Downstream Processing Research Articles

Microbial Biopolymers: From Production to Environmental Applications—A Review

Industrial evolution and agricultural expansion, explained by continuing population growth, have rendered enormous problems for the world throughout the past few decades, primarily because of waste generation. To reduce environmental impact and dependence on fossil fuels, scientists have explored replacing synthetic polymers with environmentally friendly and sustainable alternatives in many emergent applications. In this regard, microbial biopolymers have gained special attention. Many biopolymers originating from various strains of bacteria, fungi, and algae have been reported and their possible applications have increased rapidly. This review focuses on the wide range of microbial biopolymers, their characteristics, and factors influencing their production. The present study also describes the environmental applications of microbial biopolymers. The use of these biopolymers is very attractive as a value-added and sustainable approach to wastewater treatment. By acting as adsorbents, coagulants, and flocculants as well as filters in membrane processes, microbial biopolymers shine as promising solutions beyond conventional methods. They can be integrated into various stages of the treatment process, further enhancing the efficiency of wastewater treatment methods. Microbial biopolymer applications in bioremediation and soil stabilization are also reviewed. Several studies have demonstrated the strong potential of biopolymers in soil improvement due to their ability to minimize permeability, eliminate heavy metals, stabilize soil, and limit erosion. Challenges related to scaling up and the downstream processing of microbial biopolymers, as well as its future perspectives in environmental applications, are also discussed.

Physicochemical properties and stability of milk permeate as influenced by ultrafiltration processing parameters

The objective of this study was to determine the effect of membrane molecular weight cut-off (5 and 10 kDa), and filtration temperature (25, 15 and 10 °C) on the physicochemical properties of milk permeate (MP). Although MPs produced had a similar gross chemical composition, MP produced with the 10 kDa membrane at 25 °C had the lowest pH (6.64) and ionic calcium content (2.33 mm), while MP produced with the 5 kDa membrane at 10 °C had the highest levels of 6.82 and 2.85 mm for pH and ionic calcium, respectively. Incubation of MPs at 60 °C resulted in precipitation, with MPs produced with the 10 kDa membrane having larger, less soluble and thermodynamically more stable particles than MPs produced with the 5 kDa membrane. These results demonstrate that filtration parameters significantly affected the physicochemical properties of MPs, with implications for downstream processing.

Catalytic pyrolysis of torrefied biomass with molecular sieve catalysts to produce hydrocarbon rich biocrude

AbstractThe downstream processing of biocrudes obtained from direct biomass pyrolysis poses significant challenges due to stability issues, necessitating costly upgrading for further coprocessing with refinery feeds. This study examines the impact of torrefaction pretreatment on pyrolysis product distribution and biocrude composition using sawdust (SD) and groundnut shell (GS) feeds. Torrefaction was conducted at varying temperatures (200, 250 and 300°C) for 30 min under different reactor conditions. Increasing the severity of torrefaction resulted in decreased biocrude yields with reduced water content and gas formation, particularly evident with GS. A torrefaction temperature of 250°C and 30 min of pretreatment yielded higher phenolics and hydrocarbons. This increase in phenolics can be attributed to lignin enrichment during torrefaction, which, in the presence of a catalyst, undergoes deoxygenation leading to hydrocarbon formation. The influence of feed particle size, whether in powder or pellet form, on biocrude yield and composition was found to be minimal. Catalytic pyrolysis of SD using molecular sieve catalysts yielded the highest hydrocarbon (42%) and aromatic content (44%) at catalyst to biomass ratios of 1:1 and 2:3. The combination of torrefaction and pyrolysis was shown to enhance the quality of biocrude by increasing its hydrocarbon content, but at the expense of lower liquid yields. Experimental observations were supported by statistical analysis tools such as principal component analysis, which assessed pyrolysis product yields and composition.

Transformation of ABT-199 Nanocrystal Suspensions into a Redispersible Drug Product-Impact of Vacuum Drum Drying, Spray Drying and Tableting on Re-Nanodispersibility.

