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Biotechnology Progress

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https://www.readbyqxmd.com/read/28786215/a-system-identification-approach-for-developing-model-predictive-controllers-of-antibody-quality-attributes-in-cell-culture-processes
#1
Brandon Downey, John Schmitt, Justin Beller, Brian Russell, Anthony Quach, Elizabeth Hermann, David Lyon, Jeffrey Breit
As the biopharmaceutical industry evolves to include more diverse protein formats and processes, more robust control of Critical Quality Attributes (CQAs) is needed to maintain processing flexibility without compromising quality. Active control of CQAs has been demonstrated using model predictive control techniques, which allow development of processes which are robust against disturbances associated with raw material variability and other potentially flexible operating conditions. Wide adoption of model predictive control in biopharmaceutical cell culture processes has been hampered, however, in part due to the large amount of data and expertise required to make a predictive model of controlled CQAs, a requirement for model predictive control...
August 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28786214/quantitative-evaluation-of-recombinant-protein-packaged-into-outer-membrane-vesicles-of-escherichia-coli-cells
#2
Yoshihiro Ojima, Kyota Yamaguchi, Masahito Taya
Outer membrane vesicles (OMVs) are spherical bilayered proteolipids released from the cell surfaces of bacteria, which have gained traction in the biotechnology fields. Bacterial cellular machinery can be genetically engineered to produce and package heterologous enzymes into OMVs, producing nanocarriers and nanoparticle catalysts. However, the productivity or efficiency of packaging the target protein into OMVs has not been quantitatively evaluated. In this study, we packaged green fluorescence protein (GFP) into the OMVs of Escherichia coli through N-terminal fused expression to outer membrane protein W (OmpW)...
August 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28748655/high-throughput-automated-microbial-bioreactor-system-used-for-clone-selection-and-rapid-scale-down-process-optimisation
#3
M Lourdes Velez-Suberbie, John Pj Betts, Kelly L Walker, Colin Robinson, Barney Zoro, Eli Keshavarz-Moore
High throughput automated fermentation systems have become a useful tool in early bioprocess development. In this study, we investigated a 24 x 15 mL single use microbioreactor system, ambr 15f, designed for microbial culture. We compared the fed-batch growth and production capabilities of this system for two Escherichia coli strains, BL21 (DE3) and MC4100, and two industrially relevant molecules, hGH and scFv. In addition, different carbon sources were tested using bolus, linear or exponential feeding strategies, showing the capacity of the ambr 15f system to handle automated feeding...
July 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28748636/compact-cell-settlers-for-perfusion-cultures-of-microbial-and-mammalian-cells
#4
Cassandra A Freeman, Premsingh S D Samuel, Dhinakar S Kompala
As microbial secretory expression systems have become well developed for microbial yeast cells, such as Saccharomyces cerevisiae and Pichia pastoris, it is advantageous to develop high cell density continuous perfusion cultures of microbial yeast cells to retain the live and productive yeast cells inside the perfusion bioreactor while removing the dead cells and cell debris along with the secreted product protein in the harvest stream. While the previously demonstrated inclined or lamellar settlers can be used for such perfusion bioreactors for microbial cells, the size and footprint requirements of such inefficiently scaled up devices can be quite large in comparison to the bioreactor size...
July 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28734039/optimal-control-for-micro-algae-on-a-raceway-model
#5
Todd Hurst, Volker Rehbock
We apply numerical optimal control methods to an existing algae growth model with the aim to determine the best performance of the model under known conditions using a variety of decision variables. We transform the system of differential algebraic equations in the existing model to a system of ordinary differential equations which introduces dynamics for average light intensity and chlorophyll. In addition we allow for variable nitrogen concentration of the inflow as well as variable initial nitrogen concentration of the raceway...
