Isolation of a retrovirus from multiple sclerosis patients in self-generated Iodixanol gradients.
The use of Iodixanol, a relatively new iodinated gradient medium, is described for isolation of a retrovirus, which was harvested from the supernatant of lymphoid cell lines originating from patients with multiple sclerosis (MS). The virus is produced in low amounts and has been shown to be fragile, as manifested in a loss of surface glycoproteins when purified in other gradient media. The gradient fractions were analysed after centrifugation in Iodixanol by incorporation of 3H-UTP, reverse transcriptase (RT) assays and electron microscopy (EM) and it was found that Iodixanol does not cause the degree of damage to the particles observed previously. These more favourable conditions are probably due to low viscosity and almost iso-osmotic conditions even in high concentrations. Furthermore, these advantages go together with higher reproducibility in self-forming gradients, easier handling and shorter centrifugation time. Iodixanol can also be used for preparation of HTLV-1.
Møller-Larsen
A
A
Institute of Medical Microbiology, University of Aarhus, Aarhus C, Denmark.
Christensen
T
T
D002460:Cell Line; D002499:Centrifugation, Density Gradient; D015368:Human T-lymphotropic virus 1 / Q000201:enzymology / Q000302:isolation & purification / Q000648:ultrastructure; D006801:Humans; D008214:Lymphocytes; D008854:Microscopy, Electron; D009103:Multiple Sclerosis / Q000821:virology*; D012190:Retroviridae / Q000201:enzymology / Q000302:isolation & purification* / Q000648:ultrastructure; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014283:Triiodobenzoic Acids; D014776:Virus Cultivation / Q000379:methods
D014283:Triiodobenzoic Acids; C044834:iodixanol