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標題: 利用昆蟲生產禽流感疫苗流程之開發
Production of Avian Influenza Vaccines Using Insect Larvae
作者: 王敏盈
關鍵字: 生物技術, 植物保護類;avian influenza;禽流感;subunit vaccine;insect larvae;virus-like particles(VLPs);次單位疫苗;昆蟲幼蟲;似病毒顆粒;技術發展
Recently, the baculovirus/insect cell expression system has beingincreasingly used for the production of vaccine candidate. However, cultivation ofinsect cells using the conventional fermentation method was restricted by thedifficulties of the scale-up step and the high cost of culture medium. It is ofeconomically importance to minimize the cost for the production of vaccines,especially for the animal vaccines. To overcome the difficulties of fermentorscale-up and avoid the usage of expensive culture medium, larvae of Trichoplusia ni,a lepidopterous insect, is a promising host to express foreign proteins. Ourresearch proposal is to develop a production platform of animal vaccines by usingbaculovirus/insect larvae system, and to establish an efficient downstreampurification process. The avian influenza has threatened the poultry industry andeven the human beings worldwide. It has been reported that the HA protein ofavian influenza virus (AIV) produced by the baculovirus expression system canprotect avians from the disease. The HA proteins of H6 and H7 subtypes andVLPs containing H7 HA have been expressed and produced using baculovirusexpression system. Preliminary experiments demonstrated subunit vaccinescomprising VLPs can elicit an effective humoral immune response in SPF chickens.Our research mainly focuses on using insect larva instead of cell culture as afeasible and economic tool for AIV HA production. We will express H5 HA byfollowing the strategy on H7 HA in order to confirm our previous results could beapplied to other AIV subtypes. The three-year proposal will accomplish theoptimization of the infectious conditions in T. ni larva using DsRed fluorescence asa probe and the following purification processes in the first year. The second yearwill evaluate antigen immunogenecity in specific-pathogen-free chickens. Thethird years will establish the scale-up process of expression and purification of HAprotein in insect larvae and evaluate antigen immunogenecity in farm chickens.

近年來,桿狀病毒表現系統在疫苗研發上的重要性日漸增加,傳統發酵槽因為大量生產的困難性及複雜性加上商用細胞培養液的成本昂貴,且在農業應用上的成本預算遠低於人類健康醫療;因此利用具有成本低廉及操作簡便特性的桿狀病毒表現系統生產動物用疫苗就相對具有高度經濟競爭力。本計劃將開發利用昆蟲幼蟲生產動物用疫苗之高效率生產及純化流程,以便將此生產平台落實於動物疫苗之生產。由於禽流感對於台灣甚至於全世界之養雞工業威脅日益嚴重,本實驗室已成功利用昆蟲桿狀病毒的系統表現禽流感病毒弱毒株H6 及H7 亞型的HA 蛋白及H7 亞型相關似病毒顆粒,初步免疫試驗已證實了H7 亞型相關似病毒顆粒疫苗可以在SPF 雞隻中產生16 倍以上的HI 抗體以及至少40 倍以上的病毒中和能力。在這個計畫當中我們將應用先前的研究成果並以昆蟲幼蟲生產H6及H7 亞型的HA 蛋白以代替先前細胞的生產模式,發展可行且具經濟價值的生產工具,另外我們也將進行H5 亞型 HA 的表現,擴展我們的系統應用於其他亞型或綜合多價疫苗的發展。我們預計在三年內完成本計劃,第一年的工作重點為 藉由紅色螢光蛋白DsRed 在昆蟲幼蟲中的表現來偵測HA 的表現以建立蟲體感染的最適化條件,以及建立HA 之似流感病毒顆粒(influenza virus-like particles, VLPs)生產與純化方法之確立。第二年的工作重點為蟲體生產之VLPs (內含HA)的雞隻免疫效果。第三年的工作重點為大規模蟲體生產抗原流程及田間雞隻免疫效果之確立。
其他識別: NSC98-2324-B005-009
Appears in Collections:生物科技學研究所

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