Getting CHO host cell protein analysis up to speed

Aim: Host cell proteins represent a major process-related impurity in recombinant biotherapeutics. Present work reports about a fully automated platform based on a cobas instrument (Roche Diagnostic GmbH, Mannheim, Germany) allowing high-throughput determination of host cell proteins. Materials & methods: The instrument combines automated sample preparation with electrochemiluminescence-based detection technology, facilitating highly sensitive and accurate detection. Results: Our data shows that the assay performs readily comparable to conventional ELISAs, but outperforms in speed, sample throughput and by its superior linear range. Comparison of ELISA and electrochemiluminescence immunoassay validation results from more than ten independent validations of clinical product testing methods supports our comparability claim. Conclusion: The electrochemiluminescence immunoassay represents a milestone for bioprocess impurity testing and abolishes the sample throughput limitations posed by conventional ELISA.

[1]  A. Hunter,et al.  Host cell proteins in biologics development: Identification, quantitation and risk assessment , 2009, Biotechnology and bioengineering.

[2]  Griffin,et al.  Points to consider in the manufacture and testing of monoclonal antibody products for human use (1997). U.S. Food and Drug Administration Center for Biologics Evaluation and Research. , 1997, Journal of immunotherapy.

[3]  N. Hoyle,et al.  Electrochemiluminescence : Leading-edge technology for automated immunoassay analyte detection , 1996 .

[4]  J. Zhu-Shimoni,et al.  Host cell protein testing by ELISAs and the use of orthogonal methods , 2014, Biotechnology and bioengineering.

[5]  John Hickey,et al.  Profiling of host cell proteins by two‐dimensional difference gel electrophoresis (2D‐DIGE): Implications for downstream process development , 2010, Biotechnology and bioengineering.

[6]  A. Capron,et al.  Automation of enzyme-linked immunosorbent assay (ELISA). , 1979, Journal of immunological methods.

[7]  P. O’Farrell.  High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[8]  K. Champion,et al.  Proteomic studies support the use of multi‐product immunoassays to monitor host cell protein impurities , 2006, Proteomics.

[9]  C. Smales,et al.  Host cell protein dynamics in the supernatant of a mAb producing CHO cell line , 2012, Biotechnology and bioengineering.

[10]  Kathleen Champion,et al.  Defining Your Product Profile and Maintaining Control Over It , Part 2 Challenges of Monitoring Host Cell Protein Impurities , 2005 .

[11]  M. Mølhøj,et al.  Identification and Monitoring of Host Cell Proteins by Mass Spectrometry Combined with High Performance Immunochemistry Testing , 2013, PloS one.

[12]  Guillaume Rey,et al.  Full automation and validation of a flexible ELISA platform for host cell protein and protein A impurity detection in biopharmaceuticals. , 2012, Journal of pharmaceutical and biomedical analysis.

[13]  S. Kirby,et al.  A microfluidic approach to high throughput quantification of host cell protein impurities for bioprocess development , 2014 .

[14]  A. Richter,et al.  Assays for Controlling Host-Cell Impurities in Biopharmaceuticals , 2005 .