Search Results (131)

Megakaryocytes (MKs) have traditionally been viewed as terminal hematopoietic cells responsible solely for platelet production. However, recent advances in imaging and single-cell transcriptomics have revealed substantial heterogeneity among MK populations and diverse functions beyond thrombopoiesis. MKs actively participate in innate and adaptive immunity, modulate the hematopoietic stem cell (HSC) niche, and adapt to physiological and pathological stimuli. Located in distinct anatomical sites such as bone marrow and lung, MKs exhibit compartment-specific specializations that enable them to serve as critical integrators of hemostatic, immune, and regenerative processes. Experimental models using human pluripotent stem cells and inducible MKs have enhanced mechanistic insights, while innovative bioreactor platforms and xenotransplantation strategies advance translational applications in platelet production and therapy. Furthermore, immune MK subsets derived from pluripotent stem cells show promising therapeutic potential for modulating inflammation and autoimmune diseases. Continued exploration of MK diversity, tissue-specific roles, and intercellular communication will unlock new opportunities for leveraging MK plasticity in regenerative medicine, immunotherapy, and hematologic disorders, repositioning these versatile cells as central players in systemic homeostasis and defense. Full article

The increasing demand for monoclonal antibody (mAb) therapeutics has intensified the need for more efficient and consistent biomanufacturing processes. We present a data-driven, machine-learning (ML) approach to exploring and predicting upstream yield behavior. Drawing on industrial-scale batch records for a single mAb product from a contract development and manufacturing organization, we applied regression models to identify key process parameters and estimate production outcomes. Random forest regression, gradient boosting machine, and support vector regression (SVR) were evaluated to predict three yield indicators: bioreactor final weight (BFW), harvest titer (HT), and packed cell volume (PCV). SVR outperformed other models for BFW prediction (R² = 0.978), while HT and PCV were difficult to model accurately with the available data. Exploratory analysis using sequential least-squares programming suggested parameter combinations associated with improved yield estimates relative to historical data. Sensitivity analysis highlighted the most influential process parameters. While the findings demonstrate the potential of ML for predictive, data-driven yield improvement, the results should be interpreted as an exploratory proof of concept rather than a fully validated optimization framework. This study highlights the need to incorporate process constraints and control logic, along with interpretable or hybrid modeling frameworks, to enable practical deployment in regulated biomanufacturing environments. Full article

Osteoporosis is a bone disease characterized by low bone mass and changes in bone architecture, often leading to fractures and thereby decreased functional status in affected patients. About 200 million people worldwide suffer from osteoporosis, with women being affected earlier in life and more often than men. Various factors, such as genetic background, comorbidities, alcohol abuse, and medications such as glucocorticoids, are known to contribute to the development of osteoporosis. Due to the changing demographics, osteoporosis is becoming increasingly prevalent, and with this, the rate of fractures is expected to increase in the coming years. To investigate therapeutic options for treatment and to elucidate disease-causing mechanisms, various in vivo and in vitro osteoporosis models have been developed. In vivo models, in particular small animal models, remain the gold standard for osteoporosis research and the most used model to illustrate osteoporosis is the ovariectomized mouse. While in vivo models largely reflect the systemic and biological conditions, the transferability of findings to human patients is low and ethical concerns for laboratory animals must be considered. Thanks to tremendous technological improvements, such as on-a-chip platforms and high-end bioreactor systems, sophisticated in vitro models are of growing interest. These models offer the possibility of using complex cell systems, human cells from single donors, and 3D models, thus bridging the transferability gap, providing a platform for the introduction of personalized precision medicine, and ultimately replacing animal testing. Here, we summarize and discuss recent in vivo, in vitro, and in silico osteoporosis research approaches. Full article

