Simple and rapid separation of ortho- and paramyxovirus glycoproteins

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Abstract
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 non-ionic detergent octyl-Β-D-glucoside and are then applied to a column of agarose beads coupled with tyrosine-sulfanilic acid. PureHA andF are obtained in good yield in the flow-through from this column.NA andHN bind strongly and can be eluted, albeit somewhat contaminated withHA orF, by raising the pH of the column buffer. The separated non-denaturated fractions can be used for structural, functional, and antigenic studies.