Presence and quantification of cell substrate DNA in inactivated poliovirus vaccine.
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.
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
D011054:Poliovirus Vaccine, Inactivated; D014613:Vaccines, Attenuated; D004247:DNA