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Spatial Proteomics Reveals Differences in the Cellular Architecture of Antibody-Producing CHO and Plasma Cell-Derived Cells

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2022

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Kretz, Robin
Raab, Nadja
Zeh, Nikolas
Gauges, Ralph
Otte, Kerstin
Fischer, Simon
Stoll, Dieter

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Molecular & Cellular Proteomics : MCP. Elsevier. 2022, 21(10), 100278. ISSN 1535-9476. eISSN 1535-9484. Available under: doi: 10.1016/j.mcpro.2022.100278

Zusammenfassung

Most of the recombinant biotherapeutics employed today to combat severe illnesses, for example, various types of cancer or autoimmune diseases, are produced by Chinese hamster ovary (CHO) cells. To meet the growing demand of these pharmaceuticals, CHO cells are under constant development in order to enhance their stability and productivity. The last decades saw a shift from empirical cell line optimization toward rational cell engineering using a growing number of large omics datasets to alter cell physiology on various levels. Especially proteomics workflows reached new levels in proteome coverage and data quality because of advances in high-resolution mass spectrometry instrumentation. One type of workflow concentrates on spatial proteomics by usage of subcellular fractionation of organelles with subsequent shotgun mass spectrometry proteomics and machine learning algorithms to determine the subcellular localization of large portions of the cellular proteome at a certain time point. Here, we present the first subcellular spatial proteome of a CHO-K1 cell line producing high titers of recombinant antibody in comparison to the spatial proteome of an antibody-producing plasma cell-derived myeloma cell line. Both cell lines show colocalization of immunoglobulin G chains with chaperones and proteins associated in protein glycosylation within the endoplasmic reticulum compartment. However, we report differences in the localization of proteins associated to vesicle-mediated transport, transcription, and translation, which may affect antibody production in both cell lines. Furthermore, pairing subcellular localization data with protein expression data revealed elevated protein masses for organelles in the secretory pathway in plasma cell-derived MPC-11 (Merwin plasma cell tumor-11) cells. Our study highlights the potential of subcellular spatial proteomics combined with protein expression as potent workflow to identify characteristics of highly efficient recombinant protein-expressing cell lines. Data are available via ProteomeXchange with identifier PXD029115.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie

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subcellular proteomics, TMT, mass spectrometry, CHO cell line, plasma cell

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ISO 690KRETZ, Robin, Larissa WALTER, Nadja RAAB, Nikolas ZEH, Ralph GAUGES, Kerstin OTTE, Simon FISCHER, Dieter STOLL, 2022. Spatial Proteomics Reveals Differences in the Cellular Architecture of Antibody-Producing CHO and Plasma Cell-Derived Cells. In: Molecular & Cellular Proteomics : MCP. Elsevier. 2022, 21(10), 100278. ISSN 1535-9476. eISSN 1535-9484. Available under: doi: 10.1016/j.mcpro.2022.100278
BibTex
@article{Kretz2022-10Spati-59454,
  year={2022},
  doi={10.1016/j.mcpro.2022.100278},
  title={Spatial Proteomics Reveals Differences in the Cellular Architecture of Antibody-Producing CHO and Plasma Cell-Derived Cells},
  number={10},
  volume={21},
  issn={1535-9476},
  journal={Molecular & Cellular Proteomics : MCP},
  author={Kretz, Robin and Walter, Larissa and Raab, Nadja and Zeh, Nikolas and Gauges, Ralph and Otte, Kerstin and Fischer, Simon and Stoll, Dieter},
  note={Article Number: 100278}
}
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