July 22, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28726354/high-performance-of-agaricus-blazei-fungus-for-the-biological-pre-treatment-of-elephant-grass
#6
Thais Dal Picolli, Kaliane Regalin Aver, Roselei Claudete Fontana, Marli Camassola
Biological pre-treatment seems to be promising being an eco-friendly process, with no inhibitor generated during the process. The potential for elephant grass pre-treatment with white degradation fungi Pleurotus ostreatus, Agaricus blazei, Lentinula edodes, Pleurotus citrinopileatus and Pleurotus djamor, in isolated or mixed cultures of these strains, was evaluated. The highest activities of enzymes involved in the degradation of lignocellulosic biomass (laccases, endoglucanases, xylanases and β-glucosidases) were observed for A...
July 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28726331/novel-fluid-shear-based-dissociation-device-for-improved-single-cell-dissociation-of-spheroids-and-cell-aggregates
#7
Ursula L Triantafillu, Jaron N Nix, Yonghyun Kim
Biological industries commonly rely on bioreactor systems for the large-scale production of cells. Cell aggregation, clumping, and spheroid morphology of certain suspension cells make their large-scale culture challenging. Growing stem cells as spheroids is indispensable to retain their stemness, but large spheroids (>500 µm diameter) suffer from poor oxygen and nutrient diffusion, ultimately resulting in premature cell death in the centers of the spheroids. Despite this, most large-scale bioprocesses do not have an efficient method for dissociating cells into single cells, but rely on costly enzymatic dissociation techniques...
July 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28726323/characterization-of-a-cathepsin-d-protease-from-cho-cell-free-medium-and-mitigation-of-its-impact-on-the-stability-of-a-recombinant-therapeutic-protein
#8
Amareth Lim, Brandon L Doyle, Gerard M Kelly, Angelia M Reed-Bogan, Lawrence H Breen, Parviz A Shamlou, Peter K Lambooy
During purification process development of a recombinant therapeutic protein, an endoproteolytic activity endogenous to the Chinese hamster ovary (CHO) cells and leading to degradation at particular hydrophobic amino acid residues (e.g., Phe and Trp) was observed when processing at acidic pH. The presence of residual levels of protease activity in purified protein batches affected the inherent activity of the product when stored as a solution. In order to develop a robust purification strategy to minimize this undesirable impact, identification and characterization of this protease was essential to ultimately ensure that a solution formulation was stable for many years...
July 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28726315/biomineralization-of-gold-by-mucor-plumbeus-the-progress-in-understanding-the-mechanism-of-nanoparticles-formation
#9
Irena Maliszewska, Włodzimierz Tylus, Izabela Pawlaczyk-Graja, Wojciech Pusz, Anna Baturo-Cieśniewska, Jacek Chęcmanowski, Bogdan Szczygieł
This contribution describes the deposition of gold nanoparticles by microbial reduction of Au(III) ions using the mycelium of Mucor plumbeus. Biosorption as the major mechanism of Au(III) ions binding by the fungal cells and the reduction of them to the form of Au(0) on/in the cell wall, followed by the transportation of the synthesized gold nanoparticles to the cytoplasm, is postulated. The probable mechanism behind the reduction of Au(III) ions is discussed, leading to the conclusion that this process is non enzymatic one...
July 20, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28722325/polymer-mediated-flocculation-of-transient-cho-cultures-as-a-simple-high-throughput-method-to-facilitate-antibody-discovery
#10
Matthew G Schmitt, Yashas Rajendra, Maria D Hougland, Jeffrey S Boyles, Gavin C Barnard
Most biopharmaceutical drugs, especially monoclonal antibodies (mAbs), bispecific antibodies (BsAbs) and Fc-fusion proteins, are expressed using Chinese Hamster Ovary (CHO) cell lines. CHO cells typically yield high product titers and high product quality. Unfortunately, CHO cell lines also generate high molecular weight (HMW) aggregates of the desired product during cell culture along with CHO host cell protein (HCP) and CHO DNA. These immunogenic species, co-purified during Protein A purification, must be removed in a multi-step purification process...
