Comparison of batch and continuous multi‐column protein A capture processes by optimal design

Abstract

Multi-column capture processes show several advantages compared to batch capture. It is however not evident how many columns one should use exactly. To investigate this issue, twin-column CaptureSMB, 3- and 4-column periodic counter-current chromatography (PCC) and single column batch capture are numerically optimized and compared in terms of process performance for capturing a monoclonal antibody using protein A chromatography. Optimization is carried out with respect to productivity and capacity utilization (amount of product loaded per cycle compared to the maximum amount possible), while keeping yield and purity constant. For a wide range of process parameters, all three multi-column processes show similar maximum capacity utilization and performed significantly better than batch. When maximizing productivity, the CaptureSMB process shows optimal performance, except at high feed titers, where batch chromatography can reach higher productivity values than the multi-column processes due to the complete decoupling of the loading and elution steps, albeit at a large cost in terms of capacity utilization. In terms of trade-off, i.e. how much the capacity utilization decreases with increasing productivity, CaptureSMB is optimal for low and high feed titers, whereas the 3-column process is optimal in an intermediate region. Using these findings, the most suitable process can be chosen for different production scenarios.

Microheterogeneity of therapeutic monoclonal antibodies is governed by changes in the surface charge of the protein

“…Correspondence: Prof. Alois Jungbauer, Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria E-mail: alois.jungbauer@boku.ac.at Abbreviations: ADCC, antibody dependent cellular cytotoxicity; ApHGE, analytical ionexchange chromatography with pH gradient elution; CEX, Cation exchange chromatography; EGFR, epidermal growth factor receptor; mAb, monoclonal antibody; DSC, differential scanning calorimetry; SPR, surface plasmon resonance ants it is still difficult to decide, how precisely a product produced by different methods must match [14][15][16][17][18]. Cation exchange chromatography (CEX) with pH gradient elution has repeatedly been reported as a suitable method for high-resolution separation of antibody variants exhibiting different surface charge characteristics [19][20][21][22].…”

“…Either a batch process is directly converted into a continuous process or new unit operations are used in continuous manufacturing. Without knowing the biological significance of the different vari-ants it is still difficult to decide, how precisely a product produced by different methods must match [14][15][16][17][18].…”

Truly continuous low pH viral inactivation for biopharmaceutical process integration

“…The biopharmaceutical industry is following other industries in moving from discrete batch operation to integrated continuous manufacturing, especially for high demand products, such as monoclonal antibodies (MAbs; Shukla, Wolfe, Mostafa, & Norman, 2017; Walsh, 2018). The drivers for continuous processing are many‐fold; process intensification and cost savings (Baur, Angarita, Müller‐Späth, Steinebach, & Morbidelli, 2016; Hummel et al, 2018; Pagkaliwangan, Hummel, Gjoka, Bisschops, & Schofield, 2018; Pollock, Coffman, Ho, & Farid, 2017) might emerge as the most obvious ones but steady‐state operation and thus better, more reproducible quality have also been associated with continuous biomanufacturing (Karst et al, 2017; Kaufman, Wasley, & Dorner, 1988; Walther et al, 2019). While upstream processing is ahead in this transition, where chemostat and perfusion reactors are commonly employed at the manufacturing scale (Arathoon & Birch, 1986; Shukla et al, 2017; Warnock & Al‐Rubeai, 2006), downstream operations have only in recent years started this transition.…”

Optimized Continuous Multicolumn Chromatography Enables Increased Productivities and Cost Savings by Employing More Columns

“…This increase in total residence time for the load configurations with more columns results in higher capacities in the same way that increasing the residence time on a single column by slowing the load flow results in higher capacities. However, there is a diminishing return on capacity as more and more columns are added to the load zone of a capture process as demonstrated by Pfister et al …”

“…The number of mixing cells, J is related to the hydrodynamic performance of the column and depends upon the column length, the particle diameter and the quality of the packing. The mass balance equation describing the solute partitioning of the species k between the liquid and the solid phases in a given cell j is the following : where is the volumetric flow rate and is the column volume. The extragranular porosity corresponds to the volume of liquid outside the particles over the column volume.…”