2014-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/696064摘要：流行性感冒（influenza）為常見的呼吸道疾病，由流感病毒（influenza virus）所引起，近百年來已發生數次大流行，造成大量人員患病與死亡。流感病毒經由抗原微變（antigenic drift）與抗原移型（antigenic shift）等分子機制，使其主要表面抗原產生變異。因此世界衛生組織每年必需重新預測可能流行的流感病毒株以製備疫苗，而流感病毒亦逐漸對目前治療流感的藥物產生抗藥性。因此，開發通用型抗流感藥物極為重要。流感病毒表面的血球凝集素（hemagglutinin；HA）為誘發抗體產生的主要抗原之一。而HA蛋白的柄狀區域（stalk domain）序列較為保留，可做為開發通用型抗流感病毒藥物之標的。單股核酸適體（aptamer）具有專一性與親和性高、無免疫原性、體積小、容易製備、易於修飾等特性，可開發為新型的抗流感藥物。本計畫擬使用流感病毒HA蛋白的柄狀區域做為目標分子，以系統化配體篩選法（systematic evolution of ligands through exponential enrichment；SELEX）進行抗流感適體的篩選。因此，篩選出來的適體將通用於各亞型的流感病毒。篩選出通用型抗流感適體後，將分析其與流感病毒HA蛋白複合體的晶體結構，並依該結構進行適體的改良，期能提高適體的親和性與穩定性。<br> Abstract: Influenza is a respiratory illness caused by influenza viruses, which infect hundred of thousands of people every year and caused million of deaths in the past hundred years. Due to antigenic drift and antigenic shift, novel influenza viruses arise frequently and the World Health Organization (WHO) needs to recommend specific vaccine viruses for inclusion in influenza vaccines every year. In addition, more and more influenza viruses become resistant to specific antiviral drugs. Therefore, the development of novel anti-influenza drugs is an important issue. Hemagglutinin (HA) is one of the antigenic proteins of influenza virus A. The stem domain of HA is highly conserved and a potential target for novel anti-influenza drugs. Aptamers are single-stranded oligonucleotides and have some unique characteristics, like short size, high affinity to target molecule, being easily modified chemically, and can be easily synthesized. Thus aptamers have potential for development as anti-influenza drugs. In this study, we will use stem domain of HA of influenza virus as the target molecule for screening of aptamers by the systematic evolution of ligands through exponential enrichment (SELEX) process. The obtained aptamer may be able to interact with different subtypes of influenza virus A and is an universal anti-influenza aptamer. In order to realize the interaction between the aptamer and HA protein, the structure of aptamer-protein complex will be studied by using X-ray crystallography. Afterwards, the affinity and stability of the aptamer will be improved by using rational design methodology based on the structural information.流感病毒適體系統化配體篩選法X射線結晶分析法influenza virusaptamersystematic evolution of ligands through exponential enrichmentX-ray crystallography