Biochemistry & Pharmacology: Open Access
Open Access
ISSN: 2167-0501
+44-20-4587-4809
ISSN: 2167-0501
+44-20-4587-4809
Mahmoud Alebouyeh, Hossein Rastegar, Sepideh Nazerieh, Sharareh Mohammadi, Abbas Najafpour and Fariba Mohseni
Food and Drugs Control Laboratories Reference Center, Iran
Azad University Medical Sciences, Iran
Endocrinology research center, Iran
Posters & Accepted Abstracts: Biochem Pharmacol (Los Angel)
Statement of the Problem: Host cell DNA contamination occurs during the production of biopharmaceuticals and must be controlled and monitored for the purity and safety of the drug products. A dual labeled fluorescent probe Real-Time PCR assay were developed and validated for the quantitative measurement of residual host cell DNA impurity in prokaryote-based (E. coli) recombinant therapeutic proteins which have released in Iranian market. Methodology: In the present study, 48 different types of drugs including insulin, streptokinase, filgrastim, growth hormone and interferon beta 1-b which were available in Iranian market and imported or produced by Iranian companies were examined and the amounts of residual host cell DNA were measured by means of Real-Time PCR. Material & Methods: Real-Time PCR kit purchased and applied according to the manufacturer instructions, real time PCR instrument, micro-centrifuge and 2 mL microfuge tube were used. Following cycling conditions: 10 min at 95oC, one cycle followed by 45 cycles each consisting of 25 s at 95oC, 20 S at 63oC and15 s at 72oC was used. Finding: The mean amount of residual DNA in Iranian manufactured products was 3.54 pg/ml while the amount of residual DNA in imported product was 0.005548 pg/ml which was lower than World Health Organization and U.S. Food and Drug Administration guidelines (10 ng/Dose). Conclusion: The assay was demonstrated to be specific, precise, and accurate to assess residual E. coli DNA in the linear and quantitative range from 1 pg/mL to 100 pg/mL.
Mahmoud Alebouyeh has 15 years of experiences in Pharmacology and Pharmacogenetics and is working on gene-base impurities in recombinant therapeutic proteins. He has published 19 articles in scientific journals.