Scalable Production Research Articles

Rapid and Scalable Production of Functional SARS-CoV-2 Virus-like Particles (VLPs) by a Stable HEK293 Cell Pool.

At times of pandemics, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, the situation demands rapid development and production timelines of safe and effective vaccines for delivering life-saving medications quickly to patients. Typical biologics production relies on using the lengthy and arduous approach of stable single-cell clones. Here, we used an alternative approach, a stable cell pool that takes only weeks to generate compared to a stable single-cell clone that needs several months to complete. We employed the membrane, envelope, and highly immunogenic spike proteins of SARS-CoV-2 to produce virus-like particles (VLPs) using the HEK293-F cell line as a host system with an economical transfection reagent. The cell pool showed the stability of protein expression for more than one month. We demonstrated that the production of SARS-CoV-2 VLPs using this cell pool was scalable up to a stirred-tank 2 L bioreactor in fed-batch mode. The purified VLPs were properly assembled, and their size was consistent with the authentic virus. Our particles were functional as they specifically entered the cell that naturally expresses ACE-2. Notably, this work reports a practical and cost-effective manufacturing platform for scalable SARS-CoV-2 VLPs production and chromatographic purification.

Scalable fabrication of red Room-Temperature phosphorescent materials through molecular doping strategy for versatile applications

Room-temperature phosphorescence (RTP) materials renowned for their long emission lifetime have garnered tremendous research interest not only for elucidating their underlying mechanisms but also for exploring their potential applications in anti-counterfeiting, encryption, and bioimaging. Nevertheless, the RTP color of most organic materials has been limited to blue, green, and yellow. In this study, a series of red RTP materials have been successfully fabricated at a large scale through a straightforward molecular doping strategy. Remarkably, the doped materials exhibited red RTP at 622 nm and 678 nm with an ultralong lifetime of 373 ms and an efficiency of 5.21 %, which was among the best performances reported for organic RTP materials. The systematic investigation unveiled that the efficient RTP emanated from the robust interactions between the emitter and matrix molecules as well as the formation of D/A exciplexes. Furthermore, the red RTP materials could be easily manufactured into diverse forms, including fine powders, aqueous inks, and nanoparticles. Their multifunctional applications in fingerprint recognition, encryption, and live bioimaging were demonstrated. With distinctive advantages such as red RTP, ultralong lifetime, scalable production, and cost-effectiveness, these RTP materials pave the way for the extensive utilization of organic RTP materials in various engineering sectors.

Scalable production of efficient hybrid nanoadsorbent based on porous carbon nanoparticles and polyaniline nanofibers for removal of toxic methylene blue

Scalable production of efficient hybrid nanoadsorbent based on porous carbon nanoparticles and polyaniline nanofibers for removal of toxic methylene blue

Axially coordinated dual-atomic-site catalysts for nearly 100% peroxymonosulfate conversion to 1O2 in membrane filtration

Reactive membrane filtration, integrating membrane technology with catalytic processes, offers a promising avenue for efficiently generating and utilizing active species in environmental remediation. Herein, a dual-atom Co-Mn site achieved through axial coordination on N-doped graphene catalyst (sp-CoMn@NG) has been developed and applied in membrane filtration for the targeted removal of specific pollutants. Experimental investigations, alongside density functional theory (DFT) calculations, have substantiated that axial coordination can effectively reduce the adsorption energy and activation barrier of PMS, thereby facilitating nearly 100% conversion of PMS to 1O2 and enhancing catalytic performance. Mechanistic studies further elucidate the 1O2 production mechanism through a superoxide-mediated chain reaction at the axially coordinated Co-Mn center, as well as the inactivation mechanism of catalytic membranes via the addition reaction of 1O2 and phenolic-hydroxyl compounds. This research underscores the intelligent prefabrication of axial coordination in dual-atomic-site catalysts, offering an efficient approach for scalable 1O2 production in reactive membrane filtration.

Development of Efficient and Scalable Production Process of Analytical Grade Sodium Chloride at Laboratory Scale

Development of Efficient and Scalable Production Process of Analytical Grade Sodium Chloride at Laboratory Scale

Recent Advancements in Ascribing Several Platinum Free Electrocatalysts Pertinent to Hydrogen Evolution from Water Reduction.

