Host cell DNAs or residual DNAs (rDNAs) are trace/low quantity of DNA originating from the organisms used in the production process of biopharmaceutical products, which may be introduced into the final products. The rDNAs may be able to transmit viral infections, cause a potential risk for oncogenesis or adverse reactions, etc. To assure the safety and potency, rDNAs in final biopharmaceutical products must be carefully monitored and quantitated, following the requirements established by World Health Organization (WHO), the European Pharmacopeia, the US Food and Drug Administration (FDA), and other regulatory agencies. The acceptable limits have been set between 100 pg/dose and 10 ng/dose depending on the cell line used, and the mode and frequency of dosing. Hence, the sensitive, accurate, and quantitative methods must be applied to ensure the rNDA is cleared to the limits.
Creative Proteomics provides a sensitive and accurate analysis to quantitate a broad range of rDNAs in various types of samples, including monoclonal antibodies, recombinant proteins, viral products, etc. Our experts will deploy optimal method to perform rDNAs tests during all stages of drug discovery, including real-time qPCR assay, Southern Blot-like hybridization and Threshold technique and PicoGreen method, etc.
We Can Provide but Not Limited to:
- Sample preparation and DNA Extraction
- Ultra-sensitive detection of rDNAs from various species
- Quantification of total rDNAs
- Quantitation of specific rDNA
- DNA Sequencing of unknown rDNAs
Technology Platform of Host Cell DNA Analysis
Creative Proteomics provides efficient and accurate rDNAs analysis to detect and quantitate rDNAs rapidly by optimized methods and protocols depending on your specific requirement, including PCR, real-time PCR, Southern Blot, DNA sequencing, Threshold assay, Hybridization assay, etc. Threshold assay can be conducted to nonspecifically quantify the total DNA by using DNA binding-proteins with high affinity for single-stranded DNA. The most commonly used standard for rDNAs quantification is based on real-time qPCR targeting specific repetitive DNA sequences. Besides, the treatment of the initial sample plays a critical role on successful rDNAs analysis. In order to ensure suitability with the assay and sample type, different pretreatment of samples will be considered, including organic extraction, magnetic beads, etc. To increase the efficiency, a digestion-free method will be conducted to rapidly extract DNA from the samples to following real-time qPCR analysis or PicoGreen assay of rDNAs. Based on your requirements, different protocols will be employed.
Advantages of Host Cell DNA Analysis
- High sensitivity and accuracy: Detection of genomic DNA on picogram level and bacterial DNA on femtogram level, and DNA from viral origin.
- High specificity: No cross reactivity to unrelated DNA.
- Advanced system: The Applied Biosystems resDNASEQ™, etc.
- Rapid turnaround time: 5-7 days to provide comprehensive report.
- Customized service: Optimized methods will be customized based on your sample type and size.
Creative Proteomics's experienced scientists can provide extensive analysis of rDNAs, including detection, quantitation, sequencing, etc. Clear, comprehensive written reports, recommendations and protocols, and customized services will be provided to help customers solve analytical and technical problems.
- Vernay O., Sarcey E., Detrez V., et al. Comparative analysis of the performance of residual host cell DNA assays for viral vaccines produced in Vero Cells. Journal of Virological Methods, 268 (2019) 9-16.
- Wang Y., Cooper R., Kiladjian A., et al. A digestion-free method for quantification of residual host cell DNA in rAAV gene therapy products. Molecular Therapy: Methods & Clinical Development. Vol. 13 June 2019.
- Rathore A.S., Sobacke S.E., Kocot T.J., et al. Analysis of residual host cell proteins and DNA in process streams of a recombinant protein product expressed in Escherichia coli cells. Journal of Pharmaceutical and Biomedical Analysis. 32 (2003) 1199-1211.
- Ikeda Y., Iwakiri S. and Yoshimori T. Development and characterization of novel host cell DNA assay using ultra-sensitive fluorescent nucleic acid stain "PicoGreen". J Pharm Biomed Anal. 2009 May 1;49(4):997-1002.