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Biotechnology Progress

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https://www.readbyqxmd.com/read/28704889/cyclodextrins-increase-phytosterol-and-tocopherol-levels-in-suspension-cultured-cells-obtained-from-mung-beans-and-safflower
#1
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
#2
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
#3
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
#4
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
#5
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
#6
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
#7
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
#8
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
#9
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
https://www.readbyqxmd.com/read/28649776/improving-the-baculovirus-expression-vector-system-with-vankyrin-enhanced-technology
#10
Kendra H Steele, Barbara J Stone, Kathleen M Franklin, Xiufeng Zhang, Haobo Jiang, Bruce A Webb, Christoph Geisler, Angelika Fath-Goodin
The baculovirus expression vector system (BEVS) is a widely used platform for the production of recombinant eukaryotic proteins. However, the BEVS has limitations in comparison to other higher eukaryotic expression systems. First, the insect cell lines used in the BEVS cannot produce glycoproteins with complex-type N-glycosylation patterns. Second, protein production is limited as cells die and lyse in response to baculovirus infection. To delay cell death and lysis, we transformed several insect cell lines with an expression plasmid harboring a vankyrin gene (P-vank-1), which encodes an anti-apoptotic protein...
June 26, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28649725/high-throughput-screening-identifies-novel-cell-cycle-arresting-small-molecule-enhancers-of-transient-protein-expression
#11
Hermann-Josef Meyer, Rebecca Turincio, Shirley Ng, Juan Li, Blair Wilson, Pamela Chan, Mark Zak, Dorothea Reilly, Maureen H Beresini, Athena W Wong
Transient gene expression in mammalian cells is an efficient process for producing recombinant proteins for various research applications to support large molecule therapeutics development. For the first time, we report a high throughput small molecule (SM) screen to identify novel compounds that increase antibody titers after polyethylenimine (PEI) transient transfection of a HEK293 cell line. After screening 31,413 SMs in a 50 μL scaled-down process, we validated 164 SMs to improve yields by up to 2-fold...
June 26, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28649713/mechanistic-modeling-of-the-loss-of-protein-sieving-due-to-internal-and-external-fouling-of-microfilters
#12
Glen R Bolton, Alex J Apostolidis
Fed-batch and perfusion cell culture processes used to produce therapeutic proteins can utilize microfilters for product harvest. In this study, new explicit mathematical models of sieving loss due to internal membrane fouling, external membrane fouling, or a combination of the two were generated. The models accounted for membrane and cake structures and hindered solute transport. Internal membrane fouling was assumed to occur due to the accumulation of foulant on either membrane pore walls (pore-retention model) or membrane fibers (fiber-retention model)...
June 26, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28597974/a-simple-cell-transport-device-keeps-culture-alive-and-functional-during-shipping
#13
Paula G Miller, Ying I Wang, Glen Swan, Michael L Shuler
Transporting living complex cellular constructs through the mail while retaining their full viability and functionality is challenging. During this process, cells often suffer from exposure to suboptimal life-sustaining conditions (e.g. temperature, pH), as well as damage due to shear stress. We have developed a transport device for shipping intact cell/tissue constructs from one facility to another that overcomes these obstacles. Our transport device maintained three different cell lines (Caco2, A549, and HepG2 C3A) individually on transwell membranes with high viability (above 97%) for 48 h under simulated shipping conditions without an incubator...
June 9, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28593695/protozoa-inhibition-by-different-salts-osmotic-stress-or-ionic-stress
#14
Changhao Li, Jingya Li, Christopher Q Lan, Dankui Liao
Cell density and morphology changes were tested to examine the effects of salts including NaHCO3 , NaCl, KHCO3 and KCl at 160 mM on protozoa. It was demonstrated that ionic stress rather than osmotic stress led to protozoa cell death and NaHCO3 was shown to be the most effective inhibitor. Deformation of cells and cell shrinkage were observed when protozoan cells were exposed to PEG or any of the salts. However, while PEG treated cells could fully recover in both number and size, only a small portion of the salt-treated cells survive and cell size was 36% -58% smaller than regular...
June 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28593654/linkage-of-microbial-kinetics-and-bacterial-community-structure-of-mbr-and-hybrid-mbbr-mbr-systems-to-treat-salinity-amended-urban-wastewater
#15
Alejandro Rodriguez-Sanchez, Juan Carlos Leyva-Diaz, Alejandro Gonzalez-Martinez, Jose Manuel Poyatos
Three pilot-scale bioreactors were started up and operated under salinity-amended urban wastewater feeding. The bioreactors were configured as membrane bioreactor and two different hybrid, moving bed biofilm reactor-membrane bioreactor and operated with a hydraulic retention time of 9.5 h, a solid residence time of 11.75 days and a total solids concentration of 2500 mg L(-1) . The three systems showed excellent performance in suspended solids, BOD5 , and COD removal (values of 96-100%, 97-99%, and 88-90%, respectively), but poor nitrogen removal (values of 20-30%)...
