In this paper a generic process model for the simulation of various downstream processes for protein purification is presented. It consists of detailed, dynamic models for ion-exchange membrane ...adsorbers, ion-exchange and size-exclusion chromatography and ultra-/diafiltration. For the implemented ion-exchange membrane adsorber model, the steric mass action model has been applied. The isotherm parameters have been determined experimentally for bovine serum albumin (BSA) and immunoglobulin G (IgG). The comparison between simulation results of the ion-exchange membrane and experiments for two protein mixtures containing BSA and IgG shows satisfactory agreement. Two case studies based on the purification process of human serum albumin are performed to illustrate the influence of structural changes in the process flowsheet on the process performance. The simulation results show that the replacement of an anion-exchange chromatography column by an anion-exchange membrane adsorber leads to a significant reduction in overall process time at constant yield and purity. The case studies demonstrate the applicability of the generic process model for detailed process analysis.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
The authors provide their assessment of current challenges in strategic mine planning and some approaches for addressing them. Challenges covered are quantification of downstream processes and their ...integration into orebody models; spatial characterisation of geometallurgical variables and their integration into block models, scheduling and mine optimisation; spatial characterisation of new variables for new types of mining; flexibility in planning and design to manage risk and minimise its impact; IT infrastructure for rapid on-line data collection and processing. Most of these challenges require new types of data, variables, modelling and estimation methods. Foremost among new variable types are geometallurgical and dynamic rock mass characterisation variables. New types of data and data collection include rapid generation of very large amounts of on-line sensor data and the consequent need for rapid processing and modelling of these data. The paper includes examples of models and outputs to illustrate approaches and potential solutions.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
This paper focuses on a cradle-to-grave life cycle assessment (LCA) that evaluates the potential environmental impacts of the magnesium (Mg) front end auto parts over its full life cycle and ...highlights the impact of the upstream processes along the average supply-chain, auto parts manufacturing and assembly, and the downstream processes following the use stage and end-of-life value chain. Pathways for improving sustainability of magnesium use in automobiles through greening the supply chain processes, technology improvements including the end of life chain are also discussed.
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•Cysteine-modified PEG-K3PO4 ATPS was formulated for laccase purification.•Highly efficient 3D-printed micro-mixer and phase separator were fabricated.•Continuous ATPS boosts ...partition coefficient & extraction efficiency significantly.•Continuous/autonomous separation by computer vision guided interphase monitoring.•Enhanced partitioning of enzyme was depicted in low-abundance and real matrix.
Downstream processing of biomolecules, particularly therapeutic proteins and enzymes, presents a formidable challenge due to intricate unit operations and high costs. This study introduces a novel cysteine (cys) functionalized aqueous two-phase system (ATPS) utilizing polyethylene glycol (PEG) and potassium phosphate, referred as PEG-K3PO4/cys, for selective extraction of laccase from complex protein mixtures. A 3D-baffle micro-mixer and phase separator was meticulously designed and equipped with computer vision controller, to enable precise mixing and continuous phase separation under automated-flow. Microfluidic-assisted ATPS exhibits substantial increase in partition coefficient (Kflow = 16.3) and extraction efficiency (EEflow = 88 %) for laccase compared to conventional batch process. Integrated and continuous-flow process efficiently partitioned laccase, even in low concentrations and complex crude extracts. Circular dichroism spectra of laccase confirm structural stability of enzyme throughout the purification process. Eventually, continuous-flow microfluidic bioseparation is highly useful for seamless downstream processing of target biopharmaceuticals in integrated and autonomous manner.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
•Development and evaluation of three downstream process concepts.•Model-based solvent selection for efficient rhamnolipid extraction.•Evaluation of impact of pH value and phase ratio on liquid-liquid ...extraction.•Investigation of pH-shift precipitation and subsequent solid-liquid extraction.•Recoveries above 75 w% and purities above 98 w% were achieved.
Biosurfactants, such as rhamnolipids, are sustainable alternatives for petrochemically produced surfactants. Rhamnolipids can be produced based on sustainable carbon sources and merge promising properties like low toxicity, high biodegradability, and specific activities, even at extreme conditions. Despite these properties, which make rhamnolipids interesting for a wide range of applications, rhamnolipids could not be fully established in the market. One reason constitutes unreliable and cost intensive downstream processing. In the recent past, only limited effort was spent in the development, in-depth investigation, and optimisation of downstream processing concepts for rhamnolipids. Major focuses were set on production strain development and structural studies.
