Implementing Process Analytical Technology for the Production of Recombinant Proteins in Escherichia coli Using an Advanced Controller Scheme

Abstract

The performance of a bioreactor in meeting process goals is affected by the microorganism used, medium composition, and operating conditions. A typical bioreactor uses a supervisory control and data acquisition (SCADA) system for control, and a combination of software and hardware tools for real‐time data analysis. However, when the process is disrupted by utility or instrumentation failure, typical process controllers may be unable to reinstate normal operating conditions before the cells in the reactor shift to unfavorable metabolic regimes. The objective of this study is to examine how the response of a controller affects process recovery when disruptive incidences occur under a process analytical technology (PAT) framework. The process used for this investigation is the production of lethal toxin‐neutralizing factor (LTNF) by Escherichia coli (E. coli), which is controlled by a decoupled input–output‐linearizing controller (DIOLC). The performance of the DIOLC is compared to a proportional integral derivative (PID) controller subjected to the same conditions. The disruptions are introduced manually and the effect of controller action on process recovery and LTNF synthesis is measured in terms of peak purity and concentration. It is observed that DIOLC performs better after reinstating operating conditions and results in a meaningful improvement in performance.

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

“…A novel approach based on the PAT initiative was also developed for on-line estimation of µ using in-situ dielectric spectroscopy . The PAT framework was also used during the production of Lethal Toxin-Neutralizing Factor (LTNF) by E. coli, which was controlled by a decoupled input-output linearizing controller (DIOLC) (Dalal et al, 2019). LC-MS metabolomics at three bioreactor scales (10 l, 100 l, and 1,000 l) were utilized to gain insight into the basal metabolic states of the CHO cell culture during fed-batch, and this was demonstrated as a useful technique to obtain physiological information on the cell culture state during a bioprocess, regardless of scale (Vodopivec et al, 2019).…”

Biosynthesis of Lactobionic Acid in Whey-Containing Medium by Microencapsulated and Free Bacteria of Pseudomonas taetrolens

“…Utilizing this freely available waste-water along with biodiesel industry waste- crude glycerol for bio-hydrogen production is being reported by Prakash et al (2018) [ 22 ]. The study of Kumar et al (2019) [ 23 ] details a potential approach to producing biofuels from agricultural biomass, the applicability of which can be improved through up‐scaling. In the present study, an immobilized enzyme cocktail containing laccase was isevaluated in regard to its ability to enhance the saccharification and fermentation processes by reducing the amount of phenolic compounds produced.…”

Multiobjective Optimization for Enhanced Production of Therapeutic Proteins in Escherichia coli: Application of Real-Time Dielectric Spectroscopy

“…The bioreactor operation is essentially the most important step in the recombinant protein production process, as the physiological changes in the biomass growth conditions are considered to be closely related to the product quantity and quality . Harvest of a batch is typically linked to the time of induction . Rather than using time as the control criterion, basing the harvest on biomass concentration is likely to result in more consistent process performance.…”