The advancement of a sustainable and scalable catalyst for hydrogen production is crucial for the future of the hydrogen economy. Electrochemical water splitting stands out as a promising pathway for sustainable hydrogen production. However, the development of Pt-free electrocatalysts that match the energy efficiency of Pt while remaining economical poses a significant challenge. This review addresses this challenge by highlighting latest breakthroughs in Pt-free catalysts for the hydrogen evolution reaction (HER). Specifically, we delve into the catalytic performance of various transition metal phosphides, metal carbides, metal sulphides, and metal nitrides toward HER. Our discussion emphasizes strategies for enhancing catalytic performance and explores the relationship between structural composition and the performance of different electrocatalysts. Through this comprehensive review, we aim to provide insights into the ongoing efforts to overcome barriers to scalable hydrogen production and pave the way for a sustainable hydrogen economy.

Prospects of buffer layer optimization for electrodeposited CZTS-based thin-film solar cell using SCAPS-1D

Kesterite Cu2ZnSnS4 (CZTS) has recently attracted the intensive attention of researchers as a significant photovoltaic material for the scalable production of thin film solar cells. We have particularly focused on replacing the conventional CdS buffer layer with non-toxic and earth-abundant materials of zinc stannate (Zn2SnO[Formula: see text] in environmentally friendly thin-film solar cells based on CZTS. Zinc stannate (ZTO) with a wide bandgap energy of about 3.33[Formula: see text]eV can be a promising material to reduce photon absorption loss and improve the photovoltaic performance of the device. Thus, the absorber and buffer layers were practically prepared to determine the bandgaps of layers for citation in SCAPS-1D simulation program. We employed chemical methods to deposit the CdS and CZTS layers and succeeded to obtain a high-quality kesterite phase of CZTS. The common configuration of FTO/CZTS/buffer/ZnO/AZO was the basis of the simulations in which, the thicknesses of the absorber and buffer layers were optimized by using the SCAPS-1D. According to the outcome of the simulations, the ZTO buffer layer has a better performance than CdS.

Federated learning enables privacy-preserving and data-efficient dimension prediction and part qualification across additive manufacturing factories

A crucial part of quality control in additive manufacturing (AM) is the decision to accept or reject parts based on their dimensional accuracy. Machine learning (ML) models can learn nuanced relationships between process parameters and resulting geometries; however, obtaining large quantities of supervised learning data to train ML models incurs non-trivial costs. Relevant measurement data may be available at other factories or stations within the same factory but cannot be pooled together for conventional centralized learning (CL) due to privacy constraints. Here, we propose federated learning (FL) that uses private data and information from distributed sources to train ML models that predict part dimensions and inform part qualification, thereby allowing collaborative model building without compromising privacy. We manufacture and measure 405 parts having three different designs and distribute them across three to 45 factories. The performance of FL is evaluated when factories produce similar parts, parts of different designs, and parts of different qualities. When factories produce similar parts or parts of different designs, FL predicts part geometry within 10 µm of the CL benchmark and within 15 µm of the process capability limit, with as few as nine parts at each factory. FL also outperforms individual learning where factories train on their own data by up to 96% in geometry prediction, thereby providing a win-win paradigm for privacy-preserving collaborative learning. When factories produce parts of different qualities, FL outperforms individual factories only when they produce less than 15 parts locally. We show that FL performance is influenced by two factors: the quantity of data in the federation and the statistical homogeneity of data across participating factories. Overall, this research demonstrates the promise of FL for privacy-preserving, data-efficient quality control and offers key insights to design data federations in scalable AM production.

Molecularly Imprinted Polymer Biosensor Based on Nitrogen-Doped Electrochemically Exfoliated Graphene/Ti3 CNTX MXene Nanocomposite for Metabolites Detection.

