Demonstration of the use of windows of operation to visualize the effects of fouling on the performance of a chromatographic step

Abstract

The high resolution afforded by packed bed chromatography makes it an indispensable operation in the downstream processing of therapeutic molecules. Packed bed performance is however inherently susceptible to changes in feed stream characteristics and fouling processes. The impact of fouling is seldom considered during the early stages of bioprocess development which is concerned with the selection of purification conditions. Instead these are performed with rigorously clarified feeds. Under such conditions, chromatography is effectively treated as an isolated step, independent from its preceding unit operations. In this study, we demonstrate how windows of operation could be used to visualize the impact of changes in the preceding clarification step on the fouling response of a subsequent cation exchange capture step. Laboratory columns (2,5 and 12 cm height) were subjected to varying fouling challenges of Escherichia coli lysate containing different amounts of solids carried over from the previous step. Changes in trans-column pressure drop and breakthrough of the target protein (Fab') were monitored. The limits of operability of the resin were determined with respect to the process material's properties. This information was used to extract the parameters for the adsorption kinetics used in the general rate (GR) model to create windows of operation for manufacturing scale operation.

Fouling of an anion exchange chromatography operation in a monoclonal antibody process: Visualization and kinetic studies

“…Against this background, the characterization of fouling is an extremely important issue, but in spite of this, relatively few systematic investigations of packed bed chromatography column fouling have been reported in the literature. The conventional approach examines a range of performance indicators such as pressure drop profiles, dynamic capacity, and breakthrough curves where scaled down columns are repeatedly loaded with fouling material (Boushaba et al, 3 ; Bracewell et al, 4 ; Chau et al, 5 ; Shepard et al, 26 ). Rather than considering fouling directly, studies often focus on related subjects such as resin lifetime issues and CIP procedures (Muller-Spath et al, 23 ; Norling et al, 24 ).…”

Optimization‐based framework for resin selection strategies in biopharmaceutical purification process development

“…In the early stages of purification process development, different types of resins need to be tested at small scale (1.5–5000 µL) under various operating conditions, including different pH values, salt concentrations, and flow rates, to establish the resin most suited for process application at large scale . Platforms that have a capacity for high‐throughput screening (HTS) are commonly used to identify the most promising candidates for further investigation, in terms of key criteria of large scale purification, like yield, purity, and productivity . In HTS, the combination of robotic methods, parallel processing, and the miniaturization of bioprocess unit operations allows for a large number of potential process parameters to be examined within a short time, and also results in the generation of large amounts of data for evaluation.…”