Antipeptide antibodies for analysis of pathotype-specific variations in cleavage activation of the membrane glycoprotein precursors of Newcastle disease virus isolates in cultured cells.
Antipeptide antibodies have been produced which target regions either side of the cleavage activation sites of Newcastle disease virus (NDV) membrane glycoprotein precursors. Use of complementary pairs of antibodies in Western blot analysis of mercaptoethanol-reduced extracts of NDV-infected BHK-21 cells enabled analysis of the susceptibilities of NDV fusion protein precursors (Fo-proteins) to cleavage activation in these cells. In addition, it was possible to determine whether or not isolates produce haemagglutinin-neuraminidase (HN)-proteins in precursor forms (HNo-proteins). This assay system has been evaluated with a series of Australian isolates of NDV with well defined virulence properties in order to validate its use in pathotyping NDV isolates. Less well defined isolates also produced data consistent with their biological properties and an isolate was characterised which, hitherto, was not known to be present in Australian poultry. The applicability of this assay system in fundamental studies of the processes of cleavage activation of NDV Fo- and HNo-proteins and formatting of the antisera into ELISA systems are discussed.
Gorman
J J
JJ
CSIRO/Australian Animal Health Laboratory, Geelong, Vic.
Hodder
A N
AN
Selleck
P W
PW
Hansson
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D015153:Blotting, Western; D002478:Cells, Cultured; D002642:Chick Embryo; D006389:Hemagglutinins, Viral / Q000378:metabolism; D007106:Immune Sera; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009522:Newcastle disease virus / Q000145:classification / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity; D010446:Peptide Fragments / Q000276:immunology; D011498:Protein Precursors / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
D000914:Antibodies, Viral; D006389:Hemagglutinins, Viral; D007106:Immune Sera; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011498:Protein Precursors; D014760:Viral Fusion Proteins; D014764:Viral Proteins