Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/66413
標題: 禽類流行性感冒病毒H5亞型單株抗體之製備與分析
Production and characterization of monoclonal antibodies of H5 avian influenza virus
作者: 李同浩
Li, Tung-Hau
關鍵字: avian influenza virus
禽流感
monoclonal antibodies
H5
單株抗體
H5
出版社: 獸醫微生物學研究所
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摘要: 自從1997年至今,高病原性禽類流行性感冒 (Avian Influenza,AI) 病毒H5N1造成家禽工業重大的經濟損失及危害動物生命安全,H5N1目前經證實有318人感染,有192人因此死亡。高病原性禽類流行性感冒,已成為獸醫界或全球最關注的一個問題。本研究目的主要是以禽類流行性感冒病毒H5N3為目標,製備單株抗體,以進行快速診斷。BALB/c小白鼠免疫不活化H5N3全病毒,之後取脾臟細胞和骨髓瘤細胞進行融合。融合瘤篩選以血球凝集抑制作用、免疫墨點法、西方轉漬法和間接免疫螢光抗體分析。篩選出兩株對流感病毒血球凝集素有特異性反應,歷經了五次融合瘤製備,篩選了三千孔的上清液。藉由極限稀釋 (limiting dilution) 進行單株化。藉由單株抗體分型可知分別為IgA和IgG2a。
The outbreaks of highly pathogenic H5N1 influenza A virus from 1997 to the present day,The virus cancause economical losses to the poultry industry and have devastating consequences for animal health,H5N1 infections have resulted in 318 laboratory-confirmed infected people,including 192 deaths。HPAI receives immense attention in the veterinary world and is globally treated as a disease immediately notifiable on suspicion to the authorities。The purpose of this study is to prepare monoclonal antibodies against H5N3 AIV and to use these monoclonal antibodies to develop a rapid method for detection of AIV。BALB/c mice were immunized with H5N3 virus and then the splenocytes from the immunized mouse were collected and fused with NS-1 myeloma cells。 The hybridomas secreting antibodies against AIV were screened by hemagglutination inhibition、dot immunoblotting、Western blotting and immunofluorescence assay。Two hybridomas which secreted specific antibodies to AIV,were selected among 3000 hybridomas in five fusion experiments。Cloning of these established hybridomas was carried by the limiting dilution method in order to confirm their monoclonality。After two times cloning,two monoclonal antibodies were purified.Hybridomas C2-8G and D1-11G,secreting IgA and IgG2a respectively。
URI: http://hdl.handle.net/11455/66413
其他識別: U0005-3007200714381600
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-3007200714381600
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