July 19, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28722272/development-of-a-thiol-ene-based-screening-platform-for-enzyme-immobilization-demonstrated-using-horseradish-peroxidase
#11
Christian Hoffmann, Manuel Pinelo, John M Woodley, Anders E Daugaard
Efficient immobilization of enzymes on support surfaces requires an exact match between the surface chemistry and the specific enzyme. A successful match would normally be identified through time consuming screening of conventional resins in multiple experiments testing individual immobilization strategies. In this study we present a versatile strategy that largely expands the number of possible surface functionalities for enzyme immobilization in a single, generic platform. The combination of many individual surface chemistries and thus immobilization methods in one modular system permits faster and more efficient screening, which we believe will result in a higher chance of discovery of optimal surface/enzyme interactions...
July 19, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28704889/cyclodextrins-increase-phytosterol-and-tocopherol-levels-in-suspension-cultured-cells-obtained-from-mung-beans-and-safflower
#12
Lorena Almagro, Libertad Raquel Tudela, Ana Belén Sabater-Jara, Begoña Miras-Moreno, M A Pedreño
In this work, suspension-cultured cells of mung beans and safflower were used in order to analyze the effect of methyl jasmonate and/or cyclodextrins, on bioactive compound production such as phytosterols and tocopherols. The results indicated that mung bean suspension-cultured cells produced higher amount of total phytosterols and tocopherols. In particular, mung bean suspension-cultured cells produced almost 220-fold higher levels of tocopherols than safflower suspension-cultured cells in the best conditions...
July 13, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28699320/a-three-plus-three-parameters-mechanistic-model-for-viral-filtration
#13
Prakhar Misra, Abhishek Sinha, Fasil Q Mir, Anurag S Rathore, Anupam Shukla
Viral filtration is an expensive regulatory requirement in downstream processing of monoclonal antibodies (mAbs). This process step is typically operated with an overdesigned filter in order to account for any batch to batch variability in the filter, as well as the feed characteristics. Here, we propose a simple, six-parameter mechanistic model for viral filtration where three parameters are membrane-specific while the other three depend on feed characteristics and membrane-feed interactions. Viruses are considered as passive particles which are retained by the membrane on the basis of size exclusion...
July 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28699244/biophysical-characterization-of-layer-by-layer-synthesis-of-aptamer-drug-microparticles-for-enhanced-cell-targeting
#14
Kei X Tan, Michael K Danquah, Amandeep Sidhu, Sie Yon Lau, Clarence M Ongkudon
Targeted delivery of drug molecules to specific cells in mammalian systems demonstrates a great potential to enhance the efficacy of current pharmaceutical therapies. Conventional strategies for pharmaceutical delivery are often associated with poor therapeutic indices and high systemic cytotoxicity, and this results in poor disease suppression, low surviving rates, and potential contraindication of drug formulation. The emergence of aptamers has elicited new research interests into enhanced targeted drug delivery due to their unique characteristics as targeting elements...
July 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28691347/design-and-operation-of-a-continuous-integrated-monoclonal-antibody-production-process
#15
Fabian Steinebach, Nicole Ulmer, Moritz Wolf, Lara Decker, Veronika Schneider, Ruben Wälchli, Daniel Karst, Jonathan Souquet, Massimo Morbidelli
The realization of an end-to-end integrated continuous lab-scale process for monoclonal antibody manufacturing is described. For this, a continuous cultivation with filter-based cell-retention, a continuous two column capture process, a virus inactivation step, a semi-continuous polishing step (twin-column MCSGP) and a batch-wise flow-through polishing step were integrated and operated together. In each unit the implementation of internal recycle loops allows to improve the performance: a) in the bioreactor, to simultaneously increase the cell density and volumetric productivity, b) in the capture process, to achieve improved capacity utilization at high productivity and yield and c) in the MCSGP process, to overcome the purity-yield trade-off of classical batch-wise bind-elute polishing steps...
