Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system

Area of Science:

• Biotechnology
• Immunology
• Protein Engineering

Background:

• Recombinant human IgG is crucial for therapeutic applications.
• Enhancing antibody-dependent cell-mediated cytotoxicity (ADCC) is a key goal in antibody engineering.
• Current methods for producing enhanced IgG can be time-consuming.

Purpose of the Study:

• To develop a rapid method for producing recombinant human IgG with improved ADCC effector function.
• To investigate the effect of kifunensine on IgG glycosylation and FcgammaRIIIA binding.
• To optimize transient expression systems for enhanced antibody production.

Main Methods:

• Transient expression of recombinant human IgG in suspension-growing HEK-293F cells.
• Inhibition of N-linked glycosylation using the glycosidase inhibitor kifunensine.
• Assessment of IgG affinity for FcgammaRIIIA via ELISA and ADCC activity using peripheral blood mononuclear cells.

Main Results:

• A 7-day protocol for producing recombinant human IgG was established.
• Kifunensine treatment resulted in IgG with oligomannose-type glycans lacking core-fucose.
• Produced IgG demonstrated improved affinity for FcgammaRIIIA and an eightfold enhancement in ADCC.

Conclusions:

• Transient expression in HEK-293F cells with kifunensine provides a fast and effective route to engineer IgG with superior ADCC.
• This method yields IgG with enhanced FcgammaRIIIA binding, leading to significantly improved cell-mediated cytotoxicity.
• The technique offers a valuable tool for the rapid development of potent therapeutic antibodies.