Development of a high effiency baculovirus/mammalain cell expression system for recombinant protein production

• Main Authors: Chen waimei, 陳偉民
• Other Authors: Liu yu-ko
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/50849358113715763631

碩士 === 長庚大學 === 生化與生醫工程研究所 === 95 === Abstract Recently, many systems have been developed for the production of protein with medical significance. The baculovirus expression vector system is one of the best expression systems for high yield protein production in insect and mammalian cells with proper posttranslational modifications. In these experiments, baculovirus was produced in insect cell for subsequent protein production in mammalian cells. The best viral yield was achieved by virus infection at the third day and harvesting at 6 days post viral infection. Feed-batch culture yielded roughly twice viruses(ca. 3.8×108 pfu/ml) compare with that produced by batch culture. Optimal viral infection conditions for protein in mammalian cells also explored using β-galactosidase as reporter and found that a 4 fold better yields were achieved by multiple(ca. 1.265mg/L) than by single baculovirus transduction. This experiment showed that a combination of 3D-microgravity bioreactor using micro-carrier can be the best system for protein production using baculovirus in the mammalian cells.