Pressure-driven membrane processes for the recovery and recycling of deep eutectic solvents: A seaweed biorefinery case study.

Deep eutectic solvents (DES) are green alternatives for conventional solvents. They have gained attention for their potential to extract valuable compounds from biomass, such as seaweed. In this framework, a case study was developed to assess the feasibility of pressure-driven membrane processes as an efficient tool for the recovery of deep eutectic solvents and targeted biomolecules.

An inert tracer: The binding site of a fluorescent dye on the antibody and its effects on Protein A chromatography.

Fluorescently labeled antibodies are widely used to visualize the adsorption process in protein chromatography using confocal laser scanning microscopy (CLSM), but also as a tracer for determination of residence time distribution (RTD) in continuous chromatography. It is assumed that the labeled protein is inert and representative of the unlabeled antibody, ignoring the fact that labeling with a fluorescent dye can change the characteristics of the original molecule. It became evident that the fluorescently labeled antibody has a higher affinity toward protein A resins such as MabSelect Sure.

Digital twin in high throughput chromatographic process development for monoclonal antibodies.

The monoclonal antibody (mAb) industry is becoming increasingly digitalized. Digital twins are becoming increasingly important to test or validate processes before manufacturing. High-Throughput Process Development (HTPD) has been progressively used as a tool for process development and innovation.

Isolation and quantification of alginate in choline chloride-based deep eutectic solvents.

Extraction of seaweed compounds using Deep Eutectic Solvents (DES) has shown high interest. Quantification, however, is challenging due to interactions with DES components. In this research work, three chemical separation techniques were investigated to isolate and quantify alginate from a set of choline chloride-based DES.

Real-time detection of mAb aggregates in an integrated downstream process.

The implementation of continuous processing in the biopharmaceutical industry is hindered by the scarcity of process analytical technologies (PAT). To monitor and control a continuous process, PAT tools will be crucial to measure real-time product quality attributes such as protein aggregation. Miniaturizing these analytical techniques can increase measurement speed and enable faster decision-making.

Application of a fluorescent dye-based microfluidic sensor for real-time detection of mAb aggregates.

The lack of process analytical technologies able to provide real-time information and process control over a biopharmaceutical process has long impaired the transition to continuous biomanufacturing. For the monoclonal antibody (mAb) production, aggregate formation is a major critical quality attribute (CQA) with several known process parameters (i.e.

Ionic Liquid-Assisted Selective Extraction and Partitioning of Biomolecules from Macroalgae.

Macroalgae are a promising feedstock for several industries due to their large content of proteins and carbohydrates and the high biomass productivities. A novel extraction and fractionation concept based on ionic liquids (ILs) using as model organism is presented. Biomolecules are first extracted by means of IL-assisted mechanical shear, followed by two-phase partitioning or ultrafiltration in order to fractionate proteins and carbohydrates and to recover the IL.

Design of a microfluidic mixer channel: First steps into creating a fluorescent dye-based biosensor for mAb aggregate detection.

A major challenge in the transition to continuous biomanufacturing is the lack of process analytical technology (PAT) tools which are able to collect real-time information on the process and elicit a response to facilitate control. One of the critical quality attributes (CQAs) of interest during monoclonal antibodies production is aggregate formation. The development of a real-time PAT tool to monitor aggregate formation is then crucial to have immediate feedback and process control.

Small, smaller, smallest: Miniaturization of chromatographic process development.

Biopharmaceuticals are becoming increasingly important in modern healthcare. Monoclonal antibodies (mAb) are one of the most widely used therapeutic proteins and are important for the treatment of cancer and autoimmune diseases, among others. After cell culture there are still large amounts of other impurities (e.

Mild acoustic processing of Tisochrysis lutea for multiproduct biorefineries.

Cellular agriculture could represent a more sustainable alternative to current food and nutraceutical production processes. Tisochrysis lutea microalgae represents a rich source of antioxidants and omega-3 fatty acids essential for human health. However, current downstream technologies are limiting its use.

Two-Component Nanoparticle Vaccine Displaying Glycosylated Spike S1 Domain Induces Neutralizing Antibody Response against SARS-CoV-2 Variants.

Vaccines pave the way out of the SARS-CoV-2 pandemic. Besides mRNA and adenoviral vector vaccines, effective protein-based vaccines are needed for immunization against current and emerging variants. We have developed a virus-like particle (VLP)-based vaccine using the baculovirus-insect cell expression system, a robust production platform known for its scalability, low cost, and safety.

Multiproduct Microalgae Biorefineries Mediated by Ionic Liquids.

Ionic liquids (ILs) are salts with low melting points that can be used as solvents for mild extraction and selective fractionation of biomolecules (e.g., proteins, carbohydrates, lipids, and pigments), enabling the valorisation of microalgal biomass in a multiproduct biorefinery concept, while maintaining the biomolecules' structural integrity and activity.

Eutectic solvents with tuneable hydrophobicity: lipid dissolution and recovery.

Despite the promising advantages of eutectic solvents, the application of these solvents as an extraction solvent is still limited due to the challenging product recovery. Previously, it was reported that lipids could be recovered from a hydrophobic eutectic solvent with the principle of switchable hydrophobicity. However, this method still involves additional chemicals, such as polymeric amines, water, and CO, which need to be removed later.

