Foot and mouth disease virus concentration and purification by affinity chromatography.
Foot and mouth disease virus, (FMDV) from a crude cell lysate was purified in a single step by affinity chromatography with heparin as a ligand. The virus eluted from an Heparin-Ultrogel A4R column at 1M sodium chloride in 10 mM sodium phosphate buffer, pH 7.0, while most cell protein and albumin did so at lower concentrations of sodium chloride in the same buffer. Purity of the eluted fraction containing the virus was assessed by SDS-PAGE, HPLC, ultracentrifugation, and UV absorption spectrum. With this method, intact viral particles are recovered in high yield (over 90%) and specific virus purity increases nearly 1000-fold. The capacity of the chromatographic matrix for the virus was found to be 1.1 mg viral mass per mL of hydrated gel.
Navarro del Cañizo
A A
AA
Cátedra de Microbiologia Industrial y Biotecnologia, Facultad de Farmacia y Bioquimica Junin 956, Buenos Aires, Argentina.
Mazza
M
M
Bellinzoni
R
R
Cascone
O
O
D005537:Aphthovirus / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006493:Heparin; D013056:Spectrophotometry, Ultraviolet; D014461:Ultracentrifugation
D006493:Heparin