| Abstract: | A new method for the isolation of biologically active envelope antigens from paramyxo- and myxoviruses was developed using a zwitterionic detergent Empigen BB. Essentially complete recovery of hemagglutinin and sialidase from representative paramyxoviruses (PMY, NDV, and Sendai virus) and influenza virus X7 was achieved. The glycoproteins (HN and F) of PMY were purified in a DEAE-Bio-Gel A column equilibrated with bicarbonate buffer containing Empigen, Polyacrylamide gel electrophoresis analysis of pooled HN fractions revealed one band, indicating that it is a common glycoprotein. Gels of pooled F-enriched fractions revealed three bands corresponding to LGP, HN, and F. Gels of PMY virus revealed six polypeptides, and they have been tentatively designated L, LGP, HN, F, NP, and M. LGP, a large glycoprotein, was not detected in gels of NDV and Sendai virus. It has been proposed that LGP may consist of a complex of F. Antiserum was prepared against purified HN and it was found to be monospecific. The antiserum inhibited both hemagglutination and enzyme activities of PMY, which is in support of the hemagglutinin and sialidase of PMY being associated with a common glycoprotein. By enzyme inhibition analysis of PMY, NDV, and Sendai virus, it was shown that the enzymes of these viruses are antigenically distinct. The methods described for the isolation and purification of PMY glycoproteins may be useful for the preparation of myxo- and paramyxovirus subunit vaccines. |