Design of Value Chains for Microalgal Biorefinery at Industrial Scale: Process Integration and Techno-Economic Analysis.

The objective of this work was to identify industrial scenarios for the most promising microalgal biorefinery value chains on the basis of product selection, yields, and techno-economic performance, using biological characteristics of algae species. The development, value creation, and validation of several new processing routes with applications in food, aquafeeds and non-food products were particularly considered in this work. The techno-economic performance of various single product value chains (SP) and multiproduct value chains (MP) was evaluated for four industrial microalgal strains.

Combined bead milling and enzymatic hydrolysis for efficient fractionation of lipids, proteins, and carbohydrates of Chlorella vulgaris microalgae.

A combined bead milling and enzymatic hydrolysis process was developed for fractionation of the major valuable biomass components, i.e., proteins, carbohydrates, and lipids from the microalgae Chlorella vulgaris.

Mild Fractionation of Hydrophilic and Hydrophobic Components From Using Ionic Liquids.

Microalgae are a promising source for proteins, lipids, and carbohydrates for the food/feed and biofuel industry. To make microalgae production economically feasible, it is necessary to optimally use all produced compounds keeping full functionality. Therefore, biorefining of microalgae is the key to lower the cost of algal products using mild and effective processing techniques.

Forensic Nursing Education and Practice in the Netherlands: Where Are We at?

Background: Forensic nursing is a new discipline to the Netherlands. Since 2013, a program has been in place to train experienced nurses in several aspects of forensic nursing, including injury assessment and wound documentation, sexual assault examination, assessment of child abuse, death investigation, and primary care for detainees of the police.

Objective: The purpose of the study was to provide information on the working environment, self-rated competencies, and practice experiences of forensic nurses after having completed the program.

Integrated Product Recovery Will Boost Industrial Cyanobacterial Processes.

Cyanobacteria promise to be an important industrial platform for the production of a variety of biobased products such as fuels, plastics, and isoprenoids. Recent advances in synthetic biology have resulted in various cyanobacterial strain improvements. Nevertheless, these new strains are still hampered by product inhibition, resulting in low volumetric productivities, product concentrations, and yields on light.

Selective and mild fractionation of microalgal proteins and pigments using aqueous two-phase systems.

Background: Microalgal biomass is generally used to produce a single product instead of valorizing all of the cellular components. The biomass production and downstream processes are too expensive if only one product is valorized. A new approach was proposed for the simultaneous and selective partitioning of pigments and proteins from disrupted cultivated under saline and freshwater conditions.

Selective and energy efficient extraction of functional proteins from microalgae for food applications.

The use of a single controlled bead milling step of the microalga Tetraselmis suecica resulted in a soluble fraction, rich in functional proteins. This was achieved by fine-tuning the processing time, thereby exploiting the difference in rates of protein and carbohydrate release during milling. Soluble proteins were extracted under mild conditions -room temperature, no addition of chemicals, pH 6.

Techno-Functional Properties of Crude Extracts from the Green Microalga Tetraselmis suecica.

A mild fractionation process to extract functional biomolecules from green microalgae was implemented. The process includes bead milling, centrifugation, and filtration with several membrane cut-offs. For each fraction, the corresponding composition was measured, and the surface activity and gelation behavior were determined.

Biorefinery of the macroalgae : extraction of proteins and carbohydrates by mild disintegration.

The effect of osmotic shock, enzymatic incubation, pulsed electric field, and high shear homogenization on the release of water-soluble proteins and carbohydrates from the green alga was investigated in this screening study. For osmotic shock, both temperature and incubation time had a significant influence on the release with an optimum at 30 °C for 24 h of incubation. For enzymatic incubation, pectinase demonstrated being the most promising enzyme for both protein and carbohydrate release.

Chromatographic parameter determination for complex biological feedstocks.

The application of mechanistic models for chromatography requires accurate model parameters. Especially for complex feedstocks such as a clarified cell harvest, this can still be an obstacle limiting the use of mechanistic models. Another commonly encountered obstacle is a limited amount of sample material and time to determine all needed parameters.

Multi-Product Microalgae Biorefineries: From Concept Towards Reality.

Although microalgae are a promising biobased feedstock, industrial scale production is still far off. To enhance the economic viability of large-scale microalgae processes, all biomass components need to be valorized, requiring a multi-product biorefinery. However, this concept is still too expensive.

Rapid screening of IgG quality attributes - effects on Fc receptor binding.

The interactions of therapeutic antibodies with fragment crystallizable γ (Fcγ) receptors and neonatal Fc receptors (FcRn) are measured as indicators of antibody functional performance. Antibodies are anchored to immune cells through the Fc tail, and these interactions are important for the efficacy and safety of therapeutic antibodies. High-throughput binding studies on each of the human Fcγ receptor classes (FcγRI, FcγRIIa, FcγRIIb, FcγRIIIa, and FcγRIIIb) as well as FcRn have been developed and performed with human IgG after stress-induced modifications to identify potential impact .