Removing aggregates and product variants are key challenges in the development of purification processes for biotherapeutic proteins. Multi-modal resins, which contain both electrostatic and hydrophobic moieties, can be used to enhance the selectivity between monomer and aggregates over traditional IEX and HIC separations. In this study, we tested Capto Adhere and Nuvia cPrime for their ability to remove HMW species and 'clipped' variants from an Fc-fusion product.A range of kosmotropic and chaotropic molecules were tested for their ability to enhance the selective elution of intact monomer from these multi-modal resins. Using a series of high-throughput batch experiments performed in 96-well membrane plates, several promising eluents created selective windows where monomer could be separated from aggregates. Finally, the selective eluents identified in the parallel batch studies were evaluated and further optimized in a column setting to assess their ability to clear aggregates, host cell proteins and 'clipped' species in a single processing step.

Effect of mobile phase modifiers on the purification of an Fc-fusion protein using multi-modal chromatography

245th National Meeting (2013)

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