However, continuous improvements of these platforms are desirable as technologies are evolving. The use of platforms for MAb production is well-established. Downstream MAb purification platforms commonly include a protein A-based purification step followed by one or two polishing steps to remove remaining impurities (Fig 1). Technology platforms (i.e., standard sets of unit operations, conditions, and methods applied to a given class of molecules) allow for efficient processing from research and development, through clinical phase trials, to the manufacturing of the final product. MAbs, as a class of molecules, exhibit many shared properties, which make them well-suited for a platform approach to downstream processing. Improved productivity upstream has put greater demand on downstream processing to address the high titers of MAbs in harvested cell culture fluids.

As a consequence, the antibody titers in upstream cell culture processes have dramatically increased. Representing about 36% of the total biopharmaceutical market, with an annual sales growth rate of approximately 10%, MAbs are the single largest class of biological drugs today (1), The rapidly growing demand for MAbs has triggered a need for an elevated manufacturing capacity. An introduction to highthroughput process development (HTPD) is also given, together with specific case studies of purification step development. This paper is a guide to the development of MAb purification platforms and gives an overview of GE Healthcare Life Sciences offering of process chromatography media (resins) for MAb purification processes. With increased MAb titers, the cell culture supernatant might contain an elevated number of impurities (e.g., aggregates) that need to be separated from the target molecule. Increasing product titers upstream can introduce challenges in downstream purification processes. As a result, more cost-effective, efficient, and flexible process purification solutions are of high priority for MAb manufacturers. All rights reserved.The use of monoclonal antibodies (MAbs) and MAb conjugates as biopharmaceuticals have increased over the last decade. In summary, this study offers insight into the mechanism of mAb aggregation in bind-elute CEX operations, and the in-depth understanding facilitates the development of robust CEX conditions for mAb purification.Īggregate formation Cation-exchange chromatography Hydrophobicity Monoclonal antibody Structural stability.Ĭopyright © 2020 Elsevier B.V. Results suggest that the mAb-accessible hydrophobic regions of the CEX resins affect the structural stability of the bound mAbs to various degrees, leading to differences in aggregate formation upon mAb elution. Finally, the hydrophobicity of the CEX resins (Capto SP ImpRes < Fractogel EMD SE Hicap < POROS XS) was measured using a fluorescence-based method to quantitatively characterize this resin property. The interplay among these protein- and resin-related factors, together with solution conditions, ultimately dictates the aggregate formation observed. In particular, resin hydrophobicity was shown to have a critical impact. The Tm differences (∆Tm DSC (Unbound minus Bound)) between the two states correlated with the severity of mAb aggregation in CEX operations, indicating the importance of both intrinsic mAb stability and resin properties. Using differential scanning calorimetry (DSC), the measured melting temperature of the bound mAbs (Tm DSC (Bound)) was 4.5 - 6.5☌ lower than that for the unbound mAbs (Tm DSC (Unbound)) in the same solutions. Then, mAb structural stability was further investigated in the bound state on CEX surfaces. Using differential scanning fluorimetry (DSF), the measured melting temperature (Tm DSF (Unbound)) decreased from 60.7 to 52.4☌ for mAb1 and 51.5 to 45.2☌ for mAb2 when lowering pH from 6.0 to 4.5. First, mAb structural stability was studied in solutions under CEX load conditions. To gain mechanistic understanding of this phenomenon, aggregate formation in bind-elute CEX for two therapeutic mAbs (IgG1 and IgG4) was examined on three CEX resins (Capto SP ImpRes, Fractogel EMD SE Hicap, and POROS XS). High molecular weight (HMW) aggregate formation of therapeutic monoclonal antibodies (mAbs) during cation-exchange chromatography (CEX) has been frequently observed, and can be a challenge for downstream purification.

Capto impres