Purpose: To construct recombinant lentiviral vectors of bone morphogenetic protein 2(BMP-2) gene and prepare a stable lyophilized state.
Methods: The BMP-2 gene lentiviral vector was constructed by recombinant DNA technology and mixed with different stabilizers in an appropriate proportion and then lyophilized. The optimum stabilizer was selected according to the appearance and virus titer after being lyophilized. The quality of lyophilized product was measured by thermal stability of the virus, PCR and gene sequencing. SPSS13.0 software package was used for statistical analysis.
Results: Recombinant BMP-2 lentiviral vector was successfully constructed. 10% trehalose, 1% bovine serum albumin, 3%mannitol and 0.5% gelatin in group B showed good protection on BMP-2 gene lentiviral vector. The virus titer decreased 0.42 LgPFU/mL after being lyophilized, which was better than group A, C and the control group and the difference was statistically significant (P<0.05); Lyophilized protective agent made from group B still maintained a good appearance at 37 degrees centigrade after 28 days and the virus titer decreased 0.63 LgPFU/mL. The virus titer in liquid lentiviral infection control group decreased rapidly to 2.37 LgPFU/mL 1 week later and the difference between the two groups was statistically significant (P<0.05); PCR and gene sequencing showed that the target gene information after redissolve had no loss or mutation.
Conclusions: Appropriate selection of lyoprotectants can effectively protect the biological stability of recombinant BMP-2 gene lentiviral vector.