Using hydrogen peroxide to prevent antibody disulfide bond reduction during manufacturing process

Area of Science:

• Biopharmaceutical Manufacturing
• Protein Chemistry
• Process Development

Background:

• Disulfide bond reduction in monoclonal antibodies (mAbs) during large-scale manufacturing leads to low molecular weight species, causing drug substance non-compliance.
• This reduction is linked to cellular reducing agents and low dissolved oxygen levels, particularly during harvest and pre-chromatography storage.
• Existing mitigation strategies include chemical inhibition, aeration, and chilling, but a simple, effective method is needed.

Purpose of the Study:

• To evaluate hydrogen peroxide as a strategy to prevent disulfide bond reduction in mAbs.
• To determine the optimal concentration of hydrogen peroxide to prevent reduction without causing unwanted oxidation.
• To assess the impact of hydrogen peroxide treatment on mAb product quality attributes.

Main Methods:

• Laboratory-scale studies were conducted using multiple immunoglobulin G (IgG) molecules.
• Hydrogen peroxide was added to clarified harvest or cell culture fluid.
• Optimal hydrogen peroxide concentrations were determined to balance reduction prevention and oxidation avoidance.

Main Results:

• Controlled addition of hydrogen peroxide effectively prevented antibody disulfide bond reduction.
• Optimized concentrations did not negatively affect critical product quality attributes.
• Hydrogen peroxide decomposes into water and oxygen, posing no downstream purification challenges.

Conclusions:

• Hydrogen peroxide is a powerful and simple tool for quenching reducing potential in harvested cell culture fluid.
• This method offers an effective strategy to maintain mAb quality by preventing disulfide bond reduction.
• The ease of use and minimal product impact make hydrogen peroxide a valuable addition to biopharmaceutical manufacturing processes.