Elevated endoplasmic reticulum pH is associated with high growth and bisAb aggregation in CHO cells
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View abstract on PubMed
Summary
This summary is machine-generated.Intracellular pH in Chinese hamster ovary (CHO) cell culture significantly impacts therapeutic protein quality. This study reveals endoplasmic reticulum pH (pH<sub>ER</sub>) changes during bioprocessing, directly correlating with protein aggregation.
Area Of Science
- Biotechnology
- Cell Biology
- Bioprocess Engineering
Background
- Chinese hamster ovary (CHO) cell bioprocesses are crucial for therapeutic protein production.
- Intracellular conditions, particularly pH, influence protein quality but are difficult to measure.
- Understanding intracellular pH dynamics is vital for optimizing bioprocesses.
Purpose Of The Study
- To investigate endoplasmic reticulum pH (pH<sub>ER</sub>) changes in Chinese hamster ovary (CHO) cells during bioprocessing.
- To determine the impact of lactate metabolism on pH<sub>ER</sub>, cellular redox, and product quality.
- To establish correlations between intracellular pH and therapeutic protein aggregation.
Main Methods
- Integration of a fluorescent pH-sensitive biosensor into a bispecific antibody (bisAb)-producing CHO cell line.
- Monitoring of pH<sub>ER</sub>, cellular redox, and product quality in fed-batch and perfusion bioreactors.
- Analysis of lactate metabolism's influence on intracellular parameters.
Main Results
- pH<sub>ER</sub> increased during exponential growth and decreased sharply in the stationary phase.
- A loss of cellular pH regulation was observed in the stationary phase, independent of redox profiles.
- Protein aggregate levels showed the strongest correlation with pH<sub>ER</sub>.
Conclusions
- Endoplasmic reticulum pH is a critical factor affecting therapeutic protein quality in CHO bioprocesses.
- pH<sub>ER</sub> dynamics provide new insights into protein aggregate formation.
- Monitoring and controlling pH<sub>ER</sub> can improve bioprocess optimization and cell engineering strategies.
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