The present study compared vacuum drum drying (VDD) and conventional spray drying (SD) for solidifying crystalline ABT-199 nanosuspensions into redispersible oral drug products. The aim was to optimize formulation compositions and process conditions to maintain nanoparticle size after tablet redispersion. The impact of drug load (22%, 33%, 44%) and type of drying protectant (mannitol, mannitol/trehalose mix (1:1), trehalose) on redispersibility and material powder properties were investigated. Moreover, compression analysis was performed assessing the influence of compaction pressure on primary nanocrystal redispersibility and tablet disintegration. Higher drug loads and lower drying protectant levels resulted in particle growth, confirming a drug load dependence on redispersibility behavior. Notably, all drying protectants showed similar protection properties at properly chosen drying process parameters (Tg-dependent), except when VDD was used for mannitol formulations. Differences between the applied drying processes were observed in terms of downstream processing and tabletability: mannitol-containing formulations solidified via VDD showed an improved processability compared to formulations with trehalose. In conclusion, VDD is a promising drying technique that offers advantageous downstream processability compared to SD and represents an attractive novel processing technology for the pharmaceutical industry. As demonstrated in the present study, VDD combines higher yields with a leaner manufacturing process flow. The improved bulk properties provide enhanced tabletability and enable direct compression.

Data-Driven Modeling of the Spray Drying Process. Process Monitoring and Prediction of the Particle Size in Pharmaceutical Production.

Spray drying is used in the pharmaceutical industry for particle engineering of amorphous solid dispersions (ASDs). The particle size of the spray-dried (SD) powders is one of their key attributes due to its impact on the downstream processes and the drug product's functional properties. Offline and inline laser diffraction methods can be used to estimate the product's particle size; however, the final release of these ASDs is based on offline instruments. This paper presents a novel data-driven modeling approach for predicting the particle size of SD products. The model-based characterization of the process and the product's particle size, as a critical quality attribute, follows the quality by design principles. The resulting model can be used for online process monitoring, reducing the risks of out-of-specifications products and supporting their real-time release. A Tucker3 model is trained to capture and factorize the deterministic variability of the process. Subsequently, a partial least-squares regression model is calibrated to model the impact that variability in the input material properties, the process parameters, and the spray nozzle have on the products' particle size. This strategy has been calibrated and validated using large scale production data for two intermediate drug products under high sparsity of particle size data. Despite the challenges, high accuracy was obtained in predicting the median particle size (dv50) for release. The 99% confidence interval results in an error of maximum 2.5 μm, which is less than 10% of the allowed range of variation.

Fractionation of Extracellular Polymeric Substances by Aqueous Three-Phase Partitioning Systems.

Extracellular polymeric substances (EPS) are natural polymers secreted by microorganisms and represent a key chemical for the development of a range of circular economy applications. The production of EPS comes with notable challenges such as downstream processing. In this work, a three-phase partitioning (TPP) system was investigated as a fractionation technique for the separation of polysaccharides and proteins, both present in the EPS culture broth. The effect of the type of phase-forming compounds (alcohol, polymer, or ionic liquid, in combination with salt) and its concentration were evaluated and compared to the results previously obtained with model systems. The recyclability of phase-forming compounds used to form the fractionation platform was assessed by ultrafiltration. The best fractionation of EPS was achieved using a TPP system composed of 23 wt % ethanol and 25% K3C6H5O7 as 82% EPS-PS partitioned to the salt-rich/bottom phase, and 76% EPS-PN was recovered as an interfacial precipitate, which could be readily resolubilized in water. This represented an increase of 1.24 and 2.83-fold in the purity of EPS-PS and EPS-PN, respectively, in relation to the initial feed concentration. Finally, high recovery yields of phase-forming compounds (>99%) and fractionated EPS (>80%) were obtained using ultrafiltration/diafiltration (UF/DF) as the regeneration technique. The substantial fractionation yields, selectivity, and recyclability of the phase-forming compounds confirm the potential of TPP systems in combination with UF/DF as the separation method for real biopolymer mixtures. Key contributions of this study include the demonstration of the applicability of a readily scalable and cost-effective separation technique for the fractionation of EPS from real EPS-containing broths, while also the limitations of prescreening with model systems became clear through the observed deviating trends between model system studies and real broth studies.

