Online monitoring and control of upstream cell culture process using 1D and 2D‐LC with SegFlow interface

Abstract

The biopharmaceutical industry is transitioning from currently deployed batch‐mode bioprocessing to a highly efficient and agile next‐generation bioprocessing with the adaptation of continuous bioprocessing, which reduces capital investment and operational costs. Continuous bioprocessing, aligned with FDA's quality‐by‐design platform, is designed to develop robust processes to deliver safe and effective drugs. With the deployment of knowledge‐based operations, product quality can be built into the process to achieve desired critical quality attributes (CQAs) with reduced variability. To facilitate next‐generation continuous bioprocessing, it is essential to embrace a fundamental shift‐in‐paradigm from “quality‐by‐testing” to “quality‐by‐design,” which requires the deployment of process analytical technologies (PAT). With the adaptation of PAT, a systematic approach of process and product understanding and timely process control are feasible. Deployment of PAT tools for real‐time monitoring of CQAs and feedback control is critical for continuous bioprocessing. Given the current deficiency in PAT tools to support continuous bioprocessing, we have integrated Infinity 2D‐LC with a post‐flow‐splitter in conjunction with the SegFlow autosampler to the bioreactors. With this integrated system, we have established a platform for online measurements of titer and CQAs of monoclonal antibodies as well as amino acid analysis of bioreactor cell culture.

Multi‐dimensional technology – Recent advances and applications for biotherapeutic characterization

“…On the other hand, a 2D‐LC system with a post‐flow‐splitter in conjunction with the SegFlow autosampler integrated with a bioreactor was reported [63]. Protein A purification was now directly performed into the 1 st dimension of the online system and the 2 nd dimension allowed both IEX and SEC analyses to be carried out.…”

Perspective Chapter: Advanced Process Control and Automation with Special Focus on Emerging Continuous Bioprocessing

“…For online monitoring of upstream titer and nutrients as well as assessing upstream and downstream product quality of biopharmaceutical drugs with the option to perform timely process control, UPLC system interfaced with a process sample manager (PSM) can be utilized. A 1D-& 2D-LC system interfaced with an appropriate online sampling device is useful for various applications including online titer measurements, online product quality assessments and online quantitation of amino acid contents during upstream cell culture process [18]. In all cases, samples are withdrawn automatically from bioreactors through a FISP probe or equivalent device with appropriate filter discs to remove all debris from the bioreactor culture samples before introducing it into the HPLC/UPLC.…”

Advances in bioreactor control for production of biotherapeutic products

“…Guiding principles to apply MVDA on bioprocessing data have been reviewed in the literature (Rathore, 2014). In other related studies, applications of off‐gas analysis using magnetic sector mass spectrometer (Goh et al, 2020), 2D LC (Chemmalil et al, 2021) (discussed as Illustration 4), and on‐line UPLC (Chemmalil et al, 2020) have been successfully attempted. Another major advancement in regard to PAT implementation is the development of miniaturized PAT tools (Rowland‐Jones et al, 2021; Read et al, 2010a).…”

“…Other hurdles include a lack of communication protocols, limited flexibility in integrating upstream with downstream processes, lack of technical skills for troubleshooting problems or for making real-time decisions. ILLUSTRATION 4 Case Study-Process analytical technology for process control (Chemmalil et al, 2021) An interface has been developed by combining infinity 2D-LC, flow splitter, FISP probes, and SegFlow for online measurements of titre, amino acids, and quality attributes and to establish feedback control as shown in Figure 5. For evaluating product quality, eluate from 1 st dimension was injected into the 2 nd dimension to analyse the size and charge variant.…”