Degradation of 3D-printed magnesium phosphate ceramics and a prognosis on their bone regeneration potential.

Regenerative bone implants promote new bone formation and ideally degrade simultaneously to osteogenesis. Although clinically established calcium phosphate bone grafts provide excellent osseointegration and osteoconductive efficacy, they are limited in terms of bioresorption. Magnesium phosphate (MP) based ceramics are a promising alternative, because they are biocompatible, mechanically extremely stable, and degrade much faster than calcium phosphates under physiological conditions.

Quantification methods for viruses and virus-like particles applied in biopharmaceutical production processes.

Introduction: Effective cell-based production processes of virus particles are the foundation for the global availability of classical vaccines, gene therapeutic vectors, and viral oncolytic treatments. Their production is subject to regulatory standards ensuring the safety and efficacy of the pharmaceutical product. Process analytics must be fast and reliable to provide an efficient process development and a robust process control during production.

Screening for Biofilm-Stimulating Factors in the Freshwater Planctomycete to Improve Sessile Growth in a Chemically Defined Medium.

Planctomycetes such as offer a promising source of bioactive molecules, particularly when they switch from planktonic to sessile growth, but little is known about the corresponding biosynthetic gene clusters and how they are activated. We therefore screened for factors that promote sessile growth and biofilm formation to enable the cultivation of in a fixed-bed reactor. We carried out screening in microtiter plates focusing on biofilm formation and changes in optical density in response to various C:N ratios, metal ions, and oxidative stress.

A Combined Ultrafiltration/Diafiltration Process for the Purification of Oncolytic Measles Virus.

Measles virus (MV) is an important representative of a new class of cancer therapeutics known as oncolytic viruses. However, process intensification for the downstream purification of this fragile product is challenging. We previously found that a mid-range molecular weight cut-off (300 kDa) is optimal for the concentration of MV.

Statistical experimental designs to optimize the transient transfection of HEK 293T cells and determine a transfer criterion from adherent cells to larger-scale cell suspension cultures.

The transient transfection of mammalian cells is a rapid and versatile platform for the manufacture of recombinant proteins, but industrial processes depend on reliable scalability and efficient conversion from adherent to suspension cell cultures. Here we describe the optimized transfection of HEK 293T cells in both culture formats. DMEM was the best transfection medium for adherent HEK 293T cells, so we determined the kinetics of linear polyethyleneimine (LPEI) polyplex formation with plasmid DNA (pDNA) and subsequent cellular uptake.

Continuous purification of influenza A virus particles using pseudo-affinity membrane chromatography.

Robust and flexible continuous unit operations that enable the establishment of intensified bioprocesses is one of the most relevant trends in manufacturing of biopharmaceuticals, including virus-based products. Sulfated cellulose membrane adsorbers (SCMA) are one of the most promising matrices for chromatographic purification of virus particles, like influenza viruses. Here, a three 'column' periodical counter current set-up was used to continuously purify influenza A/PR/8/34 virus particles using SCMA in bind-elute mode.

In situ purification and enrichment of fructo-oligosaccharides by fermentative treatment with Bacillus coagulans and selective catalysis using immobilized fructosyltransferase.

Fructo-oligosaccharides (FOS) are prebiotic sugar substitutes that can be produced from sucrose using fructosyltransferases (FTases). However, the economic value of this process is limited by inefficient product purification and enzyme reusability. In this study, enzyme-free FOS preparations were produced by immobilizing the FTase on resin carriers.

Scaled preparation of extracellular vesicles from conditioned media.

Extracellular vesicles (EVs) especially of mesenchymal stem/stomal cells (MSCs) are increasingly considered as biotherapeutic agents for a variety of different diseases. For translating them effectively into the clinics, scalable production processes fulfilling good manufacturing practice (GMP) are needed. Like for other biotherapeutic agents, the manufacturing of EV products can be subdivided in the upstream and downstream processing and the subsequent quality control, each of them containing several unit operations.

