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03
Feb
Residual Host Cell DNA: Why It Matters in Bioprocess QC
A practical guide to residual host cell DNA monitoring in research-stage bioprocessing—specifically optimized for HEK293/293T production systems.
Understanding Residual Host Cell DNA
Residual host cell DNA (HCDNA) refers to trace amounts of genomic and process-related DNA that can carry over from the cell substrate into downstream intermediates. In workflows using human cell lines such as HEK293 or HEK293T, these sequences are critical markers for process consistency.
While regulatory requirements vary, measuring residual DNA is a cornerstone of research use only (RUO) risk assessment, helping scientists ensure process robustness before moving to larger scales.
Key Benefits of DNA Monitoring
- • Process Insight: Effectively tracks clarification and polishing efficiency.
- • Comparability: Facilitates reliable batch-to-batch comparisons.
- • Matrix Sensitivity: Identifies potential extraction inhibition early in the workflow.
Deep Dive: Understanding why specific sequences are chosen is key to assay design. Explore our technical breakdown on Why E1A and SV40LTA Are Practical DNA Marker Targets .
Quantitation via qPCR
Real-time PCR (qPCR) is the gold standard for sensitivity in HCDNA analysis. A typical absolute quantitation workflow involves:
Step 1: Extraction DNA from samples and matched controls.
Step 2: Amplification Run qPCR with a 5-point standard curve.
Step 3: Validation Verify results using inhibition controls.
Pro Tip: Matrix inhibition is a common cause of false negatives in complex bioprocess samples. Learn how to mitigate this in our guide: Implementing Internal Positive Controls (IPC) .
Streamline Your HEK293T QC
Precision duplex detection of SV40LTA & E1A with an IPC-controlled workflow.
Explore the 2G Quantitation Kit →
Biofargo team