July 10, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28675693/application-of-iterative-robust-model-based-optimal-experimental-design-for-the-calibration-of-biocatalytic-models
#16
Timothy Van Daele, Krist V Gernaey, Rolf H Ringborg, Tim Börner, Søren Heintz, Daan Van Hauwermeiren, Carl Grey, Ulrich Krühne, Patrick Adlercreutz, Ingmar Nopens
The aim of model calibration is to estimate unique parameter values from available experimental data, here applied to a biocatalytic process. The traditional approach of first gathering data followed by performing a model calibration is inefficient, since the information gathered during experimentation is not actively used to optimise the experimental design. By applying an iterative robust model-based optimal experimental design, the limited amount of data collected is used to design additional informative experiments...
July 4, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28653478/different-rna-and-protein-expression-of-surface-markers-in-rabbit-amniotic-fluid-derived-mesenchymal-stem-cells
#17
Michal Kovac, Jaromir Vasicek, Barbora Kulikova, Miroslav Bauer, Jozef Curlej, Andrej Balazi, Peter Chrenek
Over the years, there has been much confusion in defining molecular markers of mesenchymal stem cells (MSCs) for other than human species due to a lack of species-specific antibodies. Therefore, the aim of our study was to define rabbit amniotic fluid-derived mesenchymal stem cells (rAF-MSCs) and to reflect upon the current identification of AF-MSCs by providing a summary of detected surface markers in different species. The expression of rAF-MSC surface markers was analyzed at the protein and mRNA level. Flow cytometry analyses showed that rAF-MSCs were positive for CD29 and CD44, low positive for CD90, but negative for CD73, CD105 and CD166...
June 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28653476/heterotrophic-production-of-chlorella-sp-tistr-8990-biomass-growth-and-composition-under-various-production-conditions
#18
Somruethai Bouyam, Wanna Choorit, Sarote Sirisansaneeyakul, Yusuf Chisti
The green microalga Chlorella sp. TISTR 8990 was grown heterotrophically in the dark using various concentrations of a basal glucose medium with a carbon-to-nitrogen mass ratio of 29:1. The final biomass concentration and the rate of growth were highest in the 5-fold concentrated basal glucose medium (25 g L(-1) glucose, 2.5 g L(-1) KNO3 ) in batch operations. Improving oxygen transfer in the culture by increasing the agitation rate and decreasing the culture volume in 500 mL shake flasks improved growth and glucose utilization...
June 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28653475/amylosucrase-mediated-%C3%AE-carotene-encapsulation-in-amylose-microparticles
#19
Carlos Andres Morales Letona, Cheon-Seok Park, Young-Rok Kim
The beta-carotene embedded amylose microparticles (BC-AmMPs) were prepared in one-step by utilizing the unique catalytic activity of amylosucrase from Deinococcus geothermalis (DgAS), which synthesizes linear amylose chains using sucrose as the sole substrate. Synthesized amylose chains self-assembled with beta carotene to form well-defined spherical microparticles with an encapsulation yield of 65%. The BC-AmMPs produced (average diameter ∼8 µm) were bright orange due to the embedded beta-carotene, and this was confirmed by Raman analysis...
June 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28653347/preliminary-evaluation-of-pleurotus-ostreatus-for-the-removal-of-selected-pharmaceuticals-from-hospital-wastewater
#20
Laura Palli, Francesc Castellet-Rovira, Miriam Pérez-Trujillo, Donatella Caniani, Montserrat Sarrà-Adroguer, Riccardo Gori
The fungus Pleurotus ostreatus was investigated to assess its ability to remove diclofenac, ketoprofen and atenolol spiked at 10 mg/L each one in hospital wastewater. The degradation test was carried out in a fluidized bed bioreactor testing both the batch and the continuous mode (hydraulic retention time in the range 1.63-3 d). In batch mode, diclofenac disappeared in less than 24 h, ketoprofen was degraded up to almost 50% in 5 d while atenolol was not removed. In continuous mode, diclofenac and ketoprofen removals were about 100% and 70% respectively; atenolol degradation was negligible during the first 20 d but it increased up to 60% after a peak of laccase production and notable biomass growth...
June 27, 2017: Biotechnology Progress
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