Accelerate gene therapy downstream process development using scalable pre-packed chromatography columns

Accelerate gene therapy downstream process development using scalable pre-packed chromatography columns

Combination of an Optically Induced Dielectrophoresis (ODEP) Mechanism and a Laminar Flow Pattern in a Microfluidic System for the Continuous Size-Based Sorting and Separation of Microparticles.

Optically induced dielectrophoresis (ODEP)-based microparticle sorting and separation is regarded as promising. However, current methods normally lack the downstream process for the transportation and collection of separated microparticles, which could limit its applications. To address this issue, an ODEP microfluidic chip encompassing three microchannels that join only at the central part of the microchannels (i.e., the working zone) was designed. During operation, three laminar flows were generated in the zone, where two dynamic light bar arrays were designed to sort and separate PS (polystyrene) microbeads of different sizes in a continuous manner. The separated PS microbeads were then continuously transported in laminar flows in a partition manner for the final collection. The results revealed that the method was capable of sorting and separating PS microbeads in a high-purity manner (e.g., the microbead purity values were 89.9 ± 3.7, 88.0 ± 2.5, and 92.8 ± 6.5% for the 5.8, 10.8, and 15.8 μm microbeads harvested, respectively). Overall, this study demonstrated the use of laminar flow and ODEP to achieve size-based sorting, separation, and collection of microparticles in a continuous and high-performance manner. Apart from the demonstration, this method can also be utilized for size-based sorting and the separation of other biological or nonbiological microparticles.

Understanding the Manufacturing Process of Lipid Nanoparticles for mRNA Delivery Using Machine Learning.

Lipid nanoparticles (LNPs), used for mRNA vaccines against severe acute respiratory syndrome coronavirus 2, protect mRNA and deliver it into cells, making them an essential delivery technology for RNA medicine. The LNPs manufacturing process consists of two steps, the upstream process of preparing LNPs and the downstream process of removing ethyl alcohol (EtOH) and exchanging buffers. Generally, a microfluidic device is used in the upstream process, and a dialysis membrane is used in the downstream process. However, there are many parameters in the upstream and downstream processes, and it is difficult to determine the effects of variations in the manufacturing parameters on the quality of the LNPs and establish a manufacturing process to obtain high-quality LNPs. This study focused on manufacturing mRNA-LNPs using a microfluidic device. Extreme gradient boosting (XGBoost), which is a machine learning technique, identified EtOH concentration (flow rate ratio), buffer pH, and total flow rate as the process parameters that significantly affected the particle size and encapsulation efficiency. Based on these results, we derived the manufacturing conditions for different particle sizes (approximately 80 and 200 nm) of LNPs using Bayesian optimization. In addition, the particle size of the LNPs significantly affected the protein expression level of mRNA in cells. The findings of this study are expected to provide useful information that will enable the rapid and efficient development of mRNA-LNPs manufacturing processes using microfluidic devices.

Coexistence of state, choice, and sensory integration coding in barrel cortex LII/III

During perceptually guided decisions, correlates of choice are found as upstream as in the primary sensory areas. However, how well these choice signals align with early sensory representations, a prerequisite for their interpretation as feedforward substrates of perception, remains an open question. We designed a two alternative forced choice task (2AFC) in which male mice compared stimulation frequencies applied to two adjacent vibrissae. The optogenetic silencing of individual columns in the primary somatosensory cortex (wS1) resulted in predicted shifts of psychometric functions, demonstrating that perception depends on focal, early sensory representations. Functional imaging of layer II/III single neurons revealed mixed coding of stimuli, choices and engagement in the task. Neurons with multi-whisker suppression display improved sensory discrimination and had their activity increased during engagement in the task, enhancing selectively representation of the signals relevant to solving the task. From trial to trial, representation of stimuli and choice varied substantially, but mostly orthogonally to each other, suggesting that perceptual variability does not originate from wS1 fluctuations but rather from downstream areas. Together, our results highlight the role of primary sensory areas in forming a reliable sensory substrate that could be used for flexible downstream decision processes.