Machine Learning Classification of Inflammatory Bowel Disease in Children Based on a Large Real-World Pediatric Cohort CEDATA-GPGE® Registry.

The rising incidence of pediatric inflammatory bowel diseases (PIBD) facilitates the need for new methods of improving diagnosis latency, quality of care and documentation. Machine learning models have shown to be applicable to classifying PIBD when using histological data or extensive serology. This study aims to evaluate the performance of algorithms based on promptly available data more suited to clinical applications.

Influence of Ceramic Membrane Surface Characteristics on the Flux Behavior of a Complex Fermentation Broth.

The valorization of agro-industrial residues using yeasts as biocatalysts requires efficient methods for biomass separation. Filtration with ceramic membranes is suitable for this task, however, the challenge of flux decline and the unavoidable cleaning must be taken into account. We investigated the filtration of fermentation broth and its components using tubular microfiltration and ultrafiltration membranes, and hollow-fiber ultrafiltration membranes, with cut-offs of 30 and 200 nm.

Material utilization of green waste: a review on potential valorization methods.

Considering global developments like climate change and the depletion of fossil resources, the use of new and sustainable feedstocks such as lignocellulosic biomass becomes inevitable. Green waste comprises heterogeneous lignocellulosic biomass with low lignin content, which does not stem from agricultural processes or purposeful cultivation and therefore mainly arises in urban areas. So far, the majority of green waste is being composted or serves as feedstock for energy production.

Purification of Crude Fructo-Oligosaccharide Preparations Using Probiotic Bacteria for the Selective Fermentation of Monosaccharide Byproducts.

Probiotics are microbes that promote health when consumed in sufficient amounts. They are present in many fermented foods or can be provided directly as supplements. Probiotics utilize non-digestible prebiotic oligosaccharides for growth in the intestinal tract, contributing to a healthy microbiome.

Ultrasonic and enzymatic pretreatments of Monascus fermentation byproduct for a sustainable production of Bacillus subtilis.

Background: Monascus fermentation byproduct (MFB) is a biowaste generated after food colorants are extracted. Using MFB to produce probiotics (Bacillus subtilis) is a sustainable way for the entire production to be used as food or animal feed additives. However, due to the rigidity of the Monascus mycelium cell wall, B.

Enhancing the Heterologous Fructosyltransferase Activity of : Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing.

The enzymatic production of prebiotic fructo-oligosaccharides (FOS) from sucrose involves fructosyltransferases (FFTs) and invertases, both of which catalyze forward (transferase) and reverse (hydrolysis) reactions. FOS yields can therefore be increased by favoring the forward reaction. We investigated process conditions that favored transferase activity in the yeast strain GG799, which expresses a native invertase and a heterologous FFT from .

Flow-through enzymatic reactors using polymer monoliths: From motivation to application.

The application of monolithic materials as carriers for enzymes has rapidly expanded to the realization of flow-through analysis and bioconversion processes. This expansion is partly attributed to the absence from diffusion limitation in many monoliths-based enzyme reactors. Particularly, the relatively ease of introducing functional groups renders polymer monoliths attractive as enzyme carriers.

A new simplified clarification approach for lentiviral vectors using diatomaceous earth improves throughput and safe handling.

Lentiviral vectors have proven their great potential to serve as a DNA delivery tool for gene modified cell therapy and gene therapy applications. The downstream processing of these vectors is however still a great challenge, particularly because of the low stability of the virus. Harvesting and clarification are critical and until now insufficiently characterized steps for lentivirus processing.

High-Performance Polyacrylic Acid-Grafted PVDF Nanofiltration Membrane with Good Antifouling Property for the Textile Industry.

In the textile industry, a high-efficiency dye removal and low-retention of salt is demanded for recycling wastewater. In this study, polyvinylidene fluoride (PVDF) ultrafiltration membrane was transformed to a negatively charged loose nanofiltration (NF) membrane through UV-grafting of acrylic acid. At the optimal exposure of PVDF membrane in UV light for 5 min, the membrane had a high dye recovery above 99% (Congo red and Eriochrome Black T) and a low sodium chloride (NaCl) rejection of less than 15% along with pure water flux of 26 L∙m∙h∙bar.