Rapid and accurate quantification of metabolites in different bodily fluids is crucial for a precise health evaluation. However, conventional metabolite sensing methods, confined to centralized laboratory settings, suffer from time-consuming processes, complex procedures, and costly instrumentation. Introducing the MXene/nitrogen-doped electrochemically exfoliated graphene (MXene@N-EEG) nanocomposite as a novel biosensing platform in this work addresses the challenges associated with conventional methods, leveraging the concept of molecularly imprinted polymers (MIP) enables the highly sensitive, specific, and reliable detection of metabolites. To validate our biosensing technology, we utilize agmatine as a significant biologically active metabolite. The MIP biosensor incorporates electrodeposited Prussian blue nanoparticles as a redox probe, facilitating the direct electrical signaling of agmatine binding in the polymeric matrix. The MXene@N-EEG nanocomposite, with excellent metal conductivity and a large electroactive specific surface area, effectively stabilizes the electrodeposited Prussian blue nanoparticles. Furthermore, increasing the content of agmatine-imprinted cavities on the electrode enhances the sensitivity of the MIP biosensor. Evaluation of the designed MIP biosensor in buffer solution and plasma samples reveals a wide linear concentration range of 1.0 nM-100.0 μM (R2 = 0.9934) and a detection limit of 0.1 nM. Notably, the developed microfluidic biosensor offers low cost, rapid response time to the target molecule (10 min of sample incubation), good recovery results for detecting agmatine in plasma samples, and acceptable autonomous performance for on-chip detection. Moreover, its high reliability and sensitivity position this MIP-based biosensor as a promising candidate for miniaturized microfluidic devices with the potential for scalable production for point-of-care applications.

Development of a scalable recombinant system for cyclic beta-1,2-glucans production

BackgroundCyclic β-1,2-glucans (CβG) are bacterial cyclic homopolysaccharides with interesting biotechnological applications. These ring-shaped molecules have a hydrophilic surface that confers high solubility and a hydrophobic cavity able to include poorly soluble molecules. Several studies demonstrate that CβG and many derivatives can be applied in drug solubilization and stabilization, enantiomer separation, catalysis, synthesis of nanomaterials and even as immunomodulators, suggesting these molecules have great potential for their industrial and commercial exploitation. Nowadays, there is no method to produce CβG by chemical synthesis and bacteria that synthesize them are slow-growing or even pathogenic, which makes the scaling up of the process difficult and expensive. Therefore, scalable production and purification methods are needed to afford the demand and expand the repertoire of applications of CβG.ResultsWe present the production of CβG in specially designed E. coli strains by means of the deletion of intrinsic polysaccharide biosynthetic genes and the heterologous expression of enzymes involved in CβG synthesis, transport and succinilation. These strains produce different types of CβG: unsubstituted CβG, anionic CβG and CβG of high size. Unsubstituted CβG with a degree of polymerization of 17 to 24 glucoses were produced and secreted to the culture medium by one of the strains. Through high cell density culture (HCDC) of that strain we were able to produce 4,5 g of pure unsubstituted CβG /L in culture medium within 48 h culture.ConclusionsWe have developed a new recombinant bacterial system for the synthesis of cyclic β-1,2-glucans, expanding the use of bacteria as a platform for the production of new polysaccharides with biotechnological applications. This new approach allowed us to produce CβG in E. coli with high yields and the highest volumetric productivity reported to date. We expect this new highly scalable system facilitates CβG availability for further research and the widespread use of these promising molecules across many application fields.

Establishment of human hematopoietic organoids for evaluation of hematopoietic injury and regeneration effect

