loading page

Preferential precipitation for reducing acidic variant content in monoclonal antibody pools
  • +1
  • Tomasz Rumanek,
  • Michal Kolodziej,
  • Wojciech Piatkowski,
  • Dorota Antos
Tomasz Rumanek
Rzeszow University of Technology Faculty of Chemistry

Corresponding Author:[email protected]

Author Profile
Michal Kolodziej
Rzeszow University of Technology Faculty of Chemistry
Author Profile
Wojciech Piatkowski
Rzeszow University of Technology Faculty of Chemistry
Author Profile
Dorota Antos
Rzeszow University of Technology Faculty of Chemistry
Author Profile

Abstract

Microheterogeneity of mAbs can impact their activity and stability. Formation of charge variants is considered as the most important source the microheterogeneity. In particular, controlling the content of the acidic species is often a major importance for the production process and the regulatory approval of therapeutic proteins. In this study, the preferential precipitation process is developed for reducing the content of acidic variant in mAb downstream pools. The process design was preceded by determination of phase behavior of mAb variants in the presence of different precipitants. It is shown that the presence of polyethylene glycol (PEG) in protein solutions favors precipitation of acidic variants of mAbs. Precipitation yield is influenced by the variant composition in the mAb feed solutions, the concentration of the precipitant and the protein and the ionic strength of the solutions. To improve yield, the precipitate is repeatably dissolved and recycled to the precipitation process. The final product is as a mixture of supernatants pooled together from the recycling steps, in which the content of the acid variant is reduced. In addition, mAb aggregates present in the feed solutions accumulate in the precipitate., which is discarded as a waste. Such an approach can be potentially used either instead or in a combination with chromatography for adjusting the acidic variant content of mAbs, which can benefit in improvement in throughput and reduction in manufacturing costs.