Computational bioseparation process development

With the increase in computational power over the last decades, the use of modeling and simulation in process design for (petro)chemical industry has become common ground. Computational tools like ASPEN are standard in the design and operational analysis of (petro)chemical plants. However, in the bio pharmaceutical field, such modeling and simulation techniques are only recently being investigated for use and (potential) implementation. Being the workhorse of purification in the biopharmaceutical industry, chromatography is a good candidate for this modeling approach. Detailed mechanistic models describing chromatographic separation behavior are available, and software to simulate chromatography is becoming more and more available (i.e. DelftChrom, CADET, etc.). A bioseparation process normally consists of multiple chromatographic and conditioning steps, hence, an extreme large design space needs to be investigated. This may lead to prohibitive simulation times, even on state-of-the-art fast computers, when only mechanistic models are used. This presentation will show the implementation of a hybrid bioseparation process design approach using a combination of mechanistic models, artificial neural networks and high throughput experimentation for process development and optimization of the production of industrial relevant biologicals

Miniaturization of chromatographic process development: Achieving fast results with minimal costs

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Real-time process analytical technology: Fluorescent dye-based miniaturized sensor for aggregate detection

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pH-gradient chromatofocusing of proteins on a chip

We present a novel microfluidic system for the pH-gradient focusing of proteins with the integration of 16 parallel micro-mixers, a micro-column, and a multiplexer. In this work we successfully achieved the creation of 16 non-linear gradients and the generation of a solid-phase micro-column for the realization of anion exchange chromatography on a single chip. With the device we demonstrated the separation of a protein mixture of R-phycoerythrin and FITC-BSA based on pH-gradient chromatofocusing

Model-based process development for complex vaccine mixtures

The regulations, safety and purity demands are extremely high for vaccine processes and likewise reflected in process development time and cost. Reducing time-to-market is key for pharmaceutical companies, hence saving lives and money, and therefore the need raised for systematic, general and efficient process development strategies (Hanke & Ottens, 2014). Despite the tremendous variation between vaccine purification processes, platform processes for similar types of vaccines could aid to generally accelerate the process development and would be beneficial in terms of knowledge, resources, costs and regulatory aspect. High throughput process development (HTPD) approaches can be used to establish platform processes. HTPD combines high throughput technologies and statistical or mechanistic modeling in an efficient manner. In particular mechanistic models, that aim to describe the real process based upon physical processes occurring, can be of great merit to extend the level of process understanding and thereby support in making decision regarding the process design (Pirrung et al., 2019). Please click Download on the upper right corner to see the full abstract

Acceleration of vaccine development by improvement of process understanding - Analysis of the host cell proteome

While regulatory agencies require stringent product quality and safety to be upheld in biopharmaceutical products, today’s competitive biopharmaceutical market requires short process development times. The demand to accelerate especially the development of vaccines became obvious with the COVID-19 pandemic. By expanding process understanding with the use of process design tools the development time of the purification could be significantly shortened. High throughput experimentation (HTE) provides an automated experimentation platform, which minimizes the amount of used samples and saves experimental time. In this approach, HTE is used to acquire experimental data to regress parameters used as inputs for a chromatographic mechanistic model with the objective to establish an E. coli vaccine purification process development platform for a recombinant subunit vaccine. To provide a generic process development strategy that can be applied to novel antigens, the focus lies on the description of the adsorption behavior of the impurities such as host cell proteins (HCPs) during the capture step. Therefore our approach focuses on the present impurities, in specific the HCPs (Figure 1). When using the same E.coli strain the knowledge regarding the host cell proteins could be transferred to a new product. The first step is the identification of HCPs. Over a thousand HCPs are identified in the E.coli harvest sample investigated by means of mass spectrometry based proteomics. A database containing the properties of these proteins can provide assistance in the decision on chromatography resins suited for the purification process of a new developed antigen. Please click Download on the upper right corner to see the full abstract

Quantitative Determination of Glucose Transfer Between Cocurrent Laminar Water Streams in a H-Shaped Microchannel

To explore the applicability of a laminar fluid diffusion interface (LFDI) for the controlled feeding of microbioreactors, glucose diffusion experiments were carried out in a rounded H-shaped microstructure etched in a glass substrate. The diffusion channel of the microstructure had a length of 4 mm and a depth of 50 μm with a trapezoidal cross section with a width of 100 μm at the bottom and 200 μm at the surface of the channel. The microchannel was operated at residence times of less than 1 s ensuring high-mass-transfer rates. It was confirmed, both by microscopic observations as well as computational fluid dynamics (CFD) studies that the flow characteristics in the microchannel were fully laminar. Special attention was paid to flow splitting at the end of the channel, because the CFD simulations indicated that the performance of the device was sensitive to unequal flow splitting. The difference in outflow volume of the two streams was measured to be small (1.25% ± 0.6%). The measured glucose concentration in both exit ports at a fixed residence time was found to be stable in time and reproducible in multiple experiments. CFD simulation was shown to be a powerful tool for estimating the mass transfer in the LFDI, even at very short residence times. The results obtained in this work show the applicability of LFDI for the controlled diffusive supply of a solute to a water stream, with as possible application substrate and/or precursor feeding to microreactors

An adsorptive bioprocess for production and recovery of resveratrol with Corynebacterium glutamicum

BACKGROUND The growing interest in polyphenols has led to the design of industrial-scale processes able to produce them by fermentation and recover them in a more sustainable way. The goal of this work is to present two integrated approaches for the recovery of resveratrol, obtained through fermentation. The production of resveratrol using Corynebacterium glutamicum and its continuous removal using a hydrophobic resin is described. Batch production is compared with in situ product removal, where Amberlite XAD-7HP is either directly added to the medium (direct adsorption) or is present in an external column (external adsorption). RESULTS For both adsorption strategies tested, the amount of extracellular resveratrol increased from 75% to at least 90% of the total amount produced. However, lower total resveratrol concentrations were attained 3.6 and 2.2mg L-1, for the external and direct contact strategies, respectively, versus 5.3mg L-1 for batch experiments. CONCLUSIONS The proposed in situ removal strategies demonstrated the potential of increasing the excretion of resveratrol produced intracellularly. These process configurations may not only lead to a simpler downstream process design, but also to the avoidance of potential problems with the toxicity of polyphenols to the cells, especially when larger titers are obtained. © 2017 Society of Chemical IndustryWe would like to thank the European Union Framework Program 7 ‘BacHBerry’ (www.bachberry.eu), Project No. FP7- 613793 for financial support, the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, COMPETE 2020 (POCI-01- 0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145- FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio