Prevention of Stirring-Induced Microparticle Formation in Monoclonal Antibody Solutions

Abstract

Monoclonal antibodies are being widely used for the treatment of various diseases. Microparticle formation in high-concentration protein solutions is a major problem during the manufacture of therapeutic monoclonal antibodies, because aggregation leads to fouling of aseptic filters and may lead to an immunogenic reaction in patients. We found that stirring using a traditional bottom-magnetic type stirrer results in extensive and sustained formation of 500-nm diameter protein microparticles arising from shear stress on protein molecules. The antibody solution stirred for only 5 min using this type of stirrer exhibited significant fouling of aseptic filter membranes. In contrast, a top-entering type stirrer did not lead to the formation of microparticles, and the solution did not exhibit membrane fouling even after 30 min of stirring. We conclude that a top-entering type stirrer is more suited for the manufacture of concentrated therapeutic monoclonal antibody solutions.

Influence of pH on Heat-Induced Aggregation and Degradation of Therapeutic Monoclonal Antibodies

“…Previously, we reported that the stirring-induced aggregation of antibodies can be prevented by replacing bottom-magnetic type stirrers with top-entering type stirrers. 18) And our preliminary experiments indicated that heat had the most significant effects on the aggregation and degradation of antibodies among heat, vibration, and freeze-thawing (not shown). So we analyzed the effects of heat which accelerates deterioration of antibodies during storage.…”

Quality by Design for Biopharmaceutical Drug Product Development

“…Protein solutions are susceptible to particulate formation when mixed for extended times with bottom mounted mixers (Ishikawa and Kobayashi 2010). Homogeneity studies, air entrainment studies, and product impact development/characterization studies are required.…”

Evaluation of the Aggregation States of Monoclonal Antibodies by Diverse and Complementary Methods

“…20) The mobile phase contained 20 mM sodium phosphate (pH 7.0) and 500 mM sodium chloride. The experimental conditions were as follows: injected protein, 20 mg; flow rate, 0.5 ml/min; detection wavelength, 215 nm; and analysis time, 30 min.…”

“…arated from dimers and trimers. 6,20) However, SEC is not applicable to the detection of much larger aggregates. The aggregates of anti-DNP antibodies were investigated by DLS measurements, which give information on the size distribution from monomers (10 nm) to large aggregates (1000 nm).…”