June 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28593644/efficient-trna-degradation-and-quantification-in-escherichia-coli-cell-extract-using-rnase-coated-magnetic-beads-a-key-step-toward-codon-emancipation
#16
Amin S M Salehi, Mark T Smith, Song-Min Schinn, Jeremy M Hunt, Christina Muhlestein, Joann Diray-Arce, Brent L Nielsen, Bradley C Bundy
Emancipating sense codons toward a minimized genetic code is of significant interest to science and engineering. A key approach toward sense codon emancipation is the targeted in vitro removal of native tRNA. However, challenges remain such as the insufficient depletion of tRNA in lysate-based in vitro systems and the high cost of the purified components system (PURE). Here we used RNase-coated superparamagnetic beads to efficiently degrade E. coli endogenous tRNA. The presented method removes >99% of tRNA in cell lysates, while partially preserving cell-free protein synthesis activity...
June 8, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28571115/improvement-of-covalent-immobilization-procedure-of-%C3%AE-galactosidase-from-kluyveromyces-lactis-for-galactooligosaccharides-production-modelling-and-kinetic-study
#17
Flor González-Cataño, Luz Tovar-Castro, Eduardo Castaño-Tostado, Carlos Regalado-Gonzalez, Blanca García-Almendarez, Anaberta Cardador-Martínez, Silvia Amaya-Llano
No abstract text is available yet for this article.
June 1, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28556629/in-vitro-methanol-production-from-methyl-coenzyme-m-using-the-methanosarcina-barkeri-mtaabc-protein-complex
#18
Ming Dong, Tara D Gonzalez, Meghan M Klems, Lisa M Steinberg, Wilfred Chen, Eleftherios T Papoutsakis, Brian J Bahnson
Methanol:coenzyme M methyltransferase is an enzyme complex composed of three subunits, MtaA, MtaB and MtaC, found in methanogenic archaea and is needed for their growth on methanol ultimately producing methane. MtaABC catalyzes the energetically favorable methyl transfer from methanol to coenzyme M to form methyl coenzyme M. Here we demonstrate that this important reaction for possible production of methanol from the anaerobic oxidation of methane can be reversed in vitro. To this effect, we have expressed and purified the Methanosarcina barkeri MtaABC enzyme, and developed an in vitro functional assay that demonstrates MtaABC can catalyze the energetically unfavorable (ΔG° = 27 kJ/mol) reverse reaction starting from methyl coenzyme M and generating methanol as a product...
May 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28556626/media-development-for-large-scale-agrobacterium-tumefaciens-culture
#19
Ingrid K Leth, Karen A McDonald
A chemically defined media was developed for growing Agrobacterium tumefaciens at large scale for commercial production of recombinant proteins by transient expression in plants. Design of experiments was used to identify major and secondary effects of ten media components: sucrose, ammonium sulfate ((NH4 )2 SO4 ), magnesium sulfate heptahydrate (MgSO4 *7H2 O), calcium chloride dihydrate (CaCl2 *2H2 O), iron (II) sulfate heptahydrate (FeSO4 *7H2 O), manganese (II) sulfate monohydrate (MnSO4 *H2 O), zinc sulfate heptahydrate (ZnSO4 *7H2 O), sodium chloride (NaCl), potassium chloride (KCl) and a sodium/potassium phosphate buffer (Na2 HPO4 /KH2 PO4 )...
May 27, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28556621/measuring-the-aggregation-of-cho-cells-prior-to-single-cell-cloning-allows-a-more-accurate-determination-of-the-probability-of-clonality
#20
Kerensa J Klottrup, Guillermo Miro-Quesada, Layla Flack, Ivan Pereda, Pamela Hawley-Nelson
The manufacturing process for biotherapeutics is closely regulated by the Food and Drug Administration (FDA), European Medicines Agency (EMA) and other regulatory agencies worldwide. To ensure consistency of the product of a manufacturing cell line, International Committee on Harmonization guidelines (Q5D, 1997) state that the cell substrate should be derived from a single cell progenitor, i.e., clonal.Cell lines in suspension culture may naturally revert to cell adhesion in the form of doublets, triplets and higher order structures of clustered cells...
May 27, 2017: Biotechnology Progress
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