Modulating antibody N-glycosylation through feed additives using a multi-tiered approach

Area of Science:

• Biotechnology
• Biopharmaceutical Manufacturing
• Protein Engineering

Background:

• Glycosylation is a critical post-translational modification impacting protein function and stability.
• Controlling glycosylation is essential for producing high-quality biopharmaceuticals.
• Production media and feeds significantly influence protein glycosylation patterns.

Purpose of the Study:

• To evaluate the N-glycosylation-modulating effects of various compounds.
• To identify optimal modulator combinations for desired glycoforms.
• To develop a predictive model for glycosylation control.

Main Methods:

• Cultivation of Chinese hamster ovary cells producing IgG antibodies in fed-batch mode.
• One-factor-at-a-time screening to identify potent modulators.
• Response surface methodology in an Ambr® 15 bioreactor system to assess compound interactions.

Main Results:

• Significant modulation of N-glycosylation profiles was achieved.
• An up to eight-fold increase in specific glycoforms was observed.
• A predictive model for glycoform control was successfully developed.

Conclusions:

• Specific compounds effectively modulate N-glycosylation in recombinant antibody production.
• The developed model facilitates targeted glycoform engineering.
• This approach enhances control over critical quality attributes in biopharmaceutical manufacturing.