Host Cell Residual DNA Quantitation
The use of biologically derived drugs, in which host cell lines are used to express biological product, has increased dramatically in recent years. This has created a need to quantify residual host cell DNA during manufacturing and in final products.
|The removal of host cell impurities is a critical step in the purification of biopharmaceutical products. A major challenge is the accurate and sensitive quantitation of host cell DNA impurities in both the purification process and in drug substance samples.
To ensure product quality, the amount of residual DNA in a drug's final dosage form must meet regulatory guidelines established by the World Health Organization (WHO), the European Pharmacopeia, the US Food and Drug Administration (FDA), and other regulatory agencies.
A host cell quantitation system should be designed with the sensitivity to meet or exceed regulatory requirements, while enabling high throughput and cost effectiveness to enable robust process clearance study design.
Watch our free webinar to learn more about Host cell protein and residual dna.
Step-by-Step Guide to Host Cell Residual DNA Quantitation Products
The PrepSEQ™ Residual DNA Sample Preparation Kit and protocols are optimized to enable highly efficient DNA recovery from typical biopharmaceutical purification process samples. Consistent performance has been demonstrated across a wide variety of complex test sample matrices. We have also verified recovery of CHO DNA spiked into the following matrices:
- Acetate (20 mM) pH 3.5
- Acetate (500 mM) pH 3.5
- HCl (12 mM) pH 3.0
- Glycine (20 mM) pH 2.0
- Glycine (500 mM) pH 2.0
- Acetate/NaCl (300 mM/0.5 M) pH 4.5
- MES/NaCl (300 mM/0.5 M) pH 6.5
- Bis (300 mM) pH 6.5; MES (300 mM) pH 6.0
- Phosphate (300 mM) pH 5.0
- Tris (50 mM) pH 8.0
Achieve efficiencies through automation
Automation of the sample preparation procedure on the MagMAX™
Express-96 Magnetic Particle Processor helps to increase throughput and
efficiency. Using the automated system, host cell residual DNA can be
isolated from up to 24 triplicate test samples, typically in less than 2 hours.
Next, combine the purified DNA template with the assay mix included in the resDNASEQ Residual DNA Quantitation System.
The resDNASEQ® Host Cell Residual DNA Quantitation Kits exhibit a broad dynamic range and high sensitivity (Figure 2). The limit of quantitation (LOQ) of the assays is based on recovery and quantitation of standard genomic DNA recovered from a test sample matrix consisting of 50 mg/mL IgG. Typical results for these kits are:
- CHO 1.5 pg/mL
- Vero 1.5 pg/mL
- NSO 1.5 pg/mL
- E. coli 15 pg/mL
- MDCK 1.5 pg/mL
AccuSEQ™ Real-Time PCR Software is an intuitive easy-to-use analysis tool for data interpretation. The software guides the user through the experiment and plate setup and provides easy-to-use tools for defining and setting up standards, controls, and unknowns. AccuSEQ™ v2.0 software is designed for analysis of results obtained from ViralSEQ®, MycoSEQ™, resDNASEQ®, and ProteinSEQ™ kits.
- Quick Reference Card: resDNASEQ™ Quantitative CHO DNA Kit
- Protocol: PrepSEQ™ Residual DNA Sample Preparation Kit
- Protocol: resDNASEQ™ Quantitative CHO DNA
- Quick Reference Card: PrepSEQ™ Residual DNA Sample Preparation Kit
- Product Bulletin: CHO Residual DNA Quantitation
- Solutions for the Biopharmaceutical Industry
Have questions about Gibco® media, POROS® chromatography resins, Applied Biosystems® SEQ rapid contaminant tests, or our Cell Therapy products? Click below to access product brochures, training videos, mobile apps, and more.
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