A versatile design platform for glycoengineering therapeutic antibodies

Area of Science:

• Biotechnology
• Protein Engineering
• Immunology

Background:

• Glycoengineering is crucial for therapeutic antibody development, primarily targeting constant domains.
• Atypical glycosylation in variable domains of immunoglobulin G (IgG) antibodies remains largely unexplored.
• Existing strategies often focus on canonical glycosylation sites, limiting therapeutic potential.

Purpose of the Study:

• To explore the potential of atypical glycosylation in antibody variable domains for therapeutic applications.
• To develop a novel glycoengineering strategy for modulating antibody functions.
• To create and characterize antibody glycovariants with enhanced properties.

Main Methods:

• Computational tools were used to design and install N-glycosylation consensus sequences into variable domains of monoclonal antibodies.
• Two interleukin-2-binding antibodies were engineered to create glycovariants.
• Glycosylation efficiency, antibody function, and in vivo half-life were assessed.

Main Results:

• Engineered glycovariants were successfully glycosylated at novel sites in variable domains without compromising native antibody function.
• Certain glycovariants demonstrated modulated biological activities compared to the parent antibody.
• A glycovariant with dual glycosylation sites showed improved in vivo half-life when combined with a sialic acid precursor.

Conclusions:

• A versatile glycoengineering strategy leveraging variable domain glycosylation was validated.
• This approach enables modulation of antibody activity and improvement of therapeutic efficacy.
• The findings support early-stage drug development for enhanced antibody therapeutics.