Journal
BIOTECHNOLOGY JOURNAL
Volume 13, Issue 4, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201700492
Keywords
Chinese Hamster Ovary cells (CHO); flow cytometry; global protein synthesis; O-propargyl-puromycin (OP-puro); production cell line engineering
Funding
- Austrian Science Fund [FWF: W1224, I2514]
- Christian Doppler Society
- Austrian Federal Ministry of Economy, Family, and Youth
- National Foundation for Research, Technology and Development
- Federal Ministry of Science, Research and Economy (BMWFW)
- Federal Ministry of Traffic, Innovation, and Technology (bmvit)
- Styrian Business Promotion Agency SFG
- Standortagentur Tirol
- Government of Lower Austria
- Business Agency Vienna through the COMET Funding Program
- Austrian Science Fund (FWF)
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Accurate measurement of global and specific protein synthesis rates is becoming increasingly important, especially in the context of biotechnological applications such as process modeling or selection of production cell clones. While quantification of total protein translation across whole cell populations is easily achieved, methods that are capable of tracking population dynamics at the single-cell level are still lacking. To address this need, we apply O-propargyl-puromycin (OPP) labeling to assess total protein synthesis in single recombinant Chinese hamster ovary (CHO) cells by flow cytometry. Thereby we demonstrate that global protein translation rates slightly increase with progression through the cell cycle during exponential growth. Stable CHO cell lines producing recombinant protein display similar levels of total protein synthesis as their parental CHO host cell line. Global protein translation does not correlate with intracellular product content of three model proteins, but the host cell line with high transient productivity has a higher OPP signal. This indicates that production cell lines with increased overall protein synthesis capacity can be identified by our method at the single-cell level. In conclusion, OPP-labeling allows rapid and reproducible assessment of global protein synthesis in single CHO cells, and can be multiplexed with DNA staining or any type of immunolabeling of specific proteins or markers for organelles.
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