Determination of Residual Vero Cell DNA in viral vaccine products by Real-time PCR
Keywords:
Vero cell, residual DNA, Real-time PCRAbstract
Vero cells are continuous cell lines which are used as a substrate for producing many viral vaccines because of their safety, non-tumorigenicity and non-latent viruses. However, the safety of the vaccines using the Vero cell also depends on the production, purification, and quality control processes. The World Health Organization currently recommends 10 ng/dose as an upper limit of DNA residue for any vaccines produced from continuous cell lines. Institute of Biological Products had validated the Real-time PCR method to determine residual DNA of Vero cells in viral vaccine. Sixteen Vero cell-based vaccine samples were employed. The testing results showed that small amounts of residual DNA were present in all samples from 0.49 to 4.48 ng/dose, which is lower than WHO’s specification. From the analysis of linearity and range, it was found that the appropriate working range was 0.03-3,000 pg/reaction and the correlation coefficient was more than 0.990. For the accuracy, the recovery rate was 87% which was within the acceptance limit (80-120%). This method showed good repeatability when the test was conducted in the same day and at the same time and the coefficient of variance was less than 25%. For the analysis of robustness using two different analysts, it was found that the results were very close to each other and the coefficient of variance was less than 25%. Therefore, this method also showed good robustness. In conclusion, this developed real-time PCR when had been used for determination of residual Vero cell DNA in viral vaccines showed good results in linearity, accuracy, precision and robustness.
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