Economic analysis of royalactin production under uncertainty: Evaluating the effect of parameter optimization.

Royalactin is a protein with several different potential uses in humans. Research, in insects and in mammalian cells, has shown that it can accelerate cell division and prevent apoptosis. The method of action is through the use of the epidermal growth factor receptor, which is present in humans. Potential use in humans could be to lower cholesterolemic levels in blood, and to elicit similar effects to those seen in bees, e.g., increased lifespan. Mass production of Royalactin has not been accomplished, though a recent article presented a Pichia pastoris fermentation and recovery by aqueous two-phase systems at laboratory scale as a possible basis for production. Economic modelling is a useful tool with which compare possible outcomes for the production of such a molecule and in particular, to locate areas where additional research is needed and optimization may be required. This study uses the BioSolve software to perform an economic analysis on the scale-up of the putative process for Royalactin. The key parameters affecting the cost of production were located via a sensitivity analysis and then evaluated by Monte Carlo analysis. Results show that if titer is not optimized the strategy to maintain a low cost of goods is process oriented. After optimization of this parameter the strategy changes to a product-oriented and the target output becomes the critical parameter determining the cost of goods. This study serves to provide a framework for the evaluation of strategies for future production of Royalactin, by analyzing the factors that influence its cost of manufacture. © 2015 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 2015

Characterization and optimization of polymer-polymer aqueous two-phase systems for the isolation and purification of CaCo2 cell-derived exosomes

Exosomes are cell-derived vesicles that present attractive characteristics such as nano size and unique structure for their use as drug delivery systems for drug therapy, biomarkers for prognostic, diagnostic and personalized treatments. So far, one of the major challenges for therapeutic applications of exosomes is the development of optimized isolation methods. In this context, aqueous two-phase systems (ATPS) have been used as an alternative method to isolate biological molecules and particles with promising expectations for exosomes. In this work, fractionation of exosomes obtained from CaCo2 cell line and culture media contaminants were individually performed in 20 polymer-polymer ATPS. The effect of design parameters such as polymer composition, molecular weight, and tie-line length (TLL) on polyethylene glycol (PEG)-Dextran, Dextran-Ficoll and PEG-Ficoll systems was studied. After partition analysis, 4 of the 20 systems presented the best exosome fractionation from contaminants under initial conditions, which were optimized via salt addition (NaCl) to a final concentration of 25 mM, to improve collection efficiency. The PEG 10,000 gmol-1 –Dextran 10,000 gmol-1 system at TLL 25% w/w with NaCl, showed the best potential isolation efficiency. Following this proposed strategy, an exosome purification factor of 2 in the top PEG-rich phase can be expected furtherly demonstrating that ATPS have the potential for the selective recovery of these promising nanovesicles

Economic analysis of Uricase production under uncertainty: Contrast of chromatographic purification and aqueous two-phase extraction (with and without PEG recycle)

Uricase is the enzyme responsible for the breakdown of uric acid, the key molecule leading to gout in humans, into allantoin, but it is absent in humans. It has been produced as a PEGylated pharmaceutical where the purification is performed through three sequential chromatographic columns. More recently an aqueous two-phase system (ATPS) was reported that could recover Uricase with high yield and purity. Although the use of ATPS can decrease cost and time, it also generates a large amount of waste. The ability, therefore, to recycle key components of ATPS is of interest. Economic modelling is a powerful tool that allows the bioprocess engineer to compare possible outcomes and find areas where further research or optimization might be required without recourse to extensive experiments and time. This research provides an economic analysis using the commercial software BioSolve of the strategies for Uricase production: chromatographic and ATPS, and includes a third bioprocess that utilises material recycling. The key parameters that affect the process the most were located via a sensitivity analysis and evaluated with a Monte Carlo analysis. Results show that ATPS is far less expensive than chromatography, but that there is an area where the cost of production of both bioprocesses overlap. Furthermore, recycling doesn't impact the cost of production. This study serves to provide a framework for the economic analysis of Uricase production using alternative techniques. This article is protected by copyright. All rights reserved

Sustainable Strategy Based on Induced Precipitation for the Purification of Phycobiliproteins

Phycobiliproteins are fluorescent proteins mainly produced by red macroalgae and cyanobacteria. These proteins, essential to the survival of these organisms, find application in many fields of interest, from medical, pharmaceutical, and cosmetic to food and textile industries. The biggest obstacle to their use is the lack of simple environmental and economical sustainable methodologies to obtain these proteins with high purity. In this work, a new purification process is proposed based on the induced precipitation of the target proteins followed by ultrafiltration. Purities of 89.5% of both phycobiliproteins and 87.3% of R-phycoerythrin were achieved using ammonium sulfate and poly(acrylic acid) sodium salts as precipitation agents (followed by an ultrafiltration step), while maintaining high recovery yields and protein structure stability. Environmental analysis performed to evaluate the proposed process shows that the carbon footprint for the proposed process is much lower than that reported for alternative methodology, and the economic analysis reveals the cost-effective character associated to its high performance. This work is a step toward more sustainable and effective methodologies/processes with high industrial potential

