Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO...

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Line Development

Background:

• Increasing demand for monoclonal antibodies (mAb) necessitates efficient manufacturing processes.
• Transcriptional gene silencing reduces recombinant protein expression and stability in cell lines.
• Ubiquitous chromatin opening elements (UCOE) have shown potential in improving antibody expression.

Purpose of the Study:

• To investigate the impact of UCOE integration into light chain, heavy chain, or both genes on antibody expression.
• To evaluate the effect of UCOE on expression stability during cell line development.
• To compare UCOE-containing vectors with non-UCOE vectors in Chinese hamster ovary (CHO) cells.

Main Methods:

• Construction of UCOE and non-UCOE vectors.
• Generation of stable Chinese hamster ovary (CHO) cell pools using different vector combinations.
• Analysis of antibody expression levels and stability over time.

Main Results:

• All UCOE cell pools exhibited higher antibody yields than non-UCOE cells.
• Optimal expression and stability were achieved with UCOE integrated into the heavy chain gene.
• High expression levels were maintained for over four months in UCOE heavy chain cell pools.

Conclusions:

• UCOE significantly enhances both the level and stability of antibody expression.
• Integrating UCOE into the heavy chain gene yields more stable CHO cell lines with higher production capacity.
• UCOE is a valuable tool for improving mAb manufacturing processes.