Simple and rapid separation of ortho- and paramyxovirus glycoproteins.
The hemagglutinin (HA) and neuraminidase (NA) of influenza viruses, as well as the fusion protein (F) and hemagglutinin-neuraminidase (HN) of paramyxoviruses, have been separated in native form using a two-step procedure. The glycoproteins are efficiently extracted from virions using the on-ionic detergent octyl-beta-D-glucoside and are then applied to a column of agarose beads coupled with tyrosine-sulfanilic acid. Pure HA and F are obtained in good yield in the flow-through from this column. NA and HN bind strongly and can be eluted, albeit somewhat contaminated with HA or F, by raising the pH of the column buffer. The separated non-denatured fractions can be used for structural, functional, and antigenic studies.
D002846:Chromatography, Affinity; D003902:Detergents; D006023:Glycoproteins / Q000302:isolation & purification*; D006389:Hemagglutinins, Viral / Q000302:isolation & purification; D009980:Influenza A virus / Q000032:analysis*; D009439:Neuraminidase / Q000302:isolation & purification; D009522:Newcastle disease virus / Q000032:analysis*; D014764:Viral Proteins / Q000302:isolation & purification*
D003902:Detergents; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D014764:Viral Proteins; D009439:Neuraminidase