New Trends for the Processing of Poly(Methyl Methacrylate) Biomaterial for Dental Prosthodontics

Rehabilitation of masticatory function in patients with absent teeth with removable dentures is an established form of treating partial or complete dentition in edentulous patients. The developments in recent decades with dental implants dominate current dental research. However, medical contraindications, a negative attitude toward implants, or financial limitations on the part of the patients limit their universal applicability, so the rehabilitation with dental prostheses still makes up a significant portion of everyday clinical practice. Conversely, removable dentures are used in the critical conditions of the oral cavity. There are about 500 strains of microorganisms in the mouth, which form the biofilm in an acidic environment causing several issues, such as denture stomatitis, deterioration of the periodontal status of the remaining teeth, or carious lesions in the supporting teeth. Therefore, it is very important to choose a suitable material for the prosthesis. Poly(methyl methacrylate) (PMMA) is an acrylic resin usually used with a long tradition for prosthetic purposes. The aim of this chapter is to present the trends for the processing of PMMA. It includes the chemical synthesis, conventional thermal processing of this acrylic resin, the new processing technique assisted with ultrasound, the antibacterial effect on PMMA with nanoparticles, and the cytotoxicity, genotoxicity, and mutagenesis of this material

Factorial and Economic Evaluation of an Aqueous Two-Phase Partitioning Pilot Plant for Invertase Recovery From Spent Brewery Yeast

Aqueous two-phase systems (ATPS) have been reported as an attractive biocompatible extraction system for recovery and purification of biological products. In this work, the implementation, characterization, and optimization (operational and economic) of invertase extraction from spent brewery yeast in a semi-automatized pilot plant using ATPS is reported. Gentian violet was used as tracer for the selection of phase composition through phase entrainment minimization. Yeast suspension was chosen as a complex cell matrix model for the recovery of the industrial relevant enzyme invertase. Flow rates of phases did not have an effect, given that a bottom continuous phase is given, while load of sample and number of agitators improved the recovery of the enzyme. The best combination of factors reached a recovery of 129.35 ± 2.76% and a purification factor of 4.98 ± 1.10 in the bottom phase of a PEG-Phosphate system, also resulting in the removal of inhibitor molecules increasing invertase activity as reported by several other authors. Then, an economic analysis was performed to study the production cost of invertase analyzing only the significant parameters for production. Results indicate that the parameters being analyzed only affect the production cost per enzymatic unit, while variations in the cost per batch are not significant. Moreover, only the sample load is significant, which, combined with operational optimization results, gives the same optimal result for operation, maximizing recovery yield (15% of sample load and 1 static mixer). Overall res ults of these case studies show continuous pilot-scale ATPS as a viable and reproducible extraction/purification system for high added-value biological compounds

Geranium leaf-mediated synthesis of silver nanoparticles and their transcriptomic effects on Candida albicans

Candida albicans is the most predominant fungal species isolated from medical devices, including catheters, heart valves, and dental prostheses. In recent years, it has been demonstrated to be resistant to many antifungals; therefore, silver nanoparticles (AgNPs) have been proposed as an alternative. But only a handful of research is contributed to omic-based studies to study the various impacts of AgNPs on Candida species and other microorganisms. Thus, the study aims to biosynthesize AgNPs using Pelargonium-hortorum leaf and test its antifungal, cytotoxicity, and global gene expression on Candida through transcriptomic profiling. The leaf-assisted AgNPs resulted in spherical shapes with a particle size of 38 nm. The anticandidal effect demonstrated that the Minimum inhibitory concentration was 25 μg·mL−1. Later, the cytotoxicity assay reported a moderate impact on the human gingival fibroblast cells. Finally, the transcriptomic analysis demonstrated the differential gene expression of 3,871 upregulated and 3,902 downregulated genes. Thus, proving the anticandidal effect of AgNPs on Candida through RNA-seq experiments and the regulated genes is highly important to cell wall integrity, adherence, and virulence

Resistance to autosomal dominant Alzheimer's disease in an APOE3 Christchurch homozygote: a case report.

We identified a PSEN1 (presenilin 1) mutation carrier from the world's largest autosomal dominant Alzheimer's disease kindred, who did not develop mild cognitive impairment until her seventies, three decades after the expected age of clinical onset. The individual had two copies of the APOE3 Christchurch (R136S) mutation, unusually high brain amyloid levels and limited tau and neurodegenerative measurements. Our findings have implications for the role of APOE in the pathogenesis, treatment and prevention of Alzheimer's disease