Generation of Chimeric Baculovirus with Histidine-Tags Displayed on the Envelope and Its Purification Using Immobilized Metal Affinity Chromatography

• Main Authors: Chien-Tai Tsai, 蔡建泰
• Other Authors: Yu-Chen Hu
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/08192825770619701303

碩士 === 國立清華大學 === 化學工程學系 === 91 === Baculovirus has been proved to be able to infect mammalian cells and have the potential as a gene delivery vehicle. As a gene therapy vector, purity is very important. In this research, we fuse a hexahistidine (His6) tag on the virus envelope protein, gp64. By the means of immunofluorescence, western blot, and transmission electron microscope, it has been proved that it can be distributed on the surface of the infected insect cells and replicated viruses evenly and in a large quantity. This enables us to use immobilized metal affinity chromatography (IMAC) to purify the recombinant baculovirus. At present, the recovery of virus titer is 8 % and the virus structure is intact. Besides, the viruses retain the ability of infecting mammalian cells. This novel method has the potential of replacing sucrose or cesium chloride gradient ultracentrifugation and the process is more convenient. Besides, in coordination with diafiltration, this system has the possibility of dealing with large volume of viruses.