A Direct Approach for Process Development Using Single Column Experiments Results in Predictable Streamlined Multi‐Column Chromatography Bioprocesses

Abstract

An emphasis on continuous monoclonal antibody (mAb) bioprocessing in the pharmaceutical industry necessitates effective approaches for downstream process development (PD). With a PD strategy, the process parameters are optimized to directly develop streamlined three‐step continuous chromatography processes. A design of experiment (DoE) approach with single column (batch mode) is used to simulate a multi‐column (continuous mode) purification method and characterize each chromatography step: Protein A capture, anion exchange, and mixed mode cation exchange. A novel and targeted approach to quickly characterize a DoE design space was employed and empirical modeling was used to define the capacity for multi‐column chromatography to accurately transfer the batch process to continuous mode of purification. This PD approach effectively mimics the continuous mode of operation and provides the flexibility to develop multiple continuous processes with target mAb recovery and purity. By implementing this PD strategy and the process parameters defined in batch mode, two robust and predictable continuous bioprocesses were developed within 7 weeks of investigation, which resulted in a total product yield of recovery at or above 74%, host cell protein (HCP) content below 5 ppm, and aggregate content below 1%.

Optimization of Vapor Diffusion Conditions for Anti-CD20 Crystallization and Scale-Up to Meso Batch

“…It is mostly used in autoimmune diseases therapy and it is effective against CD20-expressing non-Hodgkin's lymphoma, although it is not curative [18]. Similarly to other antibodies, the high production costs of anti-CD20 ($4000/g) [21] are mainly due to the usage of traditional purification methods such as protein-A chromatography, which is very effective in mAb recovery but very expensive due to the high cost of protein-A [22,23]. Conversely, crystallization is more cost-effective because it does not require expensive chemicals, while keeping the same purity level of chromatography [24].…”

Novel anion exchange membrane chromatography method for the separation of empty and full adeno‐associated virus

“…A similar step approach had proven suc-cessful before for the identification of an elution design space for a mAb from a mixed mode sorbent. [22] To optimize the separation of empty and full capsids a range of different conductivity steps was explored with different combinations of elution duration, 5 CV up to 40 CV, and conductivity increments, 0.5 mS cm -1 versus 1.5 mS cm -1 versus 2 mS cm -1 (data not shown). As judged by UV absorbance the most successful separations were achieved with 1 mS cm -1 steps each of 10 membrane volumes, Figure 2.…”

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…A DoE approach using a single column (batch mode) was studied to simulate a multicolumn (continuous mode) purification strategy with Protein A capture, anion exchange, and MM cation exchange, and robust and predictable continuous bioprocesses were developed. The process developed yielded total product recovery at or above 74%, HCP (<5 ppm) and an aggregate content below 1% (Utturkar et al, 2019).…”