Structure-based glycoengineering of interferon lambda 4 enhances its productivity and anti-viral potency

Area of Science:

• Immunology
• Virology
• Biochemistry

Background:

• Interferon lambda 4 (IFNλ4) is implicated in hepatitis C virus (HCV) infection.
• Poor in vitro expression limits clinical applications of IFNλ4.

Purpose of the Study:

• To enhance IFNλ4 expression and potency through glycoengineering.
• To develop IFNλ4 variants for potential clinical use in HCV treatment.

Main Methods:

• Structure-based analysis to identify N-glycosylation sites on IFNλ4.
• Introduction of de novo N-glycosylation in a mammalian cell line (Expi293F).
• Assessment of receptor binding, signaling, antiviral activity, and ISG induction of engineered variants.

Main Results:

• Engineered IFNλ4 variants showed improved expression and potency compared to the E. coli version (eIFNλ4).
• Receptor binding and signaling pathways of engineered variants were comparable to eIFNλ4.
• Enhanced antiviral activity and interferon-stimulated gene (ISG) induction were observed in engineered variants.

Conclusions:

• Glycoengineering successfully improved IFNλ4 expression and functionality.
• Engineered IFNλ4 variants hold promise for clinical applications in HCV and immunological research.