BackgroundHuman hematopoietic organoids have a wide application value for modeling human bone marrow diseases, such as acute hematopoietic radiation injury. However, the manufacturing of human hematopoietic organoids is an unaddressed challenge because of the complexity of hematopoietic tissues.MethodsTo manufacture hematopoietic organoids, we obtained CD34+ hematopoietic stem and progenitor cells (HSPCs) from human embryonic stem cells (hESCs) using stepwise induction and immunomagnetic bead-sorting. We then mixed these CD34+ HSPCs with niche-related cells in Gelatin-methacryloyl (GelMA) to form a three-dimensional (3D) hematopoietic organoid. Additionally, we investigated the effects of radiation damage and response to granulocyte colony-stimulating factor (G-CSF) in hematopoietic organoids.ResultsThe GelMA hydrogel maintained the undifferentiated state of hESCs-derived HSPCs by reducing intracellular reactive oxygen species (ROS) levels. The established hematopoietic organoids in GelMA with niche-related cells were composed of HSPCs and multilineage blood cells and demonstrated the adherence of hematopoietic cells to niche cells. Notably, these hematopoietic organoids exhibited radiation-induced hematopoietic cell injury effect, including increased intracellular ROS levels, γ-H2AX positive cell percentages, and hematopoietic cell apoptosis percentages. Moreover, G-CSF supplementation in the culture medium significantly improved the survival of HSPCs and enhanced myeloid cell regeneration in these hematopoietic organoids after radiation.ConclusionsThese findings substantiate the successful manufacture of a preliminary 3D hematopoietic organoid from hESCs-derived HSPCs, which was utilized for modeling hematopoietic radiation injury and assessing the radiation-mitigating effects of G-CSF in vitro. Our study provides opportunities to further aid in the standard and scalable production of hematopoietic organoids for disease modeling and drug testing.

Continuous-flow system for loading and refining metal-supported nanofibers toward scalable production

Nanocatalysts play a significant role in various valuable reactions, but their scalable production is constrained by the cumbersome and laborious washing in refining after loading active metals species. This work firstly presented a universal continuous-flow washing for simplifying the washing protocol based on the size-matching principle. With pore-size-selected filter membranes, excessive or weakly anchored metal species can be moved away during liquid flow, while the solid nanofibers were efficiently collected. Secondly, to break through the volume limitations of the mixing kettle and realize scalable production, a home-made integrated loading-washing device was well-designed and combined the loading and washing procedures at one time. This device allowed liquid to flow through the supports, where metal species were loaded on while the excessive metal species were carried away. The optimized protocol increased the catalyst output per unit time from 10 mg/h to 500 mg/h, without losing catalytic activity and stability. This work provides a fresh idea for achieving the scalable production of nanocatalysts from the perspective of simplifying the loading and refining.

Direct Ink Writing of Low-Concentration MXene/Aramid Nanofiber Inks for Tunable Electromagnetic Shielding and Infrared Anticounterfeiting Applications.

MXene inks offer a promising avenue for the scalable production and customization of printing electronics. However, simultaneously achieving a low solid content and printability of MXene inks, as well as mechanical flexibility and environmental stability of printed objects, remains a challenge. In this study, we overcame these challenges by employing high-viscosity aramid nanofibers (ANFs) to optimize the rheology of low-concentration MXene inks. The abundant entangled networks and hydrogen bonds formed between MXene and ANF significantly increase the viscosity and yield stress up to 103 Pa·s and 200 Pa, respectively. This optimization allows the use of MXene/ANF (MA) inks at low concentrations in direct ink writing and other high-viscosity processing techniques. The printable MXene/ANF inks with a high conductivity of 883.5 S/cm were used to print shields with customized structures, achieving a tunable electromagnetic interference shielding effectiveness (EMI SE) in the 0.2-48.2 dB range. Furthermore, the MA inks exhibited adjustable infrared (IR) emissivity by changing the ANF ratio combined with printing design, demonstrating the application for infrared anticounterfeiting. Notably, the printed MXene/ANF objects possess outstanding mechanical flexibility and environmental stability, which are attributed to the reinforcement and protection of ANF. Therefore, these findings have significant practical implications as versatile MXene/ANF inks can be used for customizable, scalable, and cost-effective production of flexible printed electronics.

Generation of stable suspension producer cell lines for serum-free lentivirus production.

The production of lentiviral vectors (LVs) pseudotyped with the vesicular stomatitis virus envelope glycoprotein (VSV-G) is limited by the associated cytotoxicity of the envelope and by the production methods used, such as transient transfection of adherent cell lines. In this study, we established stable suspension producer cell lines for scalable and serum-free LV production derived from two stable, inducible packaging cell lines, named GPRG and GPRTG. The established polyclonal producer cell lines produce self-inactivating (SIN) LVs carrying a WAS-T2A-GFP construct at an average infectious titer of up to4.64×107TU mL-1in a semi-perfusion process in a shake flask and can be generated in less than two months. The derived monoclonal cell lines are functionally stable in continuous culture and produce an average infectious titer of up to 9.38×107TU mL-1in a semi-perfusion shake flask process. The producer clones are able to maintain a productivity of>1×107TUmL-1day-1for up to 29 consecutive days in a non-optimized 5L stirred-tank bioreactor perfusion process, representing a major milestone in the field of LV manufacturing. As the producer cell lines are based on an inducible Tet-off expression system, the established process allows LV production in the absence of inducers such as antibiotics. The purified LVs efficiently transduce human CD34+ cells, reducing the LV quantities required for gene and cell therapy applications.

