Extension of first principle elemental balancing soft-sensors by nonlinear reaction kinetics for increased robustness in bioprocess monitoring.

A first principle soft-sensor for biomass and substrate estimation in upstream bioprocessing based on the fusion of elemental balancing and nonlinear kinetics is presented. It aims to extend the validity range of well-established elemental balancing soft sensors to substrate saturated and overfeeding conditions that often occur in induced production phases. An experimental study with recombinant E.

Addressing raw material variability: In-line FTIR sugar composition analysis of lignocellulosic process streams.

For a sustainable economy, biorefineries that use second-generation feedstocks to produce biochemicals and biofuels are essential. However, the exact composition of renewable feedstocks depends on the natural raw materials used and is therefore highly variable. In this contribution, a process analytical technique (PAT) strategy for determining the sugar composition of lignocellulosic process streams in real-time to enable better control of bioprocesses is presented.

A Unique Response Behavior in the Dissolved Oxygen Tension in Minibioreactor Cultivations with Intermittent Feeding.

Intermittent bolus feeding for cultivations in minibioreactor systems (MBRs) profoundly affects the cell metabolism. Bolus feeding leads to temporal substrate surplus and transient oxygen limitation, which triggers the formation of inhibitory byproducts. Due to the high oxygen demand right after the injection of the substrate, the dissolved oxygen tension () signal exhibits a negative pulse.

Microbial technologies for biotherapeutics production: Key tools for advanced biopharmaceutical process development and control.

Current trends in the biopharmaceutical market such as the diversification of therapies as well as the increasing time-to-market pressure will trigger the rethinking of bioprocess development and production approaches. Thereby, the importance of development time and manufacturing costs will increase, especially for microbial production. In the present review, we investigate three technological approaches which, to our opinion, will play a key role in the future of biopharmaceutical production.

Monte Carlo-Based Error Propagation for a More Reliable Regression Analysis across Specific Rates in Bioprocesses.

During process development, bioprocess data need to be converted into applicable knowledge. Therefore, it is crucial to evaluate the obtained data under the usage of transparent and reliable data reduction and correlation techniques. Within this contribution, we show a generic Monte Carlo error propagation and regression approach applied to two different, industrially relevant cultivation processes.

Online estimation of changing metabolic capacities in continuous Corynebacterium glutamicum cultivations growing on a complex sugar mixture.

Model-based state estimators enable online monitoring of bioprocesses and, thereby, quantitative process understanding during running operations. During prolonged continuous bioprocesses strain physiology is affected by selection pressure. This can cause time-variable metabolic capacities that lead to a considerable model-plant mismatch reducing monitoring performance if model parameters are not adapted accordingly.

Establishing recombinant production of pediocin PA-1 in Corynebacterium glutamicum.

Bacteriocins are antimicrobial peptides produced by bacteria to inhibit competitors in their natural environments. Some of these peptides have emerged as commercial food preservatives and, due to the rapid increase in antibiotic resistant bacteria, are also discussed as interesting alternatives to antibiotics for therapeutic purposes. Currently, commercial bacteriocins are produced exclusively with natural producer organisms on complex substrates and are sold as semi-purified preparations or crude fermentates.

Advances in monitoring and control of refolding kinetics combining PAT and modeling.

Overexpression of recombinant proteins in Escherichia coli results in misfolded and non-active protein aggregates in the cytoplasm, so-called inclusion bodies (IB). In recent years, a change in the mindset regarding IBs could be observed: IBs are no longer considered an unwanted waste product, but a valid alternative to produce a product with high yield, purity, and stability in short process times. However, solubilization of IBs and subsequent refolding is necessary to obtain a correctly folded and active product.

Usage of Digital Twins Along a Typical Process Development Cycle.

Digital methods for process design, monitoring, and control can convert classical trial-and-error bioprocess development to a quantitative engineering approach. By interconnecting hardware, software, data, and humans currently untapped process optimization potential can be accessed. The key component within such a framework is a digital twin interacting with its physical process counterpart.

Noninvasive online monitoring of Corynebacterium glutamicum fed-batch bioprocesses subject to spent sulfite liquor raw material uncertainty.

In this study the use of a particle filter algorithm to monitor Corynebacterium glutamicum fed-batch bioprocesses with uncertain raw material input composition is shown. The designed monitoring system consists of a dynamic model describing biomass growth on spent sulfite liquor. Based on particle filtering, model simulations are aligned with continuously and noninvasively measured carbon evolution and oxygen uptake rates, giving an estimate of the most probable culture state.

Optimal process design space to ensure maximum viability and productivity in Penicillium chrysogenum pellets during fed-batch cultivations through morphological and physiological control.

Background: Biomass growth of Pencillium chrysogenum is characterised by a distinct pellet morphology consisting of compact hyphal agglomerates. Fungal pellets are advantageous in industrial process control due to rheological advantages but lead to biomass degradation due to diffusional limitations of oxygen and substrate in the pellet's core. Several fermentation parameters are known to affect key pellet characteristics regarding morphology, viability and productivity.

Generic Workflow for the Setup of Mechanistic Process Models.

Process models, consisting of transferable and applicable knowledge, can be used for various tasks such as process development and optimization, and for predicting and controlling critical process variables. In this regard, mechanistic process models, describing the mechanism of a system with a distinct model structure and characteristic parameters, are very promising.The development of a reliable and applicable model is usually the critical step, before model simulation and application show beneficial effects.

Probeless non-invasive near-infrared spectroscopic bioprocess monitoring using microspectrometer technology.

Real-time measurements and adjustments of critical process parameters are essential for the precise control of fermentation processes and thus for increasing both quality and yield of the desired product. However, the measurement of some crucial process parameters such as biomass, product, and product precursor concentrations usually requires time-consuming offline laboratory analysis. In this work, we demonstrate the in-line monitoring of biomass, penicillin (PEN), and phenoxyacetic acid (POX) in a Penicilliumchrysogenum fed-batch fermentation process using low-cost microspectrometer technology operating in the near-infrared (NIR).

Monitoring and control strategies for inclusion body production in E. coli based on glycerol consumption.

The Gram-negative bacterium E. coli is the host of choice for the production of a multitude of recombinant proteins in industry. Generally, cultivation is easy, media are cheap and a high product titer can be obtained.

Inclusion Body Bead Size in Controlled by Physiological Feeding.

The Gram-negative bacterium is the host of choice for producing a multitude of recombinant proteins relevant in the pharmaceutical industry. Generally, cultivation is easy, media are cheap, and a high product titer can be obtained. However, harsh induction procedures combined with the usage of IPTG (isopropyl β-d-1 thiogalactopyranoside) as an inducer are often believed to cause stress reactions, leading to intracellular protein aggregates, which are so known as so-called inclusion bodies (IBs).

Impact of Glycerol as Carbon Source onto Specific Sugar and Inducer Uptake Rates and Inclusion Body Productivity in E. coli BL21(DE3).

The Gram-negative bacterium is the host of choice for a multitude of used recombinant proteins. Generally, cultivation is easy, media are cheap, and a high product titer can be obtained. However, harsh induction procedures using isopropyl β-d-1 thiogalactopyranoside as inducer are often referred to cause stress reactions, leading to a phenomenon known as "metabolic" or "product burden".

Model-Based Methods in the Biopharmaceutical Process Lifecycle.

Model-based methods are increasingly used in all areas of biopharmaceutical process technology. They can be applied in the field of experimental design, process characterization, process design, monitoring and control. Benefits of these methods are lower experimental effort, process transparency, clear rationality behind decisions and increased process robustness.