Comprehensive and Automated Profiling of Host Cell Proteins Using Proteograph XT Workflow
This application note shows how the Proteograph XT platform increases discovery depth, simplifies HCP MS sample prep workflow and compares with peer-reviewed articles to benchmark the Protegraph XT performances using the NIST 8671 mAB.
Introduction
The presence of host cell proteins (HCPs) in biopharmaceuticals is a critical concern due to their potential immunogenicity and potential to diminish therapeutic efficacy, which pose detrimental risks to patients. To address this, liquid chromatography-mass spectrometry analysis (LC-MS) has emerged as a novel and versatile analytical technique for comprehensive HCP analysis.
While this approach offers exceptional sensitivity, specificity, and comprehensive accuracy in identification and quantification of HCPs, conventional LC-MS is still challenged by the inherently large dynamic range of HCPs compared to biologics. Regardless, LC-MS has the potential to detect a wide range of HCPs, delivers precise and quantitative results, and resolve the identity of individual proteins within a sample. Notably, LC-MS obviates the reliance on specific antibodies, a limitation inherent in conventional ELISA methods.
Monoclonal antibody-based detection generally requires tedious development time and resources, and while polyclonal antibody-based detection offers a broader coverage, it often lacks specificity. Nonetheless, traditional LC-MS-based HCP analysis encounters challenges stemming from the intricacy and heterogeneity of the sample matrix, as well as the prerequisite for advanced instrumentation and expertise in data analysis.
In this study we investigated the performance of the Proteograph Product Suite, an automated and standardized nanoparticle-based sample preparation and data analysis workflow, integrated with state-of-the-art microflow liquid chromatography and mass spectrometry technology. In this application note we subject the Proteograph XT workflow to rigorous evaluation of HCP identification depth, qualitative and quantitative reproducibility of the platform, and measurement reliability toward a list of common problematic HCPs. We performed the evaluation using a widely accepted community benchmark standard, the NIST monoclonal antibody (NIST Reference Material® 8671) (NIST 8671).
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