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標題: 傳染性華氏囊病病毒VP2蛋白中和性抗原決定區位之分析
Identification of the neutralization epitopes on infectious bursal disease virus capsid protein VP2
作者: 邱芳誼
Chiu, Fang-Yi
關鍵字: infectious bursal disease virus;傳染性華氏囊病病毒
出版社: 生物科技學研究所
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傳染性華氏囊病病毒(infectious bursal disease virus, IBDV)是造成雞隻傳染性華氏囊病變的病源,感染幼雞後會破壞華氏囊組織中的B淋巴細胞,造成雞隻免疫能力受損。其外鞘蛋白VP2具參與病毒接受體的結合及宿主細胞辨識的功能,其帶有能引發中和病毒之抗體的抗原決定區。若單獨表現VP2蛋白則會形成T=1 之次病毒顆粒(subviral particle, SVP)本研究將由SVP誘導出之單株抗體來找尋IBDV之中和性抗原決定區。先前研究已知VP2胺基酸序列204-344為高度變異區,本實驗依此高度變異區設計出多段VP2重組蛋白及合成胜肽(peptides),並利用單株抗體以病毒中和試驗、免疫連結吸附法、西方轉漬法及免疫沉澱等其他分析法找出IBDV之中和性抗原決定區。結果顯示由多株單株抗體中分別篩選出1株具中和效力之抗體Mab SVP-4及1株不具中和效力之抗體Mab SVP-1,其皆可辨認到VP2似病毒顆粒(SVP)及野生株(wild-type)的病毒(IBDV-P3009),而這2株單株抗體亦可辨識實驗中所構築的一段VP2重組蛋白。未來將於VP2蛋白做適當的截切,更進一步找出中和性抗原決定區。

Infectious bursal disease virus (IBDV) is the causative agent of infectious bursal disease, which is one of the most important and widespread infectious disease in commercial chickens. The structural protein VP2 spontaneously form a T=1 subviral particle (SVP). The conformational epitopes has been reported in the highly variable region of VP2 protein of IBDV. In previously study, the amino acid residues 204 to 344 of VP2 had been reported as hypervariable region, which is responsible to antibody binding. Here, we mainly use the monoclonal antibodies to identify the neutralizing epitopes. In this study, three recombinant proteins (NH61-202, CH196-329, NH291-441) of VP2 and four synthetic 16-mer peptides (PBC, PDE, PFG, PHI) were prepared and using virus neutralization assay、ELISA、western blot and immunoprepicitation to identify the neutralizing epitopes. In our results, two monoclonal antibodies (Mab SVP-1, Mab SVP-4) interact to SVP, wild-type IBDV virion. All monoclonal antibodies (Mabs) bind to the region of VP2 between amino acids ranging from 196 to 329. We investigate the possibility of the existence of linear neutralizing epitopes on IBDV in order to provide information for future development of a safe and effective synthetic peptide vaccine.
其他識別: U0005-2408200620113300
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