Vanillin is the compound widely used in the food industry as a flavoring agent. Currently, chemically synthesized vanillin provides the majority of the world’s supply. Due to the increase in consumer awareness, there is a change in preferences towards natural food additives. The main goal of this research was to obtain vanillin through Solid-State Fermentation on agri-food by-products such as brewer’s spent grain, wheat bran, and linseed oil cake. A specially designed SSF culture single-use bag bioreactor made of a poliamide-6 foil sleeve was used to conduct the process on a bench-scale (600 g of dry medium). After extraction and purification, obtained vanillin samples were subjected to sensory analysis to determine whether the origin of microbiologically obtained vanillin affects its aromatic properties. The panelists assessed that the extracts obtained from the cultures of P. chrysosporium CBS246.84 and F. culmorum MUT5855 proved to be attractive flavors as they showed more attractive sensory properties than synthetic vanillin and were comparable to commercially available vanilla bean extract. This is the first study to include sensory analysis of vanillin obtained biotechnologically by the SSF method. Full article

Anaerobic digestion (AD) plays a crucial role in renewable energy production and waste management by converting organic waste into biogas and reduces greenhouse gas emissions. Optimized bioreactor performance depends on two main categories of factors: (1) reactor and geometric factors of agitator geometry, blade configuration, rotational speed, torque, power consumption, and the impeller-to-tank ration (d/D), and (2) fluid property factors of viscosity and flow characteristics, which relates turbulence, circulation patters, and stratification. Impeller power strongly influences nutrient distribution, gas exchange, and temperature uniformity within the reactor. While higher power inputs improve turbulence and prevent stratification, they also increase energy demand. This study evaluated fifteen blade configurations to determine the optimal fluid circulation using ANSYS 2024 R1 Fluent simulations. The bioreactor tank, with a diameter of 0.130 m and a height of 0.225 m, was tested at speeds ranging from 40 to 150 RPM. Among the single-blade configurations, the curved blade achieved the highest velocity at 0.521 m/s, generating localized circulations. The Rushton blade produced strong radial flows with a velocity of 0.364 m/s, while the propeller blade reached 0.254 m/s, supporting axial flow. In double-blade arrangements, the curved-propeller combination exhibited velocities between 0.261 and 0.342 m/s, enhancing fluid motion. The three-blade configurations resulted in the highest power consumption, ranging from 1.94 W to 1.99 W, with power increasing at higher RPMs and larger impeller sizes. However, torque values decreased over time. The most efficient mixing was achieved at moderate RPMs (80–120) and an impeller-to-tank diameter ratio (d/D) of approximately 0.75. These findings highlight the significance of blade selection in balancing mixing efficiency and energy consumption for scalable AD systems. Full article

Vanillin is the compound of great interest to the industry. It is used to augment and enhance the aroma and taste of food preparations and also as a fragrance compound in perfumes and detergents. Currently, majority of the world’s supply consists of chemically synthesized or lignin-derived vanillin. The application of biocatalysis for sustainable manufacturing of food ingredients, pharmaceutical intermediates, and fine chemicals is the key concept of modern industrial biotechnology. The main goal of this research was to conduct optimization procedures aimed at intensifying the microbial hydrolysis process of the lignin-rich plant raw materials and further bioconversion of the released ferulic acid to vanillin. The tests were performed in the solid-state fermentation system with strains selected during the screening stage on agri-food by-products such as brewer’s spent grain. A specially designed single-use bag bioreactor was used to carry out the process on a preparative scale with the most effective strain. The experiment was designed using the RSM, which allowed for an increase in biosynthesis efficiency from 363 mg/kg to 1413 mg/kg (an increase of 389%). The progress of the process was controlled by the use of chromatographic techniques (HPLC) by quantitative determination of vanillin content in the obtained extracts. Full article

