Technical and Economical Evaluation of Downstream Processing Options for Monoclonal Antibody (Mab) Production - Updated downstream processes can pave the way for increased productivity, now and in the

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Technical and Economical Evaluation of Downstream Processing Options for Monoclonal Antibody (Mab) Production
Updated downstream processes can pave the way for increased productivity, now and in the future.


BioPharm International



Figure 8. The stability of the "SuRe" ligand (GE Healthcare, shown in red) and classical Protein A ligand (rPrA, shown in green) following incubation in CHO cell lysate for 18h at pH 5.0.
A novel Protein A resin, MabSelect SuRe for second generation MAb capture, has been studied from various aspects directly related to robustness: stability of the matrix under cleaning in place (CIP) conditions (NaOH), stability of the ligand on exposure to feed, and interaction with various antibodies. The resin exhibits long-term stability to CIP regimes using sodium hydroxide (Figure 7) and the ligand offers protease stability significantly higher than the level of classical Protein A (Figure 8).10 While Protein A consists of five different subunits with different binding specificities, the novel ligand is comprised of four identical subunits. This aspect has been used to explain the observation of more homogeneous elution behavior for a wide selection of antibodies and Fc fusion proteins compared with a classic Protein A resin.11 The subsequent two chromatography steps have been successfully tested for their ability to remove remaining impurities to current acceptance levels.10

Productivity and Costs

Productivity and costs will vary between individual process designs developed by different groups testing the second generation process, depending on the specific antibody for which it is applied. Therefore, we have calculated a scenario with a 10,000-L fermentation volume containing 50 kg MAb (titre 5 g/L) using conservative assumptions for capacity and volumetric flow values comparing the first and second generation processes.4 The new reference process is designed to produce up to 50 kg of antibody per batch per day of processing time. The comparison indicates that it offers 45% lower operating costs calculated for downstream steps (harvesting to the final UF/DF step), and a reduction of time needed to process the MAb by 80% compared to the first generation process.9

Process Options

Process platforms described by different companies vary somewhat in the two post-Protein A steps. Compared with the design shown in Figure 7, some processes have a flow-through anion exchange step, usually with a Q ligand in position two. Others use hdrophobic interaction chromatography (HIC) or ceramic hydroxyapatite in one of the steps. In recent years, membrane adsorbers have been reported to be useful for the flow-through anion exchange step as an alternative to Q resin chromatography. Process step comparisons published for this step variant to date all use the first generation resins and are challenged by the second generation reference process described here (Table 1).9

Conclusion


Quick Recap
Modern downstream process technology allows processing of batch sizes equivalent to ton scale production per year. There is no reason to assume that future antibody drugs cannot be produced with technology that has been introduced in recent years. This finding is not a contradiction to the permanent need for improvements of process performance and economy. Solutions presented here may provide development teams with efficient options to manufacture MAbs and sufficient time to develop a third generation technology adaptable to manufacturing needs as they evolve in the future.

Günter Jagschies, Anna Grönberg, Tomas Björkman, Karol Lacki, and Hans J. Johansson work at GE Healthcare Bio-Sciences AB, Uppsala Sweden, +46.18.6120880, fax +46.70.6121247,


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