Expression of soluble and active interferon consensus in SUMO fusion expression system in E. coli

Area of Science:

• Biotechnology
• Protein Expression
• Recombinant Protein Production

Background:

• Soluble protein expression is crucial for improved protein production.
• Interferon consensus (IFN-con) is a potent therapeutic for hepatitis C but challenging to express in E. coli.
• IFN-con exhibits superior activity compared to other interferon alpha subtypes.

Purpose of the Study:

• To develop an efficient method for expressing soluble and active recombinant IFN-con in E. coli.
• To optimize the IFN-con gene sequence for E. coli expression.
• To achieve high purity and yield of active IFN-con for therapeutic applications.

Main Methods:

• Optimized IFN-con gene sequence for E. coli expression.
• Cloned the gene into the Champion™ pET SUMO expression vector.
• Expressed the SUMO-IFN-con fusion protein in the SHuffle™ E. coli strain.
• Purified recombinant IFN-con after SUMO fusion removal using RP-HPLC.

Main Results:

• Efficient expression of soluble SUMO-IFN-con fusion protein (86-88% soluble).
• Obtained approximately 50mg of highly pure (≥98%) recombinant IFN-con per liter of culture.
• Determined specific activity of 960×10^6 IU/mg, showing 2.8 times greater antiviral activity than IFN α-2a.

Conclusions:

• Developed a robust E. coli expression system for soluble IFN-con.
• Achieved high yield and purity of active IFN-con through a simple two-step purification.
• This method offers a more efficient alternative to traditional inclusion body production for IFN-con.