吳漢忠2006-07-262018-07-092006-07-262018-07-092002http://ntur.lib.ntu.edu.tw//handle/246246/22663病毒感染後，經常會借由活化宿主的 免疫反應產生抗體，對抗病毒蛋白之抗原 決定位。這些病毒蛋白上抗原決定位的確 認，有助於病毒感染機制的暸解，對於發 展有效的疫苗及偵檢試劑也有重大的助 益。本計畫主要是要研究登革病毒的B 細 胞抗原決定位。經由ELISA, immunofluorescence 和immunoblotting 方 法，我們找出具專一性的第二型登革病毒 單株抗體(Ab4)。此抗體之抗原決定位的確 認，有助於病毒感染機制的暸解，疫苗及 偵檢試劑的研發。在此研究中，我們以噬 菌體顯現法來研究第二型登革病毒的抗原 決定位。我們也已經篩選出多株的噬菌體 可以專一性的與第二型登革病毒抗體結 合。這些專一性噬菌體所攜帶的外來 peptide，都含有His-Arg/Lys-Leu/Ile motif。 以synthetic peptide 代替噬菌體所攜帶的外 來peptide，可與第二型登革病毒抗體明顯 反應。第二型登革病毒感染之老鼠及兔子 血清，可以用此epitope-based peptide 來確 認。另外，此epitope-based peptide 可有效 的分辨登革病毒及日本腦炎病毒感染之老 鼠血清，這些抗原決定位的確認與研究， 將有助於epitope-based peptide antigen 之研 發，及了解出血性登革之病理機制。Viral infection usually results in the production of antibodies directed against the epitopes of viral proteins through the activation of the host’s humoral immunity. Identification of these epitopes on viral proteins is important for understanding the pathogenesis of viral infectious diseases as well as for developing vaccines and diagnostic reagents. Serotype-specific monoclonal antibody, Ab4, against dengue virus type 2 (DEN-2) was generated and identified by ELISA, immunofluorescence and immunoblotting analysis. The B-cell epitope of Ab4 was further identified from a random peptide library displayed on phage. The selected phage clones were specifically reacted with Ab4 and did not reacted with other monoclonal antibodies. These immunopositive phage clones displayed a consensus motif, His-Arg-Leu/Ile. Synthetic peptide corresponding to the phage-borne peptide can bind the antibody specifically. The epitope-based synthetic peptide could detect serum samples from DEN-2 immunized mice and rabbits by ELISA assay. The peptide was also able to differential very clearly between serum samples from DEN-2 and Japanese encephalitis virus (JEV) immunized mice. The results show that this monoclonal antibody and its epitope-based peptide will be useful for serologic diagnosis of DEN-2. Furthermore, the identification of the epitope of DEN-2 will permit us to functionally dissect the antibody response and to address the role of antibody in the pathogenesis of primary or secondary DEN-2 infection.application/pdf264058 bytesapplication/pdfzh-TW國立臺灣大學醫學院口腔生物科學研究所第二型登革病毒噬菌體顯現法抗原決定位登革偵檢試劑monoclonal antibodyB-cell epitopesynthetic peptideserologic diagnosis[SDGs]SDG3journal articlehttp://ntur.lib.ntu.edu.tw/bitstream/246246/22663/1/902320B002208.pdf