Characterisation of the E. coli HMS174 and BLR host cell proteome to guide purification process development

Roxana Disela, Olivier Le Bussy, Geoffroy Geldhof, Martin Pabst*, Marcel Ottens

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

1 Citation (Scopus)
26 Downloads (Pure)

Abstract

Mass-spectrometry-based proteomics is increasingly employed to monitor purification processes or to detect critical host cell proteins in the final drug substance. This approach is inherently unbiased and can be used to identify individual host cell proteins without prior knowledge. In process development for the purification of new biopharmaceuticals, such as protein subunit vaccines, a broader knowledge of the host cell proteome could promote a more rational process design. Proteomics can establish qualitative and quantitative information on the complete host cell proteome before purification (i.e., protein abundances and physicochemical properties). Such information allows for a more rational design of the purification strategy and accelerates purification process development. In this study, we present an extensive proteomic characterisation of two E. coli host cell strains widely employed in academia and industry to produce therapeutic proteins, BLR and HMS174. The established database contains the observed abundance of each identified protein, information relating to their hydrophobicity, the isoelectric point, molecular weight, and toxicity. These physicochemical properties were plotted on proteome property maps to showcase the selection of suitable purification strategies. Furthermore, sequence alignment allowed integration of subunit information and occurrences of post-translational modifications from the well-studied E. coli K12 strain.

Original languageEnglish
Article number2300068
Number of pages13
JournalBiotechnology Journal
Volume18
Issue number9
DOIs
Publication statusPublished - 2023

Keywords

  • bioprocess engineering
  • chromatography
  • e. coli
  • host cell proteomics
  • proteomics

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