Vericheck ddPCR Contaminant Testing Solutions
“Truth in Testing”
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Impurities in products and processes | Mycoplasma testing | Residual DNA Quantitation | Residual DNA Sizing Testing | Talk with a Specialist |
In cell culture and when manufacturing cell-based vaccines, biologics, and gene therapies, contaminant-free batches are critical for patient safety. Droplet Digital™ PCR (ddPCR™) provides highly precise and absolute quantification of nucleic acids by counting nucleic acid molecules encapsulated in discrete water-in-oil droplets. Digital measurement of nucleic acids using ddPCR technology is suitable for analysis of both in-process samples and final drug products due to the technology's tolerance to PCR inhibitors. The tolerance to formulation matrices enables the implementation of ddPCR technology throughout the entire manufacturing process. Bio-Rad's ddPCR systems utilize a simple workflow with minimal hands-on time while reducing sample replicates and user-to-user variability.
Impurities in products and processes
Droplet Digital PCR Applications for impurity testing- Providing Reproducible, Accurate and Sensitive results for contaminants
Cell and gene therapies have rapidly gained popularity but ensuring safety and effectiveness in patients is complex with no room for error. Research, development, and manufacturing laboratories around the world are taking advantage of the precision and convenience of Bio-Rad's Droplet Digital PCR technology as a tool to overcome challenges with existing methods for a range of applications in cell and gene therapy. The FDA requires low coefficient of variation (CV) with bioanalytical measurements. The overall accuracy and precision at each concentration level should be within ±15% of the nominal concentration.
In contrast, qPCR based QC methods are reliant on standards that are difficult to source and can be variable from batch to catch, adding time and cost. In addition, qPCR based methods are sensitive to PCR inefficiencies like secondary structure and in process impurities, leading to assay failures and expensive assay repeats. These problems also are reflected in the challenges transferring from process development to either internal manufacturing groups or externally to a contract development manufacturing organization (CDMO).
| In-Process Testing | Final QA/QC | Patient Monitoring | ||||
|---|---|---|---|---|---|---|
| Incoming Vector | Activate and Transduce T Cells | Prepare Final Cell Dose | Potency | Purity | Safety | Verify Therapeutic Delivery and Expression |
| Plasmid Quality | Transgene Expression | Vector Copy Number | Residual DNA | Mycoplasma | Gene Expression | |
-
Virus Production -
Viral Purification & QC -
Pre-clinical & Clinical -
FDA-Approved Treatment
Droplet Digital PCR Solution
Absolute quantification. Absolute confidence.
Bio-Rad's ddPCR systems eliminate the need for a standard curve, simplifying QC workflows with minimal hands-on time while reducing sample replicates and user-to-user variability. ddPCR technology is also more resistant to in process matrix components that inhibit traditional qPCR efficiency.
Mycoplasma Testing
Cell lines and patient samples are prone to outbreaks of various mycoplasma infections; therefore, it is essential to address outbreaks as soon as they occur through robust testing. The Vericheck ddPCR Mycoplasma Detection Kit is a probe-based, highly sensitive and highly specific mycoplasma detection solution. This kit is designed and validated to meet European, U.S. and Japanese pharmacopoeia requirements. Detect up to 112 mycoplasma species using Droplet Digital. Utilize the premium edition software with audit trails and tools for U.S. FDA 21 CFR Part 11 compliance to easily analyze and report your data.
Vericheck ddPCR Mycoplasma Solution
- Verify mycoplasma presence/absence with absolute quantification
- Reduce variability, plate-to-plate and user-to-user, giving highly reproducible results
- Lower cross reactivity with related species, providing low false positive results.
Residual DNA Quantification
Residual DNA content is a common impurity in cell and gene therapy manufacturing, and levels must be carefully monitored for lot release. Droplet Digital PCR is a precise and sensitive method for direct quantification of residual DNA content. Since Droplet Digital PCR does not rely on reaction efficiency for quantification, it is less prone to sample interference from complex matrices and process intermediates. Droplet Digital PCR provides a highly sensitive method for direct quantification of residual HCD to ensure adherence to the FDA guideline of <10 ng/dose.
Residual DNA Sizing Testing
Human gene therapy products are produced in immortalized cell lines, most commonly HEK293. Residual DNA from immortalized cell lines can harbor tumorigenic genetic sequences or retroviral sequences that can cause infection. The FDA recommends that manufacturers degrade any residual DNA to a size below that of a functional gene, defined as <200 base pairs. Manufacturers require an inexpensive, accurate, and reliable way of measuring the size of residual DNA, which ddPCR technology can provide. The Vericheck ddPCR HEK293 Sizing Kit is the first HEK293 specific ddPCR based sizing solution that has been designed, and validated to meet regulatory guidance requirements and capable of measuring HEK293 size without the use of reference curve.
The Residual DNA Sizing Testing is now available.