An Adsorption Based Downstream Processing Approach for Penicillin V from a Penicillium chrysogenum BIONCL I22 Culture Filtrate.

Penicillin V (phenoxy methyl penicillin) is highly sought after among natural penicillins because of its exceptional acid stability and effectiveness against common skin and respiratory infections. Given its wide-ranging therapeutic uses, there is a need to establish a greener method for its maximum recovery to reduce the carbon footprint. Here, we have identified and validated optimized operational conditions for resin-based penicillin V recovery. It was observed that Amberlite XAD4 had the highest penicillin V hydrophobic adsorption capacity among the other screened resins. Kinetic and isothermal studies using linear and nonlinear regression analysis showed that the adsorption process well fitted with pseudo-second-order kinetics (R 2 = 0.9816) and the Freundlich adsorption isotherm model (R 2 = 0.9871). Adsorption equilibrium was attained within 4 h, while maximum adsorption was observed at 3 mg/mL penicillin V concentration. Furthermore, the optimized extraction protocol was compared with the conventional butyl acetate-based downstream processing. Under optimum conditions resin-based penicillin V recovery was 2-fold higher as compared to the solvent extraction method and the resin could be reused for over six cycles without compromising the yield. These findings signify substantial progress toward the development of an environmentally sustainable approach for penicillin V recovery and a potentially viable method for extractive fermentation.

Process optimization and economic evaluation of continuous multi-column capture for monoclonal antibody – A case study

In recent years, due to significant improvements achieved in upstream processing of monoclonal antibody (mAb) manufacturing, the focus on productivity and cost-effectiveness has shifted towards downstream processing. Protein A affinity chromatography has been widely employed as the capture step for mAb purification and serves as a key driver of economic efficiency on downstream operations. To enhance the purification process and alleviate manufacturing constraints while reducing costs, continuous processing, such as multi-column capture (MCC), has gained attention. This study aimed to establish and optimize the protein A MCC process and to evaluate it with traditional single-column capture mode. Through a series of verification runs, it was determined that there was no significant difference in yield and quality attributes across different loops. Furthermore, annual cost of goods (COGs) was evaluated with the conventional single-column batch process. The results showed that MCC process reduced operation cost by 18 % and almost doubled productivity in the specific scenario.

DEVELOPING AN EXTRUDER MACHINE OPERATING SYSTEM THROUGH PLC PROGRAMMING WITH HMI DESIGN TO ENHANCE MACHINE OUTPUT AND OVERALL EQUIPMENT EFFECTIVENESS (OEE)

Designing a state-of-the-art PLC-based extrusion machine with a user-friendly HMI ensures seamless operation, enhancing Overall Equipment Effectiveness (OEE). This project focuses on automating an extrusion system with advanced technologies for optimized functionality and reliability. The architecture includes sophisticated components to boost productivity and product quality. Key aspects involve orderly control and synchronization of the extruder motor, feeder motor, lubrication pump, and vacuum pump for consistent performance with precise temperate profile. A significant innovation is the centralized blower system for machine temperature profile analysis and control, replacing individual controllers to enhance thermal management efficiency and ensure uniform temperature distribution. A high-low temperature alarm system alerts operators to deviations, maintaining process stability. Real-time data on current (Amps) and frequency (Hz) is displayed on the HMI from the inverter for monitoring and diagnostics. The system also features machine downline controlling capabilities for efficient management of downstream processes. Collectively, these innovations create a robust, efficient, and user-friendly extrusion machine that enhances OEE and product quality.

Numerical simulation of fish damage in a bulb tubular turbine with different guide vane openings

Abstract Tubular turbines have advantages in developing low-head hydropower, however, the development of hydropower shouldn’t forget to maintain a balance with ecology. In this paper, a three-dimensional model of the flow channel of a low-head power plant bulb tubular turbine unit is established using Unigraphics NX, and the hydraulic characteristics and fish damage mechanisms of the turbine under different guide vane openings of 45∼85 mm in rated operating conditions are simulated and analyzed using STAR-CCM+, and it is found that the fish mainly suffers from impact damage and pressure damage in the downstream process with the highest probabilities of 15.73% and 15.72%, respectively. Furthermore, with the increase of the opening of the guide vane, the probability of shear damage to the fish first decreases and then increases, while the probability of pressure, pressure gradient, and impact damage all gradually increase, and the probability of all four types of damage reaches the highest when the opening of the guide vane is 85 mm. Although the unit will have a large output at large opening operating conditions, from the point of view of fish protection, it is better for such turbines not to be operated with large opening of the guide vane for a long time.

