High-yield expression and purification of human interferon alpha-1 in Pichia pastoris.

Abstract

For several years, interferon alpha-1, also known as interferon alpha-D, has been studied for treatment of various viral diseases, such as hepatic fibrosis caused by hepatitis B, herpes simplex virus keratitis, and bovine respiratory diseases in calves. Currently, recombinant human interferon alpha-D (rHuIFNalphaD) is expressed intracellularly in Escherichia coli or secreted by Bacillus subtilis and Saccharomyces cerevisiae. In this report, we describe the process of obtaining a relatively high-yield secretion of biologically active recombinant rHuIFNalphaD using the Pichia pastoris system. The process produced as high as 0.7 mg of purified protein per 20 ml of shake culture of rHuIFNalphaD with better bioactivity than the commercially available rHuIFNalphaD molecule produced in E. coli.

• Authors: P T Liu, T V Ta, L H Villarete
• Journal: Protein expression and purification (Protein Expr Purif) Vol. 22 Issue 3 Pg. 381-7 (Aug 2001) ISSN: 1046-5928 [Print] United States
• PMID: 11482999 (Publication Type: Journal Article)
• Copyright: Copyright 2001 Academic Press.

Chemical References

• Antiviral Agents
• Interferon-alpha
• Recombinant Proteins
• Methanol

Topics

• Amino Acid Sequence
• Antiviral Agents (isolation & purification)
• Base Sequence
• Biological Assay
• Cloning, Molecular
• Humans
• Interferon-alpha
• Methanol (metabolism)
• Molecular Sequence Data
• Pichia (genetics)
• Recombinant Proteins (biosynthesis)