Concentration and purification of feline leukaemia virus (FeLV) and its outer envelope protein gp70 by aqueous two-phase systems.
The major protective antigens of retroviruses are considered to be their glycosylated envelope proteins. However, the methods commonly employed to enrich and purify virus from culture media such as pelleting and density-gradient centrifugation result in a low recovery of the viral external glycoproteins. This is an obvious drawback when the virus is intended for use in a vaccine. In search for alternative methods to concentrate and purify FeLV, we have attempted extraction in two-phase systems based on water-soluble polymers (Albertsson PA., Biochem Biophys Acta 1958; 27: 378-395). A variety of polymer systems was tested. Some of them seem attractive for a large-scale concentration of the virus and/or its glycoprotein. The distribution between the phases of two FeLV proteins, the outer envelope protein, gp70, and the gag protein, p27, was determined. With a system composed of dextran sulfate and polyvinyl alcohol both the glycoprotein and the gag protein were almost completely recovered in the lower phase which constitutes about 3% of the total system in weight. The two proteins were more than 40-fold purified as calculated on protein basis. The proteins can be extracted readily.
Hammar
L
L
Department of Veterinary Virology, Biomedical Center, Uppsala, Sweden.
Eriksson
S
S
Malm
K
K
Morein
B
B
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D016264:Dextran Sulfate; D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel; D015683:Gene Products, gag; D015151:Immunoblotting; D005256:Leukemia Virus, Feline / Q000032:analysis / Q000302:isolation & purification*; D008970:Molecular Weight; D012191:Retroviridae Proteins / Q000302:isolation & purification; D012995:Solubility; D014759:Viral Envelope Proteins / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D003911:Dextrans; D015683:Gene Products, gag; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins; D016264:Dextran Sulfate