Need for a next generation of chromatography models: academic demands for thermodynamic consistency and industrial requirements in everyday project work

Process chromatography modelling for process development, design, and optimization as well as process control has been under development for decades. Still, the discussion of scientific potential and industrial applications needs is open to innovation. The discussion of next-generation modelling approaches starting from Langmuirian to steric mass action and multilayer or thermodynamic consistent real and ideal adsorption theory or colloidal particle adsorption approaches is continued

Enabling total process digital twin in sugar refining through the integration of secondary crystallization influences

Crystallization is the main thermal process resulting in the formation of solid products and, therefore, is widely spread in all kinds of industries, from fine chemicals to foods and drugs. For these high-performance products, a quality by design (QbD) approach is applied to maintain high product purity and steady product parameters. In this QbD-context, especially demanded in the foods and drugs industry, the significance of models to deepen process understanding and moving toward automated operation is steadily rising. To reach these aspired goals, besides major process influences like crystallization temperature, other impacting parameters have to be evaluated and a model describing these influences is sought-after. In this work, the suitability of a population balance-based physico-chemical process model for the production of sugar is investigated. A model overview is given and the resulting model is compared to a statistical DoE scheme. The resulting process model is able to picture the effects of secondary process parameters, alongside temperature or temperature gradients, the influences of seed crystal size and amount, stirrer speed, and additives

PAT for continuous chromatography integrated into continuous manufacturing of biologics towards autonomous operation

This study proposes a reliable inline PAT concept for the simultaneous monitoring of different product components after chromatography. The feed for purification consisted of four main components, IgG monomer, dimer, and two lower molecular weight components of 4.4 kDa and 1 kDa molecular weight. The proposed measurement setup consists of a UV–VIS diode-array detector and a fluorescence detector. Applying this system, a R2 of 0.93 for the target component, a R2 of 0.67 for the dimer, a R2 of 0.91 for the first side component and a R2 of 0.93 for the second side component is achieved. Root mean square error for IgG monomer was 0.027 g/L, for dimer 0.0047 g/L, for side component 1 0.016 g/L and for the side component 2 0.014 g/L. The proposed measurement concept tracked component concentration reliably down to 0.05 g/L. Zero-point fluctuations were kept within a standard deviation of 0.018 g/L for samples with no IgG concentration but with side components present, allowing a reliable detection of the target component. The main reason inline concentration measurements have not been established yet, is the false-positive measurement of target components when side components are present. This problem was eliminated using the combination of fluorescence and UV–VIS data for the test system. The use of this measurement system is simulated for the test system, allowing an automatic fraction cut at 0.05 g/L. In this simulation a consistent yield of >99% was achieved. Process disturbances for processed feed volume, feed purity and feed IgG concentration can be compensated with this setup. Compared to a timed process control, yield can be increased by up to 12.5%, if unexpected process disturbances occur

Fast and versatile chromatography process design and operation optimization with the aid of artificial intelligence

Preparative and process chromatography is a versatile unit operation for the capture, purification, and polishing of a broad variety of molecules, especially very similar and complex compounds such as sugars, isomers, enantiomers, diastereomers, plant extracts, and metal ions such as rare earth elements. Another steadily growing field of application is biochromatography, with a diversity of complex compounds such as peptides, proteins, mAbs, fragments, VLPs, and even mRNA vaccines. Aside from molecular diversity, separation mechanisms range from selective affinity ligands, hydrophobic interaction, ion exchange, and mixed modes. Biochromatography is utilized on a scale of a few kilograms to 100,000 tons annually at about 20 to 250 cm in column diameter. Hence, a versatile and fast tool is needed for process design as well as operation optimization and process control. Existing process modeling approaches have the obstacle of sophisticated laboratory scale experimental setups for model parameter determination and model validation. For a broader application in daily project work, the approach has to be faster and require less effort for non-chromatography experts. Through the extensive advances in the field of artificial intelligence, new methods have emerged to address this need. This paper proposes an artificial neural network-based approach which enables the identification of competitive Langmuir-isotherm parameters of arbitrary three-component mixtures on a previously specified column. This is realized by training an ANN with simulated chromatograms varying in isotherm parameters. In contrast to traditional parameter estimation techniques, the estimation time is reduced to milliseconds, and the need for expert or prior knowledge to obtain feasible estimates is reduced

