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A high-level and regulatable production system for recombinant glycoproteins using a human interferon-alpha
1Fujisaki Institute, Hayashibara Biochemical Laboratories, Inc., Okayama, Japan.
Gene
|July 8, 1994
Summary
A new expression system using human interferon-alpha promoter (pIFN) in BALL-1 cells efficiently produces human erythropoietin (EPO). This novel method yields high levels of bioactive EPO, offering a promising glycoprotein production system.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Developing efficient systems for producing therapeutic glycoproteins is crucial.
- Existing methods may have limitations in yield or processing.
Purpose of the Study:
- To construct and evaluate a novel expression vector (pIFP) for producing human erythropoietin (EPO) in human cells.
- To establish a robust system for high-level, inducible glycoprotein production.
Main Methods:
- Construction of the pIFP expression vector containing human EPO cDNA under the control of the pIFN promoter.
- Transfection of human B-cell leukemia-derived BALL-1 cells and establishment of EPO-producing cell lines.
- Stimulation with Sendai virus to induce EPO production and assessment of production levels and inducibility.
Main Results:
- Transformed BALL-1 cells produced high levels of human EPO (up to 872 IU/ml) with significant inducibility (67-fold).
- The exogenous pIFN showed higher activity than endogenous IFN-alpha.
- The 'in vivo cell proliferation method' enabled large-scale production of transformed cells.
- Produced EPO exhibited proper processing and a molecular mass consistent with CHO cell-derived EPO.
Conclusions:
- The combination of human pIFN, BALL-1 cells, and the hamster method provides an effective production system for bioactive human glycoproteins.
- This system demonstrates potential for the large-scale manufacturing of therapeutic proteins like EPO.