Purification of Group V ((-)ss) RNA viruses: Arenaviruses: Lassa virus, Tacaribe virus, Junin virus, Lymphocytic choriomeningitic virus (LCV)

Dettenhoffer and Yu [1] were the first to report the use of discontinuous 6-18% (w/v) iodixanol gradient in a sedimentation velocity mode to purify HIV-1 virions without affecting the infectivity of the virus. 

This is described in Application Sheet V34. The technique was subsequently extended to the purification of Lassa virus by Lenz et al [2] and Strecker et al [3]. These workers have used the technique to study the processing of Lassa virus glycoproteins grown in Vero-E6 cells. Purification has also been carried out on an 8%/30% iodixanol discontinuous gradient [4]. 

All comparative studies between CsCl and iodixanol show that recovery of virus infectivity is much higher and the particle:infectivity ratio much lower when viruses are purified in iodixanol. 

Although sucrose is generally less deleterious to viral infectivity than CsCl, it can nevertheless also have serious effects on certain important aspects of viral function; in particular the loss of surface glycoproteins from retroviruses has been noted [5]. 

This may be related to its viscosity, which is much higher than that of iodixanol. Like CsCl, sucrose must be dialyzed before infectivity can be measured. In contrast, many add-on techniques can be performed and cells infected with virus, without dialysis of iodixanol