Mass Spectrometry for Host Cell Protein monitoring and control in pharmaceutical process development

Alphalyse
Oct 2. 2009

Mass spectrometry based Host Cell Protein identification and quantification

Here the Mass Spec Host Cell Protein (HCP) assay is applied for monitoring product purity and consistency of the pre-clinical tox batch and 2 clinical batches of a recombinant vaccine protein.

The proteins were separated by 1D SDS PAGE and visualized by silver staining. The protein bands were digested with trypsin and the proteins identified by MALDI MS/MS peptide mapping and database searching. In total, 8 specific HCPs were identified.

The protein identities and information about physical properties, such as pI, Mw, and hydrophobicity can guide further protein purification and process development for phase III. An advantage compared to ELISA-based HCP assays is that the MS HCP assay provides the identities of the contaminating HCPs and does not require time-consuming development of antibodies.

Advantages

  •  Immediate solution
  •  Alternative to ELISA when not available
  •  Identity for each HCP
  •  Relative quantity of HCPs
  •  Complementary with Western blot
  •  Fast assay setup in 2 weeks

Applications

  •  PAT compliant
  •  Monitoring process development,
  •  Comparison of pre-clinic TOX batches and clinical batches

Learn more about Mass Spec Host Cell Protein monitoring

MS Host Cell Protein Assay Principle

MS Host Cell Protein Assay Principle

Figure 1: The drug substance [DS] from different cGMP batches (samples 1-3) were loaded on a 1D SDS-PAGE gel and stained with sensitive silver stain. The gel was overloaded with DS for detection of very small amounts of contaminating proteins. Protein bands were cut out and analyzed by MALDI mass spectroscopy to identify contaminating host cell proteins. Eight specific HCP’s were identified and marked HCP1-HCP8, as well as product-related variants, marked DS.

Reference:
Mortz, E. et al. “Proteomics technology applied to upstream and downstream process development of a protein vaccine”, Bioprocess International 6, p36-43, 2008.

About Alphalyse

Alphalyse uses expertise in the field of protein chemistry and bioinformatics, combined with top-of-the-line mass spectrometry equipment, to provide protein analysis services globally.

Alphalyse aims to deliver high quality data and customer service to all of our clients in a fast and convenient way.

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