A CHO strain producing high-level human IL-6 with the 3' deletion construct.

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Culture

Background:

• Human interleukin 6 (IL-6) is a critical cytokine with therapeutic potential.
• Previous attempts to engineer cell lines for high IL-6 production faced limitations in yield and mRNA stability.
• Optimizing expression vectors and genetic constructs is key for efficient recombinant protein production.

Purpose of the Study:

• To develop a Chinese Hamster Ovary (CHO) cell strain with significantly enhanced production of human interleukin 6 (IL-6).
• To investigate the impact of deleting the 3' AU-rich region of IL-6 cDNA on mRNA stability and protein productivity.
• To establish a robust cell line for high-level biopharmaceutical manufacturing of IL-6.

Main Methods:

• Genetic engineering of CHO cells by introducing a modified human IL-6 cDNA construct.
• Deletion of the 3' AU-rich region from the IL-6 cDNA sequence.
• Cultivation of engineered CHO cells in batch and microcarrier cultures.
• Quantification of IL-6 mRNA stability and protein yield.

Main Results:

• A novel CHO cell strain was successfully established, exhibiting high-level production of human IL-6.
• The engineered IL-6 mRNA demonstrated enhanced stability due to the deletion of the 3' AU-rich region.
• Productivity was over 15 times higher compared to a control clone with intact IL-6 cDNA.
• The strain achieved IL-6 production levels of 30 µg/mL in batch culture and a continuous 20 µg/10^6 cells/day in microcarrier cultivation.

Conclusions:

• Deletion of the 3' AU-rich region in IL-6 cDNA is an effective strategy to enhance mRNA stability and protein productivity in CHO cells.
• The developed CHO cell strain represents a significant advancement for the high-level, cost-effective production of human IL-6.
• This engineered cell line holds promise for improved biopharmaceutical manufacturing processes and therapeutic applications of IL-6.