Comparison of methods for concentration and purification of bovine viral diarrhea virus.
The NADL strain of bovine viral diarrhea virus (BVDV) was concentrated by hollow fiber ultrafiltration or polyethylene glycol and purified by centrifugation through sucrose or potassium tartrate gradients. The protein content of polyethylene glycol concentrates was much lower than that of ultrafiltration concentrates. Conversely, recoveries of infectivity were greater using polyethylene glycol (100%) as compared to ultrafiltration (50%). Sucrose or potassium tartrate density gradients appeared comparable in purification of BVD virus. Peak infectivity fractions in both gradients corresponded quite well, having densities of 1.12-1.14 g/cm3, and showed a 150-fold reduction of protein when compared to crude viral supernate. Further examination by negative stain electron microscopy revealed integral pleomorphic, roughly spherical particles in both purified virus preparations. Small knob-like projections could be seen on viral particles.
Magar
R
R
Institut Armand-Frappier, Université du Québec, Laval, Canada.
Lecomte
J
J
D002499:Centrifugation, Density Gradient; D001908:Diarrhea Viruses, Bovine Viral / Q000302:isolation & purification* / Q000648:ultrastructure; D008854:Microscopy, Electron; D010576:Pestivirus / Q000302:isolation & purification*; D011092:Polyethylene Glycols; D014462:Ultrafiltration
D011092:Polyethylene Glycols