In the present contribution, this knowledge gap is addressed. Three downstream process concepts were gradually developed based on a model-based solvent screening. Influences of relevant process parameters of applied unit operations in respective downstream process concepts were evaluated with regard to the performance criteria recovery and purity. All developed and optimised downstream process concepts yielded recoveries of mono-rhamnolipids and 3-(3-hydroxyalkanoyloxy)alkanoic acids above 75 w% with purities above 98 w%.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
•Valorization of grape pomace by extracting natural pigments.•A new sustainable and economically feasible process for extracting anthocyanins.•Nicotinamide-based eutectic mixture was used to recover ...anthocyanins.•Stabilizing anthocyanins in SiO2 material approved in food and pharmaceutical sector.•Obtaining high-purity and thermal stable anthocyanins.
Concentrated in the skins of red grapes are the anthocyanins, the primary colorants responsible for the fruits' reddish-purple color. These colorants are recognized for their significant antioxidant properties and potent nutraceutical and pharmaceutical ingredients. Nevertheless, their widespread use is compromised by the (i) need for more efficient yet sustainable downstream processes for their recovery and (ii) by the challenges imposed by their poor stability. In this work, these drawbacks were overcome by applying eutectic solvents and stabilizing agents. Besides, the anthocyanins were successfully loaded into a solid host material (approved in both food and pharmaceutical sectors) based on silicon dioxide (SiO2, loading capacity: 1extract:7silica m/m). Summing up, with the process developed, the extraction yield (21 mganthocyanins.gbiomass−1) and the stability (under 55, 75, and 95 °C) of the recovered anthocyanins were over three times better than with the conventional process. Finally, the raw materials and solvents were recycled, allowing an economical and environmentally friendly downstream process.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Live virus vaccine (LVV) purification, employing chromatography, can be challenged by low binding capacities and elution yields. Alternatively, processes relying solely on enzymatic digestion steps ...and size-based membrane separations can be limited by suboptimal reduction of process related impurities and poorly scalable unit operations. Here, we demonstrate that the combination of flowthrough mode chromatography and an ultrafiltration/diafiltration (UF/DF) unit operation delivers a purification process for two different LVV candidates, V590 and Measles, expressed in adherent Vero cells. For V590, chromatography with mixed mode cation exchange resins returned final product yields of ∼50% and logarithmic reduction values (LRVs) of 1.7->3.4 and 2.5-3.0 for host cell DNA (hcDNA) and host cell proteins (HCPs), respectively. For Measles, chromatography with mixed mode anion exchange resins returned final product yields of ∼50% and LRVs of 1.6 and 2.2 for hcDNA and HCPs, respectively. For both V590 and Measles processing, the employed resins cleared a key HCP, fibronectin, which could foul the UF/DF unit operation, and thusly enabling it to further reduce HCPs and to formulate the final LVV products. This integrated purification process utilizes the complementary action of the two unit operations and its applicability across LVVs supports its consideration for their processing.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
•Evaluation and optimization of different nuclease alternatives.•MNase – the top performer nuclease– removes more than 60% of DNA.•Anion exchange chromatography is effective for extracellular ...vesicles purification.•Capto™ Q ImpRes – the best resin – has a recovery yield higher than 85%.•Isolation of purified MSC-derived extracellular vesicles, with contaminant levels below regulatory agencies's limits.
Extracellular vesicles (EVs) have emerged as a novel cell-free strategy with broad biomedical applications in disease diagnosis and prognosis as well as therapy. Moving EVs to the clinics will require the development of cost-effective manufacturing processes capable of delivering a high-quality product with high yield. In this work, we explore the purification of EVs from conditioned media derived from human mesenchymal stromal cell bioreactor cultures using a nuclease digestion followed by anion exchange (AEX) chromatography as a scalable purification platform. The performance of different nucleases was evaluated and optimized, with MNase being the top performer. Concerning the chromatographic step, all anion-exchange resins successfully isolated EVs, being Capto™ Q ImpRes the lead resin with recovery yields exceeding 85% concomitant with removal of more than 98% of proteins and host cell DNA. Overall, our results show that the purification platform proposed in this work is a viable alternative to the existing methods, capable of delivering highly purified EVs and able to meet the requirements imposed by the regulatory agencies for clinical trial testing, and that can contribute for the integration of EVs-based therapies in the clinic.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