Three‐dimensional‐printed Ni‐based scaffold design accelerates bubble escape for ampere‐level alkaline hydrogen evolution reaction

AbstractAlkaline hydrogen evolution reaction (HER) for scalable hydrogen production largely hinges on addressing the sluggish bubble‐involved kinetics on the traditional Ni‐based electrode, especially for ampere‐level current densities and beyond. Herein, 3D‐printed Ni‐based sulfide (3DPNS) electrodes with varying scaffolds are designed and fabricated. In situ observations at microscopic levels demonstrate that the bubble escape velocity increases with the number of hole sides (HS) in the scaffolds. Subsequently, we conduct multiphysics field simulations to illustrate that as the hole shapes transition from square, pentagon, and hexagon to circle, where a noticeable reduction in the bubble‐attached HS length and the pressure balance time around the bubbles results in a decrease in bubble size and an acceleration in the rate of bubble escape. Ultimately, the 3DPNS electrode with circular hole configurations exhibits the most favorable HER performance with an overpotential of 297 mV at the current density of up to 1000 mA cm−2 for 120 h. The present study highlights a scalable and effective electrode scaffold design that promotes low‐cost and low‐energy green hydrogen production through the ampere‐level alkaline HER.

Scalable Bioreactor Production of an O2‐Protected [FeFe]‐Hydrogenase Enables Simple Aerobic Handling for Clean Chemical Synthesis

AbstractThe enzyme CbA5H, a [FeFe]‐hydrogenase from Clostridium beijerinckii, has previously been shown to survive exposure to oxygen, making it a promising candidate for biotechnological applications. Thus far [NiFe]‐hydrogenases are typically considered for such applications, due to the superior O2‐tolerance and therefore simplified enzyme handling. However, methods for production of [FeFe]‐hydrogenases are generally more successful than for other classes of hydrogenases, therefore in this work we focus on demonstrating scalable CbA5H production, and report results with active enzyme prepared in bioreactors (up to 10 L) with >20‐fold improvement in purified enzyme yield. We then go on to confirm excellent H2/H+‐cycling activity of the air‐purified protein, highlighting that CbA5H can be prepared and isolated without the need for complex and expensive infrastructure. Next, we demonstrate good stability of the air‐purified CbA5H both in solution assays, and as a heterogenous catalyst system when immobilized on a carbon support. Finally, we successfully implement this enzyme within previously demonstrated biotechnologies for flavin and NADH recycling, highlighting its relevance in chemical synthesis, and we demonstrate production of an important API precursor, 3‐quinuclidinol at >0.4 g scale in standard benchtop hydrogenation infrastructure, with >100,000 CbA5H turnovers over 18 operational hours.

One-pot synthesis of interconnected carbon-coated silicon nanosheets from clay minerals for high-performance lithium-ion battery anodes

The silicon nanosheet/carbon composite emerges as a promising anode material for the next-generation commercial lithium-ion batteries. However, complex, costly, and energy-intensive synthesis procedures impede its practical application. Herein, the carbon-coated silicon nanosheets (Si/C) have been successfully synthesized via a one-pot strategy, involving a modified magnesiothermic reduction of talc followed by a reaction with CO2. The distinct chemical composition and layered nanostructure of talc enable the retention of Si nanosheets during exothermic reactions without additional templates or heat absorbents. The method shows significant potential for the scalable production of Si/C owing to the abundant silicon source, sustainable carbon source, and straightforward protocol. The resulting Si/C displays a hierarchical porous structure, with both macropores and mesopores, and an interconnected network of carbon-coated Si nanosheets. These structural characteristics facilitate rapid Li+ diffusion and enable the material to accommodate the lithiation-induced mechanical strains. As a result, the Si/C electrode exhibits outstanding electrochemical performance, including a remarkable rate capability (with a specific capacity of 1220 mAh g−1 at 10.0 A g−1) and a high initial Coulombic efficiency of 88%. The work opens a new avenue for the development of Si/C composites from natural clay minerals through an economical and scalable strategy, which would contribute to the practical application of advanced Si-based anodes in lithium-ion batteries.

