Screening of novel excipients for improving the stability of retroviral and adenoviral vectors.
In the past decade there has been an increase in the application of viral vectors in the laboratory and clinical trials of human gene therapy, retroviral and adenoviral vectors among the most used. However, the limited stability of these vectors creates problems in the design of experiments, transport, and storage. Vectors stored at -80 degrees C must be quickly shipped on dry ice, which is somewhat cumbersome. Alternatively, viral vectors can be preserved in a lyophilized form. However, loss of viral activity during lyophilization can also be a serious problem. In this report we identify novel candidate formulations containing new compatible solutes, ectoin, hydroxyectoin, and firoin, that allow better stability of retroviral and adenoviral vectors during storage. For retroviral vectors, the maximum stabilization for long-term storage was achieved through lyophilization followed by storage at -20 degrees C using a formulation of Tris buffer pH 7.2 containing firoin (0.5 M), a half-life of 340 days being obtained. Adenoviral vectors storage at -80 degrees C in solution using Tris buffer pH 8.0 with firoin was the best method for long-term storage, with a half-life exceeding 1 year.
Cruz
Pedro E
PE
IBET/ITQB, Apartado 12, P-2781-901 Oeiras, Portugal.
Silva
Ana Carina
AC
Roldão
António
A
Carmo
Marlene
M
Carrondo
Manuel J T
MJ
Alves
Paula M
PM
D000256:Adenoviridae / Q000235:genetics*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D009691:Nucleic Acid Denaturation; D012190:Retroviridae / Q000235:genetics*; D012996:Solutions; D013696:Temperature
D012996:Solutions