Journal of Advanced Biomedical Sciences
JABS
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URL: http://jabs.fums.ac.ir/article-1-237-en.html
2- Cellular and Molecular Research Center, Faculty of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran. ,
3- Molecular Medicine Department, Pasteur Institute of Iran, Tehran, Iran.
Background & Objective: Interferon belongs to a family of cytokines, which has
the most important role in the innate immune response to virus infections.
While producing recombinant interferon in biological host, some pieces of host
nucleic acids remain in product. Because of limitations in previous techniques
for detection of these impurities, the objective of this study is to use rapid
and sensitive Real time PCR method for detecting the impurities.
Materials & Methods: First, with DNA extraction from bacterial host cell and
preparation of its serial dilutions, SYBR Green-based Real time PCR reaction
was held and standard curve was plotted. After DNA extraction from interferon
and performing PCR, total DNA amount was determined using standard curve.
Results: Studies performed on some interferon samples, revealed
that the amount of DNA impurities was about 0.02 pg. per product dose. In
addition, the designed primers in the above reaction had no interaction with
each other and other interfering agents.
Conclusion: For the first time in Iran, this study was set up and it
revealed that Real time PCR can be used as a functional and accurate technique
in manufacture centers for detection of residual host cell DNA in interferon
and other recombinant pharmaceutical products.
Received: 2014/06/16 | Accepted: 2014/10/23 | Published: 2015/02/2
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