Recommendations for Artificial Intelligence Application in Continued Process Verification.

This review paper explores the transformative impact of Artificial Intelligence (AI) on Continued Process Verification (CPV) in the biopharmaceutical industry. Originating from the CPV of the Future project, the study investigates the challenges and opportunities associated with integrating AI into CPV, focusing on real-time data analysis and proactive process adjustments. The paper highlights the importance of aligning AI solutions with regulatory standards and offers a set of comprehensive recommendations to bridge the gap between AI's potential and its practical, compliant, and safe application in pharmaceutical manufacturing.

Influence of Running Surface Using Advanced Footwear Technology Spikes on Middle- and Long-Distance Running Performance Measures.

This study evaluated the effects of advanced footwear technology (AFT) spikes on running performance measures, spatiotemporal variables, and perceptive parameters on different surfaces (track and grass). Twenty-seven male trained runners were recruited for this study. In Experiment 1, participants performed 12 × 200 m at a self-perceived 3000 m running pace with a recovery of 5 min.

Continuous Process Verification 4.0 application in upstream: adaptiveness implementation managed by AI in the hypoxic bioprocess of the cell factory.

The experimental approach developed in this research demonstrated how the cloud, the Internet of Things (IoT), edge computing, and Artificial Intelligence (AI), considered key technologies in Industry 4.0, provide the expected horizon for adaptive vision in Continued Process Verification (CPV), the final stage of Process Validation (PV). producing lipase 1 under the regulation of the constitutive promoter was selected as an experimental bioprocess.

Scalable protein production by Komagataella phaffii enabled by ARS plasmids and carbon source-based selection.

Background: Most recombinant Komagataella phaffii (Pichia pastoris) strains for protein production are generated by genomic integration of expression cassettes. The clonal variability in gene copy numbers, integration loci and consequently product titers limit the aptitude for high throughput applications in drug discovery, enzyme engineering or most comparative analyses of genetic elements such as promoters or secretion signals. Circular episomal plasmids with an autonomously replicating sequence (ARS), an alternative which would alleviate some of these limitations, are inherently unstable in K.

Synergic kinetic and physiological control to improve the efficiency of Komagataella phaffii recombinant protein production bioprocesses.

The yeast Komagataella phaffii (Pichia pastoris) is currently considered a versatile and highly efficient host for recombinant protein production (RPP). Interestingly, the regulated application of specific stress factors as part of bioprocess engineering strategies has proven potential for increasing the production of recombinant products. This study aims to evaluate the impact of controlled oxygen-limiting conditions on the performance of K.

Belief That Caffeine Ingestion Improves Performance in a 6-Minute Time Trial Test without Affecting Pacing Strategy.

Unlabelled: This study aimed to analyze the placebo effect associated with caffeine ingestion on running performance.

Methods: Thirteen recreationally trained males in long-distance running (age: 38.5 ± 11.

Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries.

Cellular homeostasis requires the robust control of biomolecule concentrations, but how do millions of mRNAs coordinate their stoichiometries in the face of dynamic translational changes? Here, we identified a two-tiered mechanism controlling mRNA:mRNA and mRNA:protein stoichiometries where mRNAs super-assemble into condensates with buffering capacity and sorting selectivity through phase-transition mechanisms. Using C. elegans oogenesis arrest as a model, we investigated the transcriptome cytosolic reorganization through the sequencing of RNA super-assemblies coupled with single mRNA imaging.

Enabling growth-decoupled recombinant protein production based on the methanol-free P promoter.

The current transition towards the circular bioeconomy requires a rational development of biorefineries to sustainably fulfill the present demands. The use of () can meet this challenge, since it has the capability to use crude glycerol as a carbon-source, a by-product from the biodiesel industry, while producing high- and low-added value products. Recombinant protein production (RPP) using has often been driven either by the methanol induced promoter (P ) and/or the constitutive promoter (P ).

NOM fractionation by HPSEC-DAD-OCD for predicting trihalomethane disinfection by-product formation potential in full-scale drinking water treatment plants.

