Host cell proteins (HCPs) are a heterogeneous mixture of proteins carried over from the host expression system, and the main impurities in the process of bio-therapeutic manufacturing. The HCPs, either secreted proteins or intracellular proteins released during cell lysis, might influence the enzymatic modification and stability of products, cause failure of a certain purification step, and induce immunogenicity or decreasing activity, etc. FDA and EMA has set the acceptable limits of HCPs, but unknown HCPs at a low level might still leads to an immunogenic effect. To ensure safety and potency of biopharmaceutical products during drug discovery, detection and elimination of HCPs during bio-therapeutics purification are important and required.
Creative Proteomics provides a high-quality and high-sensitivity analysis to detect and quantify HCPs by mass spectrometry (MS), in accordance with relevant guidelines (ICH Q6B, PharmEu and USP<1047>). The HCP coverage of generic or process-specific Enzyme Linked Immunosorbent Assay (ELISA) polyclonal antibodies can also be validated. Our professional scientists and analysts will deploy optimal approaches to fully detect, characterize, and quantify HCPs in the biopharmaceutical products, and meet requirements of each stage of drug discovery.
We Can Provide but Not Limited to:
- Validation of ELISA assay
- Detection of HCPs
- Quantification of HCPs
- Individual HCP identification
- Analysis for bioprocess optimization
- Monitor and quantitate specific HCPs of concern
- Analysis of process-related residuals proteins
- HCP Antibody Coverage Analysis
Technology Platform of Host Cell Protein Analysis
Creative Proteomics provides efficient and accurate HCPs analysis to detect and characterize HCPs, through ELISA, electrophoresis, 2D Western blot and MS, according to your specific requirement. We offer rapidly comprehensive detection and quantitation of individual HCPs in one sample, based on SWATH Acquisition on a SCIEX TripleTOF system.
For MS analysis, samples will be first digested into peptides. Then the peptides mixture will be separated by two-dimensional liquid chromatography (2D-LC) or capillary electrophoresis and electrospray ionization (CESI). CESI is a process with integration of capillary electrophoresis (CE) and electrospray ionization (ESI), which can improve the sensitivity of HCP quantitation by reducing ion suppression and improving ionization efficiency. And the separated peptides will undergo MS/MS analysis. The data independent acquisition strategy can provide unbiased HCPs detection at low level, and the CESI-MS can significantly enhance sensitivity to detect low abundance HCPs.
Advantages of Host Cell Protein Analysis:
- High sensitivity: low abundance HCPs can be detected.
- Unbiased detection of unknown HCPs.
- Full analytical characterization of individual HCPs improves your Down Stream Process (DSP).
- Advanced equipment: CESI 8000 Plus, ExionLCTM AD System, and TripleTOF 6600.
- Rapid turnaround time: 5-7 days to provide comprehensive report.
- Customized service: Optimal buffers and protocols will be customized based on your project and sample type.
Creative Proteomics's analytical scientists can provide comprehensive analysis of HCPs, including detection, identification, quantitation, physico-chemical properties, etc. Clear, detailed written reports, recommendations and protocols, and customized services to will be provided to help customers solve analytical and technical problems.
References
- Bailey-Kellogg C., Gutierrez A.H., Moise L., et al. CHOPPI: a web tool for the analysis of immunogenicity risk from host cell proteins in CHO-based protein production. Biotechnology and Bioengineering. Vol. 111, No. 11, November, 2014.
- Tscheliessnig A.L., Konrath J., Bates R., et al. Host cell protein analysis in therapeutic protein bioprocessing-methods and application. Biotechnol. J. 2013, 8, 655–670.
- Valente K.N., Levy N.E., Lee K.H, et al. Applications of proteomic methods for CHO host cell protein characterization in biopharmaceutical manufacturing. Current Opinion in Biotechnology. 2018, 53:144-150.
