Design of experiments applications in bioprocessing: Chromatography process development using split design of experiments

Abstract

Development of a chromatographic step in a time and resource efficient manner remains a serious bottleneck in protein purification. Chromatographic performance typically depends on raw material attributes, feed material attributes, process factors, and their interactions. Design of experiments (DOE) based process development is often chosen for this purpose. A challenge is, however, in performing a DOE with such a large number of process factors. A split DOE approach based on process knowledge in order to reduce the number of experiments is proposed. The first DOE targets optimizing factors that are likely to significantly impact the process and their effect on process performance is unknown. The second DOE aims to fine‐tune another set of interacting process factors, impact of whom on process performance is known from process understanding. Furthermore, modeling of a large set of output response variables has been achieved by fitting the output responses to an empirical equation and then using the parametric constants of the equation as output response variables for regression modeling. Two case studies involving hydrophobic interaction chromatography for removal of aggregates and cation exchange chromatography for separation of charge variants and aggregates have been utilized to illustrate the proposed approach. Proposed methodology reduced total number of experiments by 25% and 72% compared to a single DOE based on central composite design and full factorial design, respectively. The proposed approach is likely to result in a significant reduction in resources required as well as time taken during process development. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2730, 2019

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

“…In order to minimize experimental effort in upstream process development, the Design of Experiments (DoE) approach has also been applied to investigate the various process parameters in recombinant protein production (Papaneophytou and Kontopidis, 2012;Hanke and Ottens, 2014;Shekhawat et al, 2019). Statistical experiments using various DoE approaches, such as full factorial design, fractional factorial design, Taguchi orthogonal arrays, and the response surface methodology (RSM), were used for the optimization of media to enhance protein yield because various components are present in media that interact with each other.…”

“…Statistical experiments using various DoE approaches, such as full factorial design, fractional factorial design, Taguchi orthogonal arrays, and the response surface methodology (RSM), were used for the optimization of media to enhance protein yield because various components are present in media that interact with each other. In DOE, various process parameters can be changed in a set of experimental trials, and a small number of experiments are enough to decide the effect of the various parameters and to select the most important ones (Papaneophytou and Kontopidis, 2014;Kumar et al, 2019;Shekhawat et al, 2019). In one study, RSM was applied to develop a defined medium to enhance human interferon gamma production (Unni et al, 2019).…”

“…QbD is a manufacturing principle in which product quality is integrated into the manufacturing process. The QbD method has been implemented for the process development and analytical characterization of recombinant proteins, including mAbs (Pathak et al, 2014;Yu et al, 2014;Kumar et al, 2019;Narayanan et al, 2019a;Shekhawat et al, 2019). Using the QbD approach, the effect of various media components and process parameters in Fab production was studied, resulting in a 5-fold enhancement of the target protein titer as compared to the basal medium, thus demonstrating the efficacy of QbD (Kumar et al, 2019).…”

“…A process for the purification of scFv using mixed-mode chromatography was developed using DoE, and it was found that the optimized conditions enabled binding of the scFv to Capto Adhere TM below its theoretical pI, with the majority of HCPs in the flow-through (Sakhnini et al, 2019). A split DOE approach was successfully used in HIC to remove aggregates, and CEX was used to isolate charge variants and aggregates, resulting in a reduction of the total number of experiments by 25 and 72% compared to a single DoE based on CCD and FFD, respectively (Shekhawat et al, 2019).…”

“…Continuous bioprocessing has also emerged as a novel technique and has been used in both upstream and downstream process development as well as in manufacturing for therapeutic proteins (Subramanian, 2018). The implementation of Quality by Design (QbD) and process analytical technologies (PAT) tools has improved biopharmaceutical production strategies (Kornecki and Strube, 2018;Shekhawat et al, 2019). The framework of bioprocess modeling and control also offers robust control solutions and is advantageous for optimal bioprocess design (Baumann and Hubbuch, 2017).…”