HCPs detection during DSP optimization

Multiple purification steps are employed in the Downstream Process (DSP) to eliminate HCPs, starting from the cellular system’s harvest and extending to the production of the final Drug Product. 

HCPs detection during DSP optimization

From the cellular system harvest to the final Drug Product, several purification steps are implemented through the Down Stream Process (DSP) to remove HCPs. These residual proteins are part of the Critical Quality Attribute (CQA) since it can affect product:

• quality;
• stability;
• efficacy;
• safety;
• induce or enhance immunogenicity.

Context

To ensure patient safety, HCPs evaluation contained in production batches is essential. In addition, it is needed to get a fast HCPs detection and quantification method in order to anticipate the next process developments necessary to optimize drug purification.

Given the importance of ensuring patient safety, the evaluation of HCPs in production batches becomes crucial. It is essential to have a rapid and reliable method for the identification and quantification of HCPs. This enables the anticipation of necessary process developments to optimize drug purification, thereby enhancing the overall quality and purity of the final product.

Moreover, classical analytical approaches exhibit certain technical limitations arising from immunoassay specificity issues, inadequate resolution of 2D-electrophoresis gels, and suboptimal performance in LC separation.

Anaquant’s solution for HCPs detection

Our HCP Profiler Solution enables the precise identification and quantification of Host Cell Proteins (HCPs) at every stage of the bioprocess. It supports regulatory compliance, optimizes the Downstream Process (DSP), and provides insights into the impact of the Upstream Process (USP) on HCPs. 

With its proprietary technology and universal standard, this solution allows for accurate conversion of MS signals into protein quantities. By utilizing READYBEADS™ technology, it ensures accurate quantification of all identified HCPs. Our dedicated software simplifies data analysis and facilitates sample comparison. The specific sample preparation techniques enable the detection of low-concentration HCPs. 

The HCP Profiler Solution de-risks biologic development by providing vital information about HCP identity and quantification throughout manufacturing, purification, and final product quality testing stages. It achieves this through the use of a universal calibration curve and synthetic peptides, which are accurately calibrated using READYBEADS™ technology. The calibration curve enables the conversion of peptide signals into quantitative measurements for all identified proteins. 

By normalizing the identified proteins with this calibration curve, and utilizing internal standards and dedicated software, we ensure robust data processing, facilitating the reproducible quantification of individual HCPs as an orthogonal method to complement ELISA-based HCP testing.

 

With our HCP-Profiler solution, according to MS specificity, we propose an orthogonal answer to bridge these gaps. The solution can be implemented through the whole DSP steps from harvest to final drug substance. According to its repeatability and reproducibility capabilities, it provides within a few hours, an identification and a quantification for each Host Cell Protein.

In this application note, we demonstrate the usefulness of HCP-Profiler to evaluate performances of new DSP’s purification steps.

Results

In our case study, 3 different drug substances (DS) were analyzed according to the HCP-Profiler solution and 2 major critical HCPs were identified in drug substances. These 2 major contaminants were considered as possible immunogen proteins. One year following the analysis, these three newly obtained Drug Substances (DS) were examined after undergoing an additional purification step as part of the Downstream Process (DSP).

When comparing batches from the previous and new Downstream Process (DSP), it was observed that the two critical Host Cell Proteins (HCPs) were either completely eliminated or predominantly reduced in the drug substance, possibly falling below the limit of detection. Furthermore, the total amount of HCPs was significantly reduced with the incorporation of this supplementary purification step.

 

 

 

Host Cell Proteins (HCPs) identification and quantification comparison for DSP optimization purpose

Conclusion

The HCP Profiler Solution offers a powerful tool for fast and accurate bioprocess optimization. We demonstrated several of its functions:

• Improve and forecast quickly your bioprocess;
• Gain knowledge in your HCPs (Individual Identification & Quantification);
• Support your regulatory dossier;
• Get ethical (no animal involved for HCP antibody production);
• Sample comparison possible whatever the matrix and timeslot between analysis.

The HCP Profiler Solution by Anaquant is a powerful tool that enables process optimization, regulatory support, and comprehensive HCP analysis. Its rapid and accurate identification and quantification capabilities provide valuable insights throughout the bioprocess, ensuring the production of safe and high-quality biologic products.

For more information, explore Anaquant’s HCP Profiler Solution on our website.