title: Respiratory syncytial virus. I. Concentration and purification of the infectious virus.
abstract: Respiratory syncytial (RS) virus can be purified without losing its infectivity provided that each step of purification is carried out using NT buffer containing over 20% sucrose. Firstly, the virus grown on HES cells is efficiently removed from the culture fluid by precipitating with polyethylene glycol (PEG) 6,000, and the precipitate is suspended in a small amount of 20% sucrose-NT buffer, which results in about a 24-fold concentration of the original material. Then this suspension is centrifugated through 30% sucrose-NT buffer to obtain pellets, which are again suspended in 20% sucrose-NT buffer. This suspension is further centrifuged by discontinuous and linear sucrose density gradient. Finally, the specific infectivity of the purified virus was increased about 3,000-fold over that of the original material.
mesh_terms: D002478:Cells, Cultured; D008722:Methods; D011092:Polyethylene Glycols; D012136:Respiratory Syncytial Viruses / Q000187:drug effects / Q000302:isolation & purification*; D013395:Sucrose / Q000494:pharmacology; D014461:Ultracentrifugation; D014776:Virus Cultivation / Q000379:methods
chemical_list: D011092:Polyethylene Glycols; D013395:Sucrose