Engineering a New IFN-ApoA-I Fusion Protein with Low Toxicity and Prolonged Action

Area of Science:

• Biotechnology
• Immunotherapy
• Protein Engineering

Background:

• Recombinant human interferon alpha-2b (rIFN) is a key immunotherapy agent for viral and cancer treatment.
• High efficacy of rIFN is often limited by significant side effects, necessitating the development of less toxic variants.
• Improving the pharmacokinetic profile of rIFN is crucial for enhancing therapeutic outcomes.

Purpose of the Study:

• To engineer a novel chimeric protein by fusing rIFN with apolipoprotein A-I (ApoA-I).
• To assess the biological activity, cytotoxicity, and pharmacokinetic properties of the engineered rIFN-ApoA-I.
• To evaluate the potential of rIFN-ApoA-I as a safer and more effective immunotherapy agent.

Main Methods:

• Genetic engineering to create a fusion protein of rIFN and ApoA-I.
• Biosynthesis of the chimeric protein in Pichia pastoris.
• Purification using reverse-phase chromatography.
• In vitro assessment of antiviral activity against vesicular stomatitis virus and SARS-CoV-2.
• Cytotoxicity assays on Vero cells.
• Pharmacokinetic studies in mice following subcutaneous injection.

Main Results:

• The chimeric protein ryIFN-ApoA-I was successfully produced with a yield of 30 mg/L and purified to 95-97% purity.
• ryIFN-ApoA-I retained complete biological activity of IFN against viral models.
• The chimeric protein exhibited reduced cytotoxicity towards Vero cells and enhanced cell viability under viral challenge compared to commercial IFN-a2b.
• Pharmacokinetic analysis in mice revealed a 1.8-fold increase in the half-life of ryIFN-ApoA-I compared to ryIFN.

Conclusions:

• The fusion of rIFN with ApoA-I successfully generated a chimeric protein with preserved antiviral activity.
• The engineered ryIFN-ApoA-I demonstrates reduced cytotoxicity and improved pharmacokinetics, suggesting enhanced safety and efficacy.
• This novel chimeric interferon holds promise for improved antiviral and anticancer immunotherapies.