Host cell protein networks as a novel co‐elution mechanism during protein. A chromatography
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Publication date
07.03.2024
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01A - Journal article
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Parent work
Biotechnology & Bioengineering
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Volume
121
Issue / Number
5
Pages / Duration
1716-1728
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Publisher / Publishing institution
Wiley
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Abstract
Host cell proteins (HCPs) are process-related impurities of therapeutic proteins produced in for example, Chinese hamster ovary (CHO) cells. Protein A affinity chromatography is the initial capture step to purify monoclonal antibodies or Fc-based proteins and is most effective for HCP removal. Previously proposed mechanisms that contribute to co-purification of HCPs with the therapeutic protein are either HCP-drug association or leaching from chromatin heteroaggregates. In this study, we analyzed protein A eluates of 23 Fc-based proteins by LC-MS/MS to determine their HCP content. The analysis revealed a high degree of heterogeneity in the number of HCPs identified in the different protein A eluates. Among all identified HCPs, the majority co-eluted with less than three Fc-based proteins indicating a drug-specific co-purification for most HCPs. Only ten HCPs co-purified with over 50% of the 23 Fc-based proteins. A correlation analysis of HCPs identified across multiple protein A eluates revealed their co-elution as HCP groups. Functional annotation and protein interaction analysis confirmed that some HCP groups are associated with protein-protein interaction networks. Here, we propose an additional mechanism for HCP co-elution involving protein-protein interactions within functional networks. Our findings may help to guide cell line development and to refine downstream purification strategies.
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ISBN
ISSN
0006-3592
1097-0290
1097-0290
Language
English
Created during FHNW affiliation
Yes
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Publication status
Published
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Peer review of the complete publication
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Closed
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Citation
Panikulam, S., Hanke, A., Kroener, F., Karle, A., Anderka, O., Villiger, T., & Lebesgue, N. (2024). Host cell protein networks as a novel co‐elution mechanism during protein. A chromatography. Biotechnology & Bioengineering, 121(5), 1716–1728. https://doi.org/10.1002/bit.28678