Chlorination is a common method for water disinfection; however, it leads to the formation of disinfection by-products (DBPs), which are undesirable toxic pollutants. To prevent their formation, it is crucial to understand the reactivity of natural organic matter (NOM), which is considered a dominant precursor of DBPs. We propose a novel size exclusion chromatography (SEC) approach to evaluate NOM reactivity and the formation potential of total trihalomethanes-formation potentials (tTHMs-FP) and four regulated species (i.

Screening of microplastics in water and sludge lines of a drinking water treatment plant in Catalonia, Spain.

Microplastics (MPs) are emerging pollutants detected everywhere in the environment, with the potential to harm living organisms. The present study investigated the concentration, morphology, and composition of MPs, between 20 μm and 5 mm, in a drinking water treatment plant (DWTP) located close to Barcelona (Catalonia, NE Spain). The sampling included different units of the DWTP, from influent to effluent as well as sludge line.

CPV of the Future: AI-Powered Continued Process Verification for Bioreactor Processes.

According to the standard guidelines by the FDA, process validation in biopharma manufacturing encompasses a life cycle consisting of three stages: process design (PD), process qualification (PQ), and continued process verification (CPV). The validity and efficiency of the analytics methods employed during the CPV require extensive knowledge of the process. However, for new processes and new drugs, such knowledge is often not available from Process performance qualification and Validation (PPQV).

Innovative Bioprocess Strategies Combining Physiological Control and Strain Engineering of to Improve Recombinant Protein Production.

The combination of strain and bioprocess engineering strategies should be considered to obtain the highest levels of recombinant protein production (RPP) while assuring product quality and process reproducibility of heterologous products. In this work, two complementary approaches were investigated to improve bioprocess efficiency based on the yeast . Firstly, the performance of two lipase 1 producer clones with different gene dosage under the regulation of the constitutive were compared in chemostat cultures with different oxygen-limiting conditions.

Second- and third-generation biodiesel production with immobilised recombinant Rhizopus oryzae lipase: Influence of the support, substrate acidity and bioprocess scale-up.

Rhizopus oryzae lipase immobilised onto differently functionalised polymethacrylate (Purolite®) and magnetite superparamagnetic supports was assessed as a catalyst for biodiesel production with pomace oil. The presence of surface hydrocarbon chains increased the operational stability of the biocatalysts supported on Purolite® and superparamagnetic particles up to 9 and 2 times, respectively. By contrast, the presence of functional groups had no effect on the initial transesterification rate, which was twice higher with the lipase immobilised onto Purolite®.

Scalable production and application of Pichia pastoris whole cell catalysts expressing human cytochrome P450 2C9.

Background: Currently, the numerous and versatile applications in pharmaceutical and chemical industry make the recombinant production of cytochrome P450 enzymes (CYPs) of great biotechnological interest. Accelerating the drug development process by simple, quick and scalable access of human drug metabolites is key for efficient and targeted drug development in response to new and sometimes unexpected medical challenges and needs. However, due its biochemical complexity, scalable human CYP (hCYP) production and their application in preparative biotransformations was still in its infancy.

Bioprocess performance analysis of novel methanol-independent promoters for recombinant protein production with Pichia pastoris.

Background: Pichia pastoris is a powerful and broadly used host for recombinant protein production (RPP), where past bioprocess performance has often been directed with the methanol regulated AOX1 promoter (P), and the constitutive GAP promoter (P). Since promoters play a crucial role in an expression system and the bioprocess efficiency, innovative alternatives are constantly developed and implemented. Here, a thorough comparative kinetic characterization of two expression systems based on the commercial PDF and UPP promoters (P, P) was first conducted in chemostat cultures.

Benchmarking empirical models for THMs formation in drinking water systems: An application for decision support in Barcelona, Spain.

Drinking water treatment plants (DWTPs) face changes in raw water quality, which affect the formation of disinfection by-products. Several empirical modelling approaches have been reported in the literature, but most of them have been developed with lab-scale data, which may not be representative of real water systems. Therefore, the application of these models for real-time operation of DWTPs might be limited.

