Comparative Study on Packaging and Concentration Methods of γ-Retrovirus Vector
For the optimization of packaging conditions and concentration methods for γ-retroviral vector production for CAR-T production,two packaging vectors(pMIG and pMIG Ⅱ)and different amounts of plasmids(20 μg and 40 μg)were used to produce retrovirus,respectively,and the virus titers were detec-ted by flow cytometry;Ultracentrifugation and ultrafiltration were used to concentrate the retrovirus,the titer of the two viruses and their transduction efficiency on T cells were detected by flow cytometry,the proliferation and survival rate of CAR-T cells were monitored.The data showed that the virus titer was higher using pMIG Ⅱ vector(p＜0.05),the different amounts of plasmids(20 μg and 40 μg)did not affect the virus titer(p＞0.05),ultrafiltration and ultracentrifugation concentration methods did not affect the virus titer(p＞0.05).The results showed that pMIG Ⅱ vector and the total amount of 20 μg plasmid were suitable for packaging γ-retrovirus,and ultrafiltration can be used for small-scale concentration of retrovir-us in the laboratory to content the needs of CAR-T research.
万方数据
维普