The biopharmaceutical industry is undergoing a fundamental transformation from traditional batch manufacturing to continuous manufacturing (CM) for recombinant drugs and biosimilars, driven by regulatory support through the International Council for Harmonization (ICH) Q13 guidance and compelling economic advantages. This comprehensive review examines the technical, economic, and regulatory aspects of implementing continuous manufacturing specifically for recombinant protein production and biosimilar development, synthesizing validated data from peer-reviewed research, regulatory sources, and global implementation case studies. The analysis demonstrates that continuous manufacturing offers substantial benefits, including a reduced equipment footprint of up to 70%, a 3- to 5-fold increase in volumetric productivity, enhanced product quality consistency, and facility cost reductions of 30–50% compared to traditional batch processes. Leading biomanufacturers across North America, Europe, and the Asia–Pacific region are successfully integrating perfusion upstream processes with connected downstream bioprocesses, enabling the fully end-to-end continuous manufacture of biopharmaceuticals with demonstrated commercial viability. The regulatory framework has been comprehensively established through ICH Q13 guidance and region-specific implementations across the FDA, EMA, PMDA, and emerging market authorities. This review provides a critical analysis of advanced technologies, including single-use perfusion bioreactors, continuous chromatography systems, real-time process analytical technology, and Industry 4.0 integration strategies. The economic modeling presents favorable return-on-investment profiles, accompanied by a detailed analysis of global market dynamics, regional implementation patterns, and supply chain integration opportunities. Full article

Membrane fouling remains a major challenge in full-scale membrane bioreactor (MBR) systems, reducing operational efficiency and increasing maintenance needs. This study introduces a predictive and analytic framework for membrane fouling by integrating artificial intelligence (AI)-driven feature engineering and explainable AI (XAI) using real-world data from an MBR treating food processing wastewater. The framework refines the target parameter to specific flux (flux/transmembrane pressure (TMP)), incorporates chemical oxygen demand (COD) removal efficiency to reflect biological performance, and applies a moving average function to capture temporal fouling dynamics. Among tested models, CatBoost achieved the highest predictive accuracy (R² = 0.8374), outperforming traditional statistical and other machine learning models. XAI analysis identified the food-to-microorganism (F/M) ratio and mixed liquor suspended solids (MLSSs) as the most influential variables affecting fouling. This robust and interpretable approach enables proactive fouling prediction and supports informed decision making in practical MBR operations, even with limited data. The methodology establishes a foundation for future integration with real-time monitoring and adaptive control, contributing to more sustainable and efficient membrane-based wastewater treatment operations. However, this study is based on data from a single full-scale MBR treating food processing wastewater and lacks severe fouling or cleaning events, so further validation with diverse datasets is needed to confirm broader applicability. Full article

Background: Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-immortalised cells) are already compromised in antiviral pathways, the redundancy of innate signaling complicates host cell optimization by genetic engineering. Small molecules that are hypothesized to target antiviral pathways (Viral Sensitizers, VSEs) added to the culture media offer a versatile alternative to genetic modifications to increase permissiveness and, thus, viral yields across multiple cell lines. Methods: To explore how the yield for a chimeric Zika vaccine candidate (YF-ZIK) could be further be increased in an intensified bioprocess, we used spin tubes or an Ambr15 high-throughput microbioreactor system as scale-down models to optimize the dosing for eight VSEs in three host cell lines (AGE1.CR.pIX, BHK-21, and HEK293-F) based on their tolerability. Results: Addition of VSEs to an already optimized infection process significantly increased infectious titers by up to sevenfold for all three cell lines tested. The development of multi-component VSE formulations using a design of experiments approach allowed further synergistic titer increases in AGE1.CR.pIX cells. Scale-up to 1 L stirred-tank bioreactors and 3D-printed mimics of 200 or 2000 L reactors resulted in up to threefold and eightfold increases, respectively. Conclusions: Addition of single VSEs or combinations thereof allowed a further increase in YF-ZIK titers beyond the yield of an already optimized, highly intensified process. The described approach validates the use of VSEs and can be instructive for optimizing other virus production processes. Full article

