Proteins & Peptides Residual Host Cell DNA Testing

Residual host cell DNA detection plays a key role in protein/peptide biologics quality and safety assessment, process validation, evaluation of batch-to-batch variability, and GMP batch release. With expertise in host cell protein (HCP) and DNA analysis, CD Formulation uses a range of highly sensitive methods to identify and quantify these residual host cell DNA in accordance with global regulatory requirements.

Residual Host Cell DNA & Why Detect Residual Host Cell DNA?

Residual host cell DNA refers to DNA fragments or longer molecules from host tissue cells that may appear in biological products. These DNA molecules may be introduced into the final product, causing potential risks such as viral infection, carcinogenesis, or adverse reactions. In order to monitor the safety and quality controllability of biological products, it is necessary to establish appropriate residual host cell DNA detection methods in accordance with the requirements set by the World Health Organization (WHO), European Pharmacopoeia (EP), US Food and Drug Administration (FDA) and other regulatory agencies to carefully monitor and quantify residual DNA in the final biopharmaceutical product. High-quality residual host cell DNA testing helps:

• Safety assessment: Biopharmaceutical products are usually produced by expressing and producing target proteins or biopharmaceuticals using host cells (such as bacteria or mammalian cells). During the production process, host cell DNA may be present in pharmaceutical products. Detecting host cell DNA residues can evaluate the safety of pharmaceutical products and ensure that the host cell DNA content in the product meets relevant regulations and standards.
• Quality control and consistency: The level of host cell DNA residues can be used as one of the indicators of product consistency and quality control. By detecting host cell DNA residues, consistency between production batches can be ensured and a reference can be provided for product quality assessment.
• Immunogenicity assessment: Host cell DNA residues may contain unexpressed host gene fragments that may be immunogenic. The presence of high levels of host cell DNA residues in biopharmaceutical products may trigger an immune response, which may lead to safety issues or drug failure. By detecting and controlling host cell DNA residues, immunogenicity risks can be assessed and the safety and efficacy of the product can be ensured.
• Drug registration and regulatory requirements: In many countries and regions, drug registration and regulatory agencies have strict requirements and restrictions on host cell DNA residues in biopharmaceutical products. By detecting and quantifying host cell DNA residues, data requirements in registration applications can be met and compliance with regulatory agency regulations and guidelines can be ensured.

Explore Our Proteins & Peptides Residual Host Cell DNA Testing Services

During the production and purification of various types of biological products such as recombinant proteins, antibodies, vaccines, etc., residual host cell DNA can affect the quality of biological products and cause potential hazards such as infection and carcinogenesis. As part of our quality control testing procedures, we provide our global customers unparalleled residual host cell DNA testing services to prove that the level of host cell DNA in your protein/peptide biological products doesn't exceed regulatory guidelines (such as WHO and FDA residual DNA guidelines) and is effectively purified during the manufacturing process.

Our residual DNA analysis complies with the ICH Q6B guidelines and is applicable to a range of GLP/GCP/GCMP-compliant methods. With the expertise to develop residual DNA detection for any cell type (HEK293, E. coli, and CHO cells), we can quickly detect and quantify DNA through optimized methods and protocols according to your specific requirements to provide efficient and accurate residual host cell DNA analysis results.

Available Residual Cell DNA Testing Procedures Include:

• Residual host cell DNA (HCD) detection.
• Residual host cell residual DNA fragment analysis.
• Animal origin residual cell DNA detection.
• Specific cell or vector DNA/RNA detection.

Screening of Residual Host Cell DNA Testing Analysis Methods

At CD Formulation, we provide powerful quantitative analysis using real-time qPCR, threshold assays, and digital droplet PCR (ddPCR) technologies to support process validation, monitor batch-to-batch variability, and support GMP batch release.

Real-time qPCR:

A quantitative PCR-based method is the gold standard for residual DNA analysis and is used for relative quantification of samples to detect specific DNA from a specific source or to amplify specific gene sequences.

Threshold Assay

Non-specific quantification of total DNA using DNA binding proteins with high affinity for single-stranded DNA.

Digital Droplet PCR (ddPCR)

Compared to real-time qPCR, ddPCR is used for absolute quantitative analysis of samples. This method does not require any reference or extrapolation. Once the amplification reaction is complete, the large-scale partitioning nature of ddPCR can generate up to 20,000 oil-encapsulated droplets, effectively performing 20,000 separate reactions to provide thousands of data points, thereby improving quantitative accuracy.

Fig. 1 Schematic illustration of ddPCR. (Kojabad AA, et al., 2021)

Residual Host Cell DNA Sample Preparation (Purification)

Regardless of which of the above assays are used, the success of residual host cell DNA analysis depends largely on the initial sample preparation. Our scientists will take into account the sensitivity of each assay to residuals and perform some degree of sample pretreatment to ensure the suitability of the assay and sample matrix.

Available Pretreatment Methods:

• Organic extraction.
• Proteinase K treatment.
• Magnetic beads.
• Phenol-free Wako extraction.
• Column-based nucleic acid binding methods.
• Ethanol precipitation with added co-precipitant.

Validation of Residual Host Cell DNA Testing Analysis Method

ICH guidelines state that all quantitative assays must be validated. In accordance with ICH guidelines, we validate DNA assays using standard matrices, including:

• Precision
• Accuracy
• Limit of Detection (LOD)
• Limit of Quantification (LOQ)
• Linearity
• Specificity

Why Choose Us for Proteins & Peptides Residual Host Cell DNA Testing？

• We perform residual host cell DNA analysis using qPCR and ddPCR methods to support your protein/peptide biologics development program.
• We have extensive expertise and hundreds of successful projects in performing residual host cell DNA analysis.
• Our scientists have a deep understanding of the latest regulatory developments and are able to help you meet authorization requirements in all regions around the world.
• We strictly adhere to ICH GMP/GCP/GLP guidelines to ensure that the data you need meets the requirements of the US FDA, EMA, and OECD GLP regulatory standards.
• We provide flexible experimental options and customized solutions to meet the needs of your different projects.

Publication

Residual host cell DNA testing is a critical component to ensure safety, efficacy, and consistency in the development and manufacture of therapeutic proteins and peptides. Please feel free to contact us if you are interested in our services. Learn how CD Formulation can help you conduct residual DNA analysis and verify the safety of your products in relation to regulatory requirements.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.