The expression (OS/GS)I0(hcDNA)(OS/GS)I0(hcDNA) in Eq. (1) represents the genomic mass equivalent of oncogenes in a dose. While the calculation of the safety factor is both intuitive and easy to carry out, it does not account for disruption of the oncogene sequences through enzyme digestion; neither does it take account of the sizes of the individual oncogenes. Therefore, the risk estimates derived from their method are likely to be overstated. As a remedy, we introduce a probabilistic model to mechanistically study the relationship between the risks and characteristics of the purification process

such as enzyme cutting efficiency, total amount of residual DNA in the final dose, and biological nature of the host cells including numbers and sizes of oncogenes and infectious viral DNA, amounts of oncogenes and infectious agent required Selleckchem Baf-A1 to cause oncogenic and infectious events, respectively. The method is both simple and convenient to use. It is a useful tool for residual DNA risk assessment. The use of the model is illustrated through a real application. We assess oncogenic and infective potential of residual hcDNA from a cell-based live, attenuated influenza vaccine. The product is manufactured from a production process

that uses Madin Darby Canine Kidney (MDCK) as the cell. The process employs several purification steps to remove hcDNA, which include tangential flow filtration Stem Cells antagonist (TFF) and chromatography assay. During the TFF process step, DNA is removed from the virus based on the size difference between the virus and host cell DNA. Any residual DNA is removed or reduced in size during the affinity chromatography step. DNA does not bind to the chromatography media; however, any DNA that is associated with the virus or host cell protein that

binds to the media is degraded by treating with benzonase, which is included in the chromatography buffer wash. Using a canine SINE quantitative PCR, the amount of residual hcDNA is determined to be less than 1 ng per dose. With a direct-labeling method, the size distribution of residual DNA is also examined. The median size is approximately 450 base pairs (bp); PD184352 (CI-1040) approximately 64% of residual DNA is less than 500 bp in length. The haploid genome size of the canine genome is determined to be 2.41 × 109 bp. There are approximately 200 oncogenes identified in various species [9]. Using the SOURCE (located at http://smd.standford.edu) 81 expressed human oncogenes are found in 24 different tissues [8]. The average size of human oncogenes is 1925 bp with a standard deviation of 87 bp. Because the precise number of oncogenes contained in MDCK cell genome is unknown, for the oncogenic risk analysis, we restrict our evaluation to a single oncogene presumably having a size of 1925.