Preparation of poliovirus labeled with phosphorus-33.
Phosphorus-33 ((33)P), a weak (0.25 Mev) beta-emitting isotope of phosphorus with a half-life of 25 days, has been used to label poliovirus in cell culture. HeLa cell monolayers were depleted of phosphate and then labeled by incubating at 37 C in a medium (LM) containing about 10 muCi of (33)P as orthophosphate per ml. Labeled cells were infected at a high multiplicity with poliovirus type 1 and incubated for 8 hr in LM medium. Virus from infected cells was then concentrated and purified. Virus purity was confirmed by comparison of virus infectivity and radioactivity after CsCl density gradient centrifugation and by observing purified virus preparations with electron microscopy. With the method described, yields of about 10(10) to 5 x 10(10) plaque-forming units (PFU) of highly purified poliovirus with specific activities of about 3 x 10(-4) to 10(-3) disintegrations per min per PFU have been obtained from 1.5 x 10(8) to 3.0 x 10(8) HeLa cells.
D002499:Centrifugation, Density Gradient; D002586:Cesium; D002712:Chlorides; D003470:Culture Media; D006207:Half-Life; D006367:HeLa Cells / Q000378:metabolism / Q000382:microbiology; D007553:Isotope Labeling; D008854:Microscopy, Electron; D010710:Phosphates / Q000378:metabolism; D010759:Phosphorus Isotopes; D017955:Poliovirus / Q000254:growth & development / Q000302:isolation & purification*; D014776:Virus Cultivation
D002712:Chlorides; D003470:Culture Media; D010710:Phosphates; D010759:Phosphorus Isotopes; D002586:Cesium