2-Phenylethanol (2-PE), an aromatic alcohol with a rose-like fragrance, is widely used in the food, pharmaceutical, and high-end cosmetic industries. In this study, a high-yield 2-PE-producing strain was isolated and identified as Saccharomyces cerevisiae based on morphological characterization and taxonomic identification. Fermentation medium components (carbon and nitrogen sources) were optimized through single-factor experiments in shaking flasks, and fermentation medium with 40 g/L glucose, 5 g/L malt extract, 1.75 g/L corn steep liquor, 2.5 g/L yeast extract, 5 g/L malt extract, 1.75 g/L corn steep liquor was considered suitable for 2-PE production. RT-qPCR results indicated that corn steep liquor activates expression of genes related to the shikimate pathway and Ehrlich pathway (pha2, aro4, aro8, and aro9), thereby promoting the synthesis of 2-PE through these pathways. Excess yeast extract inhibited the expression of aro8 and aro9, while enhancing the expression of tdh3 and adh2, thus promoting the de novo synthesis of 2-PE. Furthermore, fermentation in a 5 L bioreactor was applied to investigate the effects of feeding strategies, inoculum proportion, and pH on 2-PE production. With a pH of 5.5 and10% inoculum proportion, the supplementation of the substrate L-Phe led to a 2-PE production of 4.81 g/L after 24 h of fermentation. Finally, in situ product recovery (ISPR) techniques was applied to alleviate 2-PE cytotoxicity, achieving a production of 6.41 g/L. This process offers a promising strategy for producing 2-PE efficiently and naturally, paving the way for further industrial applications in food, pharmaceutical, and cosmetic sectors. Full article

In view of the growing global need for sustainable protein sources, this study explores the utilization of short-chain fatty acids into single-cell protein using the non-conventional yeast Cyberlindnera jadinii. Short-chain fatty acids can be sustainably produced via anaerobic digestion of organic waste, presenting a promising fermentation substrate for a circular bioeconomy. Cyberlindnera jadinii is demonstrated to be capable of growing on acetate, propionate and butyrate as both a carbon and energy source without strong inhibition. Bioprocess development was conducted in stirred tank bioreactors, where a fed-batch pH-stat bioprocess led to improved efficiency without substrate inhibition. The highest titer of 31.3 ± 1.0 g/L, rate of 0.67 ± 0.02 g/L/h and yield of 0.36 ± 0.01 g/g was achieved with propionate. The resulting biomass contained 41.3% crude protein, and 17.3% crude lipids with 81% unsaturated fatty acids. In contrast to acetate and butyrate, propionate as a substrate led to accumulation of 37% odd-chain fatty acids with titer, rate and yield of 1.74 ± 0.06 g/L, 0.037 ± 0.001 g/L/h and 0.020 ± 0.001 g/g. These findings confirm that short-chain fatty acids are viable fermentation substrates not only for single-cell protein, but also unsaturated and odd-chain fatty acid production with Cyberlindnera jadinii. Full article

A powerful strategy to accelerate bioprocess development is to complement parallel bioreactor systems with an automated approach, often achieved using liquid handling stations. The benefit of such high-throughput experiments is determined by the employed monitoring procedures. To gain a molecular understanding of the microbial production strains in miniaturized parallel single-use bioreactors, we extended the at-line monitoring procedures to transcriptome analysis in a parallel approach using RNA-Seq. To perform automated RNA-Seq experiments, we developed a sample preparation workflow consisting of at-line cell disruption by enzymatic cell lysis, total RNA extraction, nucleic acid concentration normalization, and Nanopore cDNA Library preparation. The pH-controlled aerobic batch growth of Saccharomyces cerevisiae was studied with six different carbon sources (glucose, pyruvate, fructose, galactose, sucrose, and mannose) on a 11 mL scale using 24 parallel stirred tank bioreactors integrated into a liquid handling station while performing at-line sample preparation for RNA-Seq on the same deck. With four biological replicates per condition, 24 cDNA libraries were prepared over 11.5 h. Off-line Nanopore sequencing yielded 20.97 M classified reads with a Q-score > 9. Differential gene expression analysis revealed significant differences in transcriptomic profiles when comparing growth with glucose (exponential growth) to growth with pyruvate (stress conditions), allowing identification of 674 downregulated and 709 upregulated genes. Insignificant changes in gene expression patterns were measured when comparing growth with glucose and fructose, yielding only 64 differentially expressed genes. The expected differences in cellular responses identified in this study show a promising approach for transcriptomic profiling of bioreactor cultures, providing valuable insights on a molecular level at-line in a high-throughput fashion. Full article