Impact of digital transformation on the automotive industry.

Digital technologies are transforming the automotive industry and disrupting traditional business models. New business opportunities related to Industry 4.0 are emerging, so companies must adapt to the new environment.

Bioreactor-scale cell performance and protein production can be substantially increased by using a secretion signal that drives co-translational translocation in Pichia pastoris.

Pichia pastoris (Komagataella spp.) has become one of the most important host organisms for production of heterologous proteins of biotechnological interest, many of them extracellular. The protein secretion pathway has been recognized as a limiting process in which many roadblocks have been pinpointed.

Assessment of dead-end ultrafiltration for the detection and quantification of microbial indicators and pathogens in the drinking water treatment processes.

A safe water supply requires distinct treatments and monitoring to guarantee the absence of pathogens and substances potentially hazardous for human health. In this study we assessed the efficiency of the dead-end ultrafiltration (DEUF) method to concentrate faecal indicator organisms (FIO) and pathogens in water samples with different physicochemical characteristics. Water samples were collected at the treatment stages of two drinking water treatment plants to analyse the concentration of a variety of 7 FIO and 4 reference microbes which have some species that are pathogenic to humans: Campylobacter spp.

Continuous Cultivation as a Tool Toward the Rational Bioprocess Development With Cell Factory.

The methylotrophic yeast () is currently considered one of the most promising hosts for recombinant protein production (RPP) and metabolites due to the availability of several tools to efficiently regulate the recombinant expression, its ability to perform eukaryotic post-translational modifications and to secrete the product in the extracellular media. The challenge of improving the bioprocess efficiency can be faced from two main approaches: the strain engineering, which includes enhancements in the recombinant expression regulation as well as overcoming potential cell capacity bottlenecks; and the bioprocess engineering, focused on the development of rational-based efficient operational strategies. Understanding the effect of strain and operational improvements in bioprocess efficiency requires to attain a robust knowledge about the metabolic and physiological changes triggered into the cells.

Implementation of an environmental decision support system for controlling the pre-oxidation step at a full-scale drinking water treatment plant.

Drinking water treatment plants (DWTPs) face changes in raw water quality, and treatment needs to be adjusted to produce the best water quality at the minimum environmental cost. An environmental decision support system (EDSS) was developed for aiding DWTP operators in choosing the adequate permanganate dosing rate in the pre-oxidation step. To this end, multiple linear regression (MLR) and multi-layer perceptron (MLP) models are compared for choosing the best predictive model.

Dissecting the Interaction Deficiency of a Cartilaginous Fish Digestive Lipase with Pancreatic Colipase: Biochemical and Structural Insights.

A full-length cDNA encoding digestive lipase (SmDL) was cloned from the pancreas of the smooth-hound (). The obtained cDNA was 1350 bp long encoding 451 amino acids. The deduced amino acid sequence has high similarity with known pancreatic lipases.

Specific growth rate governs AOX1 gene expression, affecting the production kinetics of Pichia pastoris (Komagataella phaffii) P-driven recombinant producer strains with different target gene dosage.

Background: The P-based expression system is the most widely used for producing recombinant proteins in the methylotrophic yeast Pichia pastoris (Komagataella phaffii). Despite relevant recent advances in regulation of the methanol utilization (MUT) pathway have been made, the role of specific growth rate (µ) in AOX1 regulation remains unknown, and therefore, its impact on protein production kinetics is still unclear.

Results: The influence of heterologous gene dosage, and both, operational mode and strategy, on culture physiological state was studied by cultivating the two P-driven Candida rugosa lipase 1 (Crl1) producer clones.

Rationale-based selection of optimal operating strategies and gene dosage impact on recombinant protein production in Komagataella phaffii (Pichia pastoris).

Its features as a microbial and eukaryotic organism have turned Komagataella phaffii (Pichia pastoris) into an emerging cell factory for recombinant protein production (RPP). As a key step of the bioprocess development, this work aimed to demonstrate the importance of tailor designing the cultivation strategy according to the production kinetics of the cell factory. For this purpose, K.