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利用固定化金屬親合力層析法純化基因重組昆蟲桿狀病毒
Thesis

利用固定化金屬親合力層析法純化基因重組昆蟲桿狀病毒

蔡建泰
Masters, National Tsing Hua University
2002

Abstract

昆蟲桿狀病毒固定化金屬親合力層析法病毒純化基因治療 baculovirusimmobilized metal affinity chromatography (IMAC)virus purificationgene delivery
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.

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