title: Presence and quantification of cell substrate DNA in inactivated poliovirus vaccine.
abstract: In order to follow the fate of cell substrate DNA in inactivated poliovirus vaccine (IPV), experiments simulating different steps in IPV preparation were performed. For this purpose, 3H-thymidine-labeled HeLa cell DNA released during lytic infection with Mahoney type 1 poliovirus strain was used as tracer. Low speed centrifugation (1500 xg for 15 min.) of crude virus suspension removed on the average 98.5% of the total labeled cell substrate DNA. Repeated freezing and thawing of the crude virus suspension before centrifugation did not increase the residual 1.5% radioactivity found in the supernatant. This level of contamination was not significantly influenced by filtration of the supernatant through a 0.22 micrometer pore size membrane filter. DEAE-Sepharose CL-6B chromatography of poliovirus suspensions containing large amounts of 3H-thymidine labeled HeLa cell extracts completely removed the contaminating substrate DNA. These data showed that IPV free of cell substrate DNA can easily be obtained by a usual purification procedure.
mesh_terms: D002498:Centrifugation; D002847:Chromatography, Agarose; D004247:DNA / Q000032:analysis*; D005615:Freezing; D006367:HeLa Cells; D006801:Humans; D017955:Poliovirus / Q000032:analysis*; D011054:Poliovirus Vaccine, Inactivated / Q000032:analysis* / Q000302:isolation & purification; D014613:Vaccines, Attenuated / Q000032:analysis* / Q000302:isolation & purification
chemical_list: D011054:Poliovirus Vaccine, Inactivated; D014613:Vaccines, Attenuated; D004247:DNA