Artificial neural network for fast and versatile model parameter adjustment utilizing PAT signals of chromatography processes for process control under production conditions

Preparative chromatography is a well-established operation in chemical and biotechnology manufacturing. Chromatography achieves high separation performances, but often has to deal with the yield versus purity trade-off as the optimization criterium regarding through-put. The initial trade-off is often disturbed by the well-known phenomenon of chromatogram shifts over process lifetime, and has to be corrected by operators via adjustment of peak fraction cutting. Nevertheless, with regard to autonomous operation and batch to continuous processing modes, an advanced process control strategy is needed to identify and correct shifts from the optimal operation point automatically. Previous studies have already presented solutions for batch-to-batch variance and process control options with the aid of rigorous physico-chemical process modeling. These models can be implemented as distinct digital twins as well as statistical process operation data analyzers. In order to utilize such models for advanced process control (APC), the model parameters have to be updated with the aid of inline Process Analytical Technology (PAT) data to describe the actual operational status. This updating process also includes any operational change phenomena that occur, and its relation to their physico-chemical root cause. Typical phenomena are fluid dynamic changes due to packing breakage, channelling or compression as well as mass transfer and phase equilibrium-related separation performance decrease due to adsorbent aging or feed and buffer composition changes. In order to track these changes, an Artificial Neural Network (ANN) is trained in this work. The ANN training is in this first step, based on the simulation results of a distinct and previously experimentally validated process model. The model is implemented in the open source tool CasADi for Python. This allows the implementation of interfaces to process control systems, among others, with relatively low effort. Therefore, PAT signals can easily be incorporated for sufficient adjustment of the process model for appropriate process control. Further steps would be the implementation of optimization routines based on PAT and ANN predictions to derive optimal operation points with the model

Scalable mRNA machine for regulatory approval of variable scale between 1000 clinical doses to 10 million manufacturing scale doses

The production of messenger ribonucleic acid (mRNA) and other biologics is performed primarily in batch mode. This results in larger equipment, leaning/sterilization volumes, and dead times compared to any continuous approach. Consequently, production throughput is lower and capital costs are relatively high. Switching to continuous production thus reduces the production footprint and also lowers the cost of goods (COG). During process development, from the provision of clinical trial samples to the production plant, different plant sizes are usually required, operating at different operating parameters. To speed up this step, it would be optimal if only one plant with the same equipment and piping could be used for all sizes. In this study, an efficient solution to this old challenge in biologics manufacturing is demonstrated, namely the qualification and validation of a plant setup for clinical trial doses of about 1000 doses and a production scale-up of about 10 million doses. Using the current example of the Comirnaty BNT162b2 mRNA vaccine, the cost-intensive in vitro transcription was first optimized in batch so that a yield of 12 g/L mRNA was achieved, and then successfully transferred to continuous production in the segmented plug flow reactor with subsequent purification using ultra- and diafiltration, which enables the recycling of costly reactants. To realize automated process control as well as real-time product release, the use of appropriate process analytical technology is essential. This will also be used to efficiently capture the product slug so that no product loss occurs and contamination from the fill-up phase is <1%. Further work will focus on real-time release testing during a continuous operating campaign under autonomous operational control. Such efforts will enable direct industrialization in collaboration with appropriate industry partners, their regulatory affairs, and quality assurance. A production scale-operation could be directly supported and managed by data-driven decisions

Fast and flexible mRNA vaccine manufacturing as a solution to pandemic situations by adopting chemical engineering good practice: continuous autonomous operation in stainless steel equipment concepts