Green synthesis of highly active and recyclable chromium oxide nanocatalyst for biodiesel production from novel nonedible oil seeds

AbstractThis study explores the sustainable production of biodiesel from nonedible Phyllanthus maderaspatensis seed oil (highest oil content of 35%, FFA 0.87 mg/KOH), utilizing an innovative green synthesis approach for chromium oxide nanoparticles derived from the waste fruit parts of Aubergine for the very first time in the current work. In pursuit of alternatives to fossil fuels, our research underscores the environmental and socio‐economic benefits of biofuels, particularly in reducing greenhouse gas emissions. The optimized process yielded a 92% biodiesel conversion under conditions of a 9:1 methanol‐to‐oil ratio, 0.135 wt.% catalyst concentration, and a reaction duration of 150 min at 80°C. Comprehensive analysis techniques, including XRD, FTIR, SEM, EDX, Zeta analysis, differential reflectance spectroscopy (DRS), GC–MS, and NMR (1H, 13C), were employed to characterize the synthesized nanocatalyst and biodiesel product. The biodiesel's fuel properties, such as acid value, fire point, pour point, viscosity, kinematic density, sulfur content, and cloud point, were rigorously tested, demonstrating compliance with international standards (ASTM D‐6571, EN 14214, and GB/T 20828‐2007). The use of P. maderaspatensis seed oil, an economical and environmentally friendly feedstock, in conjunction with a cost‐effective nanocatalyst, presents a viable pathway for the sustainable and scalable production of biodiesel. This study contributes to the advancement of bioproducts for a sustainable bioeconomy by demonstrating an integrated approach to bioenergy production that leverages biotechnological innovations and addresses both environmental and socio‐economic dimensions.

Flower-shaped 2D crystals grown in curved fluid vesicle membranes

The morphologies of two-dimensional (2D) crystals, nucleated, grown, and integrated within 2D elastic fluids, for instance in giant vesicle membranes, are dictated by an interplay of mechanics, permeability, and thermal contraction. Mitigation of solid strain drives the formation of crystals with vanishing Gaussian curvature (i.e., developable domain shapes) and, correspondingly, enhanced Gaussian curvature in the surrounding 2D fluid. However, upon cooling to grow the crystals, large vesicles sustain greater inflation and tension because their small area-to-volume ratio slows water permeation. As a result, more elaborate shapes, for instance, flowers with bendable but inextensible petals, form on large vesicles despite their more gradual curvature, while small vesicles harbor compact planar crystals. This size dependence runs counter to the known cumulative growth of strain energy of 2D colloidal crystals on rigid spherical templates. This interplay of intra-membrane mechanics and processing points to the scalable production of flexible molecular crystals of controllable complex shape.

Scalable Fabrication of Ionic-Conductive Covalent Organic Framework Fibers for Capturing of Sustainable Osmotic Energy.

High-performance covalent organic framework (COF) fibers are demanded for an efficient capturing of blue osmotic power because of their excellent durability, simple integration, and large scalability. However, the scalable production of COF fibers is still very challenging due to the poor solubility and fragile structure of COFs. Herein, for the first time, it is reported that COF dispersions can be continuously processed into macroscopic, meter-long, and pure COF fibers using a wet spinning approach. The two presented COF fibers can be directly used for capturing of osmotic energy, avoiding the production of composite materials that require other additives and face challenges such as phase separation and environmental issues induced by the additives. A COF fiber exhibits power densities of 70.2 and 185.3 W m-2 at 50-fold and 500-fold salt gradients, respectively. These values outperform those of most reported systems, which indicate the high potential of COF fibers for capturing of blue osmotic energy.