High throughput chromatography strategies for potential use in the formal process characterization of a monoclonal antibody

Abstract

High throughput experimental strategies are central to the rapid optimization of biologics purification processes. In this work, we extend common high throughput technologies towards the characterization of a multi-column chromatography process for a monoclonal antibody (mAb). Scale-down strategies were first evaluated by comparing breakthrough, retention, and performance (yields and clearance of aggregates and host cell protein) across miniature and lab scale columns. The process operating space was then evaluated using several integrated formats, with batch experimentation to define process testing ranges, miniature columns to evaluate the operating space, and comparison to traditional scale columns to establish scale-up correlations and verify the determined operating space. When compared to an independent characterization study at traditional lab column scale, the high throughput approach identified the same control parameters and similar process sensitivity. Importantly, the high throughput approach significantly decreased time and material needs while improving prediction robustness. Miniature columns and manufacturing scale centerpoint data comparisons support the validity of this approach, making the high throughput strategy an attractive and appropriate scale-down tool for the formal characterization of biotherapeutic processes in the future if regulatory acceptance of the miniature column data can be achieved. Biotechnol. Bioeng. 2016;113: 1273-1283. © 2015 Wiley Periodicals, Inc.

An impurity characterization based approach for the rapid development of integrated downstream purification processes

“…The product gradient elution data (both salt and pH gradients) were used in concert with this sequence information to determine the specific operating conditions for an initial process ( Figure 5). As can be seen in Figure 5, CCF was first conditioned to pH 5.0 before loading onto the Capto MMC capture step to maximize binding capacity and salt tolerance for this step (Petroff et al, 2016;Welsh et al, 2016). As can be seen in Figure 5, CCF was first conditioned to pH 5.0 before loading onto the Capto MMC capture step to maximize binding capacity and salt tolerance for this step (Petroff et al, 2016;Welsh et al, 2016).…”

“…We also chose to incorporate a moderate wash step into all of the bind-elute steps to further enhance the clearance of impurities. As can be seen in Figure 5, CCF was first conditioned to pH 5.0 before loading onto the Capto MMC capture step to maximize binding capacity and salt tolerance for this step (Petroff et al, 2016;Welsh et al, 2016). The wash condition was then set to pH 5.0, 500 mM NaCl based on the product and impurity gradient data.…”

Hybrid optimization of preparative chromatography for a ternary monoclonal antibody mixture

“…Separations with three or more similar compounds in the feed are often complex and therefore typically require considerably more optimization than the initial mAb capture step. Examples of such process development efforts to achieve sufficient clearance from antibody aggregates are reported in the literature [1][2][3][4][5] ; examples of ternary separations involving aggregates as well as other product-related impurities are less frequent. [6][7][8][9] Other examples of ternary mixtures comprising non-mAb proteins, namely BSA, ovalbumin, cytochrome C, and ribonuclease A, are commonly employed to study model-based optimization problems.…”

Development of pseudo‐linear gradient elution for high‐throughput resin selectivity screening in RoboColumn® Format

“…The range of possible applications for MCCs is broad and mostly includes sample preparation, dynamic binding capacity determination, establishing cleaning regimens, and optimization of step elution . Academic groups have proven early on that also complex separations are possible with this format employing pseudo‐linear gradients, and industry is recently following suit . Common high‐throughput technologies were extended towards the characterization of a multicolumn chromatographic purification, and the technique was found to make an appropriate scale‐down tool for the formal characterization of biotherapeutic processes …”

“…[2][3][4][5] Academic groups have proven early on that also complex separations are possible with this format employing pseudo-linear gradients, [6][7][8] and industry is recently following suit. 9 Common high-throughput technologies were extended towards the characterization of a multicolumn chromatographic purification, and the technique was found to make an appropriate scale-down tool for the formal characterization of biotherapeutic processes. 10 Process developers demand that screening results collected from gradient elution testing with MCCs on a LHS provide a meaningful basis for selection of resins or buffer conditions.…”