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標題: | 新型H7N9流感病毒血球凝集素之單株抗體的生化性質研究 Biochemical Characterization of the Mouse Monoclonal Antibodies against the Novel Influenza A(H7N9) Virus Hemagglutinin |
作者: | Heng-Yi Su 蘇恆毅 |
指導教授: | 張世宗(Shih-Chung Chang) |
關鍵字: | H7N9流感病毒,血球凝集素,單株抗體, H7N9 influenza virus,Hemagglutinin,Monoclonal antibody, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 自2013年起於中國陸續有新型H7N9禽流感病毒感染人類事件,且同年台灣也首度出現境外移入之病例。由於新型H7N9病毒的高死亡率與跨物種感染,引起全球高度關注,世界衛生組織針對新型H7N9所公布之統計資料顯示,病例與死亡人數仍持續增加,且暫無趨緩現象。相關文獻也指出新型H7N9禽流感病毒有逐漸適應人類宿主的趨勢,因此在這波疫情尚未得到控制時,對疫苗、快速檢測平台與治療性的中和性抗體的研發相當重要。本研究利用昆蟲細胞表現系統製備新型H7N9病毒之重組HA,其具備醣基化修飾與血球凝集之能力,並成功利用此蛋白製備出單株抗體1C6B。此外也鑑定出1C6B與本實驗室另外一株用HA1所製備之F3-2單株抗體的抗原結合位,並非位於HA之受體結合區,也無中和HA之血球凝集活性。然而,實驗結果顯示1C6B與F3-2皆能辨認受酸鹼調控而改變構型之HA,並能抑制HA受到蛋白酶剪切而活化。因此1C6B與F3-2極具發展潛力,應可進一步利用細胞與動物試驗,來確定其預防與治療新型H7N9流感之功效。此外,本研究也利用這兩個單株抗體來製備快篩試片,結果顯示其可以專一性地偵測出樣本中的H7N9 HA重組蛋白。 In 2013, novel H7N9 influenza virus emerged in China and caused many human infections. Furthermore, the Centers for Disease Control (Taiwan) reported that the first case of the novel H7N9 infection originates from China in the same year. Due to the high mortality rate and cross-species transmission, the novel avian influenza (H7N9) is gaining increasing global attention. WHO (World Health Organization) reported that the severe illness and death cases caused by H7N9 infection remain continuously growing. Some researches indicated the novel H7N9 virus might obtain the limited ability of sustaining human-to-human transmission. As a result, it is important for developing vaccines, rapid-test kits, and neutralizing antibodies for public health preparedness. In this study, the H7N9 recombinant hemagglutinin (rHA) proteins were expressed in the insect cells. The purified rHA proteins, containing glycosylation and hemagglutination activity, were utilized to generate the monoclonal antibody (mAb) 1C6B. The epitopes recognized by 1C6B and another mAb F3-2 which was generated by HA1 against rHA do not localize at the receptor binding sites. Therefore, they could not inhibit the hemagglutination activity of H7N9 rHA. However, 1C6B and F3-2 can recognize the rHA which has undergone pH-dependent conformational change, and prevent rHA from processing and activation by protease. These two mAbs have a great potential for analyzing the efficacy of the prevention or the treatment of H7N9 infections in cellular or animal experiments. In addition, 1C6B and F3-2 were also applied for developing the rapid-test kit. The results showed that the rapid-test strips can detect H7N9 rHA specifically. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77957 |
DOI: | 10.6342/NTU201702061 |
全文授權: | 有償授權 |
顯示於系所單位: | 生化科技學系 |
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