Development of a downstream process for the production of an inactivated whole hepatitis C virus vaccine.

There is a large unmet need for a prophylactic hepatitis C virus (HCV) vaccine to control the ongoing epidemic with this deadly pathogen. Many antiviral vaccines employ whole viruses as antigens. For HCV, this approach became feasible following the development of infectious cell culture systems for virus production.

A scalable downstream process for the purification of the cell culture-derived Orf virus for human or veterinary applications.

The large demand for safe and efficient viral vector-based vaccines and gene therapies against both inherited and acquired diseases accelerates the development of viral vectors. One outstanding example, the Orf virus, has a wide range of applications, a superior efficacy and an excellent safety profile combined with a reduced pathogenicity compared to other viral vectors. However, besides these favorable attributes, an efficient and scalable downstream process still needs to be developed.

Recent developments in the use of peroxygenases - Exploring their high potential in selective oxyfunctionalisations.

Peroxygenases are an emerging new class of enzymes allowing selective oxyfunctionalisation reactions in a cofactor-independent way different from well-known P450 monooxygenases. Herein, we focused on recent developments from organic synthesis, molecular biotechnology and reaction engineering viewpoints that are devoted to bring these enzymes in industrial applications. This covers natural diversity from different sources, protein engineering strategies for expression, substrate scope, activity and selectivity, stabilisation of enzymes via immobilisation, and the use of peroxygenases in low water media.

Selection of chromatographic methods for the purification of cell culture-derived Orf virus for its application as a vaccine or viral vector.

In recent years, the Orf virus has become a promising tool for protective recombinant vaccines and oncolytic therapy. However, suitable methods for an Orf virus production, including up- and downstream, are very limited. The presented study focuses on downstream processing, describing the evaluation of different chromatographic unit operations.

Microbial Electrosynthesis-An Inventory on Technology Readiness Level and Performance of Different Process Variants.

The objective of microbial electrosynthesis (MES) is to combine the advantages of electrochemistry and biotechnology in order to produce chemicals and fuels. This combination enables resource-efficient processes by using renewable raw materials and regenerative energies. In the last decade, different MES processes have been described, for example, MES based on biofilms or mediators, electro-fermentation, and secondary MES.

From CO to Bioplastic - Coupling the Electrochemical CO Reduction with a Microbial Product Generation by Drop-in Electrolysis.

CO has been electrochemically reduced to the intermediate formate, which was subsequently used as sole substrate for the production of the polymer polyhydroxybutyrate (PHB) by the microorganism Cupriavidus necator. Faradaic efficiencies (FE) up to 54 % have been reached with Sn-based gas-diffusion electrodes in physiological electrolyte. The formate containing electrolyte can be used directly as drop-in solution in the following biological polymer production by resting cells.

Insights in the anode chamber influences on cathodic bioelectromethanogenesis - systematic comparison of anode materials and anolytes.

Cathode and catholyte are usually optimized to improve microbial electrosynthesis process, whereas the anodic counter reaction was not systematically investigated and optimized for these applications yet. Nevertheless, the anolyte and especially the anode material can limit the cathodic bioelectrochemical process. This paper compares for the first time the performance of different anode materials as counter electrodes for a cathodic bioelectrochemical process, the bioelectromethanogenesis.

Osteoclast and osteoblast response to strontium-doped struvite coatings on titanium for improved bone integration.

To develop implants with improved bone ingrowth, titanium substrates were coated with homogeneous and dense struvite (MgNH4PO4·6H2O) layers by means of electrochemically assisted deposition. Strontium nitrate was added to the coating electrolyte in various concentrations, in order to fabricate Sr-doped struvite coatings with Sr loading ranging from 10.6 to 115 μg/cm2.