生物α或β組態的合成策略…………..….……38 3.2.3 單醣衍生物分子探針的設計與逆合成分析…..…..…41 3.2.4 單醣衍生物分子探針的合成探討…………..……….45 3.2.5 實驗結果與討論…………………………..……...…..53 3.3 以金奈米粒子修飾混合單醣基偵測與純化L-選擇素.……....64 3.3.1 α-Fuc 與sulfo-β-Gal 單醣衍生物分子探針的設計...65 3.3.2 單醣衍生物分子探針的設計與逆合成分析……...…66 3.3.3 sulfo-β-Gal 衍生物的合成探討……………………..68 3.3.4 實驗結果與討論……………………………………...74 第四章 總結………………………………………………...……81 實驗部分……………………………………………………...……83 參考文獻……………………………………………………...…..119 附錄…………………………………………………………...…..12514099927 bytesapplication/pdfen-US蛋白質純化金奈米粒子purify proteingold nanoparticles以金奈米粒子修飾混合單醣基應用在翅莢豌豆凝集素以及L-選擇素的偵測與純化Mixed-Monosaccharides Encapped Gold Nanoparticles to Detect and Purify Corresponding Binding Proteinsthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/52024/1/ntu-95-R93223031-1.pdf