Navigating the Solution to Drug Formulation Problems at Research and Development Stages by Amorphous Solid Dispersion Technology.

Amorphous Solid Dispersions (ASDs) have indeed revolutionized the pharmaceutical industry, particularly in drug solubility enhancement. The amorphous state of a drug, which is a highenergy metastable state, can lead to an increase in the apparent solubility of the drug. This is due to the absence of a long-range molecular order, which results in higher molecular mobility and free volume, and consequently, higher solubility. The success of ASD preparation depends on the selection of appropriate excipients, particularly polymers that play a crucial role in drug solubility and physical stability. However, ASDs face challenges due to their thermodynamic instability or tendency to recrystallize. Measuring the crystallinity of the active pharmaceutical ingredient (API) and drug solubility is a complex process that requires a thorough understanding of drug-polymer miscibility and molecular interactions. Therefore, it is important to monitor drug solids closely during preparation, storage, and application. Techniques such as solid-state nuclear magnetic resonance (ssNMR), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), Raman spectroscopy, and dielectric spectroscopy have been successful in understanding the mechanism of drug crystallization. In addition, the continuous downstream processing of drug-loaded ASDs has introduced new automated methods for consistent ASD production. Advanced techniques such as hot melt extrusion, KinetiSol, electro spraying, and electrospinning have gained popularity. This review provides a comprehensive overview of Amorphous Solid Dispersions (ASDs) for oral drug delivery. It highlights the critical challenges faced during formulation, the impact of manufacturing variables, theoretical aspects of drug-polymer interaction, and factors related to drug-polymer miscibility. ASDs have been recognized as a promising strategy to improve the oral bioavailability of poorly water-soluble drugs. However, the successful development of an ASD-based drug product is not straightforward due to the complexity of the ASD systems. The formulation and process parameters can significantly influence the performance of the final product. Understanding the interactions between the drug and polymer in ASDs is crucial for predicting their stability and performance.

Chinese State Capital as a Partner for Resource-Based Structural Transformation? The Belt and Road Initiative and Downstream Linkages in Bolivia and Kazakhstan

Lee argues that outbound Chinese state capital (CSC) is distinct from global private capital in terms of a greater willingness to accommodate recipient country priorities. This article uses the cases of Kazakhstan and Bolivia to explore this claim in relation to state-led efforts to foster structural transformation via upgrading in extractive industries (lithium and iron/steel in Bolivia, and petrochemicals in Kazakhstan). We focus particularly on attempts to move into domestic downstream processing. The article explores variation in the degree of accommodation with local demands on the part of CSC and proposes an explanatory framework for these differences, grounded in three axes. These are (i) mix of BRI drivers motivating a particular project from the Chinese side (export of industrial surplus, political relations with partner states and/or concern for resource security); (ii) nature of state-capital investment partnership (the mix of Chinese institutions and firms involved in negotiating and implementing the deal, as well as the time horizon implied by contract type); and (iii) a range of local contextual factors such as availability of alternative sources of capital, host state industrial capacities and local political conditions. Further research will be needed to refine and test this framework across other sectors and developmental goals beyond upgrading in extractives.

Mechanobiological insight into brain diseases based on mechanosensitive channels: Common mechanisms and clinical potential.

