Haemagglutinin of measles virus: purification and storage with preservation of biological and immunological properties.
Measles virus envelope haemagglutinin (H) was purified rapidly with Triton X-100-solubilized virions by a two-step anion-exchange chromatography using fast protein liquid chromatography. The purity of the glycoprotein in its dimeric form was demonstrated by SDS-PAGE followed by silver staining or autoradiography. The purified H glycoprotein was further freed from contaminating detergent by dialysis of octylglucoside detergent. This purification procedure, together with subsequent lyophilization and storage at -70 degrees C of the H glycoprotein which was incorporated into phospholipid vesicles allowed the full preservation of its haemagglutinating activity, its reactivity with a monoclonal anti-H antibody that recognized a conformational epitope and its capacity to elicit anti-H antibodies with haemagglutination-inhibiting and neutralizing activities.
Gerlier
D
D
INSERM U218, Centre Léon Bérard, Lyon, France.
Garnier
F
F
Forquet
F
F
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000918:Antibody Specificity; D001345:Autoradiography; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005612:Freeze Drying; D006389:Hemagglutinins, Viral / Q000276:immunology / Q000302:isolation & purification* / Q000592:standards; D007118:Immunoassay; D008081:Liposomes; D008459:Measles virus / Q000276:immunology*; D011309:Preservation, Biological; D014709:Vero Cells
D000914:Antibodies, Viral; D006389:Hemagglutinins, Viral; D008081:Liposomes