Quantitation of cell DNA in the evaluation of heteroploid cells as substrate for the preparation of killed viral vaccines.
To evaluate the risk of using heteroploid cell lines as substrates for viral vaccine production, the presence of cell DNA in poliovirus suspensions was examined. The time course of [3H]-thymidine-labeled HeLa cell DNA release during lytic infection with type 1 poliovirus was investigated. More DNA was found in filtered supernatants of poliovirus-inoculated cultures than in control cell supernatants. DNA concentration increased with time, paralleling virus release, but did not exceed 1.5% of the total DNA content of the culture. Only about 10% of this cell DNA was resistant to DNase treatment. By both ion-exchange chromatography and rate-zonal centrifugation it was possible to remove practically all cell DNA contaminating filtered poliovirus suspensions. Results obtained in this study permitted quantitative evaluation of cell substrate DNA present in poliovirus suspensions during successive steps of killed poliovirus vaccine preparation. Based on the sensitivity of our method, the amount of residual DNA was estimated at less than 0.02 pg per dose of purified vaccine.
D002501:Centrifugation, Zonal; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000032:analysis*; D005069:Evaluation Studies as Topic; D006367:HeLa Cells; D006801:Humans; D017955:Poliovirus / Q000032:analysis* / Q000254:growth & development; D011054:Poliovirus Vaccine, Inactivated / Q000032:analysis* / Q000302:isolation & purification
D011054:Poliovirus Vaccine, Inactivated; D004247:DNA