As physical signals, mechanical cues regulate the neural cells in the brain. The mechanosensitive channels (MSCs) perceive the mechanical cues and transduce them by permeating specific ions or molecules across the plasma membrane, and finally trigger a series of intracellular bioelectrical and biochemical signals. Emerging evidence supports that wide-distributed, high-expressed MSCs like Piezo1 play important roles in several neurophysiological processes and neurological disorders. To systematically conclude the functions of MSCs in the brain and provide a novel mechanobiological perspective for brain diseases. We summarized the mechanical cues and MSCs detected in the brain and the research progress on the functional roles of MSCs in physiological conditions. We then concluded the pathological activation and downstream pathways triggered by MSCs in two categories of brain diseases, neurodegenerative diseases and place-occupying damages. Finally, we outlined the methods for manipulating MSCs and discussed their medical potential with some crucial outstanding issues. The MSCs present underlying common mechanisms in different brain diseases by acting as the "transportation hubs" to transduce the distinct signal patterns: the upstream mechanical cues and the downstream intracellular pathways. Manipulating the MSCs is feasible to alter the complicated downstream processes, providing them promising targets for clinical treatment. Recent research on MSCs provides a novel insight into brain diseases. The common mechanisms mediated by MSCs inspire a wide range of therapeutic potentials targeted on MSCs in different brain diseases.

Streamlined Formation and Manipulation of Charged Polymersomes.

Charged polymersomes are attractive for advanced material applications due to their versatile encapsulation capabilities and charge-induced functionality. Although desirable, the pH-sensitivity of charged block copolymers adds complexity to its self-assembly process, making it challenging to produce charged polymersomes in a reliable manner. In this work, a flow approach to control and strike a delicate balance between solvent composition and pH for self-assembly is used. This allowsfor the identification of a phase window to reliably produce of charged polymersomes. The utility of this approach to streamline downstream processes, such as morphological transformation or in-line purification is further demonstrated. As proof-of-concept, it is shown that the processed polymersomes can be used for surface modifications facilitated by charge complexation.

Effect of Sc addition on downstream processing of twin-roll cast Al−Cu−Li−Mg−Zr-based alloys

A processing scheme for preparing strips of Al−Cu−Li−Mg−Zr-based alloys microalloyed with Sc was proposed. This scheme includes a twin-roll casting of strips with a near-net-shaped 3 mm-thick gauge and a homogenization/solution treatment processing step. Two twin-roll cast Al−Cu−Li−Mg−Zr-based alloys were studied, one with an addition of 0.17 wt.% Sc, and the other without addition of Sc. Both alloys were annealed at 300 °C for 30 min, 450 °C for 30 min, and 530 °C for 30 min and quenched into room temperature water. The first two annealing steps provide a fine dispersion of Al3Zr/Al3(Sc,Zr) particles. The final annealing step serves as homogenization/solution treatment. A fine crystallization structure of the twin-roll cast materials enables the dissolution of primary particles containing the main alloying elements during a relatively short annealing time. Shorter soaking duration limits the depletion of surface layers from Li and significant coarsening of Al3Zr/Al3(Sc,Zr) dispersoids. The Sc-containing material has significantly finer grains already in the as-cast state. The Al3(Sc,Zr) dispersoids formed during the homogenization/solution treatment step prevent grain coarsening at high temperatures. Strengthening θ′(Al2Cu), T1(Al2CuLi), and rare S’(Al2CuMg) phases are formed during subsequent artificial aging at 180 °C. The increase in yield strength was 200 MPa in the peak-aged conditions. This value is comparable to the ones reported in similar direct-chill cast materials, thus validating the proposed processing scheme.

Growth of solitary slugs in long pipes

We show that a pipe transporting gas and liquid can be operated at conditions where both stratified flow and slug flow are stable. In this bi-stable flow regime, it is possible to observe stratified flow or regular hydrodynamic slug flow, depending on the inlet conditions. However, it is also possible to get one single slug at a time in the pipe. The distinctive characteristic of such “solitary” slugs is that they grow continuously as they travel along the pipe and therefore, in long pipelines, they can reach lengths up to thousands of pipe diameters and flood the downstream process installations. We first show how the length of solitary slugs can be predicted using slug tracking simulations. Based on those transient simulation results, we provide an explanation for the slug growth and we propose a simple quasi-steady-state model allowing us to predict the slug length in good agreement with the transient simulations. Finally, we explain how we can determine the boundaries of the bi-stable flow regime in which solitary slugs occur.