Production and application of infectious bursal disease virus VP2-formed particles using insect larvae

• Main Authors: Dau-Shiang Yang, 楊道翔
• Other Authors: 王敏盈
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/36505538212151481299

碩士 === 國立中興大學 === 生物科技學研究所 === 93 === The aim of this study was to develop an optimal process for the production of IBDV VP2-formed particles in Trichoplusia ni (T. ni) larvae and to display a foreign protein on the surface of these particles for its future application. In the first of this study, the optimal condition for protein production could be reached when the early third instar T.ni larvae were reared at the density of 10 larvae per 7 cm2 area, infected with 1.5x109 pfu of recombinant baculovirus per larva, and harvested at 4 days post infection (dpi). The production level of VP2-formed particles was estimated to be approximately 3 mg per gram larvae. We scale up the rearing scale under the same condition, in which the total VP2-formed particles yield could be reached at 4 mg (70 larvae) and 9 mg (140 larvae). Furthermore, the melanogenesis of crude larvae extract was conquered by adding with 1% ascorbic acid. The second part of this study, we successfully constructed a chimeric protein, mVP2E1, in which a peptide fragment of E proein from SARS coronavirus was fused at the C-terminus of VP2 protein. The mVP2E1 could forming virus-like particles (VLPs) spontaneously. However, our AC-ELISA analysis of the purified particles indicated that the E1 peptide could not be displayed on the surface of VLPs.