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Process characterization and viral clearance validation are pivotal studies in biopharmaceutical manufacturing, particularly for monoclonal antibodies (mAbs) produced in mammalian systems, which carry inherent risks of viral contamination.
As the demand for these therapeutics continue to grow, these studies have become increasingly important for ensuring product safety and regulatory compliance. However, conventional column packing and validation methods often create bottlenecks in downstream workflows, driving the need for more efficient and scalable solutions.
In a recent webinar, Matt Taylor, senior process development associate, Downstream Process Development at KBI Biopharma, discussed how pre-packed chromatography columns can provide reliable small-scale models. These formats fit seamlessly into process development workflows and reduce preparation time, freeing teams to work on value-added activities, while adhering to ICH Q5A (R2) guidelines.
Demonstrating feasibility
In the case study Taylor presented, KBI used OPUS® ValiChrom® Pre-packed Chromatography Columns (Repligen Corporation) to execute a scale-down model in viral clearance spiking studies for a clinically relevant mAb purification process entering Phase III studies. The goal was to verify the small-scale model for use in process characterization studies and a viral clearance study fully compliant with ICH Q5A (R2), using four viruses.
Scale-down models typically reduce column sizes by a factor of ≤ 1/700 and reduce product requirements, resin consumption, and viral input. The customizable OPUS ValiChrom Pre-packed Chromatography Columns are designed to match the OPUS production-scale columns in chromatographic efficiency and bed height.
In order to qualify a small-scale model, a series of chromatography runs is executed, maintaining scale-independent process parameters: column bed height, linear flow rates, mobile phase column volumes, buffer composition, pH and conductivity, load material composition, product collection criteria, and operational temperature. Comparability is assessed via qualitative and quantitative analyses.
Performing the study
Two small-scale models, 0.8 cm and 1.1 cm inner diameter columns, were assessed across three chromatography steps: one capture and two polishing operations. Each model underwent six replicate runs per unit operation, using material sourced from large-scale campaigns.
The comparability assessment was done through a qualitative comparison of the chromatographic profiles and a statistical analysis of process performance outputs. Performance outputs included eluate (or flowthrough) concentration, eluate (or flowthrough) volume, and step yield, in addition to a product quality (PQ) profile.
“Qualitatively, we’re looking for no split peaks or flowthrough peaks, no significant tailing or fronting on the elution peak and no significant product breakthrough prior to elution,” said Taylor. The quantitative analysis used a two one-sided test (TOST) analysis to assess whether two populations, in this case the large- and small-scale process data sets, meet a specified equivalence margin.
Reproducible results
Chromatographic purifications were completed and post-purification intermediates submitted for a PQ testing panel. Each small-scale set was determined to be acceptable based on the compilation of the large-scale dataset.
In one unit operation, eleven (1.1 cm ID) and twelve (0.8 cm ID) out of fifteen attributes demonstrated full equivalency. The on-the-edge remaining attributes were found to be practically insignificant or not at risk of failing the product specification. Similar results were observed in the other chromatography unit operations.
Once the model was qualified, mock spikes and viral clearance studies were performed at the 0.8 cm ID scale. “We were able to qualify both small-scale models. We chose the 0.8 cm ID for our viral clearance studies to lower material consumption,” Taylor said. All three chromatography steps were included in the viral clearance evaluation, with both new and aged resins. Duplicate evaluations were done for each virus, at each resin condition.
The data show that the process consistently generated reproducible results across the duplicate runs for each virus, exceeding the industry benchmark for a retroviral clearance and achieving suitable clearance for the other three viruses with both new and aged resins.
Enhancing productivity
Implementation of the scalable OPUS line provided benefits throughout the mAb manufacturing campaign. Pre-packed columns offer a simpler asset induction process, along with minimalization of packing activities and frequently bottlenecking documentation (packing protocols, IQ/OQ, document review). At a large scale, pre-packed columns can save up to 72 man hours per column.
At a small scale, utilization of the pre-packed columns for process characterization and viral clearance studies can save up to 116 man hours per unit operation. The flexibility of the OPUS line in column packing parameters and the precision in the packed bed heights ensure a consistent residence time and column loadings across all scales.
Additionally, outsourcing column packing alleviates consequences of an initial packing failure, permits a wider catalog of resin choices without reconfiguring packing protocols, and offers no operational discrepancies to a traditionally packed column.