SARS-COVID-19 vaccine supply for the total worldwide population has a bottleneck in manufacturing capacity. Assessment of existing messenger ribonucleic acid (mRNA) vaccine processing shows a need for digital twins enabled by process analytical technology approaches in order to improve process transfer for manufacturing capacity multiplication, a reduction in out-of-specification batch failures, qualified personal training for faster validation and efficient operation, optimal utilization of scarce buffers and chemicals and speed-up of product release by continuous manufacturing. In this work, three manufacturing concepts for mRNA-based vaccines are evaluated: Batch, full-continuous and semi-continuous. Technical transfer from batch single-use to semi-continuous stainless-steel, i.e., plasmid deoxyribonucleic acid (pDNA) in batch and mRNA in continuous operation mode, is recommended, in order to gain: faster plant commissioning and start-up times of about 8–12 months and a rise in dose number by a factor of about 30 per year, with almost identical efforts in capital expenditures (CAPEX) and personnel resources, which are the dominant bottlenecks at the moment, at about 25% lower operating expenses (OPEX). Consumables are also reduceable by a factor of 6 as outcome of this study. Further optimization potential is seen at consequent digital twin and PAT (Process Analytical Technology) concept integration as key-enabling technologies towards autonomous operation including real-time release-testing

Digital twin of mRNA-based SARS-COVID-19 vaccine manufacturing towards autonomous operation for improvements in speed, scale, robustness, flexibility and real-time release testing

Supplying SARS-COVID-19 vaccines in quantities to meet global demand has a bottleneck in manufacturing capacity. Assessment of existing mRNA (messenger ribonucleic acid) vaccine processing shows the need for digital twins enabled by process analytical technology approaches to improve process transfers for manufacturing capacity multiplication, reduction of out-of-specification batch failures, qualified personnel training for faster validation and efficient operation, optimal utilization of scarce buffers and chemicals, and faster product release. A digital twin of the total pDNA (plasmid deoxyribonucleic acid) to mRNA process is proposed. In addition, a first feasibility of multisensory process analytical technology (PAT) is shown. Process performance characteristics are derived as results and evaluated regarding manufacturing technology bottlenecks. Potential improvements could be pointed out such as dilution reduction in lysis, and potential reduction of necessary chromatography steps. 1 g pDNA may lead to about 30 g mRNA. This shifts the bottleneck towards the mRNA processing step, which points out co-transcriptional capping as a preferred option to reduce the number of purification steps. Purity demands are fulfilled by a combination of mixed-mode and reversed-phase chromatography as established unit operations on a higher industrial readiness level than e.g., precipitation and ethanol-chloroform extraction. As a final step, lyophilization was chosen for stability, storage and transportation logistics. Alternative process units like UF/DF (ultra-/diafiltration) integration would allow the adjustment of final concentration and buffer composition before lipid-nano particle (LNP) formulation. The complete digital twin is proposed for further validation in manufacturing scale and utilization in process optimization and manufacturing operations. The first PAT results should be followed by detailed investigation of different batches and processing steps in order to implement this strategy for process control and reliable, efficient operation

Artificial Neural Network for Fast and Versatile Model Parameter Adjustment utilizin PAT signals of Chromatography Processes for Process Control under Production Conditions

Preparative chromatography is a well-established operation in the chemical and biotechnology manufacturing. Chromatography achieves high separation performances but often has to deal with the yield versus purity trade-off as the optimization criterium regarding through-put. The initial trade-off is often disturbed by the well-known phenomenon of chromatogram shifts over process lifetime and has to be corrected by operators via adjustment of peak fraction cutting. Nevertheless, with regards to autonomous operation and batch to continuous processing modes, any advanced process control strategy is needed to identify and correct shifts from the optimal operation point automatically. Previous studies already presented solutions for batch-to-batch variance and process control options with the aid of rigorous physico-chemical process model-ling. These models can be implemented as distinct digital twins as well as statistical process op-eration data analysers. In order to utilize such models for advanced process control, the model parameters have to be up dated with aid of inline PAT data to describe the actual operational status. Also including any occurring operational change phenomenon and its relation to their physico-chemical root cause. Typical phenomena are fluid dynamic changes due to packing breakage, channelling or compression as well as mass transfer and phase equilibrium related separation performance decrease due to adsorbent ageing or feed and buffer composition changes. In order to track these changes an Artificial Neural Network (ANN) is trained in this work. The ANN training is in this first step based on the simulation results of a distinct and pre-viously experimentally validated process model. The model is implemented in the open source tool CasADi for python. This allows the implementation of interfaces to e.g. process control systems with relatively low effort. Therefore, PAT signals can easily incorporated for the suffi-cient adjustment of the process model for appropriate process control. Further steps would be the implementation of optimization routines based on the PAT and ANN predictions to derive optimal operation points with the model