Single-use bioreactors (SUBs) are revolutionizing biotechnology and biopharmaceutical manufacturing by offering cost-efficient, flexible, and scalable alternatives to traditional reusable systems. These bioreactors, made from disposable and pre-sterilized materials, streamline cell cultivation for biological production while minimizing the need for complex cleaning and sterilization. A critical aspect of SUB performance lies in optimizing hydrodynamic parameters flow field, power consumption, mixing time, and energy efficiency, which directly influence process outcomes. This study investigates the hydrodynamic performance of an SUB system through stereo Particle Image Velocimetry (PIV) to analyze flow fields, Planar Laser-Induced Fluorescence (PLIF) for mixing time, and Electro Resistance Tomography (ERT) for further insights into mixing dynamics. The results, evaluated at varying impeller speeds and fill heights, provide a comprehensive understanding of flow behavior, mixing efficiency, and power requirements. This work highlights the importance of hydrodynamic characterization in optimizing SUB design and operation, contributing to more sustainable and efficient biopharmaceutical production. Full article

Cupriavidus necator can produce single-cell proteins (SCPs) using electrons produced by hydrogen as energy, oxygen as electron acceptors, and CO₂ as carbon sources. Gas fermentation is a process of microbial fermentation that uses gas substrates (such as hydrogen, carbon dioxide, etc.) which faces several challenges, mainly including the low solubility of gas substrates, the danger of hydrogen and oxygen mixing, and the optimization of fermentation conditions. To overcome these challenges, this article explores a variety of strategies—including the design of a self-developed bioreactor—to reduce the risk of static electricity. Without the addition of filler material, the results showed that the maximum cell dry weight (CDW) of 30% secondary seed inoculation was 20.41% higher than that of 10% secondary seed inoculum, and 5.99% higher than that of 20% secondary seed inoculum. Combined with the filler material and with the use of high-efficiency continuous fermentation technology, the average yield of continuous fermentation was 23.31 g/day, while the average yield of batch fermentation was 14.33 g/day. The daily yield of continuous fermentation is 1.63 times that of batch fermentation. These efforts are aimed at improving the efficiency and safety of gas fermentation. Full article

Dissolved hydrogen is a critical factor in maintaining the delicate balance among microbial species that drive anaerobic digestion. Since previous findings have demonstrated a correlation between dissolved hydrogen concentration and volatile fatty acid (VFA) levels, we propose to evaluate the use of dissolved hydrogen concentration in digestate as an alternative to traditional VFA measurements. The aim is to determine whether dissolved hydrogen could serve as a faster, more accurate, and more efficient indicator of process instability in anaerobic digestion. An integral part of this task also involves addressing the technical challenge of identifying a suitable sensor that meets our requirements. In this study, we evaluated the ratio of dissolved hydrogen concentration to Total Inorganic Carbon as a potential alternative to the traditional stability indicator, Volatile Fatty Acids/Total Inorganic Carbon (VFA/TIC), also referred to as Flüchtige Organische Säuren/Totales Anorganisches Carbonat (FOS/TAC). The single-stage anaerobic digestion process was carried out in a Terrafors IS rotary drum bioreactor for 150 days at an average temperature of 40 °C and an organic volatile load of 0.092 kg m⁻³ d⁻¹. Corn silage was dosed on weekdays as the substrate. With a theoretical retention time of 45 days, a biogas production of 0.219 $\mathrm{N}{\mathrm{m}}^3\mathrm{k}\mathrm{g}\mathrm{V}{\mathrm{s}}^{-1}$ with a CH₄ content of 31.6% was achieved. The values of the determined VFA/TIC stability indicator ranged from 0.22 to 5.66, with the highest values obtained when the reactor was overloaded. The dissolved hydrogen concentration ranged 0.005–0.196 $\mathrm{m}\mathrm{g}\mathrm{d}{\mathrm{m}}^{-3}$. The Pearson correlation coefficient was 0.337 and the Spearman correlation coefficient was 0.468. The amperometric microsensor has proven to be unsuitable for field applications due to its lack of sensitivity and short lifetime. The proposed ratio of dissolved hydrogen concentration to TIC did not prove to be significantly more effective than the established VFA/TIC indicator. Full article